Rotifer Symposium V

Developments in Hydrobiology 52

Series editor
H. J. Dumont
 Rotifer Symposium V
Proceedings of the Fifth Rotifer Symposium,held in Gargnano, Italy,
September 11-18, 1988

Edited by
C. Ricci, T. W. Snell and C. E. King

Reprinted from Hydrobi%gia, vo/s. 186/187 (1989)

....
"
Kluwer Academic Publishers
Dordrecht / Boston / London
Library of Congress Cataloging-in-Publication Data

International Rotifer Symposium (5th: 1988 : Gargnano, Italy)

Rotifer Symposium V I edited by C. Ricci, T.W. Snell, and C.E.
King.
p. cm. -- (Develop_ents in hydrobiology ; 52)
"Reprinted fro. Hydrobiologia."
1. Rotifera--Congresses. I. Ricci, C. (Claudia) II. Snell,
Terry W. III. King, C. E. (Charles E.) IV. Title. V. Series.
OL391.R8157 1988
595. 1 '81--dc20 89-17596

ISBN-13: 978-94-010-6694-5 e-ISBN-13: 978-94-009-0465-1

DOl: 10.1007/978-94-009-0465-1
Printed on acid-free paper

Kluwer Academic Publishers incorporates the publishing programmes of Dr W. Junk Publishers,

MTP Press, Martinus Nijhoff Publishers, and D. Reidel Publishing Company.

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for all other countries: Kluwer Academic Publishers Group, P.O. Box 322, 3200 AH Dordrecht,
The Netherlands

Copyright

© 1989 Kluwer Academic Publishers, Dordrecht

Softcover reprint ofthe hardcover 1st edition 1989
All rights reserved. No part ofthis publication may be reproduced, stored in a retrieval system,
or transmitted in any form or by any means, mechanical, photocopying, recording, or otherwise,
without the prior written permission from the copyright owners.
 v
Contents

Preface.......................................................................... IX
List of participants. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . X
Josef Donner, in memoriam ........................................................ XVII

Rotifer study as a way of life

by W. T. Edmondson............................................................ 1

PART ONE: ECOLOGY

Food
Food limitation and body size in the life cycles of planktonic rotifers and cladocerans
by A. Duncan .................................................................. 11
Is food availability the main factor controlling the abundance of Euchlanis dilatata lucksiana Hauer
in a shallow, hypertrophic lake?
by J. Ejsmont-Karabin, R. D. Gulati and J. Rooth ................................... 29
Influence of cyanobacterial diet on Asplanchna predation risk in Brachionus calyciflorus
by P. L. Starkweather and E. J. Walsh. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Nutritional effect of freshwater Chlorella on growth of the rotifer Brachionus plicatilis
by K. Hirayama, I. Maruyama and T. Maeda ....................................... 39
Life history characteristics of Brachionus plicatilis (rotifera) fed different algae
by J. Korstad, Y. Olsen and O. Vadstein ........................................... 43
Feeding kinetics of Brachionus plicatilis fed Isochrysis galbana
by J. Korstad, O. Vadstein and Y. Olsen ........................................... 51

Competition
Epizoic and parasitic rotifers
by L. May..................................................................... 59
Interrelations of rotifers with predatory and herbivorous Cladocera: a review of Russian works
by L. K. Matveeva .............................................................. 69
Competitive interactions between the rotifer Synchaeta oblonga and the cladoceran Scapholeberis
kingi Sars
by J. J. Gilbert. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75

Autoecology
Salinity and temperature influence in rotifer life history characteristics
by M. R. Miracle and M. Serra ................................................... 81
Empirical evidence for a complex diurnal movement in Hexarthra bulgarica from an oligotrophic
high mountain lake (La Caldera, Spain)
by P. Carrillo, L. Cruz-Pizarro and R. Morales-Baquero .............................. 103
VI

Morphological variation in Kellicottia longispina

by W. T. Edmondson and A. H. Litt ............................................... 109
Morphological structure and functional patterns of Keratella cochlearis (Gosse) populations in
stratified lakes
by G. A. Galkovskaya and I. F. Mityanina ......................................... 119
The development of Hexarthra spp. in a shallow alkaline lake
by A. Herzig and W. Koste . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 129
On choice of substrate and habitat in brachionid rotifers
by B. Pejler and B. BerziJ;ls ....................................................... 137
Temporal analysis of clonal structure in a moss bdelloid population
by C. Ricci, M. Pagani and A. M. Bolzern .......................................... 145
Horizontal distribution of the plankton rotifers Keratella cochlearis (Bory de St Vincent) and
Polyarthra vulgaris (Carlin) in a small eutrophic lake
by A. P. Saunders-Davies ........................................................ 153
Oviposition behavior of the littoral rotifer Euchlanis dilatata
by E. J. Walsh .................................................................. 157
Developmental times of Synchaeta oblonga eggs from the Danube (Austria)
by W. Zoufal ................................................................... 163

Communities
Community structure and coexistence of the rotifers of an artificial crater lake
by G. Bogaert and H. J. Dumont. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 167
The rotifer communities of acid-stressed lakes of Maine
by M. T. Brett .................................................................. 181
Composition and distributional patterns in arctic rotifers
by R. Chengalath and W. Koste ................................................... 191
Abundance and diversity of planktonic rotifers in the Po River
by I. Ferrari, A. Farabegoli and R. Mazzoni ........................................ 201
Rotifer distribution in relation to temperature and oxygen content
by E. Mikschi .................................................................. 209
Patterns in the composition of the rotifer communities from high mountain lakes and ponds in Sierra
Nevada (Spain)
by R. Morales-Baquero, L. Cruz-Pizarro and P. Carrillo .............................. 215
Rotifer associations of some wetlands in Ontario, Canada
by T. N ogrady .................................................................. 223
Opportunist rotifers: colonising species of young ponds in Surrey, England
by R. M. Pontin ................................................................ 229
Percentage of rotifers in spring zooplankton in lakes of different trophy
by S. Radwan and B. Popiolek .................................................... 235
Tasmania revisited: rotifer communities and habitat heterogeneity
by R. J. Shiel, W. Koste and L. W. Tan ............................................ 239
Occurrence of Rotifera in the field under natural and intentionally-changed conditions. II. Lake
Numasawa
by M. Sudzuki, T. Matsumoto and K. Narita ....................................... 247
 VII

PART TWO: TAXONOMY AND MORPHOLOGY

The skeletal muscles of rotifers and their innervation

by P. Clement and 1. Amsellem ................................................... 255
Classical taxonomy and modern methodology
by W. Koste and R.I. Shiel ...................................................... 279
Mastax morphology under SEM and its usefulness in reconstructing rotifer phylogeny and syste-
matics
by G. 1. Markevich and L. A. Kutikova ............................................ 285
Problems in taxonomy of rotifers, exemplified by the FiUnia longiseta-terminalis complex
by A. Ruttner-Kolisko ........................................................... 291
Systematics, reproductive isolation and species boundaries in monogonont rotifers
by T. W. Snell .................................................................. 299
Phylogenetic relationships within phylum Rotifera: orders and genus Notholca
by R. L. Wallace and R. A. Colburn ............................................... 311
A re-appraisal of two members of the genus Notholca from the Andes, with a note on the fme
structure of the lorica of N. foliacea (Ehrenberg)
by E. D. Hollowday and C. G. Hussey ............................................. 319

PART THREE: PHYSIOLOGY

Protein patterns in rotifers: the timing of aging

by M. 1. Carmona, M. Serra and M. R. Miracle ..................................... 325
Brachionus plicatilis tolerance to low oxygen concentrations
by A. Esparcia, M. R. Miracle and M. Serra ........................................ 331
Analysis of protein, carbohydrate and lipid in rotifers
C. Guisande and L. Serrano ...................................................... 339
A laboratory study of phosphorus and nitrogen excretion of Euchlanis dilatata lucksiana
by R. D. Gulati, 1. Ejsmont-Karabin, 1. Rooth and K. Siewertsen ...................... 347
A new method to estimate individual dry weights of rotifers
by H.-R. Pauli .................................................................. 355
Temperature aspects of ecological bioenergetics in Brachionus angularis (Rotatoria)
by N. Walz, T. Gschloessl and U. Hartmann. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 363
Biomass and elemental composition (C.H.N.) of the rotifer Brachionus plicatiUs cultured as larval
food
by M. YUfera and E. Pascual ..................................................... 371

PART FOUR: GENETICS

Molecular genetics of rotifers: preliminary restriction mapping of the mitochondrial genome of

Brachionus plicatiUs
by C. E. King .................................................................. 375
Size variation in Brachionus plicatiUs resting eggs
by L. Serrano, M. Serra and M. R. Miracle ......................................... 381
VIII

PART FIVE: AQUACULTURE

Rotifers as food in aquaculture

by E. Lubzens, A. Tandler and G. Minkoff ......................................... 387
Nitrogen flow through a Brachionus/Chlorella mass culture system
by W. D. Nagata. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 401
Kinetics of n-3 fatty acids in Brachionus plicatilis and changes in the food supply
by Y. Olsen, J. Rodriguez Rainuzzo, O. Vadstein and A. Jensen ....................... 409
Effect of incubation and preservation on resting egg hatching and mixis in the derived clones of
the rotifer Brachionus plicatilis
by A. Hagiwara and A. Hino ..................................................... 415
Intensive rotifer cultures using chemostats
by C. M. James and T. Abu Rezeq ................................................ 423

Author Index 431

Subject Index 433

 IX

Preface

The Fifth International Rotifer Symposium was organized by Dr. Claudia Ricci and held in the northern
Italian town of Gargnano (Brescia) from September 12-17, 1988. Through the generosity of the Rector
of Milano University, a beautiful villa on the shores of Lake Garda was made available to the 83 people
from 20 countries who attended the symposium. Ten of these rotifer workers had attended the four
previous meetings.
Such symposia serve three major functions, the results of which will be apparent in the papers contained
in this volume. First, because of the heterogeneity of interests and absence of concurrent sessions, the
attendees are exposed to an unusually large variety of research problems, approaches and modes of
interpretation. Bridges are thus built between one's own investigations and developments in the field as
a whole. Second, the extensive informal interactions that occur outside of the meeting room during coffee
breaks, dinners and excursions provide remarkable opportunities for research planning and sharing of
results of work in progress. Third, the acquaintances established at these meetings have facilitated
interactions during the three-year intervals between symposia. The result has been that visits between
laboratories, acquisition of research materials from distant sources and coordination of related investi-
gations have all been greatly enhanced.
A description of the week's events may serve to convey the ambience of the meeting. The tenor of the
symposium was set on Monday morning with a delightful reflection by Tommy Edmondson on a lifetime
of rotifer research. Twelve papers followed focusing on competition, parasitology, biogeography and
systematics. Each session was interrupted by at least one break where coffee and tea were served on the
veranda overlooking the lake. A full day of eleven papers was presented on Tuesday covering physiologi-
cal ecology, life history traits, interactions with cladocera and muscle ultrastructure. That night, a poster
session with ten papers was presented stimulating enthusiastic discussion. Wednesday morning consisted
offour papers on species concepts, molecular evolution and genetics. Wednesday afternoon, attendees
were treated to a two-hour boat ride to the northernmost shore of Lake Garda and the medieval town
of Riva. After a few hours of exploration, everyone climbed back aboard for a splendid return cruise which
included an elegant dinner. Thursday included seven papers on taxonomy, morphological variation and
cladistics along with an afternoon workshop on systematics. The conference ended on Friday with fifteen
papers focusing on aquaculture and bioenergetics. Many participants attended a post-conference
excursion to the beautiful old city of Verona.
On behalf of all participants, we wish to thank Dr. Giulio Melone, Dr. Mauro Mariani and his
colleagues at the Civico Acquario e Stazione Idrobiologica of Milan for their invaluable organizational
help both before and during the meeting. We are also grateful to Prof. Paolo Mantegazza, Rector of the
State University of Milan, and to Prof. Giulio Lanzavecchia, past head of the Department of Biology,
for their support of the meeting. In addition, grants-in-aid were obtained from the Italian Ministry of
Public Education (MPI), the National Council of Research (CNR), and from ENEA. Facilities were
provided by Comunita del Garda and by Direzione N avigazione Laghi.

CLAUDIA RICCI
TERRYW. SNELL
CHARLES E. KING
x
List of participants CLEMENT, Pierre
Equipe Neuro-Ethologie Univ. Lyon 1
43 Boulevard du 11 nov.
BARBATO, Fabio
69622 Villeurbanne Cedex, France
Via Monti di Creta 47
00167 Roma, Italy
COUTTEAU, Pierre
Artemia Reference Center
BELLELLI, Enrico
Rozier 44
Dip. BioI. Anim., Univ. La Sapienza,
9000 Gent, Belgium
00185 Roma, Italy
CRUZ-PIZARRO, Luis
BERNER-FANKHAUSER, Heidi
Catedra de Ecologia
Othmarsingerst. 18
Facultad de Ciencias
5600 Lenzburg, Switzerland
18071 Granada, Spain
BIANCHI, Maria Irene
DE MANUEL, J ordi
Via Appiani 24
Dpt. Ecologia, Facult. Biologia
20121 Milano, Italy
Av. Diagonal 645
08028 Barcelona, Spain
BOGAERT, Geert
Kerrebroek 83
DE RIDDER, Margaretha
9850 Nevele, Belgium
V. van Malderlaan 37/3
1710 Dilbeek, Belgium
B0RSHEIM, Knut Yngve
Dep. Microbiology Plant Physiol. DUMONT, Henri J.
Allegt 70 Inst. Ecology, State Univ. Ghent
5007 Bergen, Norway K.L. Ledeganckstraat 35
9000 Gent, Belgium
BRAIONI, Gianna
DUNCAN, Annie
Dip. BioI. Animale [Padova]
Dept. Zoology
Padua, Italy
Royal Holloway College
Englefield Green, Surrey TW20 9TY, U.K.
BRETT, Michael
Uppsala University, Limnology Institute, EDMONDSON, W. T.
Box 557, [15233] Dept. Zoology NJ-15
Uppsala, Sweden Univ. Washington
Seattle, Washington 98195, U.S.A.
CARMONA NAVARRO, Maria Jose
Depto. EcoI., Univ. Valencia EJSMONT-KARABIN, Jolanta
46100 Burjasot, Valencia, Spain Polish Acad. Sci., Inst. Ecology
ul. Lesna 13
CHENGALATH, Rama 11-730 Mikolajki, Poland
National Mus. Nat. Sciences,
P.O. Box 3443, ESPARCIA COLLADO, Angeles
Station'D' Depto Ecol., Univ. Valencia
Canada-Ontario-Ottawa, KIP 6P4 46100 Burjasot, Valencia, Spain
 XI

F ARABEGOLI, Alessandra HERZIG, Alois

Istituto di Zoologia Biologische Stat. Neusiedlersee
Via Borsari 46 7142 Illmitz, Austria
44100 Ferrara, Italy
HIRAYAMA, Kazutsugu
FERRARI, Ireneo Fac. Fisheries, Nagasaki Univ.
Istituto di Zoologia 1-14 Bunkyo Machi
Via Borsari 46 Nagasaki 852, Japan
44100 Ferrara, Italy
HOLLOWDAY, Eric D.
FRANCEZ, Andre Jean 45 Manor Road
INRA Stat. Agronomie Aylesbury, Bucks HP20 UB, U.K.
12 Avenue du Brezet
63039 Clermont-Ferrand, France J AMES, Charles M.
Kuwait Inst. Scient. Res.
GALKOVSKAYA, Galina P.O. Box 1638
Inst. ZooI., Acad. Sci. BSSR 22017 Salmiya, Kuwait
Academytcheskaya 27
Minsk 220072, U.S.S.R KING, Charles E.
Dept. Zoology, Oregon State Univ.
GILBERT, John J. Corvallis, Oregon 97331, U.S.A.
Dept. BioI. Sci.
Dartmouth College KORSTAD, John
Hanover, New Hampshire 03755, U.S.A. Dept. Biology, Oral Roberts Univ.
Tulsa, Oklahoma 74171, U.S.A.
GUEVARA ESPINOSA, Maria Dolores
Sagasta 100, 50 D. KOSTE, Walter
35008 Las Palmas G.c., Spain Rontgenstrasse IjOfen.
2903 Bad Zwischenahn, F.RG.
GUISANDE GONZALEZ, Castor
Department of Ecology, Faculty of Biology KUHN, Jurgen
41080 Sevilla, Spain Zoologische Institut
Berliner Strasse 28
GULATI, Ramesh 3400 Gottingen, F.RG.
Limnol. Inst. Koninklijke
Rijkestraatweg 6 KUNICKI-GOLDFINGER, Wladys1aw J. H.
3631 AC Nieuwersluis, The Netherlands Warsaw University, Inst. Microbiology
Nowy Swiat 67
HAGIWARA, Atsushi 00-046 Warsaw, Poland
Graduate School of Marine Science and
Engineering, University of Nagasaki KUTIKOV A, Ludmilla A.
Bunkyo 1-14 ZooI. Inst. Acad. Sciences
Nagasaki City 852, Japan Leningrad B 34, 199034, U.S.S.R.

HERNANDEZ CRUZ, Carmen Maria LAXHUBER, Ruth

Centro Tecnologia Pesquera Zool. Inst. LMV-Munchen
Apdo 56 Seidlstrasse 25
35200 Telde Las Palmas G.c., Spain 8000 Munchen 2, F.R.G.
XII

LUBZENS, Esther MORALES BAQUERO, Rafael

Israel Oceanogr. LimnoI. Res. Catedra Ecologia, Fac. Cienc., Univ. Granada
Tel-Shikmoma, P.O. Box 8030 18071 Granada, Spain
Haifa, Israel
NAGATA, Warren D.
MARIANI, Mauro Malaspina College, 900 Fifth Street
Acquario Civico e Stazione Idrobiologica Nanaimo, B.C., V9R 5S5, Canada
V.le Gadio 2
20121 Milano, Italy NOGRADY, Thomas
Dept. BioI. Queen's Univ.
MASSUTI, Sofia Kingston K7L 3N6, Canada
G.E.S.A. Cta II "Es Murterar"
Alcudia- Mallorca, Spain OLSEN, Yngvar
SINTEF Div. Applied Chemistry
MATVEEVA, Lilian K. 7034 Trondheim, Norway
Severtsov Inst. EvoI. MorphoI.
Leninskii pro 33 PAGANI, Manuela
Moscow 117071 U.S.S.R. Dip. Biologia Universita di Milano
Via Celoria 26
MAUTI, Rita 20133 Milano, Italy
Acquario Civico e Stazione Idrobiologica
V.le Gadio 2 PAULI, Hans-Rainer
20121 Milano, Italy Limnologisches Inst.
7750 Konstanz, F.R.G.
MAY, Linda
PEJLER, Birger
Inst. Terrestrial EcoI.
Inst. Limnology, Box 557
Bush Estate, Penicuik
75122 Uppsala 1, Sweden
Midlothian EH26 OQB, U.K.
PEREGO, Cristina
MELONE, Giulio Acquario Civico e Stazione Idrobiologica
Dip. BioI. U. Milano V.le Gadio 2
Via Celoria 26 20121 Milano, Italy
20133 Milano, Italy
PONTIN, Rosalind M.
26 Hermitage Woods Crescent
MIKSCH!, Ernst
St. John's, Woking, Surrey GU211UE, U.K.
Goethegasse 4
2380 Perchtoldsdorf, Austria
RADWAN, Stanislaw
Acad. Agric. Dept. ZooI. HydrobioI.
MINKOFF, Gidon Akademicka 13
Tinamenor S .A. Pesues 20-934 Lublin, Poland
Cantabria, Spain
RICCI, Claudia
MIRACLE, Maria Rosa Dip. BioI. An., Un. Torino
Dept. EcoI., Fac. Cien. BioI., Univ. Valencia Via Accademia Albertina 17
46100 Burjasot, Valencia, Spain 10123 Torino, Italy
 XIII

ROCHE, Kennedy SNELL, Terry W.

Inst. Zool., State Univ. Ghent Div. Sci. Math., Univ. Tampa
K.L. Ledeganckstraat 35 Tampa, Florida 33606, U.S.A.
9000 Gent, Belgium
STARKWEATHER, Peter L.
ROTH HAUPT, Karl. O. Dept. BioI. Sci., Univ. Nevada
Max Planck Inst. fur Limnologie Las Vegas, Nevada 89154, U.S.A.
Postfach 165
2320 Plan, F.R.G. SUDZUKI, Minoru
Nihon Daigaku
RUTTNER-KoLISKO, Agnes Higashi-Arai 557, Omiya-shi
Biologische Station Saitama-Ken, 330 Japan
3293 Lunz am See, Austria
VADSTEIN, Olav
SAUNDERS-DAVIES, Anthony SINTEF Div. Applied Chemistry
33 Park Road 7034 Trondheim, Norway
Esher, Surrey KTI0 8NP, U.K.
VELLANO, Camillo
SCHABER, Peter Dip. BioI. Animale
Abt. Limnol. Inst. Zool. Via Accademia Albertina 17
Technikerstrasse 13 10123 Torino, Italy
6020 Innsbruck, Austria
WALLACE, Robert Lee
SCHMID-ARAYA, Jenny M. Dept. Biology, Ripon College
Dep. Biology RHBNC Ripon, Wisconsin 54971, U.S.A.
Egham, Surrey TW20 9TY, U.K.
WALSH, Elizabeth J.
SCHRIMPF, Andreas
Dept. Zoology OSU
Wasserwirtschaftsamt Rosenheim
Corvallis OR 97331, U.S.A.
Konigstrasse 19
8200 Rosenheim, F.R.G.
W ALZ, Norbert
SERRA, Manuel Zool. Inst., Univ. Milnchen
Dept. EcoI., Fac. Cien. BioI., Univ. Valencia Seidlstrasse 25
46100 Burjasot, Valencia, Spain 8000 Miinchen 2, F.R.G.

SERRANO MARTIN, Laura WILMS, Anja

Department of Ecology, Faculty of Biology Limnol. Inst. Koninklijke
41080 Sevilla, Spain Rijkestraatweg 6
3631 AC Nieuwersluis, The Netherlands
SHIEL, Russell J.
Murray Darling Freshwater Res. YUFERA, Manuel
P.O. Box 921, Albury N.S.W. Inst. Cien. Mar. Andalucia CSIC
2640 Australia Poligono S. Pedro sin Apdo oficial
Puerto Real, Cadiz, Spain
SKJOLDAL, Lillian
Insitute of Marine Research ZOUFAL, Wolfgang
Division of Aquaculture Schweglerstrasse 4214
Austevol, 5392 Storeb0, Norway 1150 Wien, Austria
XIV

Plate 1 (see facing page):

Upper panel: Giulio Melone, Eric Hollowday, Henri
Dumont, Claudia Ricci, Manuela Pagani; Mila Kutikova,
Minoru Sudzuki, Mila Kutikova, Rama Chengalath.
Upper middle panel: Terry Snell, Davi Ehlert, Charles King,
Lilian Matveeva.
Lower middle panel: Peter Starkweather, Manuela Pagani,
Claudia Ricci, Giulio Melone; Tom Nogrady, Alois Herzig,
Walter Koste.
Lower panel: Walter Koste, Tommy Edmondson, Yvette
Edmondson; Heidi Berner-Fankhauser, Birger Pejler; John
Gilbert.

Plate 2 (see p. xvi):

Upper panel: coffee break in the garden of Palazzo Feltrinelli;
Excursion to Verona.
Upper middle panel: Hans-Rainer Pauli, Atsushi Hagiwara,
Linda May, Manuel Serra, Claudia Ricci, Nan Duncan,
Manuel Yufera, Luis Cruz-Pizarro, Maria Rosa Miracle,
Tom Nogrady; Charles King, Linda May, Bob Wallace, Eric
Hollowday, Tony Saunders-Davies, Rosalind Pontin.
Lower middle panel: Henri Dumont, Agnes Ruttner-Kolisko,
Peter Schaber, Wolfgang Zoufal, Pierre Clement, Rama
Chengalath; Claudia Ricci, Gala Galkovskaya, Tommy
Edmondson, Birger Pejler, Giulio Melone, Maria Rosa
Miracle, Gideon Minkoff, Henri Dumont, Ruth Laxhuber.
Lower panel: Back row: Esther Lubzens, Warren Nagata,
Russ Shiel, Bob Wallace. Centre row: Kennedy Roche,
Margaretha De Ridder, Ruth Laxhuber, Walter Koste. Front
row: Mauro Mariani, Claudia Ricci, Giulio Melone, Maria
Rosa Miracle; Pierre Clement, Terry Snell, Manuel Yufera.
Plate 1
Plate 2
 XVII

Pater Dr.h.c. Josef Donner (28.2.1909-8.1.1989) in memoriam

Suddenly and unexpectedly the "father of the bdelloids" (in Claudia Ricci's words) died in January 1989
at the age of eighty years. His life was divided between religious vocation and scientific research. He was
able to fulfill both tasks to a high degree without conscientious scruple.
Josef Donner spent his youth and school years in Northern Bohemia (at that time part of the Austrian
monarchy) and after ordination as Redemptorist priest he taught biology at the school of his convent.
During World War II he was transferred to a small parish in Southern Bohemia. During that time he
became fascinated with the microscopical life in ponds and streams. He bought a microscope of his own,
which he considered his most valuable worldly possession. The first publications dating from those years
deal mainly with monogonont rotifers.
After the end of the war Pater Donner - being German and a priest - was evicted from his parish,
now situated in Communist Czechoslovakia. Carrying a small rucksack and his microscope he arrived
on foot at the Biological Station, Lunz to meet his friend and teacher, Prof. V. Brehm, who - also
a refugee - had found a simple shelter here. That was the occasion when I met Pater Donner first; the
story of his escape is still vividly in my mind.
A short stay at the monastery Admont (situated in a boggy region) initiated Josef Donner's interest
in the fauna of humic soils, where bdelloid rotifers are particularly abundant, a group which was
practically unknown so far; he specialized and published his first papers on bdelloids.
After the interlude in Admont Pater Donner settled for several years at an extension of his own order
in Vienna, where he was again teaching and supervising schoolchildren. Even here, in the surroundings
of a big city, he continued his investigations on bdelloids. The result was a book "Die Ordnung
Bdelloidea" (Best. Bucher zur Bodenfauna Europas, 1965), which made him renowned worldwide.
XVIII

Already several years earlier a very useful little booklet "Radertiere" had appeared in the Kosmos' Series
(Einfiihrung in die Kleinlebewelt, 1956), which was translated into English by H.G.S. Wright in 1966.
His final homestead found Pater Donner in the Redemptorist-Convent in Katzelsdorf (Lower Austria),
where he taught biology in the private high school of this convent. But, after a few years, in recognition
of his scientific fame, he was released from teaching by his superiors. In conversations with scientific
colleagues he always very thankfully acknowledged this concession. Now completely dedicated to
research, this fact became visible very soon in Josef Donner's output of papers; altogether they amount
to approximately fifty. * Although he regretted losing contact with young people, he now concentrated
intensively not only on taxonomy of rotifers, but included also ecological and coenological aspects. It was
a lucky coincidence that he found at the same time a connection with the newly founded working group
investigating the Danube (I.A.D.). Extensive papers on the litoral zone and the river meadows of the
Danube, and also on the reed belt of the Neusiedlersee were produced in cooperation with this working
group.
These broader aspects of Pater Donner's work were the main, but not the only reason, that he was
elected in 1975 Doctor honoris causa by the University of Salzburg. Rightly, Josef Donner considered the
election the crowning of his scientific life.
While compiling Dr. Donner's life history and his own personal notes, it became very obvious to me,
how intensely physical fate, deep religious vocation and scientific aspiration have been woven together
in the modest, kind and pleasant personality of our dear colleague Pater Dr. phil. h.c. Josef Donner.

A. RUTTNER-KoLISKO

* A complete list of publications appears in "Wasser und Abwasser', Bd. 32.

Hydrobiologia 186/187: 1-9, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 1
© 1989 Kluwer Academic Publishers.

Rotifer study as a way of life

W.T. Edmondson
Department of Zoology, University of Washington Seattle, WA 98195, USA

Introduction

When Claudia Ricci asked me to present a review of my research program on rotifers at the Fifth
International Rotifer Symposium I was puzzled about what to do because I did not think I had a program.
All my life various opportunities have come up and I have gone along step-by-step, doing whatever seemed
most interesting at the time, within the normal constraints of existence. In fact, when I look back now
I can see a sort of pattern, but it was not the result of planning. Apparently there was some selection
on my part from a random assortment of opportunities.
A review of one's work necessarily involves some autobiography. To understand what I tell you about
my research requires some information about my early activities. Investigation is an integral part of a
scientist's life, and there is no way to discuss it intelligibly without the biographical background. Chance
happenings in one's personal life can change the direction of his professional experience.

Learning A turning point came when, at about age 12, I

was given a toy microscope, built something like
I was born in Milwaukee, Wisconsin, on the shore Leeuwenhoeck's, which permitted me to see what
of Lake Michigan, on April 24, 1916 and lived those minute gliding points looked like. Eventually
there for two years. As soon as I could walk my I was able to recognize that the first rotifer I had
mother would take me on strolls along the lake- seen was Scaridium longicaudum. (The first sight
shore every afternoon. We then moved to of a rotifer must have been an important moment
southern Indiana where there were no lakes, only in the life of every rotiferologist.) An even more
ponds, creeks and rivers. They were very different, decisive point came a year later when I learned
but I was fascinated by them and liked to look about Ward and Whipple's Freshwater Biology.
into them to see what was going on. Fish bored My grandmother gave me money for it as a birth-
me, but crayfish were exciting, fairy shrimp beauti- day gift and sewed a cloth carrying case because
ful, and Chaoborus was mysterious. There seemed it was so heavy. Of course that wonderful book
to be other things, barely visible, gliding around opened the world of rotifers for me, and a lot
and generating wonder. more. You can imagine my pleasure when, some
At home I had natural history books and read 20 years later, in 1949, I was asked to edit a
everything I could find about freshwater animals second edition. That finally appeared in 1959, one
in the public library. Several books mentioned year past the 50th anniversary of the first edition.
rotifers, sometimes with illustrations, but gave no
real information. The most that any book did was
to emphasize how beautiful and interesting High School
rotifers were, making them seem most desirable of
all. This all gave me a yearning for real informa- A major step forward came when we moved to
tion about them. New Haven, Connecticut, in 1930,just in time for
2

me to start high school. Hillhouse High School and Gosse and had been eager to see one (one!
was an excellent institution with a staff of highly - hundreds of colonies in view at once).
professional teachers. Many of them were taking Hutchinson invited me to use his microscopes.
courses at Yale. I was a freshman in high school at the time and
One day I went to the biology teaching rooms, happily took him up on the invitation. At some
entered the first one where there was a teacher, time he suggested that I write to F. J. Myers and
and asked whether I could use a microscope after I entered into a long correspondence and ex-
school to look at pond water. By accident I had change of slides with Myers. Hutchinson taught
found the best possible person, Miss Ruth Ross, me how to make slides with the glycerine and
for she let me use the microscopes and helped me melted paraffin technique. I would send Myers a
in many other ways. slide of a difficult specimen labelled with the name
For months I spent at least an hour after school of what I thought it was, and he would send it
almost every day looking at samples from ponds back with a corrected label or with congratula-
around New Haven. My interest was not limited tions for having got it right, often accompanied by
to rotifers. The filamentous algae were very attrac- some slides of his own. He had trouble with my
tive (Spirogyra, Mougeotia, Bulbochaete), the pro- handwriting so he gave me a typewriter to make
tozoa and crustaceans fascinating. The only our correspondence easier. During this period I
requirement was that it be small and beautiful. So was concentrating on learning the rotifers and
with the help of Ward and Whipple I found my how to study them.
way around in the pond world. I got more and I read Hudson and Gosse intently, going so far
more interested in the things that attached to as to memorize this part of a wonderful passage
plants, and the sessile rotifers were best. All in the Introduction:
through this I think I must have been as motivated
by esthetics as by any scientific consideration; I , '" But if, retaining sense and sight, we could
certainly was not generating any hypotheses. shrink into living atoms and plunge under the
Miss Ross was taking a course in histology water, of what a world of wonders should we
from a very kindly, grandfatherly professor at then form part! We should find this fairy king-
Yale, Wesley Roswell Coe, and he agreed me to dom peopled with the strangest creatures: -
let me attend his lectures, which I enjoyed. How- creatures that swim with their hair, that have
ever, my interests were obviously more toward the ruby eyes blazing deep in their necks, with tele-
invertebrates, so Dr. Coe took me around to meet scopic limbs that now are withdrawn wholly
Professor Lorande Loss Woodruff, a proto- within their bodies and now stretched out to
zoologist. He was interested in the history of many times their own length. Here are some
science and had collected an amazing number of riding at anchor, moored by delicate threads
classical early works which he let me browse in. spun out from their toes; and there are others
He had the wonderful Ehrenberg book of 1838, flashing by in glass armour, bristling with sharp
Hudson & Gosse, and many others. Once he spikes or ornamented with bosses and flowing
realized what I was interested in, he took me a few curves; while, fastened to a green stem, is an
doors down the hall to introduce me to the animal convolvulus that by some invisible
youngest instructor, G. Evelyn Hutchinson. As power draws a never-ceasing stream of victims
soon as I mentioned rotifers a dramatic thing into its gaping cup, and tears them to death with
happened. Hutchinson turned, muttering about hooked jaws deep down within its body.
having just been collecting, poured some water Close by it, on the same stem, is something
from a thermos bottle into a finger bowl and put that looks like a filmy heart's-ease. A curious
it under a dissecting microscope. It was the wheelwork runs round its four outspread
densest group of Conochilus hippocrepis that I have petals; and a chain of minute things, living and
ever seen. I had read about this animal in Hudson dead, is winding in and out of their curves into
 3

a gulf at the back of the flower. What happens what they were, and noticed fascinating objects
to them there we cannot see; for round the stem swimming past; rotifers, of course. After a period
is raised a tube of golden-brown balls, all regu- of exploring local ponds, he wrote to H. S.
larly piled on each other. Some creature dashes Jennings at the University of Pennsylvania for
by, and like a flash the flower vanishes within help in identification. Jennings had trouble
its tube. because he was familiar with the fauna of alkaline
We sink still lower, and now see on the waters only, and Myers had been collecting
bottom slow-gliding lumps of jelly that thrust a in Sphagnum bogs. Jennings recommended H.K.
shapeless arm out where they will, and, grasp- Harring as a better source of information.Harring
ing their prey with these chance limbs, wrap was a specialist in precise screw instruments at
themselves round their food to get a meal; for the U. S. Bureau of Standards in Washington.
they creep without feet, seize without hands, eat Somehow he became interested in rotifers and
without mouths, and digest without stomachs. built a microscope. According to Myers, Harring
Time and space, however, would fail me to completed his monumental study of the literature,
tell of all the marvels of the world beneath the 'Synopsis of the Rotatoria' before seeing a live
waters. They would sound like the wild fancies rotifer. Chancey Juday at Wisconsin learned
of a child's fairy tale, and yet they are all literally about the work of Harring, and invited him to
true; and, moreover, nearly all of them are true work on the rotifer fauna of Wisconsin, leading to
of that rotiferous world which it is my purpose the set of five papers in collaboration with Myers.
to describe.' In 1932 Hutchinson was Biologist of the First
Yale North India Expedition. When he came
In high school I took two years of German and back to New Haven in January, 1933, he had a
practiced by translating many pages of Remane's fabulous collection of material, from protozoa to
section on rotifers in Bronn's Klassen und fish, from many ponds, lakes and streams in a
Ordnungen des Tier-Reichs. I was particularly range of altitude from 528 m to 5334 m in the
fascinated by the retrocerebral organ and struck Himalayas and adjacent regions. He sent out
by a seeming anatomical correlation: rotifers with most of the collections to specialists for identifi-
the best developed retrocerebral-subcerebral sys- cation but kept the rotifers and asked me to
tem tended to have the simplest protonephridial identify them. He cleared off one end of a bench
system (compare Remane's figs. 119 and 266). I in his laboratory and set me up there with a couple
never pursued the point. of microscopes for my own use to do the work on
In 1932, Miss Ross took me down to meet Mr. his collections. For the next six years this comer
Myers at the American Museum of Natural His- of his laboratory was the center of my existence.
tory in N ew York where he was an honorary Hutchinson's substitute while he was away was
Research Associate. Later Myers invited me Dr. Richard M. Bond who had a lot of material
several times to visit him in his home in Ventnor, from Hispaniola, and he, too, asked me to identify
New Jersey, which is near the Pine Barrens his rotifers. So my first published paper, on the
region, full of interesting ponds, including rotifers of Hispaniola, came out in the spring of
Sphagnum bogs with a remarkably diverse rotifer my last year in high school, and the second, writ-
fauna. ten jointly with Hutchinson, came out later in the
Myers was independently wealthy, and was year, just after I entered college.
able to spend all his time on his hobby, rotifers. The lab was a busy place. Hutchinson's repu-
He had been an art student in college, which tation in limnology was spreading, and it was
accounts for the excellent illustrations in his becoming widely understood that there was a cen-
papers. His interest in rotifers started when he ter in New Haven quite different intellectually
noticed strange spots on some tropical fish he from those in Wisconsin and Michigan. This dif-
had. He bought a microscope so that he could see ference is made clear in Beckel's (1987) account
4

oflimnology in Wisconsin (see pp. 24-25) and by tology. In addition to auditing Histology I had
Cook's (1977) review of a controversy about a taken Hutchinson's course in Invertebrate Zoo-
paper by Lindeman (1942). As I was sitting at one logy (with examinations!) while I was in high
end of the room looking at rotifers, Hutchinson school. This time I enrolled formally so that they
would be running chemical analyses at the other would appear on my college record. I liked his-
end, calculating the data, and expressing surprise tology, probably because of the interesting
or pleasure at some new finding. It was interesting functional patterns of the cells in the tissues and
to read the formal publication of the outcome as, the colorful stains. I still treasure my slide of
for instance, in the 1941 Ecological Monographs Amphiuma spleen, vastly overstained with
or the 1938 Proceedings of the National Academy Mallory triple stain. Each student chose a topic of
of Sciences. Many visitors, foreign and domestic, special interest to him as part of the laboratory
went out of their way to visit Hutchinson's labora- work. I made serial sections in paraffin of Lindia
tory. tecusa. This was easier than it may sound because
the specimens, supplied by Myers, were a full
millimeter long. For orientation, I laid the animal
College along one of my hairs while the paraffin was still
soft and transparent. I was then told that histo-
All during my undergraduate years I spent as logists went to great lengths to keep such objects
much time in Hutchinson's laboratory as possi- out of the blocks because they caused the paraffin
ble, working on local material and collections ribbon to tear and my project would fail. Fortu-
from other places, publishing a few new species. nately my knife was sharp, and I have a nice clear
I continued to collect in ponds because of my section of hair next to each slice of rotifer.
interest in sessile rotifers. Inevitably I was sub-
consciously absorbing information about charac-
teristics of the best ponds for various kinds of Graduate school
rotifers and what plants were best for the sessile
ones. I helped Hutchinson's first two graduate When I was about to graduate, Hutchinson sug-
students, Gordon A. Riley and Edward S. gested that it would be good to get experience
Deevey, Jr., with their field work, thus gaining somewhere else, so I went for one year to the
experience with lake plankton and broadening my University of Wisconsin, primarily to take Juday's
interests to population and community ecology. limnology course (Birge was no longer teaching).
I entered into correspondence with other rotifer Another graduate student in Juday's course,
investigators including E. H. Ahlstrom, P. M. de Yvette Hardman, helped me collect rotifers and I
Beauchamp, B. Carlin, J. Hauer, M. Neal, A. helped her with her work on surface tension of
Ruttner-Kolisko, J. Wizniewski and K. Wulfert. lakes (Hardman, 1941). We were married in 1941.
We exchanged drawings and slides of new and By then I had decided to write a taxonomic mono-
unusual species. Much later, in 1959, my wife and graph on sessile rotifers (I still have 5 notebooks
I visited de Beauchamp in Paris. He showed us of information) and to do a Ph. D. on some aspect
his collection of brightly colored wooden working of their ecology. Through the kindness ofProfes-
models of different types of trophi. Each of us sor Juday I had an assistantship to work at the
could read the other's writing in his own language Trout Lake Laboratory in northern Wisconsin for
but not understand the other's speech in any a full summer. When I asked what my assignment
language, so that the conversation was conducted would be, he said 'Study the sessile rotifers'. I
mostly on paper with an occasional oral assist collected lots of taxonomic and ecological data
from my wife. having to do with environmental factors (1944
I took courses in a wide spread oftopics includ- paper).
ing Hutchinson's invertebrates and Coe's his- I went back to Yale in 1939, and as a graduate
 5

student had my own space so was no longer able the war, I stayed at Woods Hole for a year
to eavesdrop on Hutchinson's fascinating conver- working on fertilization of salt ponds with
sations with visitors. nutrients.
My Ph.D. dissertation might have been limited
to the kind of distributional problem covered in
Harvard University
my 1944 paper but for a chance observation. One
day I took a collection of Utricularia covered by
In 1946 I joined the Department of Biology at
Floscularia conifera with a striking appearance. All
Harvard to help teach a new General Education
of the animals except the small ones had a dark
course. While I was in Cambridge, Andre Burger
band across the tube, much as in the figure on
came from Switzerland for a year to work on
Plate V in Hudson and Gosse. I suddenly realized
bdelloids and discovered the interesting anatomi-
that it had rained heavily the day before, and the
cal structure of the stomach of Habrotrocha. The
dark bands consisted of pellets made while the
work became widely known when Libbie Hyman
water was mUddy. It occurred to me that this way
included it in the third volume of her treatise on
of marking tubes could tell how fast the animals
invertebrates (1951). He also found, at my sug-
were growing and settling, because the clear part
gestion, that the growth of some bdelloids in cul-
above the dark band would show how much the
ture was increased by adding little chips of glass
animals had grown between the time and the mud
because they improved conditions for egg-laying.
settled and the time they were collected. The
small, young ones with no dark band must have
attached after the water had cleared. I was plan- University of Washington
ning to go out with a stick and stir the mud up
again, when I thought of a better way: pouring in When I was invited to move to the University of
a cloud of carmine. As recorded in my 1945 paper, Washington in 1949 I expected to resume work on
this permitted getting several kinds of population rotifers, and did so, but the emphasis of the work
dynamics data. Although Hutchinson was at first changed from studying them for themselves to
skeptical, it worked, and he later included the studying them as parts of communities in the
experiment in an entry on populations in the context of general ecological problems.
Encyclopedia Britannica. During this work I I re-established communication with my pre-
realized that the eggs held inside the tube were the war rotifer correspondents and soon added over
next generation of animals. This led to the idea a dozen more including B. Berzins, J. Donner, J.
that a count of eggs could be used for calculating Evans, A. L. Galliford, E. D. Hollowday, T.
the birth rate, an idea I was not able to implement Nogradi, B. Pejler, R. Pourriot, C. Rudlin, V.
immediately, but did for planktonic rotifers more Sladecek, M. Sudzuki and H. G. S. Wright.
than fifteen years later. (My official retirement in Some of the British workers formed a Rotifer
1986 was marked by the presentation by my Society in 1949. The number of new people inter-
colleagues of an elegant bronze Floscularia, ested in rotifers was growing and included a large
standing 21 cm high and complete with a band on proportion with academic positions where they
the tube). were able to apply advanced techniques to prob-
lems of systematics, physiology, morphology and
ecology. This development eventually made pos-
World War II sible the remarkable series ofInternational Rotifer
Symposia, five to date.
During the war I worked as a civilian for the U. S. The first summer in Seattle my wife and I drove
Navy, mostly at Woods Hole Oceanographic allover western Washington collecting in lakes to
Institution, on a variety of marine problems, so see what we had to work with. I had wanted to
rotifers were not a large part of my thinking. After continue with carmine marking experiments, but
6

did not find suitable populations. There were ses- could eat, accompanied by samples in which I
siles, but there was no Utricularia near Seattle. could count rotifer eggs. By applying the informa-
There was considerable interest at that time in tion gained at Pallanza, I could calculate birth
Gause's work on competition. I had read some- rates and relate them to food supply. This resulted
where that Asplanchna priodonta and A. brightwelli in a long paper in the 1965 Ecological Mono-
did not occur together, but I noticed that the two graphs. This method of calculation can be applied
did in fact occur together in a collection from Hall to any species of animal that carries its eggs.
Lake. When I went back to the lake to begin a Many people have used it for Cladocera and
study of the situation there was no Asplanchna to Copepoda, and the methods for calculating the
be found at all, but I discovered that the lake was rates have been improved.
meromictic, a condition that interested me greatly, After returning to Seattle I had several visitors
so I started what turned out to be a three-year who came to do research on rotifers. The first was
study. We also found the saline lakes in the Lower Agostino Parise whom I had met at Pallanza. He
Grand Coulee to be interesting. Soap Lake and arrived soon after we returned and worked for a
Lake Lenore had an usual Hexarthra, which year, mostly on population dynamics of
Koste much later named H. polydonta soaplakeien- Euchlanis. (When he returned to Italy in 1961 he
sis [sic], based on my material. Through unusual left some cultures which were soon found by
circumstances one of my graduate students was Charles King. Charles came to work as a graduate
able to spend two years at the Arctic Research student with a marine invertebrate zoologist,
Laboratory at Point Barrow, Alaska, incidentally but fortunately he discovered rotifers and did a
providing the material on Kellicottia for a paper Ph.D. study of population dynamics of
included in this volume. Euchlanis.) John Gilbert finished his Ph.D. with
In 1954 the Coulee lakes suddenly gained new Hutchinson and came here in 1963 with a Post-
interest when we found that they were being doctoral fellowship from the Public Health Ser-
diluted by irrigation water and the plankton was vice [sic]. Starting from observations by de Beau-
changing. Likewise, the next year Lake Washing- champ, Gilbert carried out his magnificent set of
ton showed signs of eutrophication and the plank- experiments on the induction of spines on
ton changed. We started studies of the lakes as Brachionus calycif/orus by Asplanchna brightwell.
ecosystems, with emphasis on the plankton com- Robert Wallace finished a Ph.D. on sessile rotifers
munities. Since then, much of my time and atten- with Gilbert in 1974 and came to Seattle for two
tion has been given to studies of situations that are years. He took a unique approach to substrate
accidental whole-lake experiments. While the selection that successfully combined his knowl-
entire community is the object of study, I look for edge of larval behavior and plant physiology. All
interesting rotifer conditions, such as that along, as these things were happening, I con-
reported for Conchilus at the fourth rotifer sym- tinued to work on the plankton ecology of lakes.
posium. The studies of the diluted saline lakes, the
In 1959-60, I had a Senior Postdoctoral Fel- eutrophied Lake Washington and others have
lowship from the National Science Foundation been fascinating. I have paid a great deal of atten-
for study in Europe. I wanted to develop the idea tion of Daphnia which is a wonderful animal, but
about using the counts of the eggs of planktonic I have never lost sight of the rotifers. Every time
rotifers to calculate birth and death rates. We my assistants make a sampling trip on any lake
went to the Instituto Italiano di Idrobiologia on they bring back a live plankton haul so that I can
Lago Maggiore in Italy to get data on the rate of enjoy looking at the rotifers in it. I keep my eyes
egg development as a function of temperature. open for anything unusual, and sometimes collect
Armed with that information, we went on to extra material for future attention. I have a num-
Windermere where I found that John Lund had a ber of small unfinished projects similar to the one
long set of counts of phytoplankton that rotifers with Kellicottia, which I hope to be finishing up in
due time.
 7

References 9. Edmondson, W. T., 1939. New species of Rotatoria with

notes on heterogonic growth. Trans. Am. Micro. Soc. 58:
Beckel, A. L. 1987. Breaking new waters. A century of 459-472.
limnology at the University of Wisconsin. Trans. Wis. 10. Edmondson, W. T., 1940. The sessile Rotatoria of
Acad. Sci. Arts & Lett. Special Issue. Wisconsin. Trans. Am. Micr. Soc. 59: 433-459.
Burger, A. 1948. Studies on the moss dwelling bdelloids 11. Edmondson, W. T., 1944. Ecological studies of sessile
(Rotifera) of eastern Massachusetts. Trans. am. Micro. Rotatoria. Part I. factors affecting distribution. Eco!.
Soc. 67: 111-142. Monogr. 14: 31-66.
Cook, R. E. 1977. Raymond Lindeman and the trophic- 12. Edmondson, W. T., 1945. Ecological studies of sessile
dynamic concept in ecology. Science. 198: 22-26. Rotatoria. Part II. Dynamics of populations and social
Gilbert, J. J. Asplanchna and postero-lateral spine production structures. Eco!. Monogr. 15: 141-172.
in Brachionus calyciflorus. Arch. Hydrobio!. 64: 1-62. 13. Edmondson, W. T., 1946. Factors in dynamics ofrotifer
Hardman, Y. 1941. The surface tension of Wisconsin lake populations. Eco!. Monogr. 16: 357-372.
waters. Trans. Wis. Acad. Sci. Arts Lett. 33: 395-404. 14. Clarke, G. L., W. T. Edmondson & W. E. Ricker, 1946.
Hudson, C. T. & P. H. Gosse. 1886. The Rotifera; or wheel Mathematical formulation of biological productivity.
animalcules. Longmans, Green. (Part of a symposium on dynamics of production in
Hutchinson, G. E. 1941. Limnological studies in Connecticut aquatic populations). Eco!. Monogr. 16: 336-337.
IV. Mechanism of intermediary metabolism in stratified 15. Edmondson, W. T. & Yvette H. Edmondson, 1947.
lakes. Eco!. Monogr. 11: 21-60. Measurements of production in fertilized salt water.
Hutchinson, G. E. 1938. Chemical stratification and lake Sears J. Mar. Res. 6: 228-246.
morphology. Proc. natn. Acad. Sci. U.S.A. 24: 63-69. 16. Bigelow, H. B. & W. T. Edmondson, 1947. Wind waves
King, C. E. 1967. Food, age and the dynamics of a laboratory at sea, breakers and surf. U.S. Hydrographic Office Pub.
population of rotifers. Ecology. 48: 111-128. 602 xi + 177.
Lindeman, R. L. 1941. The trophic-dynamic aspect of 17. Edmondson, W. T., 1948. Rotatoria from Penikese
ecology. Ecology. 23: 399-418. Island, Mass., with description of Ptygura agassizi, n. sp.
Biological Bulletin 94: 169-173.
18. Edmondson, W. T., 1948. Two new species Rotatoria
from sand beaches. Trans. Am. Micro. Soc. 67: 149-152.
Bibliography of W. T. Edmondson
19. Edmondson, W. T., 1948. Ecological applications of
Abstracts and book reviews are omitted. For a Lansing's physiological work on longevity in Rotatoria.
complete list see Limnology and Oceanography Science 108: 123-126.
Vol. 33, Part 1. 20. Edmondson, W. T., 1949. A formula key to the
Rotatorian genus Ptygura. Trans. Am. Micro. Soc. 68:
1. Edmondson, W. T., 1934. Investigations of some His- 127-135.
paniolan lakes. (Dr. R.M. Bond's Expedition) 1. The 21. Edmondson, W. T., 1950. Centrifugation as an aid in
Rotatoria. Arch. Hydrobio!. 26: 465-471. examining and fixing rotifers. Science 112: 49.
2. Edmondson, W. T. & G. E. Hutchinson, 1934. Yale 22. Comita, G. W. & W. T. Edmondson, 1953. Some aspects
North India Expedition. Article IX. The Rotatoria. of the limnology of an Arctic Lake. Stanford Univ. Pub.
Mem. Conn. Acad. Arts. Sci. 10: 153-186. Univ. Ser. BioI. Sci. 11: 7-13.
3. Edmondson, W. T., 1935. Some Rotatoria from Arizona. 23. Edmondson, W. T., 1955. Seasonal life history of
Trans. Amer. Micr. Soc. 54: 301-306. Daphnia in an Arctic Lake. Eco!. 36: 439-455.
4. Edmondson, W. T., 1936. Fixation of sessile Rotatoria. 24. Nelson, P. R. & W. T. Edmondson, 1955. Limnological
Science 84: 444. effects of fertilizing Bare Lake, Alaska. U.S. Fish &
5. Edmondson, W. T., 1936. New Rotatoria from New Wildlife Service. Fishery Bull. 102: 413-436.
England and New Brunswick. Trans. Am. Micro. Soc. 55: 25. Edmondson, W. T., 1955. Factors affecting productivity
214-222. in fertilized salt water. Deep Sea Res. 3: 451-464. (supp!.
6. Edmondson, W. T. & John L. Fuller, 1937. Food condi- for H. B. Bigelow)
tions in some New Hampshire lakes. New Hampshire 26. Edmondson, W. T., 1956. The relationship of photo-
Fish and Game Department Survey Report No.2. synthesis by phytoplankton to light in lakes. Eco!. 37:
Biological Survey of the Androscoggin, Saco and 161-174.
Coastal Watersheds. 95-99. 27. Edmondson, W. T., 1956. Measurement of conductivity
7. Edmondson, W. T., 1938. Notes on the plankton of some of lake water in situ. Eco!. 37: 201-204.
lakes in the Merrimack watershed. New Hampshire Fish 28. Edmondson, W. T., G. C. Anderson & Donald R.
and Game Department Survey Report No.3. Biological Peterson, 1956. Artificial eutrophication of Lake
Survey of the Merrimack Watershed. 107-210. Washington. Limno!. Oceanogr. 1: 47-53
8. Edmondson, W. T., 1938. Three new species of 29. Edmondson, W. T., 1956. Biological aspects of the
Rotatoria. Trans. Am. Micr. Soc. 57: 153-157. problem. (In A new critical phase of the Lake
8

Washington problem, by R. O. Sylvester, W. T. 45. Edmondson, W. T., 1968. A graphical model for evaluat-
Edmondson and R. H. Bogan). Trend in Engineering 8: ing the use of the egg ratio for measuring birth and death
11-13. rates. Oecologia 1: 1-37.
30. Edmondson, W. T., 1957. Trophic relations of the 46. Edmondson, W. T., 1969. Eutrophication in North
zooplankton. Trans. Am. Micr. Soc. 76: 225-245. America. Eutrophication: Causes, Consequences, Cor-
31. Edmondson, W. T., 1959. Ward and Whipple's Fresh- rectives. pp. 124-149. National Academy of Sciences
water Biology. Editor of second edition; author of Pre- Publication No. 1700.
face, Introduction and Chapters on Rotifera and 47. Edmondson, W. T., 1969. Cultural eutrophication with
Methods. special reference to Lake Washington. Mitt. int. Ver.
32. Edmondson, W. T., 1960. Reproductive rate ofrotifers in Limnol. 17: 19-32.
natural populations. Me. ist. Ita!. Idrobio!. 12: 21-77. 48. Edmondson, W. T., 1969. The present condition of the
33. Edmondson, W. T., 1961a. Changes in Lake Washington saline lakes in the Lower Grand Coulee, Washington.
following an increase in the nutrient income. Verh. Inter- Verh. int. Ver. Limnol. 17: 447-448.
nat. Verein. Limno!. 14: 167-175. (Reprinted 49. Griffiths, M., P. S. Perrott & W. T Edmondson, 1969.
pp. 364-372 in Ford, R. F. and W. T. Hazen, eds. 1972. Oscillaxanthin in the sediment of Lake Washington.
Readings in aquatic ecology, Saunders). Limnol. Oceanogr. 14: 317-326.
34. Edmondson, W. T., 1961b. Secondary production and 50. Edmondson, W. T., 1970. Phosphorus, nitrogen and
decomposition. Verh. int. Ver. Limnol. 14: 316-339. algae in Lake Washington after diversion of sewage.
35. Edmondson, W. T., 1962. Food supply and reproduction Science 196: 690-691. (Reprinted pp. 372-374 in Ford,
of zooplankton in relation to phytoplankton population. R. F., W. E. Hazen, eds. 1972. Readings in aquatic
Rapp. et Proc.-Verb. Cons. Internat. Explor. de la Mer. ecology, Saunders.).
153: 137-141. 51. Edmondson, W. T. & David E. Allison, 1970. Recording
36. Edmondson, W. T., G. W. Comita & G. C. Anderson, densitometry of X-radiographs for the study of cryptic
1962. Reproductive rate of copepods in nature and its laminations in the sediment of Lake Washington. Limnol.
relation to phytoplankton population. Ecology 43: Oceanogr. 15: 138-144.
625-634. 52. Shapiro, J., W. T. Edmondson & D. E. Allison, 1971.
37. Edmondson, W. T., 1963. Pacific Coast and Great Basin, Changes in the chemical composition of sediments of
pp. 371-392 In D. G. Frey (ed.) Limnology in North Lake Washington, 1958-1970. Limno!. Oceanogr. 16:
America. University of Wisconsin Press, Madison, 437-452.
Wisconsin. 53. Edmondson, W. T. & G. G. Winberg (editors), 1971. A
38. Edmondson, W. T., 1964. The rate of egg production by manual on methods for the assessment of secondary
rotifers and copepods in natural populations as con- productivity in fresh waters. IBP handbook No. 17.
trolled by food and temperature. Proc. int. Ass. Theor. Blackwell, Oxford.
and Appl. Limnol. 15: 673-675. 54. Edmondson, W. T., 1972. Nutrients and phytoplankton
39. Edmondson, W. T., 1965. Reproductive rate of plank- in Lake Washington. pp. 172-193 in Nutrients and
tonic rotifers as related to food and temperature in Eutrophication. American Society of Limnology and
nature. Eco!. Monogr. 35: 61-111. Oceanography, Special Symposia No.1. G. Likens (ed.).
40. Edmondson, W. T. & G. C. Anderson, 1965. Some fea- 55. Edmondson, W. T., 1972. The present condition of Lake
tures of saline lakes in Central Washington. Limnol. WAshington. Verh. int. Ver. Limnol. 18: 284-291.
Oceanogr. 10: (suppl.): R87-R96. 56. Edmondson, W. T., 1972. Instantaneous birth rates of
41. Edmondson, W. T., 1966. Changes in the oxygen deficit zooplankton. Limnol. Oceanogr. 17: 792-795.
of Lake Washington. Verh. int. Ver. Limnol. 16: 153-158. 57. Beeton, A. M. & W. T. Edmondson, 1972. The
42. Oglesby, R. T. & W. T. Edmondson, 1966. Control of eutrophication problem. J. Fisheries. Res. Bd. Canada.
eutrophication. Jour., Water Pollution Control Federa- 19: 673-682.
tion. 1966: 1452-1460. 58. Edmondson, W. T., 1973. Lake Washington.
43. Edmondson, W. T., 1967. Why study blue-green algae? pp. 281-298. In Environmental quality and water devel-
pp. 1-6 (introductory remarks) in Symposium: Environ- opment. Ed. C. R. Goldman, James McEvoy III and
mental Requirements of Blue-Green Algae. Pub. Pacific Peter J. Richerson. Freeman.
Northwest Water Laboratory, Corvallis, Oregon. 59. Edmondson, W. T., 1974. Secondary production. Mitt.
44. Edmondson, W. T., 1968. Water quality management int. Ver. Limnol. 20: 229-272. (See 1974 in Part II).
and lake eutrophication: The Lake Washington Case. 60. Edmondson, W. T., 1974. The sedimentary record of the
Water Resources Management and Public Policy. eutrophication of Lake Washington. Proc. Nat. Acad.
pp. 139-178. T. H. Campbell and R. O. Sylvester (eds.) Sci. USA 71: 5093-5095.
University of Washington Press, Seattle. 61. Edmondson, W. T., 1975. Microstratification of Lake
Note:This paper has been translated into Hungarian by Washington sediments. Ver. into Ver. Limnol. 19:
Dr Olga Sebestyen. Visminosegi ugyvetes es a 770-775.
tavak eutrofikacioja: A Lake Washington iigy. 62. Griffiths, M. & W. T. Edmondson. 1975. Burial of
 9

oscillaxanthin in the sediment of Lake Washington. 74. Edmondson, W. T. 1984. Volcanic ash in lakes. North-
Limno!. Oceanogr. 20: 945-952. west Env. Jour. I: 139-150.
63. Edmondson, W. T., 1977. Recovery of Lake Washington 75. Edmondson, W. T. 1985. Reciprocal relations between
from eutrophication. pp. 102-109 in Recovery and resto- Daphnia and Diaptomus in Lake Washington. Arch.
ration of damaged ecosystems. ed. J. Cairns, Jr., K. L. Hydrobio!. Beih. Ergeb. Limno!. 21: 475-481.
Dickson & E. E. Herricks, University Press of Virginia. 76. Infante, A. & W. T. Edmondson. 1985. Edible phyto-
64. Edmondson, W. T., 1977. Lake Washington in North plankton and herbivorous zooplankton in Lake
American Project - A study of United States Water Washington. Arch. Hydrobio!. Ergebn. Limno!. 21:
Bodies. A Report of the Organization for Economic 161-171.
Cooperation and Development. (EPA-600/3-77-086) 77. Edmondson, W. T. 1985. Recovery of Lake Washington
Published by Environmental Research Laboratory, from eutrophication. Proc. Internat. Congress Lake Pol-
Environmental Protection Agency, Corvallis, Oregon. lution and Recovery, Rome, April, 1985. pp.308-314
65. Edmondson, W. T., 1977. Trophic equilibrium of Lake (228-234 in Preprints).
Washington. Final Report on EPA Project R 8020 78. Wallace, R. L. & W. T. Edmondson. 1986. Mechanism
82-03-1. (EPA-600/3-77-087). Published by Environ- and adaptive significance of substrate selection in the
mental Research Laboratory, Environmental Protection sessile rotifer Collotheca gracilipes. Ecology 67: 314-323.
Agency, Corvallis, Oregon. 79. Edmondson, W. T. & A. H. Litt. 1987. Conochilus in
66. Edmondson, W. T., 1977. Population dynamics and Lake Washington. Proc. Fourth Internat. Rotifer Sym.
secondary production. In First International Symposium Hydrobio!. 147: 157-162.
on Rotifers. Ed. C. E. King. Arch. Hydrobio!. Beih. 80. Edmondson, W. T. & Sally E. B. Abella. 1988. Un-
Ergebn. Limno!. 8: 56-64. planned biomanipulation in Lake Washington. Limno-
67. Edmondson, W. T., 1979. Lake Washington and the logic a 19: 73-79.
predictability of limnological events. (Symposium on 81. Edmondson, W. T. 1987. Daphnia in experimental
Lake Metabolism and Lake Management celebrating the ecology: notes on historical perspectives. Me. 1st. Ita!.
500th Anniversary of the University of Uppsala). Arch. Idrobio!. 45: 11-30. (Actually published in 1988).
Hydrobio!. Beih. Ergebn. Limno!. 13: 234-241. 82. Edmondson, W. T. 1988. On the modest success of
68. Edmondson, W. T., 1979. Problems of zooplankton Daphnia in Lake Washington in 1965. p.223-243 in
popUlation dynamics. In R. de Bernardi (Ed.) Proc. Algae and the aquatic environment. Contributions in
Symp. Biological and mathematical aspects in popula- honour of1. W. G. Lund, C.B.E., F.R.S., ed. F. E. Round.
tion dynamics. Mem. 1st. Ital. Idrobio!. Supp!. 37: 1-11. Biopress.
(Actually appeared in 1980). 83. Edmondson, W. T. (in press). Lessons from Washington
69. Edmondson, W. T., 1980. Secchi disc and chlorophyl!. Lakes. International Mountain Watershed Symposium,
Limno!. Oceanogr. 25: 378-379. Lake Tahoe.
70. Edmondson, W. T. & J. T. Lehman. 1981. The effect of 84. Edmondson, W. T. & Y. H. Edmondson. (in press).
changes in the nutrient income on the condition of Lake Pallanza as a haven for visiting limnologists. Me. ist. Ita!.
Washington. Limno!. Oceanogr. 26: 1-29. Idrobio!.
71. Edmondson, W. T. & A. H. Litt. 1982. Daphnia in Lake 85. Edmondson, W. T. (in press). Perspectives in plankton
Washington. Limno!. Oceanogr. 27: 272-293. research. Me. ist. Ita!. Idrobio!.
72. Lehman, 1. T. & W. T. Edmondson, 1983. The 86. Edmondson, W. T. 1989. Rotifer study as a way of life.
seasonality of phosphorus deposition in Lake Proc. Fifth Internat. Rotifer Symp. Hydrobiologia
Washington. Limno!. Oceanogr. 18: 796-800. 186/187: 1-9.
73. Edmondson, W. T. & A. H. Litt. 1984. Mt. St. Helens ash 87. Edmondson, W. T. & Arni H. Litt. 1989. Morphological
in lakes in the Lower Grand Coulee, Washington State. variation in Kellicottia /ongispina. Fifth Internat. Rotifer
Verh. int. Ver. Limno!. 22: 510-512. Symp. Hydrobiologia 186/187: 109-117.
Hydrobiologia 186/187: 11-28, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 11
© 1989 Kluwer Academic Publishers.

Food limitation and body size in the life cycles of planktonic rotifers
and c1adocerans

Annie Duncan
Department of Biology, Royal Holloway & Bedford New College, University of London, Egham, Surrey
TW20 OEX, U.K.

Key words: food limitation, body size, duration of development, fecundity, life cycle studies, rotifers,
cladocerans

Abstract

This review considers what is known about the effects offood limitation upon the life cycle characteristics
of rotifers and planktonic cladocerans. The characteristics considered in rotifers are the size of eggs,
juveniles and adults and the durations of the juvenile phase and period of egg production. In cladocerans,
the life history features dealt with are their length-weight relationships, the body size, instar stage, age
and fecundity of the primiparous female and their fecundity-adult size relationship. The influence of
limiting food conditions is demonstrated for these characteristics by comparison with the situation in
non-limiting circumstances; the comparison is confined to experiments where food concentrations are
quantified. A direct comparison is made between rotifers and cladocerans in conditions of defined food
resource availability in terms of their length-weight relationships, the daily allocation of adults or
near-adults to growth and reproduction and their threshold food concentrations. These comparisons are
discussed in relation to the following topics: the high cost of cumulated respiration resulting from
prolongation of the juvenile phase of body growth; the fundamentally different nature of growth in the
two taxonomic groups; the body size of species and the size that must be attained for reproduction; the
ecological implications of the very different threshold food concentrations.

Introduction edged than the more thoroughly investigated

effects of temperature on the development of
Herbivorous zooplankton from large water rotifers, cladocerans and copepods (Bottrell
bodies often have to face food limitation for as et aI., 1976; Herzig, 1983a, b). For example,
long as a generation. Such limiting food con- King's (1967, Fig. 2) study on Euchlanis dilatata
ditions can affect several life cycle characteristics shows both a doubling of the juvenile phase and
which are important ecologically: the duration of a reduction in the body length of the primipara
development, the body size at which certain female in animals grown in ten times less algal
stages of development are attained and repro- food (16.4 and 1.64 jlgDW ml- 1). Pilarska's
duction begins and the fecundity of adults (King, (1977) eco-physiological investigation on Brachio-
1967; Hrbackova, 1971; Weglenska, 1971 ; nus rubens presents evidence of similar size and
Hrbackova & Hrbacek, 1979). Developmental time effects, provided that the range of test food
effects of food limitation are much less acknowl- concentrations include some limiting ones. This
12

was not so in Leimeroth's (1980) energy budget relation to geographical or seasonal morphologi-
for juvenile B. calyciflorus but is so in the unpub- cal variation within species and to whether this
lished study by Skrdla & Starkweather on the has a genetic origin (Pejler, 1980; King, 1977;
same species (Starkweather, 1987). Similarly in Snell, 1977). In aquaculture, variation in the body
crustaceans, Fig. 6 in Weglenska's (1971) paper size of Brachionus plicatilis is of direct interest for
shows that both body length and the age at which finding suitable strains to feed growing fish larvae
four species of Cladocera from Mikolajskie Lake (Snell & Carrillo, 1984; Lubzens, 1987) and there
attained maturity were changed when reared on is evidence that body size and biochemical com-
natural food manipulated to provide a wide range position of this species vary with food availability
in food concentration from 0.5 to 0.02 mgC L - 1 when reared in mass culture. There is little infor-
(converted approximately from 10 to mation on the ecological significance of changes
0.41 mgWW L -1). in body size, stage or duration of development
There is confusion in the literature on ecologi- within the life cycle that might be induced environ-
cal studies on body sizes in cladoceran popu- mentally.
lations because changes in the size distribution The aim of this paper is to review the evidence
detected in the field are usually attributed to the for the effects offood limitation upon the life cycle
presence of size-selective predation but may also characteristics of planktonic rotifers and clado-
result from food limitation. Similar uncertainty cerans. The cladoceran review is based upon a
may occur with rotifer populations, as, for exam- comparable series of unpublished studies of tem-
ple, in Parakrama Samudra whose small-size perate and tropical species of different body size
rotifer community was thought to be due to fish and in which the experiments were designed to
predation and not to the effects of food limitation clarify the food effects. There appears to be only
(Duncan, 1984). In planktonic communities, one comparable published study for rotifers, that
these two pressures often occur simultaneously so ofPilarska (1977) for B. rubens, although another
that there is a need to find techniques to dis- one, Skrdla & Starkweather on B. calyciflorus
tinguish between the two effects, considering the (Starkweather, 1987), will be available in future.
difficulty of quantifying food availability in the Later in the paper, some comparisons will be
field. For field populations of Daphnia species, made on the threshold food concentrations of
Duncan (1985) proposed the use oflength-carbon rotifers and cladocerans as well as on how the two
weight regressions whose elevations decrease groups allocate scarce food resources to growth
significantly as the food levels become limiting. and reproduction.
The duration of the period of food limitation can In this paper, the condition of food limitation
therefore be defined by regular monitoring of the is not taken to be equivalent to starvation, which
length-weight regressions of field populations exists below the threshold food concentration for
(Santos, 1989). In experimental and field studies growth and reproduction and which may lead to
on rotifers, the species' responses to limiting food some form of metabolic depression or diapause.
conditions is usually measured in terms of There may be a series of limiting concentrations
changes in density (as the instantaneous rate, r or from the optimal down to the threshold value
as the net reproductive rate per generation (Ro), which, in long-term life cycle studies, can taken to
which takes account of numbers of individuals be the minimal food level at which the life cycle
but not their body size or nutritive state. Changes can be completed (i.e. reproduction can occur).
in rotifer body size, shape and spines have been This differs from Lampert's (1977) definition that
reported and have generated considerable interest it is the concentration at which an animal just
in relation to avoidance of predation (Gilbert & balances its metabolic losses so that it cannot and
Stemberger, 1984) or cladoceran interference does not grow but does not lose weight. This
(Gilbert, 1985), in relation to the influence of definition, which implies different threshold
abiotic variables (Serra & Miracle, 1987) or in values for different stages in the life cycle, is the
 13

one adopted by Schiemer (1986) in his compari- Fenchel (1980) used another measure of food
son of food threshold for different aquatic inver- thresholds for ciliates, which was the minimal
tebrates but is not so easy to use ecologically. The food particle concentration needed to cover the
definition being used in this paper is rather similar metabolic costs when clearing water maximally at
to that employed by Sternberger & Gilbert (1985, low food concentrations.
1987) which is the food concentration at which
the instantaneous 'r' is zero or slightly above zero.
Body size of rotifers and c1adocerans
WEIGHT (,tl9 C)
300 1.2.3
Although rotifers and cladocerans differ greatly in
their definitive body sizes (Fig. 1), there is a size
200
of overlap between the two groups. The length-
carbon weight regressions for the three large tem-
100
perate species of Daphnia and for the smaller
tropical Daphnia gessneri and Moina reticulata
4 were measured by Santos (1989) and Hardy
(1989) under a range of known food conditions
and cover the full size range of the species. These
regressions overlap with the lengths and dry
10 weights of fourteen species of adult rotifers
measured by Sternberger & Gilbert (1987) and,
for this figure, converted to carbon (0.5 DW =
carbon weight). These rotifers included three
species of Asplanchna, two of Brachionus, one of
Keratella, two of Polyarthra and three of
Synchaeta. Also included in this figure are the
1 length-dry weights (converted to carbon) of adult
Keratella quadrata (Doohan & Rainbow, 1971)
and juvenile and adult Brachionus rubens
(Pilarska, 1977) as well as the carbon weights for
• Brachionus angularis and Keratella cochlearis
(Walz, 1983). The rotifer lengths and weights are
0,1
o 0"
• .. • largely for well-fed adults, apart from those for
B. rubens from Pilarska (1977) which consist of
,;
• "0
o litre (I), at 1 mgC per litre (2) and at 0.1 mgC per litre (3).
· 0
•• Regression 4 is for Daphnia magna reared in 0.01 mgC per
litre. Regressions 5 and 6 are for the tropical Daphnia gessneri
••• reared in food concentrations of 0.5 (5) and 0.05 (6) mgC per
litre. Regressions 7 and 8 are for the tropical Moina reticulata
0,01 • reared in food concentrations of 0.5 (7) and limiting food
levels (8). Black dots: adult rotifers from Sternberger &
0,05 0,1 1 2 3 4
Gilbert (1987). Empty circle: adult Brachionus angularls;
LENGTH (mm)
black square: adult Keratella cochlearls; from Walz (1983).
Empty square: adult Keratella quadrata from Doohan &
Fig. 1. The length-weight relationship in rotifers and tem- Rainbow (1971). Triangles: three ages of juveniles and adults
perate and tropical planktonic cladocerans. Regressions 1, 2 of Brachionus rubens from Pilarska (1977); the empty
and 3 are pooled regressions for Daphnia magna, pulicaria triangles are for 'deficient' diets and the black triangles for
and hyalina at food concentrations of greater than 1 mgC per 'optimal' diets.
14

the sizes for three ages of juveniles and for adults the lengths and weights of the cladoceran species
reared at her 'deficient' and 'optimal' food levels. is 9 times for length and 800 times in weight. The
Although it is hazardous to extrapolate beyond definitive body size of species is taken to be an
the measured ranges of the regressions, the higher inherited trait, presumably constrained by the
elevations of the rotifer points in Fig. 1 imply physiological possibilities ofthe taxonomic group,
heavier animals per unit length. This implication and is the result of an evolutionary 'decision' to be
is true for the two species measured directly in large or small relative to the rest. The genetic basis
carbon weight (Walz, 1983) as well as the other for the definitive size is unknown, jUdging from
species determined as dry weight, with the excep- King (1977): 'to my knowledge not one single
tion of the three points for Asplanchna spp., and morphological feature has ever been associated
may be associated with the lorica weight. with a specific single allele in (rotifers),. Since
Guisande (this symposium) found that rotifers body size seems to be a phenotypic variable
and Daphnia magna fell on the same length- responding to physiological conditions in the
protein weight regressions. environment, it is interesting to explore whether
The range of adult sizes belonging to the rotifers being large or small is a 'trade-off' in food limiting
in Fig. 1 can be compared with the sizes of well conditions which are common in the open water
fed adults instars (3rd or 4th) of the species of of large water bodies.
planktonic cladocerans subjected to food limi-
tation studies. These are given in Table 1, together
with their site of origin and names of the authors Food limitation and life cycle studies in Cladocera
who studied them. The span in the size range
In Cladocera, the species' life cycle characteristics
amongst the two groups of species is rather
which are affected when an individual is reared
similar. For the rotifers, there is a 10 times span
from neonate to some known adult instar under
in length and a 74 times span for weight or, if the
food limiting conditions appear to be that:
campanulate form of A. silvestrii is included, the
span in length and weight comes to 27 times and (1) the body size is reduced, particularly in
350 times, respectively. The comparable range for weight;

Table 1. The species of planktonic Cladocera subjected to food limitation studies. Body sizes refer to third or fourth adult instars.

Species Author l Origin 2 Body sizes

Length Weight
(mm) (J-LgC)

Daphnia magna Straus R LR 4.5 322

Daphnia pulicaria Forbes R LR 2.8 85
Daphnia pulicaria Forbes D LW 2.5 36
Daphnia hyalina Leydig R LR 2.1 31
Daphnia thorata Forbes D LW 1.7 19
Daphnia gessneri Herbst H U 1.5 7
Daphnia lumholzi Sars J KR 1.4 6
Diaphanosoma excisum Sars J KR 1.2 11
Diaphanosoma brachyurum Lieven H U 1.2 11
Moina micrura Kurz J KR 0.8 2
Moina reticulata Daday H U 0.8 2
Ceriodaphnia cornuta Sars J KR 0.5 0.4

1 Authors: Duncan (in prep.), Hardy (1989), Jayatunga (1986), Rocha (1983)
2 Sites of origin: KR - Kalawewa Reservoir, Sri Lanka; U - Lake Jacaretinga, Brazil; LR - London Reservoirs, u.K.; LW -
Lake Washington, U.S.A.
 15

(2) the number of instars to a certain point in the country of origin and close to the water body from
life cycle, such as the onset of reproduction, which they came as one aim of all the studies was
increases; to interpret field biological interactions by appli-
(3) the duration of development is prolonged; cation of experimental results. Table 2 provides
(4) fecundity is reduced. information on the temperature conditions for
each of the food concentrations tested by each
Thus, if the primiparous female is taken as a author, using algal food organisms which were
comparable point in the life cycle which is at the either the single-celled Scenedesmus acutus
end of juvenile growth and at the beginning of (Chlorophyceae) or the flagellate Cryptomonas
reproductive growth, then she is smaller in size, sp., both of which were adequate food for all the
appears at a later instar, is older and has reduced species, except for C. cornuta.
fecundity under limiting food conditions. These All the cladoceran results come from long-term
effects appear simultaneously when individuals life cycle studies where the development of a
are reared under food limitation and differ in neonate is followed daily throughout its life his-
degree according to the severity of the limitation tory to a reproducing adult whilst being
down to the threshold food level for growth and maintained in constant conditions of food and
reproduction. temperature. One of the greatest problems in such
These effects will be illustrated for the species long-term experiments is to maintain constancy of
listed in Table 1 which come from various tem- food conditions, especially in batch culture which
perate (the London reservoirs and Lake is the commonly adopted culture technique. It is
Washington, USA) and tropical (Kalawewa and commonly found that published results on how
Lake J acaretinga) sites and for all of which there food quantity affects the life cycles of cladocerans
is a background of ecological information. Apart are not comparable for methodological reasons.
from the Amazonian species which were studied Incomparability of results can arise for various
in the UK, the species were studied in their reasons: too small experimental volumes; using

Table 2. The concentrations of food used in cladoceran studies on food limitation.

Food levels Food species Authors

(mgC L -1)
Scenedesmus acutus Cryptomonas sp.

5.0 yes R
4.5 yes D
1.5 yes D
1.0 yes yes R,D,H,J
0.5 yes yes D.H,J
0.25 yes yes D,H,J
0.125 yes D
0.10 yes R
0.06 yes D
0.05 yes H,J
0.03 yes yes D,H,J
0.01 yes R,J
0.005 yes R,D

Key
R O. Rocha (1983) D. magna. D. pulicaria. D. hyalin a 5,10,15 & 20°C
D A. Duncan D. pulicaria. D. thorata 9,14 & 19°C
H E. R. Hardy (1989) D. gessneri. D. brachyurum. M. reticulata 22,27 & 32 °C
J Y. N. A. Jayatunga (1986) D. lumholzi. D. excisum. M. micrura. C. cornuta 22,27 & 32°C
16

un stirred or rotated vessels; putting too many stages. This size effect is illustrated in Fig. 1 for
animals in one container; too infrequent a chang- the large temperate Daphnia magna (regressions 1
ing of the food medium; too high a concentration and 4) and the two tropical species, the larger
of food to show food limiting effects. The most Daphnia gessneri (regressions 5 and 6) and the
serious causes of incomparability of results is that smaller Moina reticulata (regressions 7 and 8). In
either the experimental food levels have not been all cases, the slopes for the paired regressions of
quantified or they have been quantified in terms each species are parallel (not significantly dif-
which cannot be converted into common units. ferent) but the elevations are significantly dif-
For example, many authors gives food levels as ferent. Details of the food concentrations in which
cells/ml, without indicating the food cell dimen- the animals were reared are given in the legend for
sions or volume or dry weight or carbon weight. Fig. 1. The significant lowering of the elevations
The experimental details for this comparable of regressions 4, 6 and 8 occurred in food concen-
series of life cycle studies are given in Rocha trations of 0.01, 0.05 and 0.03 mgC L - 1, respec-
(1983), Duncan (1985a, b), Jayatunga (1986) & tively. In all species, non-limiting levels were at
Hardy (1989). concentrations greater than 0.1 mgC L - 1 and,
under such conditions, the regression 1,2 and 3
for the three temperate species of Daphnia (magna,
Body size pulicaria and hyalina) were not different and could
be pooled.
Figure 1 shows how the length-carbon weight Table 3 shows the extent to which the body size
regressions for a cladoceran species has a signifi- of the primipara female can be reduced when she
cantly reduced elevation when it has been reared is reared in the near-threshold food concentra-
throughout its life cycle in a limiting compared tions of 0.01-0.05 mgC L -1 from her size in non-
with a non-limiting food concentration. These limiting food levels. The size change is presented
regressions involve, of course, all the life cycle for six species of Cladocera of different size (col-

Table 3. The ranges in length, weight, instar and fecundity of the primiparous female in temperate and tropical species of
cladocerans reared under various limiting and non limiting food conditions and at various temperatures. The values in brackets
refer to 20-22 °C only.

Species Range in Non-limiting to limiting food concentrations 2

length 1
mm Primipara Primipara Primipara Primipara Primipara
length weight instar age fecundity
mm jJ.gC days eggs brood - 1

Daphnia 0.90-4.50 3.70-2.20 190-12 (V-IX) (7-16) 21-1

magna (111-17)
Daphnia 0.80-2.90 2.45-1.60 53-4 (V-XII) (8-26) 8-1
pulicaria (31-6)
Daphnia 060-2.20 1.90-1.40 8-4 (VI-IX) (8-16) 6-1
hyalin a (13-3)
Daphnia 0.50-1.50 1.30-1.10 5-1 (V-VI) (6-10) 4-1
lumholzi (4-1)
Diaphanosoma 0.47-1.30 1.10-0.85 7-2 (XII-XIII) (5-8) 4-1
excisum (5-1)
Moina 0.45-0.81 0.70-0.50 1.5-0.5 (III-V) (2+ -4+) 4+-3
reticulata

1 Neonate to 3rd or 4th adult instar.

2 See table 2. Limiting food concentrations were 0.06 mgC L - 1 or less.
 17

umns 1 and 2) in terms oflength (column 3) and Fecundity

carbon weight (column 4). In column 4, the values
in brackets refer to experiments conducted at the Associated with an increase in the age of the
comparable temperatures of 20-22 °C. The con- primipara female is a marked reduction in the size
sistent result is that the primipara female is of her first brood, which is shown in column 7 of
smaller in length and weight when reared in low Table 3. However, a better assessment of the
food compared with optimal levels. The degree of effect oflow food concentrations upon the fecun-
reduction differs with size of species, which prob- dity of cladoceran species is given by linear regres-
ably reflects the difficulty of detecting small sion of eggs per brood against adult length. In the
change in small organisms. Weight is the better life cycle experiments, these could be calculated
measure and is reduced by three to five times in from the lengths and fecundities of the first three
smaller species compared with a thirteen to fifteen or four adult instars. An increasing fecundity with
times reduction in larger ones. increase in body size of female has been reported
by many workers (see review by Ivanova et al.,
1987) and the regressions plotted in Fig. 2 for five
Instar of primipara species of Cladocera show very steep slopes in
optimal non-limiting food concentrations. The
Column 5 in Table 3 gives the instar stage at slope of the regression flattens as the food level is
which the onset of reproduction occurs in the lower until, at extreme food limitation, no signifi-
various cladoceran species under the experi- cant regression can be calculated as all sizes of
mental food conditions and at 20-22 ° C. In all the female are able to produce no more than one or
species, the primiparous female produces her first two eggs in a brood. The steeper slopes of Moina
brood of eggs at an earlier instar when reared are probably associated with her small size rather
under optimal food conditions but is forced to than high temperature since 32 ° C is as high a
pass through several later instars (from one to temperature for tropical species as is 20 ° C for
seven more) before she can achieve this as food temperate ones. These fecundity-length regres-
availability approaches more severely limiting sions are as good ecological indicators of the
levels (0.01-0.05 mgC L - 1 ). nutritive state of field populations of cladocerans
as the length-weight regressions considered
earlier.
Age of primipara

The age of the primipara female under the dif- Food limitation and life cycle studies in rotifers
ferent experimental conditions is given in column
6 of Table 3 and, again, refers to the 20-22 °C Four periods in the rotifer life cycle are dis-
experiments only. This represents the end of the tinguished by King (l969): (l) the embryonic
juvenile period of body growth and the beginning period from egg formation to egg hatching, (2) the
of the mature phase in which reproductive as well pre-reproductive period from egg hatching to first
as body growth occurs. It is an ecologically rele- reproduction, (3) the period of fecundity and
vant temporal point in the life cycle for detecting (4) the post-reproductive period from cessation of
any prolongation of the juvenile phase under food reproduction to death. As in the cladocerans, the
limitation. Table 3 shows that the prolongation of main periods in the rotifer life cycle which are
this phase is quite marked in severely limiting vulnerable to the environmental stress of a sparse
levels of 0.01-0.05 mgC L -1 in all the species, food supply are the juvenile phase (2) and the
from 1.5 to 3.3 times longer compared. period of adult reproduction (3) although there is
some evidence that egg size and so the duration
of embryonic development may be affected by
18

."0
0
0
20
L.
J:J

'"
~
E
VI
a:

J
~
15 (
VI A
~
!::i
::>
Cl
<
a: 10 B
::>

/7
0
LL..

/
~
VI
Q;
LL..
LL..
0 5
>-
I
II lI~l
2
cz
~

fo
::>
u
u.I
LL..
3I1i ~ lIn 3
0
(}O (}5 1·0 1·5 2·0 2·5 3·0 3-5
BODY LENGTH OF FIRST ADULT INSTARS (mmJ

Fig. 2. Length-fecundity regressions for several species of temperate and tropical cladocerans which had been reared throughout
their life cycle in different concentrations of food. Species: A - Moina micrura (32 °C); B - Diaphanosoma excisum (32°C); C
- Daphnia thorata (14 °C); D - Daphnia pulicaria (14 °C); E - Daphnia magna (20 °C). Food level: 1- 0.5-1.0 mgCI-1;
2- 0.1 mgCI - I; 3- 0.01-0.05 mgCI- I

maternal diet (Yufera, 1987; see also Duncan, ml- I. The rates were more or less similar in 16.4
1983, 1984). The postembryonic growth in rotifers and 49.2 jlgDW ml- I. His Fig. 2 shows that the
is limited to expansion of cells or syncytial tissues primipara female became both older and smaller
since there is no proliferation of cells beyond the in food below 16.4 jlgDW ml- 1 so that, for this
number fixed during the organ primordia species at 22 0 C, food appears to become limiting
(Starkweather, 1987). As Starkweather com- below this concentration. Figures 3, 4 and 10 in
ments, this cytological constraint sets close Robertson & Salt (1981) illustrate a reduced net
bounds on adult body dimensions. Various reproductive rate, a prolongation of the age offirst
authors have divided the juvenile period into an reproduction and a reduction in the body volume
active feeding phase followed by a period of of the primiparous female of Asplanchna girodi
enlargement to an adult size by stretching and when fed low numbers of Paramecium per indi-
incorporation of assimilated material (King, vidual per day.
1967; Pourriott, 1973; Ruttner-Kolisko, 1974) There is only one published series of experi-
but it is not clear whether the primiparous female ments which studied the influence of a wide range
is capable of body growth as well as egg produc- of food concentrations upon the life cycle charac-
tion. teristics of a rotifer species and which is com-
There are not many published papers on rotifer parable with the cladoceran experiments. This is
life cycle studies involving different food levels. the eco-physiological study of Pilarska (1977) on
The findings of King (1967) were that the growth Brachionus rubens cultured throughout its life
rate in length of individual Euchlanis dilatata cycle at 20 0 C and at ten concentrations of
depends on food concentration but only in his Chlorella vulgaris. Her food levels ranged from
three lower food levels from 1.64-16.4 jlgDW 0.12.106 to 10.106 cells ml- 1 which, if one accepts
 19

100
-I
"0

10 ,r;::,¥-
r;::,<"'~'\
4 ~\.\.

..c::
• r;::,fc
VJ~\.
~\):
+- ,¥-~
~
....o 1·0 • \.

Cl

CII
>
+-
u
~
"0
0·1 R= 1-63. GO' 92 (1,13; P = 0·0000)
o 2 Daphnia hyalina: R=2·14.GO' 99 (1,14; P=O·OOOO)
L-
a.
CII
0::
3 Diaphanosoma excisum: R=5·1l.G1-06 ( 1,9; P = 0·004)
4 Moina micrura: no significaBt regression.
0·01...j<:------.-----------.-----=-----.---=---..,
0·01 0·1 1·0 10 100
-1 -1
Body growth (JIg C. indo d )
Fig. 3. The allocation offood resources to reproductive or body growth in adult animals of two temperate and two tropical species
of c1adocerans. The regressions quantify the daily growth rates of the adults under different temperature conditions and limiting
and non-limiting food concentrations. The values in brackets are degrees of freedom and probability.

her very high cell weight for C. vulgaris ments. Three food levels are compared which are
(29.10 - 6 ,ugDW cell- 1), can be converted to 0.35 characteristic of Pilarska's 'deficient', 'optimal'
to 290 mgDW L - 1 or to 0.18 to 145 gC L - 1, and 'excessive' diets; these are 0.012.10 6 , 1.0.106
assuming carbon is 50% of dry weight. Similar and 10.106 cells ml- 1.
published studies which used comparable batch Table 4 shows that the largest juvenile (less
culture rather than chemostat techniques either than 12 more than 24 hours old) and the largest
did not define the food concentration provided amictic mature adult were those grown under
(Doohan, 1973) or used food levels which were optimal food conditions and that these larger
too high to show food limitation effects females also produced the largest eggs. Her
(Leimeroth, 1980). Figure 3 in Starkweather papers show that this is so in linear dimensions as
(1987) shows that there is another unpublished well as in dry weight. In food deficient concentra-
study by Skrdla & Starkweather on the combined tions, the juveniles and the females were about
influences of temperature and food density on 30 % smaller by weight and the diploid eggs were
Brachionus calyciflorus fed on Euglena gracilis smaller by 15-20%. There was a prolongation of
which used a wide range of food levels (0.1 to the juvenile period to 2.8 days in 'deficient' food
100 ,ug ml- 1). This should provide a further com- compared with 1.26 days in 'optimal' food.
parison in future with both Pilarska's investi- Pilarska also found a marked shortening of the
gation and with the cladoceran ones. period of egg production to 0.73 days in the lowest
Pilarska's methods and results on the life cycle food level compared with 6.8 days in the 'optimal'
energy balance for individual B. rubens have been food. Instantaneous rates of body growth in
published fully in a set of three papers (1977) and weight can be calculated from the sizes and dura-
have been re-worked by Duncan (1984) to pro- tion of the juvenile phase; these were 0.21 day - 1
duce Table 4 which will form the basis for detect- and 0.35 day- 1 for 'deficient' and 'optimal' food
ing any food effects on life cycle characteristics respectively. This work demonstrates that food-
and for comparison with the cladoceran experi- deficient juveniles grow slowly into small adults
20

Table 4. Energy budget of Brachionus rubens at 20°C for the post-embryonic period and for the period of egg production and
some resulting efficients, re-calculated from Pilarska (1977).

Food concentration Period of growth Period of egg production

Deficient Optimal Excessive Deficient Optimal Excessive

Food concentration
106 cell ml- I 0.012 1.0 10.0 0.012 1.0 10.0
j.LgCml-l(a) 0.18 15.0 150.0 0.18 15.0 150.0
Body size (j.LgDW ind - I)
Egg (2n) 0.04 0.06 0.06
Juvenile (young to old) 0.06-0.11 0.09-0.14 0.08-0.10
Amictic female 0.13 0.17 0.14
Duration of period of body growth or egg 2.84 1.26 1.43 0.73 6.77 3.80
production (days)
Energy parameters
(cal 103 ind - I period - I)
Consumption 30.55 58.47 73.15 0.35 13.18 9.06
Production 4.26 8.00 3.96 0.05 2.34 0.32
Respiration 13.08 5.38 5.21 0.19 1.79 1.03
Efficiencies for the period
Assimilation/consumption 0.57 0.23 0.13 0.68 0.31 0.15
Respiration/assimilation 0.75 0.40 0.57 0.80 0.43 0.77
Respiration/consumption 0.43 0.09 0.07 0.54 0.14 0.11

(a) Cell weight (29 x 10- 6 j.LgDW cell-I (Pilarska, 1977) and carbon weight is 50% of dry weight.

which (if they can) produce small eggs but only for reproductive periods of life cycle are better dem-
a very limited period. The food concentration at onstrated by cumulative energy budgets, which
which this occurred in B. rubens was she illustrated in her Fig. 3 but did not tabulate.
0.35 mgDW L - 1 and this appears to be very near These can be re-calculated for the two periods of
to the species' threshold for concentration for growth and egg production and are presented in
growth and reproduction. With a more usual cell Table 4. This shows that, in 'deficient' food, a
weight of 2.5 pgC cell- 1 for Chlorella vulgaris higher proportion of consumed food is required to
(Rocha & Duncan, 1985), the threshold concen- cover the respiration of the juvenile phase than in
tration for the species would be lower 'optimal' food (due to its prolongation), despite a
(0.02-0.06 mgC L - 1). If we accept Pilarska's higher assimilation efficiency. It is therefore
results, rotifers appear to respond phenotypically understandable why the oldest juvenile is small
to limiting food stress similarly as cladocerans, by (0.1 jlgDW compared with 0.14 jlgDW). The
reducing the body size at first reproduction, energetic situation in cumulative terms is similar
prolonging development and reducing fecundity. for the periods of egg production in the adults: in
Because Pilarska measured the daily feeding 'deficient' food, a greater proportion of consumed
rates and hourly respiratory rates of overigerous food is needed for respiration and less is available
amictic females and three ages of juveniles at for egg production; this accounts for the short-
various food levels, she was able to calculate daily ness of the period of egg production and the
instantaneous energy budgets for each of these life smallness of the eggs and neonates. That all the
cycle stages for deficient, optimal and excessive life cycle stages measured were smaller in condi-
diets (her Table 1, page 345). However, the stress tions of food deficiency is not surprising and nor
that food deficiency imposes upon the growth and is it surprising that food deficient juveniles and
 21

adults assimilate more efficiently (57 %and 68 %) female; knowing the body weight, her absolute
than the same stages in optimal food (23 % and daily growth rate could be computed. The daily
31%). reproductive rate was calculated from the number
The value of Pilarska's research in the present of eggs in the first brood, egg weight and the
context is to show that a high cost of cumulated embryonic development duration for the experi-
respiration has to be paid in order to complete the mental temperature involved. Rocha's growth
life cycle at threshold food levels and that this rates, therefore, apply to the primiparous female.
appears to result from the food-limited prolon- Jayatunga (1986) also found that the Richard's
gation of the rotifer juvenile growth phase. growth function, as modified by Schnute (1981),
Another life cycle study on a species of benthic provided good fits to the life cycle growth patterns
nematode demonstrates the same result of her tropical species. However, she calculated
(Schiemer et al., 1980). That the same bioenerge- the daily growth rates for all the first four adult
tic pressures at threshold food concentrations instars so that her results are likely to demonstrate
may cause similar life cycle effects in cladocerans slightly lower body growth rates and slightly
seems very likely, judging from the results of higher reproductive rates, because of the body
Bohrer & Lampert (1987) when they measured size effect.
simultaneously the effect of food concentration on It is proved possible to obtain significant
assimilation and respiration in adult Daphnia double logarithmic regressions for reproductive
magna. growth on body growth in three of the above four
species and these are illustrated in Fig. 3. All three
regressions lie above the line of equal allocation
Allocation of food resources into body growth and and more or less parallel to it; the slopes of the
reproductive growth equations are close to one and are not signifi-
cantly different (P = 0.71). As the regressions are
As the allocation of food resources into body based on all the replicates of all the food-tempera-
growth or reproduction in different species is of ture treatments that could provide growth and
ecological and evolutionary interest (Calow, reproductive rates (Table 2), this result suggests
1978; Lynch, 1980, 1985; Sibly & Calow, 1986), that these three species are able to invest more
an attempt was made to illustrate the investment food resource into reproduction than body
allocation in a species of rotifer and in large and growth at all the food-temperature conditions that
small, temperate and tropical species of permit them to complete their life cycle. The
Cladocera when supplied with non-limiting levels particular food-temperature condition merely
of food or when faced with continuously scarce determines the point along the regression line
food resource. where the species population exists (low food-low
It proved possible to define the relationship temperature at its lower end, for example). The
between the daily rates of body and reproductive higher elevation of the tropical Diaphanosoma
growth in adults of four of the planktonic species excisum may come from Jayatunga's inclusion of
of cladocerans, using the rates calculated by older adult instars in her calculations and this
Rocha (1983) for Daphnia magna and Daphnia needs to be checked by a calculation for the
hyalina and for Diaphanosoma excisum and Moina primiparous female only.
micrura by J ayatunga (1986). Rocha was able to A significant relationship between reproductive
fit the Chapman-Richards growth function and body growth could not be obtained for Moina
(Richards, 1959) to the life cycle data of daily micrura but the points for the seven experimental
weights and ages for each temperature-food treat- treatments that produced positive body growth
ment and use the parameters of the curvilinear rates are plotted in Fig. 3. These lie at a higher
growth equation to calculate the relative growth elevation than the regression for Diaphanosoma
rate of an animal which had the age of a primipara excisum, the larger of the two tropical species. In
22

only 12 of the 16 different experimental treat- optimal or excessive diet. Figure 4 shows that,
ments to which Moina micrura was subjected in unlike the cladoceran plot, the points for food-
order to determine its threshold food concen- limited rotifers lies well below the line of equal
tration did the species attain a reproductive size allocation into juvenile growth and adult repro-
and, in several of these food levels at high tem- duction so that more of the scarce food is being
peratures (32 DC), the very high reproductive put into enhancing juvenile body growth and
rates were accompanied by negative body growth. much less into supporting reproductive growth.
At 32 DC, a possible water temperature in the However, as the available food approaches the
tropics, Moina seems to have invested in repro- optimal level, the rotifers invest more equally into
duction at the expense of body growth. growth and reproduction. Within the group of
Because of the nature of growth in rotifers, it is optimal food concentrations, all the points lie well
impossible to produce a comparable figure show- above the line of equal allocation; both rates are
ing the allocation of food resources into growth or high and probably maximal so that they change
reproduction of adults. In Figure 4, a compromise little with food level. In excessively high food con-
was reached by comparing the daily growth rates centrations, the unfavourable circumstances are
of the oldest juvenile Brachionus rubens with the shown by the reduced body growth and a sharp
daily reproductive rate of adults when reared in decline in adult egg production.
ten different food concentrations (Pilarska, Since the growth of the oldest juvenile under
1977); these food concentrations have been allo- different food conditions is likely to be affected by
cated in Fig. 4 into those representing a deficient, its earlier growth, a comparison was made of the
cumulated body growth for the whole juvenile
Q
period and the cumulated reproductive growth for
'I the period of egg production under the different
Q
"0
Q 1:1 experimental conditions. This is plotted in Fig. 5.
'I 1.
-t:i In this plot, the points for food-deficient rotifers
c • lie on the line of equal allocation. When the food
'"' 0
conditions are optimal, the cumulated growth for
0
rou • the juvenile phase is high, probably maximal but
~
changes little with more food; an additional
availability of food is chanelled directly into ever
~
0
<...
• 0
higher cumulated egg production. In the poor
C71

c:u conditions of excessive food levels, the cumulated

+-
>
0·1 0 juvenile growth is low and changes very little with
u
:::J
"0
increase in food level; as the available food
0
<... becomes ever more excessive, the cumulated egg
a.
c:u
<...
production declines more and more.
+- o deficient diet Figures 3 and 4 show rather different daily allo-
~ o
Q optimal diet cations into growth and reproduction by adult
:::J
"0
« • excessive diet cladocerans and near-adult B. rubens. They illus-
trate how the allocations differ in each taxonomic
group when food resources span from non-limit-
0·0 1 ~----,r---""---'r--..---""'-..,....,---r-o ing ('excessive' and 'optimal') to limiting ('defi-
01 }O
-3 -1-1 cient' and 'near-threshold') food levels. In
Body growth of oldest juvenile. cal.10. indo d.
B. rubens (Fig. 4), a more scarce resource is put
Fig. 4. The allocation of food resources to the body growth
of the oldest juvenile Brachionus rubens and to the egg pro-
into body growth than into egg production in
duction of the adults under various conditions of food limi- deficient food conditions, whereas, once an opti-
tation: the daily picture. Data of Pilarska (1977). mal juvenile growth rate is achieved, additional
 23

, 0·4 above the threshold to optimal, the ratio of adult

allocation into growth: reproduction seems to
~

'0
.~
... remain constant. This difference in allocation pat-
~
-c:i terns seems to be a consequence of the funda-
...
0
Cl. mental difference in the nature of growth in
~
,.
'0
0·3
1:1
rotifers and crustaceans, since the latter can con-
tinue to grow as adults by cell proliferation.
.E
", . According to Snell & King (1977), a basic
,,
'0
.-:
~

ro assumption within the area of trade-offs between

u

.s:. 0·2
0 life history characteristics is 'that of resource limi-
I
+-
tation: resources directed towards reproduction
...
~ I
0 I
C\ I
I
0 reduce those available for growth and main-
tenance. Thus one consequence of reproduction
QJ
> I
:<: I
::l
'0
u
I is presumed to be decreased survival.' In indi-
I.
...
0
Cl.
0·1 I
I
vidual cladocerans reared in food concentrations
...
QJ
0 deficient diet that allow them to complete their life cycle and
:=::l
'0
<
-I
I
'f
Q

•
optimaL diet
excessive diet
reproduce, the conflict is different for the juvenile
and reproductive phases of the life cycle. In the
juvenile phase, it lies between the highly cumulat-
ing metabolic costs due to food-limited prolon-
0·1 0·2 0·3
gation of the phase and the body size at which it
Body growth of oLdest juveniLe. caL10- 3.ind-',[juv.periodf'
can mature. The latter characteristic is a species
Fig. 5. The allocation of food resources to juvenile body
characteristic. In the reproductive phase, the con-
growth and adult reproduction cumulated for the periods of
body growth and egg production and when reared under flict is between growth plus reproduction and
various conditions of food limitation. Brachionus rubens maintenance, where the allocation of growth:
(Pilarska, 1977). reproduction is kept constant.
On the basis of results on cladocerans reared in
food resource does not alter this and most goes to undefined food conditions, Lynch (1980) grouped
increase egg production. It seems that there are 'well-fed' animals into (1) the larger species which
close bounds set on adult body dimensions of mature later in their life cycle at a size close to the
rotifers (Starkweather, 1987) but only in optimal maximal size and the channel most of their availa-
food conditions and not in limiting or deficient ble food into reproduction and (2) small-sized
food levels perhaps because maximal body dimen- species which mature early and continue to grow
sions were not achieved. In cladocerans (Fig. 3), after the onset of maturity, thus channelling a
the pattern of allocating slightly more to reproduc- larger proportion into body growth and a lesser
tive than body growth remains the same for the proportion into reproduction. Although it is true
full span of non -limiting to limiting food resource. that well-fed cladocerans oflarger species mature
This is the implication of the slopes of the three at a later instar, older age and larger but not
significant regressions being near to one. When maximal body size, it is not at all established that
body growth is about 1 j.lgC ind - 1 day - 1 in good a larger proportion of the food is channelled into
food conditions, the ratios of the reproductive reproduction in larger compared with smaller
ratios of the three species are 1.63: 2.14: 5.27 (or species. The two regressions for the larger
1 : 1.3 : 3.2 for D. magna, D. hyalina and D. magna and smaller D. hyalina in Fig. 3 are not
D. excisum). When the body growth rate is lower significantly different, can be pooled and show a
at about 0.5 j.lgC ind - 1 day - 1, these ratios similar slightly higher investment into repro-
remain much the same (1 : 1.3: 2.9). Irrespective duction than body growth at all food-temperature
of the level offood resource within the range from treatments. The main difference lies in the smaller
24

and larger absolute daily rates of the smaller and of 17 times, and revealed a strong relationship
larger species, respectively. The higher investment with the body weight of the rotifer species: larger
into reproduction in the two smaller species which species had higher threshold than smaller species.
are also different in being tropical is interesting A data point has been added to Fig. 6 for
but not yet established and needs re-calculation Pilarska's (1977) Brachionus rubens reared at the
for a comparable adult stage. If one takes well-fed lowest of the deficient food levels (0.35 mg DW
rotifer individuals as representatives of even L - 1) of Chlorella vulgaris (29 pgDW cell- 1) and
smaller zooplankton species, they allocate much this fits well into the series. When compared with
more into reproduction once juvenile growth has the threshold values for temperate species of
been satisfied but the reverse is true in 'deficient' Daphnia (0.01-0.02 mgDW L - 1) and for tropical
food levels (Figs. 4 and 5). species of cladocerans (0.06-0.1 mgDW L - 1 at
27°C and 32 °C; 0.1-0.2 mgDW L -1 at 22°C,
which is a low temperature for tropical species),
Comparison of threshold food concentrations in this series of threshold values for rotifers seem
rotifers, c1adocerans and ciliates very high. Only a few of the smaller species of
rotifers have threshold values as low as tropical
The series of papers by Sternberger & Gilbert cladocerans and none as low as temperate
(1985a, b; 1987) on rotifer threshold food levels, Daphnia. The threshold concentration cited by
determined as the minimum food at which the Schiemer (1985) for a temperate and a tropical
population's instantaneous rate of increase (r) is species of freshwater calanoid copepods (less
more or less zero, permitted a comparison with than 0.1 mgDW L - 1) fit the cladoceran picture.
the threshold concentrations for cladoceran These results appear to indicate a significant
which were determined experimentally and difference in the minimal food requirements
defined as the minimum food at which the life between rotifers and planktonic crustaceans. It
cycle could be completed. These two sets of data was fortunate that further data points for this
are plotted in Fig. 6, with food level on a log scale diagram were provided during the rotifer sym-
and in terms of dry weight as this was the measure posium at Gargnano by K.O. Rothhaupt from his
used by Sternberger & Gilbert. Where the algal PhD dissertation on Brachionus rubens and
concentration was measured in carbon weight, B. calyciflorus, by P Starkweather from unpub-
the values were doubled, assuming that 0.5 dry lished research with B Skrdla on B. calyciflorus
weight is carbon. and by T W Snell from recently published
The threshold food values tabulated in research on B. plicati/is (Snell & Boyer, 1988).
Sternberger & Gilbert (1987) for ten species of The threshold value of 0.1 mgDW L - 1 published
rotifers were determined using algal species in Starkweather (1987) for B. calyciflorus when
whose cell sizes and types were most highly reared on Euglena gracilis is lower than the value
preferred or efficiently harvested by the rotifer of 0.38 mgDW L -1 given by Sternberger &
species studied in order that the r-values would be Gilbert (1987) with Cryptomonas erosa as the food
comparable. They used Cryptomonas erosa var. organism. The threshold of 0.56 mgDW L -1 for
reflexa (850 Jim 3 ; equivalent spherical diameter amictic B. plicatilis cultured on Dunaliella tertio-
ESD of 6.5 Jim) as food for the larger rotifer lecta (370 pgDW cell- 1) is higher than Dewey's
species and a series of smaller cryptomonads for value of 0.40 (Sternberger & Gilbert, 1987).
the smaller species: Cryptomonas ovata (160 JiM 3 ; Rothhaupt determined the food concentrations at
ESD 3.7 Jim), Cryptomonas spp. (130 Jim 3 ; ESD which the instantaneous rate (r) is more or less
3.5 JiM) and Rhodomonas minuta (85 Jim; ESD zero for the two brachionid species when they
3.0 Jim). The threshold concentrations ranged were reared on a series of algal species of different
from 1.03 mgDW L - 1 for Keratella crassa to cell and size and of different 'harvestability' effi-
0.06 mgDW L - 1 for Keratella eochlearis, a span ciencies (which were tested). He defines the size
 25

FOOD SIZE
SPECIES .urn
•
~ ~:
en Cc •
Pc CILIATES

Sc
L
~ Bt

t
I
Kcr •
i Ap •
I Sp •
I Bp •
cao o
Be ROTIFERS
[ Br o 00 .0 0

I Kb •
•
~I ~~
..,. Kco •
•
•

Pa.As ....... ]
TROPICAL AND TEMPERATE
FRESHWATER CALANOID

Dg. 01. Db. De. Mm. Mr

] TROPICAL CLADOCERANS

Om. Op.Oh. Oth ] TEMPERATE CLADOCERANS

1 I 1 I
0,001 0,01 0,1 10
FOOD LEVEL mg DW.C'

Fig. 6. A comparison of the threshold food concentrations of rotifers, planktonic cladocerans and copepods and ciliates. Key:
Ciliates: Cg Cyclidium glaucoma, Gs Glaucoma scintillans, Cc Colpidium colpoda, Pc Paramecium caudatum, Sc Stentor
coerulens, L Lembadion, Bt Bursaria truncatella. Rotifers: Kcr Keratella crasse, Ap Asplanchna priodonta, Sp Synchaeta
pectinata, Bp Brachionus plicatilis, Bc Brachionus calyciflorus, Br Brachionus rubens, Kb Kellicottia bostoniensis, Ke Keratella
earlinae, So Synchaeta oblonga, Kco Keratella cochlearis. Copepods: Pa Phyllodiaptomus annae, As Arctodiaptomus spinosus.
Cladocerans: Dg Daphnia gessneri, Dl Daphnia lumholzi, Db Diaphanosoma brachyurum, De Diaphanosoma excisum, Mm
Moina micrura, Mr Moina reticulata, Dm Daphnia magna, Dp Daphnia pulicaria, Dh Daphnia hyalina, Dth Daphnia thorata.
Sources: Fenchel (1980); Stemberger & Gilbert (1987); Pilarska (1977); Starkweather (1987); Rothhaupt (pers. comm.);
Schiemer (1985); Taylor (1978), Snell & Boyer (1988).

of his food cells as the equivalent spherical diame- (12 Jlm; 0.31 ± 0.013; 0.185 ± 0.010). These data
ter (ESD). In the following list of algal species points, doubled to give dry weight, have been
used by Rothhaupt, the values in brackets are first plotted in Figure 6, with Rothhaupt's permission,
the cell ESD, then the threshold concentration in and show that a range of threshold concentrations
the original mgC L - 1 for Brachionus rubens fol- exist for anyone species according to the suita-
lowed by that for B. calyciflorus: Chlorella minutis- bility of the food species. The most divergent
sima (2 Jlm; 0.215 ± 0.014; 1.958 ± 0.217), points for B. calyciflorus are for the small-sized
Monorhaphidium minutum (3.5 Jlm; Chlorella minutissima (2 JLm) which gave a very
0.071 ± 0.013; 0.236 ± 0.027), single-celled high threshold value and for Euglena gracilis
Scenedesmus obliquus (5.5 JLm; 0.074 ± 0.009; whose threshold was the lowest; the values for
0.195 ± 0.011). Chlamydomonas reinhardii which other food cells are rather similar to that for
was found to be nutritively poor (6 Jlm; Cryptomonas erosa (Sternberger & Gilbert, 1987).
0.091 ± 0.011; not used for B.c), Cyclotella Rothhaupt's results gave both higher and lower
meneghinianae (8.5 JLm; 0.107 ± 0.039; threshold values for B. rubens compared with
0.193 ± 0.011) and Chlamydomonas sphaeroides Pilarska's point with Chlorella vulgaris; the higher
26

threshold concentrations were given by both the might help in the interpretation of the rotifer
smallest (c. minutissima, 2 f.Lm) and the largest thresholds shown in Figure 6. Are the smaller
tested algal species (Chlamydomonas sphaeroides, rotifer species with low thresholds planktonic
12 f.Lm). An additional point of some significance forms, like the cladocerans, and are the larger
is provided by Snell & Boyer (1988) who found species with high threshold inhabitants of particle-
that the threshold concentration for mictic repro- rich waters such as ponds and activated sludge?
duction in B. plicatilis was 5.7 mgD W L - 1, which
is ten times the value for amictic animals; this
point has not been plotted in Fig. 6. Acknowledgements
For interest, threshold food concentrations for
various kinds of ciliates have been included in I am grateful to Dr. O. Rocha, Dr. Y.N.A.
Fig. 6. These come from Fenchel (1980), who cal- J ayatunga, Dr. E.R. Hardy and Mr. L.c. dos
culated the minimum food particle concentration Santos for use of their data without which this
needed to cover metabolic costs. This comes from paper would not have been possible. I thank Dr.
the volume of water cleared per ml of oxygen C. Ricci for splendid hospitality at the Rotifer
respired, a value derived from two relationships Symposium in Gargnano and Royal Holloway &
generated from published data, namely, the rela- Bedford New College for some technical assist-
tionship between the maximal clearing rate at low ance with the production of this paper.
food concentration and the ciliate cell volume and
the relationship between oxygen consumption
and cell volume. An assumption of a respiratory Conclusions
quotient of the one allowed Fenchel to calculate
the minimum weight of food needed to cover (1) Our understanding of the effects offood limi-
respiration and, from the volume of water cleared, tation upon the life cycle characteristics of rotifers
the concentration per unit volume of water. The and planktonic cladocerans is dependent upon
threshold values are only very approximate the results of experiments in which the food con-
because of the scatter in the published respiratory centrations are quantified and do not vary too
data and because of the existence of two different much and in which something is known about the
R/V regressions for ciliates. However, his calcu- size, ingestibility and food value of the food orga-
lations demonstrate that the larger ciliates feeding nism offered. The results from many published
on larger food particles (1-5 f.Lm) are similar to studies are incomparable because this kind of
planktonic Daphnia in their low food threshold information is not available.
whereas the smaller ciliates which specialize on (2) Although rotifers and cladocerans differ in
very small particles (0.2-1 f.Lm) need much higher their adult body sizes, there is an overlap between
concentrations of suspended particles. The two the two groups, particularly with the small, rarely
points added for Cyclidium glaucoma and studied tropical cladocerans. The elevation of the
Colpidium colpoda come from Taylor (1978) and length-carbon weight regression of a cladoceran
fall in the middle of Fenchel's ranges. The ecology species reared in near-threshold food concentra-
of these two groups of ciliates differs; as Fenchel tions (0.01-0.05 mgC L - 1) is significantly lower
comments, the dependence of small-particle than that for animals cultured in optimal concen-
feeding bacterivorous ciliates upon the presence trations. This occurs in both large and small
of high concentrations of suspended bacteria (10 9 cladoceran species and can be used as a useful
cells ml- 1) precludes their colonisation of the ecological indicator of the nutritive state of field
open waters of lakes or the sea where bacteria are popUlations.
sparse (10 5 cells ml- 1). The planktonic ciliates (3) Evidence from one study and on one
(like tintinnids and oligotrich ciliates) feed on species of rotifer showed that the weight of eggs,
large particles and have lower thresholds. These juveniles and adults were smaller, that the dura-
insights into ciliate thresholds and trophic ecology tion of the juvenile phase was longer and that the
 27

period of egg production was shorter when it was a species but varies with the suitability of the algal
reared in near-threshold concentrations of food food species.
compared with optimal food levels. There is a
need to confirm these results with other life cycle
References
experiments on rotifers under defined conditions
of food limitation. Similar effects were demon- Bottrell, H. H., A. Duncan, Z. M. Gliwicz, E. Grygierek, A.
strated for six species of temperate and tropical Herzig, A. Hillbricht-Ilkowska, H. Kurasawa, P. Larsson
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Bohrer, R N. & W. Lampert, 1988. Simultaneous measure-
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under food limiting conditions. There is a conflict and respiration in Daphnia magna. Functional Ecology. 2:
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Doohan, M. & V. Rainbow, 1971. Determination of dry
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mature. Doohan, M., 1973. An energy budget for adult Brachionus
(4) Allocation of food resource into growth and plicatilis Muller (Rotatoria). Oecologia (Berl.). 13:
reproduction was compared in the two taxonomic 351-362.
Duncan, A., 1983. The influence of temperature upon the
groups, using the primiparous female in the clado-
duration of embryonic development of tropical Brachionus
cerans and the oldest juvenile-adult female in the species (Rotifera). IN: Schiemer, F. (ed.), Limnology of
rotifer species studied. The pattern of allocation Parakrama Samudra - Sri Lanka. Junk. 107-115.
differed. In the rotifer, scarce food resource was Duncan, A., 1984. Assessment offactors influencing the com-
invested more into body growth than egg produc- position, body size and turnover rate of zooplankton in
Parakrama Samudra, an irrigation reservoir in Sri Lanka.
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Hydrobiologia. 113: 201-215.
went to increase egg production. In cladocerans, Duncan, A., 1985. Body carbon in daphnids as an indicator
the pattern of allocating slightly more to repro- of the food concentration available in the field. Arch.
duction than to body growth remained the same Hydrobiol. Beih. Ergbn. Limnol. 21: 81-90.
for all conditions of food resource. Moreover, the Duncan, A., 1985b. Carbon weight on length regressions of
Daphnia spp. grown at threshold food concentrations.
ratio of adult allocation into growth: reproduc-
Verh. Int. Ver. Limnol. 22: 3109-15.
tion remained constant. It is suggested that these Fenchel, T., 1980. Suspension feeding in ciliate Protozoa:
differences are the consequence of the funda- feeding rates and their ecological significance. Microb.
mentally different nature of growth in the two Ecol. 6: 13-25.
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Daphnia. Ecology 66(6): 1943-1950.
(5) Threshold food concentrations for eleven
Gilbert, 1. 1. & R S. Sternberger, 1984. Spine development
species of rotifers show that, for the group, they in the rotifer Keratella cochlearis: induction by cyclopoid
are much higher than for temperate planktonic copepods and Asplanchna. Freshwat. BioI. 14: 639-647.
cladocerans and that only the thresholds for Hardy, E. R, 1989. Effect of temperature , food concentration
smaller rotifers overlap with the small tropical and turbidity on the life cycle characteristics of planktonic
cladocerans in a tropical lake, Central Amazon: a field and
species of cladocerans. Larger rotifer species have
experimental study. PhD Thesis. University of London
as high threshold food concentrations as (RHBNC). 337 pp.
Fenchel's small-particle feeding bacterivorous Hrabackova, M., 1971. The size of primipara and neonates
ciliates, and, like them, must be confined ecologi- of Daphnia hyalina Leydig (Crustacea: Cladocera) under
cally to non-pelagic particle-rich environments. natural and enriched food conditions. Vest. Ces. Spol.
Zool. 38(2): 98-105.
Planktonic ciliates feed on larger particles and
Hrabackova, M. & J. Hrbacek, 1979. Rates of post-
have lower threshold food concentrations. The embryonic development in several popUlations of the
work of K.O. Rothhaupt demonstrated that the group of species Daphnia hyalina Leydig at various concen-
threshold food concentration is not a constant for trations of food. Vest. Ces. Spol. Zool. 43(4): 253-259.
28

Herzig, A., 1983a. The ecological significance of the relation- Salonen, K., 1979. A versatile method for the rapid and
ship between temperature and duration of embryonic accurate determination of carbon by high temperature
development in planktonic freshwater copepods. Hydro- combustion. Limno!. Oceanogr. 24: 177-183.
biologia. 100: 65-91. Santos, L. C. dos., 1989. The effects offood limitation on the
Herzig, A., 1983b. The ecological significance of the relation- population dynamics, production and biological inter-
ship between temperature and duration of embryonic actions of three Daphnia species, co-existing in a London
development of rotifers. Hydrobiologia. 104: 237-46. reservoir. PhD Thesis. University of London (RHBNC).
Ivanova, M. B. & S. V. Vasilenko, 1987. Relationship 236 pp.
between number of eggs, brood weight and female body Schiemer, F., 1985. Bioenergetic niche differentiation of
weight in Crustacea. Int. Revue ges. Hydrobio!. 72(2): aquatic invertebrates. Verh. Int. Ver. Limno!. 22:
147-169. 3014-3018.
Jayatunga, Y. N. A., 1986. The influence of food and tem- Schiemer, F., A. Duncan & R. Z. Klekowski, 1980. A bio-
perature on the life cycle characteristics of tropical energetic study of a benthic nematode, Plectus palustris de
cladoceran species from Kalawewa Reservoir, Sri Lanka. Man 1880, throughout its life cycle. Oecologia (Ber!.). 44:
PhD Thesis. University of London (RHBNC) 410 pp. 205-212.
King, C. E., 1967. Food, age and the dynamics ofalaboratory Schnute, 1., 1981. A versatile growth model with statistically
population of rotifers. Ecology. 48( 1): 111-128. stable parameters. Can. J. Fish. Aquat. Sci. 38: 1128-1140.
King, C. E., 1969. Genetics of reproduction, variation and Serra, M. & M. R. Miracle, 1987. Biometric variation in three
adaptation in rotifers. Arch. Hydrobio!. Beih. Ergebn. strains of Brachionus plicatilis as a direct response to abio-
Limno!. 8: 187-201. tic variables. Hydrobiologia 147: 83-89.
Lampert, W., 1977. Studies on the carbon balance of Daphnia Sibly, R. M. & P. Calow, 1986. Physiological Ecology of
pulex De Geer as related to environmental conditions. IV. Animals. Balckwell Sci. Pub!. 179 pp.
Arch. Hydrobio!. Supp!. 48: 361-8. Snell, T. W., 1977. Clonal selection, competition among
Leimeroth, N., 1980. Respiration of different stages and clones. Arch. Hydrobio!. Beih. Ergbn. Limno!. 8: 202-204.
energy budget of juvenile Brachionus calyciflorus. Hydro- Snell, T. W. & E. M. Boyer, 1988. Thresholds for mictic
biologia. 73: 195-197. female production in the rotifer Brachionus plicatilis
Lubzens, E., 1987. Raising rotifers for use in aquaculture. (Muller). J. expo Mar. Bio!. Eco!. 124: 73-85.
Hydrobiologia. 147: 245-255. Snell, T. W. & K. Carrillo, 1984. Body size variations among
Lynch, M., 1980. Predation, enrichment and the evolution of strains of the rotifer Brachionus plicatilis. Aquaculture. 37:
cladoceran life histories: a theoretical approach. IN: W. C. 359-367.
Kerfoot (ed.). Evolution and ecology of zooplankton com- Snell, T. W. & c. E. King, 1977. Lifespan and fecundity pat-
munities. 367-376. Univ. Press, New Hampshire. terns in rotifers: the cost of reproduction. Evolution. 31(4):
Lynch, M., 1985. Elements of a mechanistic theory for the life 882-890.
history consequences offood limitation. Arch. Hydrobiol. Starkweather, P. L., 1987. Rotifera. chap. 5. IN: Pandian &
Beih. Ergebn. Limno!. 21: 351-362. Vernberg (eds.) Animal Energetics. 1: 159-183.
Pejler, B., 1980. Variation in the genus Keratella. Hydro- Sternberger, R. S. & J.1. Gilbert, 1985a. Body size, food
biologia. 73: 207-213. concentration and population growth in planktonic
Pilarska, 1., 1977. Eco-physiological studies on Brachionus rotifers. Ecology. 66(4): 1151-1159.
rubens Ehrbg. (Rotatoria). Po!. Arch. Hydrobio!. 24: Sternberger, R. S. & J. J. Gilbert, 1985b. Assessment of
319-354. threshold food levels and population growth in planktonic
Pourriott, R., 1973. Rapports entre la temperature, la taille rotifers. Arch. Hydrobio!. Beih. Ergebn. Limno!. 21:
des adultes, la longuer des reufs et Ie taux de developpe- 269-275.
ment embryonnaire chez Brachionus calyciflorus. Pallas Sternberger, R. S. & 1. 1. Gilbert, 1987. Rotifer threshold
(Rotifere). Ann. Hydrobio!. 4: 103-115. food concentrations and the size efficiency hypothesis.
Richards, F. J., 1959. A flexible growth function for empiracle Ecology. 68(1): 181-187.
use. J. expo Botany. 10: 290-300. Taylor, W. D., 1978. Growth response of ciliate Protozoa to
Robertson, 1. R. & Salt, G. W., 1981. Responses in growth, the abundance of their bacterial prey. Microb. Eco!. 4:
mortality and reproduction to variable food levels by the 207-214.
rotifer Asplanchna girodi. Ecology. 62(6): 1585-1596. Walz, N., 1983. Continuous culture of the pelagic rotifers,
Rocha, 0., 1983. The influence of food-temperature combi- Keratella cochlearis and Brachionus angularis. Arch. Hydro-
nations on the duration of development, body size, growth bio!. 98(1): 70-92.
and fecundity of Daphnia species. PhD Thesis, Royal Hol- Weglenska, T., 1971. The influence of various food concen-
loway College, University of London. 337 pp. trations of natural food on the development, fecundity and
Rocha, O. & Duncan, A., 1985. The relationship between cell production of planktonic crustacean filtrators. Eko!. Pol.
carbon and cell volume in freshwater algal species used in 19(30): 427-473.
zooplanktonic studies. J. Plankton. Res. 7(2): 279-294. Yufera, M., 1987. Effect of algal diet and temperature on the
Ruttner-Kolisko, A., 1974. Plankton Rotifers. Biology and embryonic development time of the rotifer Brachionus
Taxonomy. Die Binnengewasser. 26: 1-146. plicatilis. Hydrobiologia. 147: 319-322.
Hydrobiologia 186/187: 29-34, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 29
© 1989 Kluwer Academic Publishers.

Is food availability the main factor controlling the abundance of

Euchlanis dilatata lucksiana Hauer in a shallow, hypertrophic lake?

J. Ejsmont-Karabin,l R. D. Gulati 2 & J. Rooth 2

1 Polish Academy of Sciences, Institute of Ecology, Hydrobiological Station, ul. Lesna 13,
11-730 Mikolajki, Poland; 2 Limnological Institute, Vijverhof Laboratory, Rijksstraatweg 6,
3631 A C Nieuwersluis, The Netherlands

Key words,' Euchlanis dilatata lucksiana, blue-green algae, threshold concentrations of food, trophic
conditions, predation

Abstract

Visual observations and experiments on food preference of Euchlanis dilatata lucksiana show that this
euchlanid can feed on blue-green algae not consumed by the most planktonic animals. Nevertheless, even
in lakes with blooms of blue-green algae, E. d. lucksiana occur infrequently and generally in low numbers.
The paper is an attempt to explore into the causes for the rare occurrence of Euchlanis in the pelagial.
A comparison of threshold food concentrations calculated from N and P excretion rates (Gulati et aI.,
this volume) with the concentrations of seston in the Lake Loosdrecht shows that the latter were several
times higher during study period in 1984. This implies that the food requirements of Euchlanis were always
satisfied in this lake. The time needed for the consumption of the total food fraction in a liter oflake water
by a concentration of 50 Euchlanis 1- 1 was also calculated. This time varied from 70 to 200 days, so a
Euchlanis population even at its maximum density will not cause major changes in blue-green algae
biomass by grazing. Thus, food limitation cannot be viewed as a factor controlling the Euchlanis densities
in Loosdrecht Lakes. There is some evidence that Euchlanis is heavily predated in Loosdrecht Lakes,
losses in its biomass accounting for 126 % of the production. Adaptation of this species to the littoral
zone, as expressed by the deposition of eggs on plants, can also limit the occurrence of the lucksiana form
to water bodies with blooms of blue-green algae.

Introduction dance. This species may, however, rarely domi-

nate in the pelagial like in Glubokoe Lake
Unlike other forms of Euchlanis dilatata which are (Matveeva, 1986), coinciding with blooms of the
littoral, E. dilatata lucksiana is a pelagic form blue-green alga Aphanizomenon. Also, Carlin
(Kutikova, 1970; Koste, 1978). In fact, this (l943) noted a high correlation between the occur-
euchlanid is rarely observed also in pelagial and rence of Euchlanis and blue-green algae (Oscilla-
usually in small numbers. For example, Karabin toria and others). Even if observations of this kind
(l985a), who studied summer communities of can be viewed as sufficient to say that larger
Rotifera in 64 lakes of north-eastern Poland, most appearances of E. d. lucksiana in the pelagic zone
of which had blue-green algae (Spodniewska, occur only during blooms of filamentous algae,
1983), did not find Euchlanis dilatata in abun- there is some evidence that the reverse statement
30

is not true. Blooms of blue-green algae are rarely rates of nitrogen and phosphorus excretion
combined with the appearance of Euchlanis. (Gulati et al., this volume).
The present paper is an attempt to explore the The formula given by Rooth (1985) was used to
factors responsible for the rare occurrence of calculate a relationship between the rate of food
E. d. lucksiana in pelagial of lakes. assimilation by E. d. lucksiana and food concen-
tration:

Material and methods A = 0.98 _ 0.51 , (1)

FC
Information used in this paper comes from 3 main
sources: 1) literature data, 2) unpublished results where: A = assimilation in JigC indo - I d - I
of field studies, and 3) partly published results FC = food concentration in JigC ml- I.
(Rooth, 1985; Gulati et al., 1987) of laboratory
experiments on the rate of food assimilation and The minimum metabolism of Euchlanis was deter-
the rate of phosphorus and nitrogen excretion by mined by using two indices: the rate of phos-
E. d. lucksiana. phorus or nitrogen excretion by starved animals,
The study was conducted in Lake Loosdrecht, assuming that the rate of food assimilation corre-
a shallow hypertrophic lake with a mean depth of sponding to this excretion rate is the minimum
1.85 m. Except in early spring, the phytoplankton that Euchlanis requires to survive. QIO values of
of this lake was dominated by filamentous blue- 2.6 for phosphorus and 2.4 for nitrogen
green algae (mainly Oscillatoria redekei, O. agard- (Ejsmont-Karabin, 1984) were used to relate the
hii and Aphanizomenonjios-aquae) (Bosewinkel-de rate ofP and N excretion to ambient temperature.
Bruyn et al., 1986). U sing the rate of nutrient excretion, the corre-
Zooplankton samples were collected every four sponding rate offood assimilation, E PorN (C : P or
weeks or fortnightly from January to October, C : N), was calculated. This takes into account the
from a station in the lake. Data on carbon, phos- C : P or C : N ratio by weight in the food fraction.
phorus and nitrogen concentrations in the seston Thus, the threshold concentration was calculated
fraction < 3 Jim taken concurrently with the from the formula:
zooplankton samples were provided by Gulati (in
press). TFC = 0.51 , (2)
The females of E. d. lucksiana were obtained 0.98 - EPorN/C: P or C : N)
from a culture developed from ten individuals
taken from Loosdrecht Lakes. Cultured rotifers where: TFC = threshold concentration of food
were fed on the lake seston fraction < 33 Jim, i.e. in JigC ml- I
in lake water filtered using 33 Jim mesh. This E PorN = rate of phosphorus or nitrogen
fraction comprised mostly blue-green algae which excretion in Jig indo - I d - I
were the main food of the cultured animals. The C : P or C : N = carbon to phosphorus or
optimum rotifer density maintained at a constant nitrogen ratio by weight
level in the culture was about 40 indo ml- I. Light in food.
in the laboratory corresponded to natural condi- The time needed for the consumption of the
tions prevailing during the same period. total content of carbon contained in 1 ml of lake
Food requirements of Euchlanis, both con- water of the seston fraction < 33 Jim by one
sumption and assimilation rates, in relation to the individual was calculated using the equation given
available food supply, have been analysed in the by Rooth (1985):
laboratory using C I4 technique (Gulati et ai.,
1982; Gulati et ai., 1987). Threshold food require- CON = 1.17 _ 0.92 , (3)
ments of Euchlanis were determined from the FC
 31

where: CON = consumption in J-lgC indo -1 d- 1 These densities are very low, however, compared
FC = as in formula 1. with those of the dominant rotifers like
Anuraeopsis fissa (Gosse), with which the
Euchlanis maxima coincided.
Results and discussion Trophic conditions. Food availability and abun-
dance are viewed as among the most important
The rotifer community of Loosdrecht Lakes was factors determining numbers and the structure of
dominated by small species, mostly detrito- zooplankton (Benndorf & Horn, 1985; Karabin,
phagous Anuraeopsis, Keratella (Fig. 1). The 1985b). However, visual observations and experi-
highest densities occurred from June to early ments on food preference (Gulati et al., 1987)
October. Euchlanis dilatata lucksiana appeared in show that E. dilatata lucksiana can feed on blue-
small numbers (1 indo 1- 1) in April, but reached green algae not consumed by most planktonic
about 50 indo 1- 1 in mid-August and September. animals (Dumont, 1977). Nevertheless, even in

11
"".......
I

IV)
8
~
"ti 7
.~
1.1)"' 6
~j::;: 5
<::>
t:I:: 4
I.!..
~
3
V:l
Q::
~
ttl
2
~~ 1

JAN FfB MAR APR MAY JUN JUL AUG SfP OCT
'tooo
\
.......
~ 50
.~

~' 40
~
~
;t 30
~
~ 20

~ 10
">
~
~
JAN fEB MAR APR MAY JUN JUL AUG SIP OCT
~
~
Fig. 1. Changes in numbers of the main genera of rotifers and Euchlanis dilatata lucksiana in Loosdrecht Lakes in 1984.
32

lakes with blooms of blue-green algae, E. d. luck- 0.53 to 0.73 ,ugC ml- 1) than for N excretion. A
siana occur infrequently and generally in low comparison of threshold concentrations with the
numbers. Is such a high density of algae insuf- actual concentration of seston in Loosdrecht
ficient to meet the food requirements of this Lakes (Fig. 2) shows that the latter were several
rotifer? To test this hypothesis, food consumption times higher during study period. This implies that
of Euchlanis was compared with food supply in in 1984 there was no situation in which food
water in an attempt to determine a critical food requirements of Euchlanis could not be satisfied in
concentration (equations 1 & 2) below which this lake.
Euchlanis would not be able to meet their metabo- The effect of Euchlanis on the phytoplankton
lic needs. depends on rotifer densities. The time needed for
The ratio by weight of the nutrients excreted by consumption of the total carbon content of 1 ml
Euchlanis was C: N: P = 25: 4: 1 (Gulati et aI., lake water of the seston fraction < 33 ,urn by one
this volume). Seasonal mean ratio of these individual was calculated using the equation 3.
nutrients in seston < 3 ,urn was 113 : 17: 1, respec- The calculated turnover time varied from 3 to 6
tively. Thus, compared with phosphorus there days in spring and autumn (Table 1) and 7 to 10
was relatively more nitrogen in food than in the days in summer. Thus, a Euchlanis population
excretion products. Therefore, the threshold con- comprising 1000 indo 1- 1 (a concentration 2
centrations of food calculated from the rate of N orders of magnitude higher than normally
excretion are lower than from P excretion (Fig. 2). encountered in the lake) would not cause any
Also, they were almost stable during the season .. major change in blue-green algal biomass by
The range of the threshold concentrations calcu- foraging. The 10-20 %daily loss due to grazing by
lated from the rate ofP excretion was higher (from Euchlanis could have been easily compensated for

11
10
9

-
.....
I

'-.J
~
8
7
6 c
~
':{5

4
3
2
1

FEB MAR APR MAV JUN JUL AUG ,rEP OCT NOV
Fig. 2. Changes in actual concentrations of33 pm seston carbon (C), and its threshold concentrations for Euchlanis, as calculated
from the rate of phosphorus (P) and nitrogen (N) excretion.
 33

Table 1. Seasonal changes in the concentration of 33 /lm ture at which it occurred was 12 to 21 ° C, with
seston, consumption rate of algae by Euchlanis dilatata two maxima at 13°C and 21 °C. Its low numbers
lucksiana and turnover time of the seston C as a result of
Euchlanis foraging.
at intermediate temperatures (14-20 0c) in late
spring and early summer cannot be explained.
Date Seston Consumption Turnover
/lgC ml- 1 rate time Predation. The fact that in the cultures Euchlanis
/lgC indo - 1 d - 1 (days) reached very high densities means that in labora-
84.02.15 3.24 0.88 4 tory the factors limiting the occurrence of these
03.14 2.85 0.85 3 animals in the lake were relieved. The absence of
04.11 4.63 0.97 5 predators, such as fish, may contribute to their
04.24 7.48 1.05 7 high densities in the cultures.
05.09 6.03 1.02 6 E. d. lucksiana is a relatively large rotifer, which
06.06 7.85 1.05 7
06.20 8.98 1.07 8
reaches the maximum 400 J.l.m in length in Lake
07.04 8.51 1.06 8 Loosdrecht. Their size is thus within the range of
07.18 9.51 1.07 9 sizes of Bosmina coregoni Baird and cyclopoids,
08.01 10.66 1.08 10 which are heavily predated by bream in this lake
08.15 10.77 1.08 10 (Van Densen et al., 1986). Smaller individuals of
08.29 9.56 1.07 9
09.12 8.69 1.06 8
Euchlanis may also be predated by invertebrate
09.26 8.98 1.07 8 predators, such as Leptodora kindtii (Focke)
10.24 7.36 1.06 7 (Dumont, 1977).
11.21 6.36 1.03 6 To test this hypothesis, a total loss of rotifers
(D) in the water column was estimated using the
method described by Duncan & Gulati (1983),
due to primary production. Therefore, food avail- which is based on the formula:
ability cannot be the factor limiting Euchlanis den-
sities in Loosdrecht Lakes. AN
D=B-- ,
At

Other factors where: B = recruitment rate

Temperature. The fact that peak numbers of
-AN = ch ange m
. stand'mg crop.
Euchlanis in Loosdrecht Lakes were noted in late At
summer suggests that ambient temperature
should be considered among the factors con- In a field study lasting 29 h on 11-12 September,
trolling the occurrence of this rotifer. E. dilatata Euchlanis was found to be among the most heavily
can occur over a large range of ambient tempera- predated rotifers; only for the genus Polyarthra the
tures, from 0.1 to 31 ° C, and is considered as an elimination rates were higher. Losses in Euchlanis
eurythermic species (Lair, 1980; Bazilevitsch, biomass accounted for 126 % of its production.
1985; Kosova, 1985). There is no information of This result seems to support the hypothesis that
this kind on its pelagic form. Its clear peaks in late the Euchlanis population is under heavy predation
summer in both Glubokoe Lake (Matveeva, pressure.
1986) and Lake Loosdrecht suggest that this may
be a thermophilous form. This suggestion is not
confirmed, however, by the analysis of the occur- Conclusion
rence of Euchlanis in Lake Loosdrecht in the
annual cycle. Presence of this species in the lake The adaptation of Euchlanis to living in the littoral
was observed in April and the range of tempera- zone, as expressed by the deposition of eggs on
34

plants, can limit the occurrence of its lucksiana natural food particles on tropical species of planktonic
rotifers. In F. Schiemer (ed.), Limnology of Parakrama
form to the bodies of water with abundant blooms
Samudra - Sri Lanka: a case study of an ancient man-made
of blue-green algae. This form lay eggs on tight lake in the tropics. Developments in Hydrobiology 12:
clusters of filamentous blue-green algae. How- 117-125.
ever, Euchlanis females raised in aquaria did not Ejsmont-Karabin, J., 1984. Phosphorus and nitrogen excre-
use blue-green algae as a substratum, but attached tion by lake zooplankton (rotifers and crustaceans) in
relationship to individual body weights of the animals,
their eggs to the aquarium walls. It is expected
ambient temperature and presence or absence of food.
that there is a threshold of the abundance of fila- Ekol. pol. 32: 3-42.
mentous algae below which searching for dense Gulati, R. D. (in press). Zooplankton structure in
clusters of algae and the deposition of eggs on Loosdrecht Lakes in relation to the trophic status and the
them cannot be worth the energy invested. recent restoration measures. Hydrobiologia.
The dominance of blue-green algae was noted Gulati, R. D., J. Rooth & J. Ejsmont-Karabin, 1987. A labo-
ratory study of feeding and assimilation in Euchlanis
mostly in hypertrophic lakes dominated by small
dilatata lucksiana. Hydrobiologia 147: 289-296.
forms of zooplankton. Euchlanis, which is rela- Gulati, R. D., K. Siewertsen & G. Postema, 1982. The
tively large and conspicuous, is likely to be subject zooplankton: its community structure, food and feeding
to a high predation pressure and, because of its and its role in the ecosystem of Lake Vechten. Hydro-
size, it can be attacked by invertebrate predators biologia 95: 127-163.
Gulati, R. D., J. Ejsmont-Karabin, J. Rooth & K. Siewertsen,
as well as by early developmental stages of fish.
1989. A laboratory study of phosphorus and nitrogen
Due to their ability to feed on blue-green algae, excretion of Euchlanis dilatata lucksiana. this volume.
which are not often consumed by other zoo- Karabin, A, 1985a. Pelagic zooplankton (Rotatoria +
plankters, Euchlanis can occasionally reach den- Crustacea) variation in the process oflake eutrophication.
I. Structural and quantitative features. Ekol. pol. 33:
sities of the order of several ten individuals per
567-616.
liter (as in Lake Loosdrecht) or even several Karabin, A., 1985b. Pelagic zooplankton (Rotatoria +
hundred individuals (as in Glubokoe Lake; Crustacea) variation in the process oflake eutrophication.
Matveeva, 1986), although it is not well adapted II. Modifying effect of biotic agents. Ekol. pol. 33: 617-644.
to the life in the pelagic zone. Kosova, A A, 1985. Ecological characteristics of Rotifera of
reservoirs of the Volga Delta. In L. A. Kutikova (ed.),
Rotatoria, Proceedings of the National Rotifer Sympo-
sium, Nauka, Leningrad: 199-204. (in Russian).
References Koste, W., 1978. Rotatoria, Monogononta. Die Radertiere
Mitteleuropas. Gebriider Borntraeger, Stuttgart, Vols 1 &
Bazilevitsch, V. M., 1985. Rotifera of some canals of 2, 673 pp., 234 pis.
southern Ukraine. In L. A Kutikova (ed.), Rotatoria, Kutikova, L. A., 1970. Rotifer Fauna USSR. Fauna USSR.
Proceedings of the National Rotifer Symposium. Nauka, 104. Acad. Nauk. SSSR, Leningrad, 744 pp. (in Russian).
Leningrad: 205-208. (in Russian). Lair, N., 1980. The rotifer fauna of the river Loire (France),
Benndorf, J. & W. Horn, 1985. Theoretical considerations on at the level ofthe nuclear power plants. Hydrobiologia 73:
the relative importance offood limitation and predation in 153-162.
structuring zooplankton communities. Arch. Hydrobiol. Matveeva, L. K., 1986. Pelagic rotifers of Lake Glubokoe
Beih. Ergebn. Limnol. 21: 383-396. from 1897 to 1984. Hydrobiologia 141: 45-54.
Boesewinkel-de Bruyn, P. J., L. van Liere & B. Z. Salome, Rooth, J., 1985. Een laboratoriumstudie van het metabolisme
1984. Phytoplankton species composition. In S. Parma & van de rotiferen - soort Euchlanis dilatata lucksiana uit de
R. D. Gulati (eds), Limnological Institute, Progress Report Loosdrechtse Plassen. Limnologisch Instituut Nieuwer-
1983. Verh. Kon. Ned. Akad. Wet., Afd. Nat., Tweede sluisjOosterzee, Studentenverslag nr. 1985-5, 50 pp. (in
Reeks, 82. Dutch).
Carlin, B., 1943. Die Planktonrotatorien des Motalastrom. Spodniewska, I., 1983. Ecological characteristics oflakes in
Medd. Lunds Univ. Limnol. Inst. 5: 260 pp. north-eastern Poland versus their trophic gradient. VI.
Dumont, H. J., 1977. Biotic factors in the population dynam- The phytoplankton of 43 lakes. Ekol. pol. 31: 353-382.
ics of rotifers. Arch. Hydrobiol. Beih. Ergebn. Limnol. 8: Van Densen, W. L. T., C. Dijkers & R. Veerman, 1986. The
98-122. fish community of the Loosdrecht Lakes and the perspec-
Duncan, A. & R. D. Gulati, 1983. Feeding studies with tive for biomanipulation. Hydrobiol. Bull. 20: 147-163.
Hydrobiologia 186/187: 35-38, 1989.
C. Ricci. T. W. Snell and C. E. King (eds), Rotifer Symposium V. 35
© 1989 Kluwer Academic Publishers.

Influence of cyanobacterial diet on Asplanchna predation risk in

Brachionus calycijlorus

Peter 1. Starkweather & Elizabeth J. Walsh'

Department of Biological Sciences, University of Nevada, Las Vegas, Las Vegas, Nevada 89154 U.S.A.;
'Department of Zoology, Oregon State University, Corvallis, Oregon 97331 U.S.A.

Key words.' Anabaena, Asplanchna, Brachionus, rotifer, cyanobacteria, feeding behavior, predator-prey
interaction

Abstract

Asplanchna sylvestrii does not discriminate between groups of Brachionus calyciflorus fed either the
cyanobacterium Anabaenaflos-aquae or a control diet of Euglena gracilis. We based our analysis on the
observed probabilities of attack, capture and ingestion during encounters between predator and prey.
While A. sylvestrii was very sensitive to brachionid size, we found no significant affects of prey diet on
predatory behavior. Thus, cyanobacterial diet did not influence the short-term predation risk of
B. calyciflorus exposed to an effective predator. On the other hand, matched cohorts of A. sylvestrii fed
B. calyciflorus cultured on the cyanobacterium reproduced more slowly than those fed the same prey
cultured on the control food. With prolonged sympatry, therefore, the long-term risk of Asplanchna
predation may be reduced for Brachionus by the latter's consumption of cyanobacteria.

Introduction biomass and the productivity of several sus-

pension-feeding rotifers.
The monogonont rotifer Brachionus calyciflorus On the other hand, if grazers on cyanobacteria
consumes several species of cyanobacteria, with were avoided by predatory species, the energetic
particularly well-documented benefit derived impact of 'blue-greens' would be restricted. They
from the filamentous Anabaenaflos-aquae (Stark- would contribute to secondary production (of
weather, 1981; Starkweather & Kellar, 1983). brachionid rotifers, for instance), but would
Thus, in freshwater systems in which this rotifer represent a less-than-proportional contribution to
or ecologically similar taxa are present, a fraction the energetics of higher trophic levels.
of cyanobacterial primary production may be In this paper we report the first experiments to
directed into conventional grazing food webs. The determine whether or not a prey diet of
more traditional view that cyanobacterial cyanobacteria may deter immediate predation for
biomass is largely relegated to detrital energetic individuals of a suspension feeding species or
pathways may, therefore, be subject to some mo- diminish predator effectiveness in the longer term.
dification. These findings support the conclusions For rotifer predator-prey systems, both short and
of field studies (cf. Lewis, 1979) which have long term dietary influences are important. In the
shown correlations between cyanobacterial former case, single prey may benefit from reduced
36

individual predation risk; in the latter, prey vation we pipeted several Brachionus into the
(clonal) survival is enhanced by inhibition of vessel containing a single Asplanchna. We ob-
predator population growth. The intent of the served and recorded the following predator-prey
work is both to examine the behavioral implica- interactions: encounter, attack, capture and in-
tions of predator functional response as in- gestion, in each instance noting the success or
fluenced by prey diet and to determine if a failure of sequential steps in the functional pro-
moderately toxic prey diet may have a negative cess. To avoid any influence of satiation, we trun-
impact on predator population growth. cated observation of each predator after ingestion
of three prey.
To determine the influence of prey diet on
Methods and materials predator population dynamics we established
matched cohorts of A. sylvestrii neonates drawn
We cultured the Brachionus calyciflorus used in from cultures fed Brachionus grown on Euglena.
this study on pure cultures of either Euglena gra- We supplied each Asplanchna with 10 prey per
cilis (UTEX-753) or Anabaenajlos-aquae (NRC- day, a rate above the threshold for sustained
44-1). Both foods allow sustained population population growth, but well below saturation
growth for the rotifer (Starkweather & Kellar, (Sternberger & Gilbert, 1984). We observed each
1983), the latter despite its persistent toxicity (as animal in each cohort (12-18 animals per cohort,
measured in mammalian bioassays; W. Carmi- n = 3 for each of two treatments) daily and
chael, pers. commun.). Before use in predation recorded individual survival and reproduction.
trials, we selected individual animals and washed All steps in this study, from food culture through
them free of residual food materials and culture direct observation, were performed at 20 0 C.
medium. The predator in this system, Asplanchna
sylvestrii, had been previously isolated from Soda
Lake, Nevada, U. S .A.; we cultured individuals Results
and their immediate progeny in tissue culture well
plates using excess numbers of Euglena-fed B. ca- Our ftrst observations of predator-prey inter-
lyciflorus as their only food. For each experiment, actions determined that prey reproductive state,
we isolated a single Asplanchna and placed her in perhaps reflecting age and certainly body size, has
fresh rotifer medium. Predators were maintained a substantial influence on Brachionus susceptibili-
without food for between 15 min to 4 h; however, ty to predation by these Asplanchna. Table 1
the measurements we report here were drawn shows that smaller non-ovigerous (and likely ju-
largely from observations of 85 animals held venile) B. calyciflorus have both signiftcantly
without food for less than 2.5 h. For each obser- greater likelihood of attack after encounter and

Table 1. Influence of prey (Brachionus calyciflorus fed Euglena gracilis) reproductive condition on predatory responses of adult
Asplanchna sylvestrii. Values shown are mean probabilities ± s.d. for all observed predator-prey encounters; parenthetical values
indicate ranges for individual animals. Differences between groups evaluated using the Kruskal-Wallis ranks test (* = p < 0.05).

Prey Attack after Capture after Ingestion after Ingestion after

condition encounter attack capture encounter

Non-ovigerous 0.80 ± 0.32 0.71 ± 0.28 0.69 ± 0.30 0.44 ± 0.37

(0.19-1.00) (0.20-1.00) (0.20-1.00)
Ovigerous 0.56 ± 0.24 0.56 ± 0.27 0.39 ± 0.31 0.14 ± 0.16
(0-1.00) (0-1.00) (0-1.00)

p(K.-W.) * n.s. * *
 37

Table 2. Influence of prey diet on predator responses of adult A. sylvestrii to ovigerous B. calyciflorus cultured on dits of either
E. gracilis or Anabaena fios-aquae. Numerical values and statistical treatment as in Tab. 1.

Prey Attack after Capture after Ingestion after Ingestion after

diet encounter attack capture encounter

Euglena 0.56 ± 0.24 0.56 ± 0.27 0.39 ± 0.31 0.12 ± 0.16

(0-1.00) (0-1.00) (0-1.00)
Anabaena 0.58 ± 0.31 0.66 ± 0.35 0.37 ± 0.43 0.14 ± 0.38
(0.19-1.00) (0.14-1.00) (0-1.00)

p(K.-W.) n.s. n.s. n.s. n.s.

ingestion after capture than do ovigerous adults. affect on B. calyciflorus vulnerability to predation
The two reproductive classes do not differ in their by A. sylvestrii. In neither composite (ingestion
probability of capture after attack. The combined after encounter) nor subsidiary behaviors was
probabilities for the entire functional sequence there any indication that the predator detected or
(ingestion after encounter) indicates that, in a responded to a factor correlated with prey diet. It
given interaction, ovigerous animals are at least is also important to note here that we observed no
three times less likely to be ingested than the differences in prey size, shape or behavior based
non-ovigerous and smaller individuals. on their original diets. Thus, the lack of predator
These results led us to use only ovigerous (and response was not due to confounding factors
therefore somewhat larger and older) prey when which might mask an influence based on prey
examining the effect of prey diet on susceptibility diet.
to Asplanchna predation. We wanted to maximize Asplanchna sylvestrii cohorts fed Brachionus cul-
the opportunity for the Brachionus to accumulate tured on Euglena have instantaneous population
any chemical 'signal' which might influence growth rates approximately twice those of match-
A. sylvestrii behavior and to extend the period of ed groups fed Anabaena-cultured prey (Table 3).
handling of prey (due to their larger size) by This result appears to be due almost exclusively
individual predators - again, to maximize the to the differential in net fecundity (Ro), with no
possibility of the predators detecting and re- significant differences between the two groups in
sponding to any such factor. median survivorship (O.5Ix ) or calculated gener-
Table 2 shows that in these experiments prey ation time (T).
diet (Euglena vs. Anabaena) had no significant

Table 2. Influence of prey diet on predator responses of adult A. sylvestrii to ovigerous B. calyciflorus cultured on dits of either
E. gracilis or Anabaena fios-aquae. Numerical values and statistical treatment as in Tab. 1.

Prey Attack after Capture after Ingestion after Ingestion after

diet encounter attack capture encounter

Euglena 0.56 ± 0.24 0.56 ± 0.27 0.39 ± 0.31 0.12 ± 0.16

(0-1.00) (0-1.00) (0-1.00)
Anabaena 0.58 ± 0.31 0.66 ± 0.35 0.37 ± 0.43 0.14 ± 0.38
(0.19-1.00) (0.14-1.00) (0-1.00)

p(K.-W.) n.s. n.s. n.s. n.s.

38

Summary and conclusions upon the species of rotifer present, the type of
cyanobacteria available in suspension and the
In these experiments, cyanobacterial diet had no period of sympatry of all involved taxa.
effect on the short-term susceptibility of prey
(B. calyciflorus) to predation by an expectedly
selective predator (A. sylvestrii). The expectation
Acknowledgements
here was based on literature reports which have
shown species of Asplanchna to be extraordinarily
We extend sincere appreciation Penelope E.
sensitive to the nature of their prey, distinguishing
Kellar for expert technical assistance and to Dr.
even between alternate con specific clones (see
Wayne Carmichael for providing the
Gilbert, 1980). Accordingly, we conclude that
cyanobacterial culture. Supported by the Na-
cyanobacterial production in a pelagic system
tional Science Foundation (U.S.) DEB-8105724.
containing appropriate rotifers can be converted
to animal biomass throughout the grazing food
web.
As noted earlier, Brachionus calyciflorus can References
utilize the current strain of A.flos-aquae as a sole
or complementary food source, despite its toxic Gilbert, J.1., 1980. Feeding in the rotifer Asplanchna: Be-
havior, cannibalism, selectivity, prey defenses and impact
properties (Starkweather & Kellar, 1983). While
on rotifer communities. pp. 158-172. In: W. C. Kerfoot
the cyanobacterial diet does not protect the (ed.). Evolution and Ecology of Zooplankton Commu-
rotifers from individual predator attack, including nities. University Press of New England. Hanover, NH.
A. flos-aquae in the diet may have a beneficial Lewis, W. M. Jr., 1979. Zooplankton Community Analysis.
influence beyond nutrition. The results described Springer Verlag. New York.
show that long-term population growth of a Starkweather, P. L., 1981. Trophic relationships between the
rotifer Brachionus calyciflorus and the blue-green alga
predatory species may be reduced when prey diet Anabaena jlos-aquae. Verh. into Ver. Limnol. 21:
includes 'blue-greens'. This type of effect may be 1507-1514.
particularly significant in zooplankton predator- Starkweather, P. L. & P. E. Kellar, 1983. Utilization of
prey systems involving two rotifers, since gener- cyanobacteria by Brachionus calyciflorus: Anabaena jlos-
ation times are short (2-5 d) and overlapping. It aquae (NRC-44-1) as a sole or complementary food
source. Hydrobiologia 104: 373-377.
remains to be seen if such processes operate in Sternberger, R. S. & J. J. Gilbert, 1984. Body size, ration
natural pelagic communities; we would predict level and population growth in Asplanchna. Oecologia. 64:
that the influences described here would depend 355-359.
Hydrobiologia 186/187: 39-42, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), RotiJer Symposium V. 39
© 1989 Kluwer Academic Publishers.

Nutritional effect of freshwater Chlorella on growth of the rotifer

Brachionus plicatilis

K. Hirayama 1, 1. Maruyama 1, 2 & T. Maeda 2

1Graduate School of Marine Science and Engineering, Nagasaki University, Nagasaki, Nagasaki, 852
Japan; 2Chlorella Industry Co., Ltd., Chikugo, Fukuoka, 833 Japan

Key words: Freshwater Chlorella, nutritional effect, Brachionus plicatilis, Vitamin B 12 , supplementary
food

Abstract

Mass production of Brachionus plicatilis is usually accomplished by feeding so-called marine Chlorella
(Nannochloropsis oculata) to the rotifers in marine fish hatcheries. If the marine Chlorella are in short
supply, baker's yeast is usually used as a supplementary food. Recently, a condensed suspension of
freshwater Chlorella (Chlorella vulgaris, k-22) was commercially developed as another supplementary
food. We have evaluated the dietary value of this freshwater Chlorella for growth of the rotifer by means
of individual and batch cultures. Rotifers cultured with the freshwater Chlorella suspension under almost
bacteria-free conditions, showed very suppressed growth. However if the ChIorella was supplemented
with vitamin B 12 by adding the vitamin solution into the suspension or by culturing the ChIorella in a
medium containing vitamin B 12 , the nutritional value of freshwater Chlorella was greatly improved and
almost at the same level as that of marine ChIorella. Condensed Chlorella may therefore be effective as
a supplementary food if vitamin B 12 is supplied.

Introduction (Hirayama & Funamoto, 1983). There have been

many attempts to develop another supplementary
In Japan, mass production of the rotifer, food or substitute for marine Chlorella. Con-
Brachionus plicatilis is usually conducted by densed suspensions of freshwater Chiorella have
feeding so-called 'marine Chlorella', which have been developed as commercially available supple-
been taxonomically transferred from Chlorella to mentary foods. However, some culturists have
Nannochloropsis oculata (Maruyama et aI., 1986). complained that Chlorella suspensions some-
In marine hatcheries, a much greater area is pro- times produce suppressed or unstable growth. In
vided for mass production of marine Chlorella this study, we evaluate the nutritional effect of
than for the rotifer. At times, especially in the freshwater ChIorella on rotifer growth under al-
summer, rotifer production exceeds the required most bacteria-free conditions. We report that
supply of marine Chlorella for food. At that time, freshwater Chlorella is nutritionally deficient for
baker's yeast is usually supplied as a supplemen- rotifer growth unless the food suspension
tary food for the rotifer. However, baker's yeast contains vitamin B 12 •
is nutritionally incomplete for rotifer growth
40

Materials and methods fIrst-laid eggs were placed in the food suspension
to be tested and after 4-7 days the number of
A freshwater Chlorella (Chlorella vulgaris k-22) individuals was compared with that in control
was cultured in 100 rnl of a medium which does food suspensions. In the individual cultures, the
not contain vitamin B 12 (Table 1), by using 500 ml fIrst-laid eggs were cultured individually in the test
glass flasks with continuous shaking (25 0 C, 4000 suspension. We followed the growth of each
lux). After harvesting at (5-10) x 108 cells ml- 1, individual by observation of daily survival and
we washed this suspension with sterile water by number of eggs laid. These data were used to
centrifugation and then resuspended it at calculate net reproduction rate (Ro) and intrinsic
10 x 108 cells ml- 1. This treatment made the sus- rate of population increase (r) on the basis of
pension almost bacteria free. Marine ChIorella Birch's computational method (Birch, 1948).
(Nannochloropsis oculata) used as a control was Details of the individual culture method are
cultured under bacteria-free conditions in explained elsewhere (Hirayama & Funamoto,
Erdschreiber medium supplemented with vitamin 1983).
B 12 at 0.5 f.,lg 1- 1. Additionally, freshwater
ChIorella was cultured in medium supplemented
with vitamin B 12 at 50 f.,lg 1- 1. Baker's yeast was Results and discussion
cultured under aeration in Yeast Nitrogen Base of
Wako Company plus saccharose. Rotifer eggs Results of these experiments are presented in
used for the experiments were sterilized by using Fig. 1. Growth of the rotifer on freshwater
an antibiotic mixture (AM9) (Provasoli et al., ChIorella was suppressed and did not change with
1959). Throughout our experiments, we used bac- increase of Chlorella density. Both r and Ro were
teria-free techniques so as to avoid the excess
contamination.
In order to evaluate the nutritional effects of
0
foods or supplementary nutrients, the fIrst-laid
eggs collected from an actively growing culture of 10 0
rotifers were divided into several groups. These
groups were cultured in the suspension to be
tested and in control food suspensions, by means 5 Ro <>
of batch culture or individual culture. For the 0
\l
batch culture method, offspring hatched from
0
340 681 1362 _10 4
(cells/ml)
Table 1. Composition of culture medium for freshwater
Chlorella.
• •
Composition

Glucose
(NH 2 hCO
Amount

109
1.2 g
0.5

0
•
~

A
KH 2 P0 4 1.2 g
MgS04 '7H 2 0 0.6 g
Fe.EDTA 15 mg - 0.5L------<>-----<------
A5 solution 1 mg 340 681 1362 (b~ls/ml)
Tap water 1000 ml Fig. 1. Net reproduction rates (Ro) and intrinsic rates of
population increase (r) of the rotifers cultured by individual
pH 6.6 cultures method in freshwater Chlorella suspensions at
various cell densities. The same symbol is used to represent
A5 solution contains metals of Mn, Zn, Cu, Mo and B. values from the same experiment.
 41

quite variable. These observations suggest that Ro r

the nutritional value of freshwater Chlorella is ...
usually very low and that different lots of fresh-
"
.20 1.
water Chlorella varied in nutritional value even
under similar conditions. We suggest that this
may be due to the growth phase at harvest of the
Chlorella culture.
16 0.5
Figure 2 presents the nutritional effects of
marine and freshwater Chlorella on growth of the
rotifer and also the effects of supplementation
with baker's yeast. Freshwater Chlorella had very
low nutritional value compared with marine
Chlorella. Supplementation of low densities of
0 1 I.
M.Chlor<l'lIa F.Chtor<l'lIa
(1) (1)
Y<l'as\
1.
Y<l'ast
M.Chlor<l'lIa F.Chlor<l'lIa
o
(0.03) ( 0.03)
both Chlorellas with baker's yeast had little effect
on growth of the rotifer. Fig. 2. Nutritional effects of marine and freshwater Chlorella
Figure 3 gives the results of 4 day batch cultures on rotifer growth and effects of supplementation of both
Chlorella with baker's yeast. The number in parentheses
in the freshwater Chlorella suspensions with and
presents the cell density of Chlorella. Number one is used to
without supplementation by vitamin B 12 solution indicate a density of 681 x 104 cells ml- I. The density of
at 1.4,ug ml- 1 and baker's yeast. Growth on a baker's yeast is 200 J.Lg ml- I.
marine Chlorella suspension is presented as a
control. Vitamin B 12 substantially improved the
nutritional value of the freshwater Chlorella sus- tation of the rotifer medium or use of marine
pension. Chlorella cultured in a medium con- Chlorella. Effects of supplementation by vitamin
taining vitamin B12 produced almost as many B12 using the two methods with individual cul-
rotifers as either direct vitamin B12 supplemen- tures are presented in Fig. 4. These results con-

cuture (includ 5)
Supplement
40 100 200 300 (num b er) 400
F.Ch 110
death
F.Chlorello ( 1) surv ival
+ VB 12

l nit ial number

Fig. 3. Numbers of individuals after 4 day batch cultures of the rotifer in marine and freshwater Chlorella suspensions and in
vitamin B12 supplemented freshwater Chlorella suspensions. Explanations ofthe numbers in parentheses and yeast density are
in Fig. 2. F. Chlorella (V B 12) indicate freshwater Chlorella cultured in media containing vitamin B 12 at 50 J.Lg 1- I. Chlorella + V
B 12 indicates addition of vitamin B 12 to the rotifer medium at 1.4 j.tg ml- I.
 42

Ro r even ifbaker's yeast is supplemented with vitamin

o • B 12 , its nutritional value is still much lower than
20 that of marine Chlorella. Freshwater Chlorella
also has low nutritional value for rotifer growth.
However, if vitamin B12 is used to supplement the
freshwater Chlorella suspension by either ad-
10 0.5 dition to the algal or to the rotifer medium, rotifer
growth is improved to the level of marine
Chlorella which is considered as the best food for
o o the rotifer.

F.Chlor~lIa F.Chlor~lIa( 1) F.Chlor~\la(l)

OJ ( V.B12 )
V. 811
References
Fig. 4. Effects on rand Ro of vitamin B12 supplementation.
Explanations of abscissa labels are in Fig. 3. Birch, L. c., 1948. The intrinsic rate of natural increase of an
insect population. J. Anim. Ecol. 17: 15-26.
Hirayama, K. & H. Funamoto, 1983. Supplementary effect
of several nutrients on nutritive deficiency of baker's yeast
firm effects reported from our batch culture expe- for population growth of the rotifer Brachionus plicatilis.
riments. Bull. Japan. Soc. Sci. Fish. 49: 505-510.
Maruyama, I., T. Nakamura, T. Matsubayashi & T. Maeda,
1986. Identification of the alga known as 'marine Chlorella'
as a member of the Eustigmatophyceae. Jap. 1. Phycol. 34:
Conclusion 319-325.
Provasoli, L., K. Shiraishi & J. R. Lance, 1959. Nutritional
Vitamin B 12 is an essential nutrient for rotifer idiosyncrasies of artemia and tigriopus in monoxenic cul-
ture. Ann. N. Y. Acad. Sci. 77: 250-261.
growth (Scott, 1981). The nutritional deficiency of Scott, J. M., 1981. The vitamin B12 requirement of the marine
baker's yeast suspension is mainly due to vitamin rotifer Brachionus plicatilis. 1. mar bioI. Ass. U.K. 61:
B12 (Hirayama & Funamoto, 1983). However, 983-994.
Hydrobiologia 186/187: 43-50, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 43
© 1989 Kluwer Academic Publishers.

Life history characteristics of Brachionus plicatilis (rotifera) fed

different algae

J. Korstad,l Y. Olsen & O. Vadstein

SINTEF Div. of Applied Chemistry, Aquaculture Group, 7034 Trondheim, Norway; 1 Present Address:
Department of Biology, Oral Roberts University, Tulsa, Oklahoma 74171, USA

Key words: Brachionus plicatilis, Rotifera, life history, food

Abstract

A detailed study of the life history of the rotifer Brachionus plicatilis was done at 20 0 C, 20 ppt salinity,
and 90 mg C 1- 1 food concentration. Rotifers were grown individually in culture plate wells (150 J.ll
culture volume) and fed Isochrysis galbana Tahiti, Tetraselmis sp., Nannochloris atom us, or a 1 : 1 mixture
(weight) of two of the algae. Observations were made every 2-8 hr and rotifers were sized and transferred
to new food daily. A total of 19 different parameters were compared. Rotifers fed Isochrysis averaged 21
offspring per female, a 6.7 day reproductive period, a lifespan of 10.5 days and a mean length of 234 J.lm.
After Isochrysis, the foods giving the highest growth, survival, and reproduction in decreasing order were
Isochrysis + Nannochloris, Nannochloris, Isochrysis + Tetraselmis, Tetraselmis + Nannochloris, and
Tetraselmis. Although the small volume culture system used in this study seems appropriate for studying
life history of B. plicatilis, the results cannot always be directly applied to larger cultures.

Introduction B. plicatilis. Most other studies of various aspects

of the life history of B. plicatilis have investigated
Brachionus plicatilis MUller is a euryhaline rotifer the influence of food and temperature on popula-
commonly used as initial food for a variety of tion growth rates (Snell et al., 1983; Yufera,
marine fish larvae. Several reviews have discussed 1987).
its biological characteristics and its importance in More information on the growth, survival, and
aquaculture (Walker, 1981; Lubzens, 1987, reproduction of B. plicatilis fed different diets is
1989). needed before optimization of the culture of this
Although much is known about the biology of rotifer can take place. Mathematical models of
B. plicatilis, relatively few researchers have inves- the production process of live feed for marine fish
tigated the life history of this rotifer. The only larvae (e.g., Slagstad et al., 1987) might also be
detailed studies of the life history of individually rermed by this information.
cultured B. plicatilis were done by Ruttner- This study examines the size at birth, size of
Kolisko (1972), who tested the influence oftem- first egg, size at first newborn, maximum size,
perature and salinity on several important life his- juvenile period, time to last egg, time between
tory parameters, and King & Miracle (1980), who egg-laying, egg development time, total number of
tested the influence of temperature on rates of eggs, total number of eggs hatching, percent eggs
survivorship and reproduction in three clones of hatching, maximum number of eggs carried at one
44

time, time to fIrst newborn, time to last newborn, selected from each stock culture and individually
time between newborn, reproductive period, life- micro-pipetted into 150,u1 tissue culture plate
span, post-reproductive period, and percent of wells (Nundon) containing the same algal culture
lifespan post-reproductive of individually cul- at a concentration of 90 mg C I - I. These concen-
tured B. plicatilis fed single and mixed diets of trations were determined spectrophotometric ally
Isochrysis galbana Tahiti, Tetraselmis sp., and using an established carbon versus absorption
Nannochloris atomus. relationship. These rotifers were observed every
1-2 hr under a Wild dissecting microscope at
25 x magnillcation, and as soon as neonates
Methods appeared, the mother was removed. Thereafter,
observations were made every 2-8 hr for the
Brachionus plicatilis were obtained from the Insti- appearance of eggs and neonates which were
tute of Marine Research in Bergen, Norway. counted and removed. Rotifers were sized (under
Separate cultures were maintained in 4 I beakers 50 x magnifIcation) and transferred to new food
with continuous aeration at 20 0 C and 20 ppt daily. Between observations, the culture plates
salinity. Sea water was obtained from direct pipe- were kept inside a large beaker immersed in a
line supply to the laboratory from the nearby water bath at 20 0 C. Light intensity was approxi-
fjord, diluted to the desired concentration, and mately 6 ,uEin m -2 sec- I (12L: 12D).
fIltered through a Whatman GFjC glass-fIber A separate experiment was done to study the
fIlter. Rotifer cultures were daily fed one of the low survival and reproduction of B. plicatilis
following algae: Isochrysis galbana Tahiti, which were fed Tetraselmis. Individual rotifers
Tetraselmis sp., and Nannochloris atomus (here- were cultured under the same conditions as the
after called Iso, Tetra and N anno, respectively), fIrst study. Four groups of six rotifers each were
or a 1: 1 mixture (weight) of two of the algae: Iso fed Iso diluted with different solutions to a fInal
+ Tetra, Iso + Nanno, and Tetra + Nanno. concentration of 60 mg C I - I. One group was
Algae were obtained from the culture collections diluted with fIltrate from dense cultures of Tetra
at the Institute of Microbiology and Plant (Tetra grown in stock culture at a concentration
Physiology in Bergen (Iso and N anno) and the of 240 mg C I - 1 and fIltered through a Whatman
Laboratory of Biotechnology, University of GFjC fIlter). Another group was diluted with
Trondheim (local strain of Tetra; species Tetra fIltrate at a concentration of 90 mg C I - 1
unknown). fIltered as above and a third group was diluted
Algal cultures were grown in semi-continuous with f media to check if some compound in the
cultures diluted at 0.33 day- 1 using f media media not metabolized by Tetra was toxic to
(Guillard 1975) and constant light (160 ,uEin m - 2 Brachionus. A fourth group was diluted with
sec - I). About 500 ml (13 %) of the water in the 20 ppt sea water (control). All rotifers were
rotifer cultures was removed each day and replen- observed and transferred to new food daily as in
ished by new sea water with algae. Rotifer cul- the other study.
tures were physiologically adapted to the experi- Differences between means for each life history
mental conditions for a minimum of 10 days prior parameter were analyzed with one-way ANOVA.
to the life history experiments to assure complete Signillcant differences (p < 0.05) were further
acclimation to the respective food (Korstad et aI., analyzed with the Least Signillcant Difference
1989). In another life history study, we deter- multiple comparisons test (Statgraphics, Statisti-
mined that there are no signifIcant differences cal Graphics Corp.).
among all life history parameters measured for
four generations (F I, F 2 , F 3 , and F 4 generations),
indicating that 10 days acclimation is sufficient.
Twelve egg-bearing females were randomly
 45

Results and Iso + Nanno reached reproductive maturity

in the shortest time, followed by animals fed Iso,
A total of 19 different parameters were deter- Tetra + Nanno, Tetra, and Iso + Tetra. Time to
mined for each rotifer (Table 1). Size at birth, time last egg and last newborn, reproductive period,
to fIrst newborn, and post-productive period were and lifespan were longest for rotifers fed Iso and
not signifIcantly different between foods. Other shortest for those fed Tetra and Tetra + Nanno.
life history traits showing signifIcant dietary Percent of lifespan post-reproductive was lowest
effects included size at fIrst egg, size at fIrst new- for animals fed Iso, Iso + Tetra, and Iso +
born, and maximum size, all of which were larger Nanno and highest for those fed Tetra, Nanno,
for rotifers fed Iso and Iso + Nanno. Rotifers and Tetra + Nanno.
were smallest when fed Tetra. Rotifers fed Nanno Rotifers fed Iso had the highest reproductive

Table 1. Life history parameters for Brachionus plicatilis fed different algae. When one-way ANOVA for diet effects had
significance values of p < 0.05, a Least Significance Difference (LSD) multiple comparison test was done. Numbers under
Multiple Comparison indicate sample means which are similar (same number) or different (different number).
A. Size parameters.

Food species Size at Size at Size at Max. size

birth (/lm) first egg first (/lm)
(/lm) newborn (/lm)

Iso: N= 12 12 12 12
mean = 130.00 190.00 207.50 234.17
SE = 1.18 0.00 1.25 4.32
Tetra: N= 12 5 0 11
mean = 128.18 174.00 165.45
SE = 1.73 2.19 4.52
Nanno: N= 12 12 10 12
mean = 128.33 189.17 202.00 208.33
SE = 1.08 0.80 1.90 3.50
Iso + Tetra: N= 9* 4 4 8
mean = 126.67 187.50 197.50 183.75
SE = 1.57 2.17 6.50 7.06
Iso + Nanno: N= 12 12 12 6
mean = 128.33 194.17 210.00 230.00
SE = 1.08 1.42 2.04 6.24
Tetra + Nanno: N= 12 7 0 8
mean = 125.83 184.29 180.00
SE = 1.42 2.75 6.12

Anova: p= 0.37 <0.001 0.014 <0.001

LSD multiple comparison:

Iso 1
Tetra 2 2
Nanno 1 2 3
Iso + Tetra 1 3 4
Iso + Nanno 3
Tetra + Nanno 4 4

* Only 9 newborn were obtained.

46

Table 1. (continued). B. Egg parameters.

Food species Time to Time to Time Egg Total Total Percent Max. no.
first egg Last between devel. number number hatching eggs
Uuvenile egg egg-laying time eggs hatching carried at
period) (days) (days) (days) (Ro) one time
(days)

Iso: N= 12 12 12 12 12 12 12 12
mean = 1.39 8.11 0.33 0.41 22.42 21.17 0.93 4.00
SE = 0.07 0.32 0.02 0.04 1.46 1.66 0.03 0.26
Tetra: N= 5 5 0 12 12 12 0.12
mean = 1.80 2.21 2.04 0.50 0 0 0.50
SE = 0.09 0.29 0.19 0 0 0.19
Nanno: N= 12 12 11 10 12 12 12 12
mean = 1.23 4.92 0.79 1.97 5.75 3.08 0.41 2.50
SE = 0.09 0.98 0.10 0.32 1.28 1.12 0.07 0.22
Iso + Tetra: N= 4 4 4 4 9 9 9 9
mean = 1.84 4.26 0.39 0.78 3.22 3.00 0.39 1.56
SE = 0.19 0.44 0.02 0.21 1.39 1.37 0.15 0.59
Iso + Nanno: N= 12 11 12 11 11 11 11 12
mean = 1.19 6.73 0.42 0.42 17.36 16.00 0.92 4.33
SE = 0.04 0.57 0.07 0.02 1.62 1.56 0.02 0.27
Tetra + Nanno: N= 7 7 4 0 12 12 12 12
mean = 1.57 2.20 0.89 1.00 0 0 0.92
SE = 0.09 0.26 0.07 0.29 0 0 0.25

Anova: p= <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001

LSD multiple comparison:

Iso
Tetra 2 2 2 2 2 2 2
Nanno 3 3 3 2 3 3 3 3
Iso + Tetra 2 3 1 3 4 3 3 4
Iso + Nanno 3 4 1 1 5 4 1 1
Tetra + Nanno 4 2 3 2 2 2 2

output both in terms of eggs and newborn. Those Iso lived a minimum of 8.5 days and a maximum
fed Iso and Iso + Nanno had the highest per- of 12.5 days, whereas rotifers fed Tetra and Tetra
centage of viable eggs and carried the most eggs + Nanno lived a minimum of about 1.0 day and
at one time. Rotifers fed Tetra and Tetra + a maximum of 5.4 days.
N anno produced a few eggs but none were viable. The effects of Tetra filtrate on reproductive rate
Time between egg-laying, egg development time, are presented in Table 2. The highest repro-
and time between newborn were shortest for ani- duction was achieved with rotifers fed a diet ofIso
mals fed Iso, Iso + Nanno, and Iso + Tetra. cells and Tetra filtrate and lowest for the control
Percent survival was highest for rotifers fed Iso, (sea water) and f media dilutions. Differences
followed by Iso + Nanna, Nanno, Iso + Tetra, between the dilute and concentrated Tetra filtrate
Tetra, and Tetra + Nanna (Fig. 1). Rotifers fed were not statistically significant.
 47

Table 1. (continued). C. Newborn, reproduction, lifespan, and post-reproductive parameters.

Food species Time to Time to Time Repro. Life- Post- Percent of

first last between period span repro. lifespan
newborn newborn newborn (days) (days) period post-
(days) (days) (days) (days) repro.

Iso: N= 12 12 12 12 12 12 12
mean = 2.25 8.88 0.36 6.72 10.46 2.35 0.22
SE = 0.07 0.39 0.04 0.34 0.41 0.39 0.03
Tetra: N= 0 0 0 12 12 5 4
mean = 0.17 2.47 1.65 0.39
SE = 0.16 0.40 0.46 0.08
Nanno: N= 10 10 7 12 12 12 12
mean = 2.66 5.47 1.06 3.70 7.92 3.00 0.39
SE = 0.42 1.08 0.20 0.99 1.11 0.70 0.05
Iso + Tetra: N= 4 4 4 9 9 4 4
mean = 2.81 5.07 0.33 1.08 4.40 1.52 0.24
SE = 0.31 0.44 0.11 0.48 0.60 0.60 0.07
Iso + Nanno: N= 12 11 11 11 11 12 11
mean = 1.96 7.21 0.36 5.54 8.87 1.96 0.20
SE = 0.05 0.59 0.02 0.56 0.86 0.76 0.05
Tetra + Nanno: N= 0 0 0 12 12 7 7
mean = 0.37 2.71 2.71 1.29
SE = 0.16 0.32 0.32 0.28

Anova: p= 0.12 0.006 <0.001 <0.001 <0.001 0.58 0.049

LSD multiple comparison:

Iso 1 1 1
Tetra 2 2 2
Nanno 2 2 3 3 2
Iso + Tetra 2 4 4 1
Iso + Nanno 3 5 3 1
Tetra + Nanno 2,4 2 2

Discussion and Iso + Nanno and about 38 percent for

rotifers fed the other algal combinations.
The life history parameters of rotifers fed Iso in The three clones of B. plicatilis used by King &
our study are similar to those for B. plicatilis fed Miracle (1980) had a wider range of mean lifespan
excess Dunaliella at 20 ° C reported by at 20 °C than those used in our study. Clone SP
Ruttner-Kolisko (1972), indicating that the food had the longest lifespan (16.5 days), while rotifers
value of these two algae is similar. Walker (1981) fed Iso, which had the longest lifespan in our
analyzed Ruttner-Kolisko's (1972) data and study, lived 11 days. Clones LA and MC in King
found that the rotifers spent approximately one- & Miracle's study had fairly short lifespans
third of their lifespan in the post-reproductive (6-8.5 days), which were similar to rotifers fed all
stage at all temperatures tested. In our study, other algal types in our study (2.5-8.9 days).
however, we observed a wider range, averaging Yufera (1987) measured the effect of algal diet
about 22 percent for rotifers fed Iso, Iso + Tetra, and temperature on egg development time in two
48

Q.
:r:
(/J
a: 50
o
>
>
a:
::J
(/J
•
*""*
o -+---r----..----...........
o
~5 I T .......---..,----.,..........
10 15
AGE ,N DAYS

Fig. 1. Survivorship (fraction surviving over time) of Brachionus plicatilis fed Iso (I), Tetra (T), Nanno (N), Iso + Tetra (IT),
Iso + Nanno (IN), and Tetra + Nanno (TN).

strains of B. plicatilis. He found the slowest devel- higher on the mixed diet than on either unialgal
opment time in rotifers fed Nannochloropsis diet. Other researchers have reported similar find-
gaditana, which was most similar to rotifers fed ings with yeast and algae as food (Hirayama &
Nannochloris atom us in our study. Watanabe, 1973; Yufera & Pascual, 1980).
Snell et al. (1983) studied the effects of unialgal Yufera and Pascual, however, found highest
and mixed diets of Chlorella sp., Dunaliella growth rates on rotifers fed Tetraselmis suecica.
tertiolecta, and Schizothrix calcicola on the repro- The results from our study are quite different from
ductive rate of B. plicatilis. They found that the those reported above. Rotifers fed Tetra and
reproductive rate was an average of 2.7 times Tetra + Nanno had the lowest reproductive

Table 2. Reproductive rate of Brachionus plicatilis fed a diet of Iso diluted to 60 mg C I-I with 20 ppt sea water (Control), f
Media, dilute Tetra filtrate (DTF), and concentrated Tetra filtrate (CTF). See text for details. Numbers represent mean ± (SE);
n = 6 for each treatment. Description of statistical analyses same as for Table 1.

Dilution Total Newborn Percent

no. eggs ~ hatching

Control 7.50 3.33 38

( 1.45) (1.10) (14)
f Media 6.17 2.33 38
(1.04) (0.38) (3)
DTF 16.00 13.00 81
( 1.65) (1.43) (2)
CTF 17.33 15.17 89
(3.04) (2.69) (2)

Anova: p= 0.002 <0.001 <0.001

LSD multiple comparison:

Control 1 1 1
f Media 1 1 1
DTF 2 2 2
CTF 2 2 2
 49

rates, while those fed only Iso had the highest heating on the Tetra filtrate in our study, our
reproductive rate. results were different. Reproductive rate of
The results of the experiment testing the effect Brachionus did not significantly increase from the
of Tetra filtrate on rotifers suggest that Tetra cells, dilute to the concentrated Tetra filtrate. There
but not filtrate, have an inhibitory effect on was, however, a nearly five-fold increase in Ro
rotifers. However, we have also maintained larger between rotifers fed Iso in control (sea water) and
volume aerated batch cultures of B. plicatilis fed f media treatments and those in the two Tetra
the same algae used in this study (Korstad et ai., filtrate treatments. Rotifers may possibly have
in prep.). Rotifers had somewhat similar growth obtained extra nutritional benefits from the chemi-
rates on each food, indicating that the effects of cals released by the Tetra cells and present in the
Tetra are different in the culture plate wells than filtrate.
in the larger volume batch cultures. Tetra cells are
motile but tend to readily attach to culture vessel
Acknowledgements
surfaces, while Iso cells tend to remain motile
(personal observation). Relatively more Tetra
This study was supported by the Royal Nor-
cells may therefore have been attached to the wegian Council for Scientific and Industrial
walls of the culture plate wells than in the batch Research (NTNF), and forms part of a research
culture vessels because of the larger surface-to- project on live feed for marine fish larvae financed
volume ratio in the smaller containers. This might
by NTNF. The first author also gratefully ack-
account for the lower life history values for
nowledges support from the Royal Norwegian
B. plicatilis fed Tetra in our study. Marshall Fund and Oral Roberts University.
We have observed B. plicatilis feeding on Iso
and Tetra. They seem to be able to ingest Iso cells
more readily than Tetra. Whether this is because References
Iso cells move rapidly in the water while Tetra
cells tend to adhere to surfaces is not clear. The Fukusho, K., M. Okauchi, H. Tanaka, S.1. Wahyuni, P.
Iso cells are also smaller than Tetra (approxi- Kraisingdecha & T. Watanabe, 1985. Food value of a
mately 11 and 250 ,um3 , respectively), which may rotifer Brachionus plicatilis, cultured with Tetraselmis tetra-
thele for larvae of a flounder Paralichthys olivaceus. Bull.
affect ingestion. Another possibility is that Tetra
Natl. Res. Inst. Aquaculture 7: 29-36.
may be assimilated with lower efficiency than Iso. Guillard, R. R. L., 1975. Culture of phytoplankton for feeding
It's also probable that different species of Tetra marine invertebrates. In Smith, W. L. & Chanley, M. H.
may have differences in afTmity to attachment, (eds) Culture of Marine Invertebrate Animals. Plenum
nutritional quality, ingestion by Brachionus, and Press, New York: 29-60.
Hirayama, K. & K. Watanabe, 1973. Fundamental studies
other characteristics. Tetra has been commonly on physiology ofrotifer for its mass culture. IV. Nutritional
used as a nutritious food for B. plicatilis by many effect of yeast on population growth ofrotifer. Bull. Japan.
researchers (Trotta, 1983; Okauchi & Fukusho, Soc. Sci. Fish. 39: 1129-1133.
1984; Fukusho et aI., 1985). King, C. E. & M. R. Miracle, 1980. A perspective on aging in
Snell et al. (1983) reported that the enhance- rotifers. Hydrobiologia, 73: 13-19.
Korstad, 1., O. Vadstein & Y. Olsen, 1989. Feeding kinetics
ment of the reproductive rate of B. plicatilis fed a
of Brachionus plicatilis fed Isochrysis galbana. Hydro-
combination of Chlorella end Schizothrix was not biologia, this volume.
dependent on ingestion of the Schizothrix cells. Lubzens, E., 1987. Raising rotifers for use in aquaculture.
The reproductive rate of B. plicatilis increased in Hydrobiologia, 147: 245-255.
direct proportion to the amount of the Schizothrix Lubzens, E., 1989. Rotifers as food in aquaculture. Hydrobio-
logia, this volume.
filtrate added to the Chlorella. Further investi-
Okauchi, M. & K. Fukusho, 1984. Food value of a minute
gation revealed that the enhancing factor was a alga, Tetraselmis tetrathele, for the rotifer Brachionus plica-
heat labile substance deactivated at 100 0 C. tilis culture. I. Population growth with batch culture. Bull.
Although we did not investigate the effects of Nat!. Res. Inst. Aquaculture 5: 13-18.
50

Ruttner-Kolisko, A., 1972. The metabolism of Brachionus Walker, K. F., 1981. A synopsis of ecological information on
plicatilis (Rotatoria) as related to temperature and chemi- the saline lake rotifer Brachionus plicatilis Muller, 1786.
cal environment. Dt. Zool. Ges. 65: 89-95. Hydrobiologia 81: 159-167.
Slagstad, D., Y. Olsen & S. Tilseth, 1987. A model based Yufera, M., 1987. Effect of algal diet and temperature on the
system for control of live feed level for larval fish. embryonic time ofthe rotifer Brachionus plicatilis in culture.
Modeling, Identification and Control 8: 51-60. Hydrobiologia 147: 319-322.
Snell, T. W., C. J. Bieberich & R. Fuerst, 1983. The effects of Yufera, M. & E. Pascual, 1980. Estudio del rendimiento de
green and blue-green algal diets on the reproductive rate cultivos del rotifero Brachionus plicatilis O. F. Muller ali-
of the rotifer Brachionus plicatilis. Aquaculture 31: 21-30. mentados con levadura de panificaci6n. Inves. Pesq. 44:
Trotta, P., 1983. An indoor solution for mass production of 361-368.
the marine rotifer Brachionus plicatilis Muller fed on the
marine microalga Tetraselmis suecica Butcher. Aquacult.
Engin. 2: 93-100.
Hydrobiologia 186/187: 51-57, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 51
© 1989 Kluwer Academic Publishers.

Feeding kinetics of Brachionus plicatilis fed Isochrysis galbana

John 1{0rstad, 1 ()lav Vadsteln &, Yngvar ()lsen

SINTEF, Division of Applied Chemistry, Aquaculture Group, N-7034 Trondheim, Norway; 1 Present
Address: Department of Biology, Oral Roberts, University, Tulsa, Oklahoma 74171 USA

Key words: Brachionus plicatilis, feeding kinetics, clearance rates, ingestion rates, Isochrysis

Abstract

Clearance and ingestion rates of Brachionus plicatilis were measured using 14C-labeled Isochrysis galbana
Tahiti. Experiments were conducted at 20-22 °C, 20 ppt salinity, and algal concentrations ranging from
0.13-64 mg C 1- 1. Clearance rates were constant and maximal at concentrations < 2 mg C 1- 1, with
maximum rates ranging from 3.4-6.9/l1 indo - I hr-l. The ingestion rate varied with food concentration,
and was described by a rectilinear model. The maximum ingestion rate varied considerably, and was
dependent on the growth rate of the rotifers. Depending on the pre-conditions, B. plicatilis ingested about
0.5 to 2 times its body carbon per day at saturating food concentrations.

Introduction using radio-actively labeled algae to measure

clearance and ingestion rates of B. plicatilis. The
A wealth of information has been published on results demonstrate the effect of food concen-
feeding in the rotifer genus Brachionus, primarily tration and growth rate on rotifer feeding kinetics.
in B. calyciflorus (see Starkweather, 1980).
Although many feeding studies have also been
done on B. plicatilis, most have employed metho- Materials and methods
dology requiring several hours duration which can
result in spurious results (cf. Dewey, 1976; Yufera Brachionus plicatilis was obtained from stock cul-
& Pascual, 1985; Yamasaki & Hirata, 1986). tures at SINTEF Aquaculture Center in Trond-
Doohan (1973) applied the more sensitive radio- heim, and maintained on a diet of Isochrysis
tracer technique to measure clearance and in- galbana (Tahiti) for at least one week before
gestion rates in this species. Her results, however, experiments commenced. Stock rotifer cultures
were quite variable and the ingestion rates were were continuously aerated and kept at 20 ° C and
probably underestimated because of the long in- 20 ppt salinity. The average length of adults of this
cubation time used. Thus, knowledge of the strain is about 150/lm. Isochrysis cultures were
feeding kinetics of B. plicatilis is incomplete. grown in semi-continuous culture diluted at 0.33
Because B. plicatilis is cultured as start food for day-l using f media (Guillard, 1975). Radio-
marine fish larvae (Howell, 1973; Gatesoupe, actively labeled algae were prepared as follows:
1987; Robin, 1987; Theilacker, 1987), this infor- 20/lCi NaH I4C0 3 , unlabeled NaHC0 3 corre-
mation would ultimately aid aquaculturists. sponding to 50 mg C 1- 1, and about 25 ml new f
This paper describes a series of experiments media were added to approximately 25 ml Iso-
52

chrysis from the stock culture (ca. 120 mg C 1- 1 suspensions without added animals and process-
before dilution) in a 125 ml glass-stoppered ed samples as above. These controls were not
bottle. The bottle was placed in constant light at significantly different from zero time samples with
about 60 JiEin m - 2 sec - 1. animals. Heat-killing of the rotifers before fil-
B. plicatilis used for the feeding experiments tration yielded irreproducible results because the
was taken from the stock culture, placed in a algae stuck to rotifers and netting.
50 Jim sieve in a beaker, and concentrated by Three different types of experiments were con-
carefully pipeting most of the excess water out of ducted to describe the feeding kinetics of B. plica-
the beaker. The rotifers were pre-acclimated with !ilis. A time course experiment was conducted at
non-radio-active Isochrysis at the desired concen- two food concentrations (5 and 15 mg C 1- 1) to
tration for 1-2 hours. All experiments were con- determine the gut passage time of the rotifer.
ducted at 20-22 °C, 20 ppt salinity, and about Twenty measurements between 0 and 60 min
15 JiEin m - 2 sec - 1light intensity. Sea water used were taken. Following the results of this experi-
for dilution and rinsing had been previously filter- ment, subsequent feeding studies were conducted
ed through a Whatman GF/C filter, autoclaved, for 15-20 min. Two functional response experi-
cooled to experimental temperature, and aerated. ments were done to determine the clearance and
After the pre-feeding incubation, the rotifers ingestion rates of the rotifers at different food
were concentrated as before and added to radio- concentrations. Rotifer stock cultures used in
active algae in flasks. Volume and incubation time these experiments were fed daily in the first expe-
depended upon the type of experiment and the riment and every other day in the second experi-
flasks were agitated periodically during incu- ment. Finally, maximum ingestion rates were de-
bation (Schlosser & Anger, 1982). At the end of termined at 30 mg C 1- 1 throughout a batch cul-
the experiment, replicate 3-5 ml samples were ture experiment to observe variability in the in-
taken and filtered by gravity through 50 Jim Nitex gestion rates following different long term pre-
netting in 2.5 cm diameter Millipore filtering acclimation times. To obtain rotifers with close to
units, and rinsed with 50 ml20 ppt sea water. The zero growth rate, they were starved for 3 days
rotifers were maintained in about 3 ml of water prior to starting this experiment.
throughout the rinsing. The Nitex netting was
then placed in a scintillation vial. It took about
2 min from taking the sample from the flask to Results
placing the Nitex netting in the scintillation vial.
Replicate samples were taken for determination The results of the time series experiment (Fig. 1)
of specific activity (3 x 100 Jil) and for rotifer revealed that the gut passage time of B. plicatilis
counts (5 x 200 Jil). One hundred Jil H 2 0 2 was was dependent on food concentration. As indi-
added to each vial to remove color. After 2 hr, cated by the abrupt change in the slope of the
20 Jil 0.5 N HCI was added to the specific activity regression lines, the gut passage time was approx-
samples to remove inorganic 14C and 5 ml scintil- imately 30 and 45 min at 5 and 35 mg C 1- 1,
lation cocktail (Optifluor, Packard) was thereafter respectively. We therefore chose a feeding interval
added to each vial. Samples were counted with a of 15-20 min for the other experiments.
Nuclear Chicago Isocap 300 liquid scintillation The clearance rates for both functional re-
counter for 10 min using the external standard sponse experiments (Fig. 2) were highest and con-
ratio. Control experiments revealed similar stant at food concentrations below 1-2 mg C 1- 1
counts and counting efficiencies of rotifer samples and decreased with increasing food concen-
treated as above with the inclusion of adding tration. Ingestion rates for both functional re-
tissue solubilizer (200 Jil Soluene-350, Packard). sponse experiments increased linearly with in-
To estimate background radioactivity on the creasing food concentration up to a maximum
screens we used the radioactive algal feeding and thereafter remained constant at higher food
 53

'i
c 5
Q)

•
Q)
ti
V)

4
•
•
3
•
2

I I I
o 20 40 60

Time (min)

Fig. 1. Accumulation of radioactive food C4 C I. ga1bana) by B. plicatilis over time at two food concentrations. Gross CPM
per Nitex screen is given on the ordinate.

8 t A 4
I B

1
0

1 0 T

2
T

1 j
0 T

--
0
6 01 3 .J.

.., 0
.t;.

".;;
"0
4 2 T
:i. 0
.J. Q
a: 0
()
2

a a
0
0
0
0 0
0
0.5 1 5 10 50 0.1 05 5 10 50

Food conc. (mgc·r1)

Fig. 2. Clearance rates (CR) of B. plicatilis for functional response experiment 1 (A) and 2 (B). Standard errors indicated when
larger than symbol.
54

concentrations (Fig. 3). Ingestion and clearance

rates were higher in the first experiment than in
25 the second experiment at all food concentrations.
The ingestion data could be described by recti-
T
20 T1
3-t.~ T
linear, Ivlev and Michaelis-Menten models
(P < 0.001). In Table 1 results from the rectilinear
T
... regressions are presented. We have chosen this
"'..c
0 l~
1 ..L model in the further treatment because of its
'".f;"0 15 simplicity (zero order approximation), and the
U
, . - --
...- -- - - - - FR1- biologically relevant parameters in the model- the
maximum clearance rate (CRmax ) and maximum
..1..
OJ
c 10
ingestion rate (ImaJ. A 2.6 and a 2.0 times dif-
...
2-t. ference was recorded in Imax and CRmax , respec-
I i ! ~t tively, between the two experiments and the dif-
5 ············· FRJ,·
Ii ferences were significant (Table 1). The incipient
7-t. • limiting concentration (ILC) was, on the other
o hand, not significantly different in the two experi-
o 20 40 60 ments.
Food conc. (mgc·r1 ) In the batch experiment the rotifers were grow-
ing exponentially from day 1 to day 4 when the
Fig. 3. Ingestion rates of B. plicatilis as a function of concen- food was depleted, and the culture reached
trations of I. galbana: Time Series experiment (calculated stationary phase at day 5-6 (Fig. 4). The maxi-
from the slope of the curve before the inflection points shown mum ingestion rate (Imax) was intermediate on
in Fig. 1)(<», both Functional Response experiments
day 2, increased by a factor of 2.7 on day 3,' and
(FR 1 • and FR 2 .), and the Batch experiment ("*). Day
number indicated for the batch experiment. Error bars indi- decreased dramatically upon entering stationary
cate ± SE. Lines represent model curves for the two phase (Fig. 3). Overall, a 6 fold difference was
functional response experiments (cf. Table 1). recorded in Imax during this experiment. No sig-

Table 1. Results from fitting a rectilinear model to the data of ingestion rate (I) as a function offood concentration (C) in the
two functional response experiments. Maximum ingestion rate (lmax) and maximum clearance rate (CRmax) was fitted by
zero-order approximation, and incipient limiting concentration (ILC) was calculated as Imax/CRmax. Parameters given with one
standard error. Differences between parameters were evaluated using the Student's t-test. For comparison Dewey's (1976) data
for B. plicatilis fed I. galbana, are included.

1= CRmax'C (C:::; ILC)

1= Imax (C ~ ILC)

Exp. CRmax Imax ILC p

(Ill indo~ 1 hr~ I) (ng C indo ~ 1 hr~ I) (mg C l~ I)

6.81 ± 0.68 12.93 ± 0.63 1.90 ± 0.21 <0.001

(n = 12) (n = 36) (n = 46)
2 3.38 ± 0.31 4.93 ± 0.22 1.46 ± 0.15 <0.001
(n = 18) (n = 54) (n = 70)

P< 0.001 P< 0.001 P> 0.05

Dewey 6.01 ± 0.38 19.1 ± 1.9 3.18 ± 0.37

(n = 10) (n = 6) (n = 16)
 55

0 30
\
\

A
80 A
25
b
\ A

40 \
\
\

20
\

0
20

'I
"a.
0
0

E A , 15
\ n
C/)
10 0
::J
'- (")
Q)
.....
:;::;
~
0 3
a: , 10 «)

.... 5 n
A Q
~
\
,
,
\ 5
\
\

t t \
\

b
t
I 0
I I I I I I I I
0 1 2 3 4 5 6 7 8

Day

Fig. 4. Number of rotifers and I. galbana concentration versus day for the batch experiment. Arrows indicate days on which
ingestion rates were measured.

nificant differences were recorded in the length of bert, 1977b). Our results with B. plicatilis are in
the animals, that averaged 155 ± 4 J..l.m (± SD). agreement with the higher values of this range.
It has been suggested that filter feeders would
benefit by reducing their filtering rate at very low
Discussion food concentrations to ensure energy optimi-
zation (Mullin et a!., 1975; Lehman, 1976). No
The gut passage time is known to vary with reduction in clearance rate was found in the
species, temperature, food concentrations, and present study for food concentrations down to
the nutritional state of the animals (Starkweather 0.13 mg C 1- 1. Our results are in accordance with
& Gilbert, 1977b). It probably also varies for most other studies on rotifer feeding (e.g. Erman
foods with different assimilation efficiencies. Re- 1956; Dewey, 1976; Starkweather & Gilbert,
ported gut passage times in other species of 1977a; Chotiyaputta & Hirayama, 1978). The
rotifers under varying conditions ranges from maximum clearance rates reported by Dewey
2-20 min (cf. Resvoi, 1926; Starkweather & Gil- (1976) for B. plicatilis fed Isochrysis galbana were
56

difference in [max which was recorded in the two

functional response experiments. The rotifers us-
25
ed in experiment 2 were fed every second day,
which caused the rotifers to experience large oscil-
20 lations in the food concentration and periods of
....c starvation. We found no significant correlations
...
"0 15
between maximum ingestion rates and size, swim-
.S ming speed, egg ratio, or the abundance of rotifers
U in the experimental containers.
OJ
C
>< 10 Dewey (1976) has reported similar variability in
'"
E Imax= 35.8 fJ + 6.0 I max and attributed it to size differences in the
rotifers. Her results with B. plicatilis fed Dunaliella
5 2 r2=0.969
tertiolecta can be re-interpreted, and in Fig. 5 we
have plotted Imax as a function of the specific
growth rate. There is a significant linear relation
o
0.0 0.1 0.4 0.5 between the two parameters (P < 0.005), which
supports our hypothesis that the feeding kinetics
of B. plicatilis is affected by the physiological
status of the animals. We therefore conclude that
Fig. 5. Maximum ingestion rate (Imax) at saturating food the feeding rate of B. plicatilis is not only a
concentrations as a function of specific growth rate (J.!) for
B. plicatilis fed Dunaliella tertiolecta, assuming J.! = 0 for starv-
function of the food concentration, but also of the
ed animals. Data are from Dewey (1976) and 95 % CI are growth rate of the rotifer. Further research into
given for Imax' the feeding biology of B. plicatilis and other roti-
fers should take the long term (days) pre-feeding
history of the animals into account.
intermediate to those estimated for the two
functional response experiments (Table 1). The
ingestion rates obtained on days 2 and 7 in the Acknowledgements
batch culture experiment were similar to [max of
functional response experiment 2, whereas those This study was supported by the Royal Norwe-
obtained on day 3 of the batch experiment and for gian Council for Scientific and Industrial Re-
the two time series experiment were comparable search (NTNF), and forms part of a research
to that obtained in functional response experiment project on live feed for marine fish larvae financed
1. The results presented in Fig. 3 indicate that by NTNF. The first author also gratefully ack-
individual B. plicatilis can ingest 0.5-2 times their nowledges support from the Royal Norwegian
body carbon per day at saturating food concen- Marshall Fund and Oral Roberts University.
trations (assuming 0.23 J,lg C rotifer- l ).
The results obtained in the batch culture experi-
References
ment suggest that the maximum ingestion rates
were related to the physiological status of the Chotiyaputta, C. & K. Hirayama, 1978. Food selectivity of
rotifers. Starved rotifers exhibited 6 times lower the rotifer Brachionus plicatilis feeding on phytoplankton.
[max than well fed rotifers, although the starved Mar. BioI. (Berl.) 45: 105-110.
rotifers were allowed to equilibrate their gut before Dewey, J. M., 1976. Rates offeeding, respiration and growth
the experiment (see methods). At least three days of the rotifer Brachionus plicatilis and the dinoflagellate
Noctiluca miliaris in the laboratory. Ph. D. Thesis Univ.
at saturating food concentrations are needed Wash. 117 pp.
before the rotifers are capable of maintaining the Doohan, M., 1973. An energy budget for adult Brachionus
highest [max' This also agrees with the 2.6 times plicatilis MUller (Rotatoria). Oecologia 13: 351-362.
 57

Erman, L. A., 1956. Quantitative aspects of the feeding of Aquaculture Europe '87 International Conference (in
rotifers. Zoo!. Zh. 35: 965-971. (In Russian). press).
Gatesoupe, F. l, 1987. Further advances in the nutritional Schlosser, l l & K. Anger, 1982. The significance of some
and antibacterial treatment of rotifers as food for turbot methodological effects of filtration and ingestion rates on
larvae, Scophthalmus maximus L. Aquaculture Europe '87 the rotifer Brachionus plicatilis. HelogHinder wiss. Meersun-
International Conference (in press). ters. 35: 215-225.
Guillard, R R L., 1975. Culture of phytoplankton for feeding Starkweather, P. L., 1980. Aspects of the feeding behavior
marine invertebrates. In Smith, W. L. & Chanley, M. H. and trophic ecology of suspension-feeding rotifers. Hy-
(eds) Culture of Marine Invertebrate Animals. Plenum drobiologia 73: 63-72.
Press, New York: 29-60. Starkweather, P. L. & J. J. Gilbert, 1977a. Feeding in the
Howell, B. R, 1973. Marine fish culture in Britain. VIII. A rotifer Brachionus calyciflorus. II. Effect of food density on
marine rotifer, Brachionus plicatilis Miiller, and the larvae feeding rates using Euglena glutinis. Oecologia 28: 133-139.
of the mussel, Mytilus edulis L., as foods for larval flatfish. Starkweather, P. L. & J. J. Gilbert, 1977b. Radiotracer deter-
Cons. int. Explor. Mer 35: 1-6. mination of feeding in Brachionus calyciflorus. The impor-
Lehman, l T., 1976. The filter-feeder as an optimal forager, tance of gut passage times. Arch. Hydrobio!. Beih. Ergebn.
and the predicted shape of feeding curves. Limno!. Ocean- Limno!. 8: 261-263.
ogr. 21: 501-516. Theilacker, G. H., 1987. Feeding ecology and growth ener-
Mullin, M. M., E. F. Stewart & F. l Fuglister, 1975. In- getics oflarval Northern Anchovy, Engraulis mordax. Fish.
gestion by planktonic grazers as a function of concen- Bul!. 85: 213-228.
tration of food. Limno!. Oceanogr. 20: 259-262. Yamasaki, S. & H. Hirata, 1986. Food consumption rates of
Resvoi, P., 1926. Observations on the feeding of rotifers. two types of rotifer Brachionus plicatilis. The Aquiculture
Trav. Soc. Nat. Lenin. 56: 73-89. (In Russian; English 34: 137-140.
translation provided by Dr. John Gilbert). Yufera, M. & E. Pascual, 1985. Effects of algal food concen-
Robin, l H., 1987. The quality ofliving preys for fish larval tration on feeding and ingestion rates ofBrachionus plicatilis
culture: preliminary results of mineral supplementation. in mass culture. Hydrobiologia 122: 181-187.
Hydrobiologia 186/187: 59-67, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 59
© 1989 Kluwer Academic Publishers.

Epizoic and parasitic rotifers

Linda May
Institute of Freshwater Ecology, Edinburgh Research Station, Bush Estate, Penicuik,
Midlothian EH26 OQB, Scotland, UK

Key words: epizoic, parasitic, rotifers

Abstract

Many rotifer species live in close association with plants or other animals. Most of these associations
are of a commensal or synoecious nature, some rotifer species having lost the ability to live independently.
Few rotifers are true parasites, actually harming their hosts.
The Seisonidae, Monogononta and Bdelloidea include epizoic and parasitic species. The most widely
known are probably the parasites of colonial and filamentous algae (e.g. Volvox, Vaucheria). However,
rotifers are also found on a wide range of invertebrates: colonial, sessile Protozoa; Porifera; Rotifera;
Annelida; Bryozoa; Echinodermata; Mollusca, especially on the shells and egg masses of aquatic
gastropods; Crustacea, including the lower forms (e.g. Daphnia, Asellus, Gammarus) and in the gill
chambers of Astacus and Chasmagnathus; the aquatic larvae of insects. There appear to be few records
of epizoic or parasitic rotifers among vertebrates, apart from Encentrum kozminskii on carp, Limnias
ceratophylli on the Amazonian crocodile, Melanosuchus niger, and an unidentified Bdelloid apparently
living as a pathogenic rotifer in Man.

Introduction This paper reviews the literature which de-

scribes parasitic or epizoic associations between
Rotifers have long been known to live in close rotifers and other organisms. Much of this litera-
association with other organisms, but little is ture was published in the late 1800's and early
known of the types of relationships involved. It is 1900's, when naturalists spent many hours ob-
difficult to judge from the literature whether these serving the behaviour of live specimens. Many of
associations are parasitic, symbiotic, commensal, the species names given by the original authors
epiphytic or epizoic. Many of the records are are now out-of-date; these have been updated, as
simply anecdotal and, while some authors have far as possible, according to Harring (1913),
guessed at the type of relationship involved, few Bartos (1951), Kutikova (1970) and Koste (1978).
have based their guesses on careful biological This review is not exhaustive and, in many
observation. For the purposes of this paper, a ways, adds little to the information given by
parasitic rotifer is considered to be one which Budde (1925) because the topic has received so
lives in or on a plant or animal species, feeding on little attention since that time. Hopefully, how-
its host and completing its life cycle in that envi- ever, the details given below, organised according
ronment. In contrast, an epizoic rotifer is defined to the type of host, will renew interest in this much
as a rotifer which lives on another animal for all neglected area of rotifer research. The text is
or part of its life, but does not feed on its host. divided into sections according to the kingdom
60

and, in some cases, the phylum (after Bames 1984) by P. werneckii has been recorded in a variety of
to which the various 'host' organisms belong. Vaucheria species, including V. prona, V. gemina-
ta, V. dillwynii, V. erythrospora, V. ?racemosa and
V. canalicularis (Davis & Gworek, 1973; Ott,
Parasitic and epizoic associations 1977; Christensen, 1987). In general, the rotifer
enters the developing gametophore of the Vauche-
Protista and Monera ria filament where it induces gall formation. Here
it feeds on the cytoplasm of the host by breaking
Rotifer parasites are common among colonies of the tonoplast and ingesting the cytoplasmic orga-
Volvox, especially V. globator (Williams, 1852; nelles. The rotifer deposits its eggs within the
Gosse, 1852; Hood, 1895; Galliford, 1946), galls; these hatch and the emergent rotifers escape
V. aureus (Rousselet, 1911; Sauer, 1978), and to parasitise more filaments (Davis & Gworek,
V. tertius (Ganf et al., 1983). These rotifers have 1973; Ott, 1977).
been identified as Proales parasita (Ehr.) (Wil- Many rotifers are thought to be parasitic on
liams, 1852; Thompson, 1892; Rousselet, 1911, protozoa, including Dicranophorus difflugarium
1914; Harring & Myers, 1922; Rich & Pocock, (Penard) on Difflugia acuminata var. inflata (Pen-
1933; Hollowday, 1949; Wulfert, 1960; Sauer, ard, 1914; Koste, 1978), Brachionus rubens Ehrb.
1978), Ascomorphella volvocicola (Plate) (Gosse on colonies of Carchesium, Proales parasita and
1852; Hood, 1895; Murray, 1906; Galliford, Pleurotrocha petromyzon on colonies of Ophridium,
1946; Ganf et aI., 1983) and, more rarely, Cepha- and Albertia vermisculus Dujardin on Limax
lodella catel/ina volvocicola (Zawadovsky) (Ed- species (Koste, 1978). In contrast, Eosphora gibba
mondson & Hutchinson, 1934). Parasitism in Gamer seems to have a commensal association
Ascomorphella volvocicola seems to be specific to with colonies of Carchesium and Vorticella, the
Volvox species, as this rotifer has never been ob- rotifer benefiting from the food carried towards it
served on other phytoplankton species, or living in the strong water currents generated by the
freely (Ganf et al., 1983). The mature rotifer enters protozoa (Hollowday, 1949).
the Volvox colony and feeds on the cells causing
extensive damage (Hood, 1895; Hollowday,
1949; Ganf et al., 1983). Whether this actually Fungi
causes the Volvox population to decline is un-
certain (Ganf et al., 1983). Gamer (1937) records Macrotrachela fungicola
Rotifers have also been recorded as parasites Gamer living in the gelatinous, orange-yellow
on other colonial algae, Ptygura melicerta (Ehrb.) fungus Dacrymyces deliquescens on decayed
on Gloeotrichia (Edmondson, 1940; Koste, 1978), wood, in Britain. M. fungicola is probably an
Proales parasita on Uroglenopsis americana (Car- ectoparasite of this fungus, but the exact relation-
lin, 1939), Proales uroglena and P. parasita on ship between the species is unclear.
Uroglena (Koste, 1978). Little is known of the
relationships between these species.
Rotifers have long been known to parasitise Plantae
Vaucheria filaments (Lister, 1884; Thompson,
1892; Brain, 1894; Wollny, 1879), although these Bryophyta
early authors were unsure of the identification of Although bdelloid rotifers are often found living
the rotifer species involved. Later descriptions among mosses, few could be thought of as para-
showed that Vaucheria filaments are usually para- sites. Habrotrocha roeperi (Milne) and Habrotro-
sitised by Proales werneckii (Ehrb.) (Jennings, cha reclusa (Milne) may be the exceptions, as they
1894; Harring & Myers, 1922; Davis & Gworek, actually live inside the outer cells of submerged
1973; Ott, 1977; Christensen, 1987). Parasitism branches of Sphagnum (Milne, 1888; Bartos,
 61

1951). Whether they simply live inside the empty trophi to suck out the body fluids. Among these
cells, filtering food from the surrounding water parasites are Cephalodella parasitica (Jennings),
(Milne, 1888), or whether they are true parasites, found on a variety of freshwater oligochaetes (e.g.
is unknown. Vejdovkyella comata, Stylaria lacustris, Chaetogas-
ter spp., Nais spp.) (Jennings, 1894; Beauchamp,
1905; Koste, 1972, 1978), and Drilophaga buce-
Animalia phalus Vejdovsky, found on oligochaetes (e.g.
Lumbriculus variegatus, Rhynchelmis sp., Stylodril-
Porifera us sp., Nais elinguis) (Vejdovsky, 1883; Koste,
Several rotifer species have been found living in 1978) and the body and posterior sucker of
close association with the freshwater sponge leeches (e.g. Herpobdella octoculata, H. nigricollis,
Spongilla lacustris in lakes of southern Sweden H. testacea and Hirudo medicinalis) (Pawlowski,
(Berzins, 1950). The first, Ptygura melicerta 1935; Koste, 1978). Murray (1906) also records
(Ehrb.) does not seem to injure the sponge in any Proales daphnicola as living epizoically on oligo-
way and appears to have a commensal asso- chaetes. However, his identification of the rotifer
ciation with its host. This species is invariably species in thought to be incorrect (Dr W Koste,
found around the pores and oscula of the sponge, pers. comm.).
taking advantage of the increased food supply Endoparasitic rotifers of worms were originally
carried in the water currents generated by its host. discovered by Dujardin (1838) who found Albertia
However, Lecane clara (Bryce) and Lepadella vermisculus in the expressed body fluids of
triba Myers, do appear to be true parasites. They earthworms and slugs. The parasites live in the
graze on the sponge itself, leaving furrows up to gut of their host and most belong to the genera
50 Jlm deep in the surface tissues. Neither of these Albertia (A. vermisculus, A. crystallina SchUltze,
species appears to be an obligate parasite, as both A. naidis Bousfield, A. reichelti Koste, A. bernhar-
are also found swimming freely and have no di Hlava) and Balatro (B. calvus Claparede, B. fri-
particular adaptation to a parasitic way of life. dericiae Kunst, B. anguiformis IsseI, B. aciliatus
(Radkevitsch)). The hosts of Albertia species in-
Rotifera clude the freshwater oligochaetes Stylaria spp.
Records of rotifers parasitising other rotifer (e.g. S. proboscidae, S. lacustris) (Levander, 1894;
species are rare. However, it has been noted that Hlava, 1905; Murray, 1906; Koste, 1969, 1970),
Acyclus inquietus Leidy is parasitic on colonies of Nais species (Hudson & Gosse, 1886; Bilfinger,
Sinatherina socialis (Linne) (Koste, 1968), while 1894; Koste, 1969, 1978), Paranais litoralis, Ripis-
Proales decipiens (Ehrb.) attacks Stephanoceros tis parasita (Koste, 1978), and the earthworm
jimbriatus (Goldfusz) (Stevens, 1907). In the for- Allolobophora caliginosa (Rees, 1960), while Bala-
mer case, the parasite feeds on the newly hatched tro species are more commonly found in the guts
young of its host. In the latter, P. decipiens, only of Henlea ventriculosa, H. perpusila, Fridericia bul-
one tenth the size of Stephanoceros, enters the bosa, F. perrieri, Trichodrilus sp., Buchholzia ap-
protective tube and feeds on the adult and its pendiculata, Enchytraeus buchholzi and E. albidus
developing eggs. The parasite lays its own eggs (Claparede, 1868; Hudson & Gosse, 1886;
within the tube and these subsequently hatch, and Koste, 1978). The rotifers attach themselves firm-
leave in search of new hosts. ly to the intestine of their host by grasping, with
their trophi, the small papillae of the intestinal
Annelida mucosa (Rees, 1960). A firm attachment is essen-
Segmented worms and leeches have both ecto- tial to prevent the rotifer being dislodged by the
and endo-parasitic rotifers. The ecto-parasites peristaltic movements of the intestine, and the
live on the epidermis of the host, feeding on the pressure of food moving along the gut. A detailed
epidermal cells and piercing the skin with their study of Albertia naidis in the gut of Nais elinguis
62

has shown that the rotifer feeds on cells from the tomentosa (Pfeiffer) in Australia. This author con-
intestine wall of its host and does not ingest any cluded that the many rotifer species found in the
of the algal remains passing through the worms natural habitats, and in laboratory cultures, of
digestive tract (Coineau & Kunst, 1964). these snails had no harmful effects. Had he
looked more closely at the egg masses, his con-
Mollusca clusions may well have been different.
One of the first records of rotifers parasitising Freshwater snails (e.g. Lymnaea tomentosa,
freshwater snails was published by Glascott Biomphalaria alexandrina, B. galbrata, B. pfeifferi,
(1893), who observed the rotifer Notommata gi- Bulinus truncatus) are of particular interest to
gantea Glascott, inside the eggs of the common researchers because of their associations with
water snail. Stevens (1912) also observed this parasitic infestations such as schistosomiasis, bil-
rotifer in the eggs of Limnaea auricularia L. and harzia, and liver flukes. Many workers in this area
Paludina vivipara L., although he concluded that have complained of infestations of rotifers in their
it belonged to another genus and renamed it experimental cultures which have caused a variety
Proales gigantea (Glascott). Stevens (1912) de- of problems (Stirewalt & Lewis, 1980; Hassan
scribed the rotifer in detail, including an interest- et aI., 1985; 1.M. lewsbury, pers. comm.; G.l.
ing account of its life history. The adult nibbles Greer, pers. comm.). Stirewalt & Lewis (1980),
through the outer shell of the snail egg, wriggles for example, describe how Rotaria rotatoria Pallas
through the hole and enters the shell. It then and Philodina acuticornis Murray colonize the
begins to feed on the fluid surrounding the snail snail shells, beginning at the centre of the whorl
embryo, killing the embryo within a few days. The and developing over the entire surface. A serious
adult rotifer lays up to 13 eggs, one at a time, infestation can result in snails looking as though
inside the snail egg. She then moves on to parasi- they are 'wearing fur coats' (l.M. lewsbury, pers.
tise a new egg, or dies and is eaten by her progeny. comm.). These rotifers seem to live commensally
The developing eggs hatch and the first to hatch on the adult snails, but, like Proales gigantea, their
feed on the remains of the snail egg before escap- main effect is on the egg masses. Hassan et al.
ing to parasitise another egg. Those which hatch (1985) have recorded up to 100% mortality
later tend to starve. P. gigantea seems to be a true among young snails (up to the blastula stage) in
parasite, spending most of its life cycle inside the rotifer-infested cultures. The reason for this is
snail egg. It was only found outside an egg for unclear, but the authors suggest that it may be due
short periods as it travelled through the jelly-like to an irritating effect of the rotifer, or to the effect
substance of the egg mass to parasitise another of its excretory products on the ootheca of the egg
egg. mass and on the embryo it contains. These results
Similar observations were made by Giard have led the authors to propose that rotifers could
(1908) & Nekrassow (1928), who also noticed be used in the biological control of these snails.
that the eggs of several other snail species (e.g. A further effect of parasitic rotifers on disease-
Lymnaea stagnalis L., L. auricularia L., L. ovata spreading snails is described by Stirewalt & Lewis
Drap., Myxas glutinosa Mull., and Physafontinalis (1980). They found that, in their studies of
L.) were similarly parasitised by P. gigantea. Nek- Biomphalaria galbrata infected with Schistosoma
rassow (1928) also noted that L. stagnalis eggs mansoni, cercariae production fell from 3123 cer-
were parasitised less often than those of other cariae/snail/day to 591 cercariae/snail/day when
species, and suggested that this may be because their cultures became infested with Rotaria rota-
the outer shell of its eggs is tough and difficult to toria and Philodina acuticornis. They also found
penetrate. that the cercariae which did emerge showed re-
These early observations on Proales gigantea duced motility and infectivity. This effect occur-
appear to be in contrast to the later findings of red not only in the presence of rotifers, but also
Boray (1964), from ecological studies of Lymnaea in rotifer-conditioned water from which the roti-
 63

fers themselves had been removed. The swimming water currents generated during swimming and
activity (speed and distance) of the cercariae was filter feeding (Viaud, 1947). The rotifer attaches
greatly depressed and their ability to penetrate the itself by its foot to the carapace of its host, and
skin of their potential hosts (mice) was reduced by apparently benefits from this association by
50%, compared to the penetration rate of 'non- feeding on the excrement of the cladoceran
rotifer cercariae'. (Schluter, 1984).
Another rotifer which is often found on the
Bryozoa carapace of Daphnia is Proales daphnicola
Philodina megalotrocha Ehrb. is the only rotifer (Thompson, 1892; Harring & Myers, 1922; Beau-
which has been recorded as living on Bryozoa. champ, 1923; Galliford, 1946; Hollowday, 1949;
This species was found in clusters around the Koste, 1968). This rotifer attaches itself firmly to
upper parts of Lophopus crystallinus by Pittock its host by means of its pedal cement glands; it
(1894). The authors notes that even newly hatch- also fixes its eggs to the carapace. P. daphnicola
ed young polypides were heavily infested. Col- appears to gain nothing from the association,
onies of this rotifer seemed to establish themselves apart from transportation (Hollowday, 1949).
on the shoulders of the bryozoan in order to take Other rotifers which are also epizoic on Clado-
advantage of the plentiful food supply drawn into cera, but occur less frequently, include Testudinel-
the area by the bryozoan's feeding activity; a la epicopta Myers on Acantholeberis curvirostris
colony of Lophopus creates a strong vortex in the (Myers, 1934), Brachionus variabilis (Hempel) on
surrounding water which brings food particles Daphnia longispina and Ceriodaphnia longispina
towards it. There was no evidence that Philodina (Hempel, 1896; Ahlstrom, 1940), Collotheca volu-
was actually parasitic on the Lophopus itself, but tata var. sessilis Sebestyen on Monospilus dispar
the rotifer seemed to gain considerable benefit (Sebestyen, 1937) and Brachionus novaezealandiae
from the association (Pittock, 1894). (Morris) on Pseudomonia lemnae, Daphnia carina-
ta and Ceriodaphnia species.
Crustacea Rotifers also occur as ectoparasites on species
Epizoic and parasitic rotifers have often been of the marine opossum shrimp Nebalia. Koste
found on marine and freshwater Crustacea. They (1975) found Seison annulatus Claus among the
are particularly common among freshwater Cla- thoracic appendages which the shrimp uses to
doc era, where some rotifer species occur on a filter organic detritus. The rotifer also attaches its
wide variety of hosts. Brachionus sessilis Varga has eggs to the base of the gills of its host. Paraseison
been found on Diaphonosoma excisum Sars in nudus Plate, P. proboscideus Plate and P. ciliatus
India and Sri Lanka (Chengelath et aI., 1973; Plate have also been found attached to the bran-
Sharma, 1979), on Diaphanosoma sarsii Riahrd in chial lamellae of Nebalia species (Plate, 1888).
India (Sharma, 1979) and on Diaphanosoma bra- These rotifers attach themselves by means of
chyurum Lieven in Hungary (Varga, 1931); this mucus secreted from their pedal gland.
rotifer species has always been found on the same Rotifers have been found on the thoracic and
genus of cladoceran over a wide geographical area abdominal appendages, and the branchial plates,
(Chengelath et al., 1973). In contrast, Brachionus of the freshwater louse Asellus aquaticus (Giglioli,
rubens is more widely distributed among the Cla- 1863; Hudson & Gosse, 1886; Bilfinger, 1894;
docera. Although mostly found on Daphnia Hood, 1895; Rousselet, 1898; Murray, 1906;
species (e.g. D. magna, D. pulex, D. longispina) Hofsten, 1909; Galliford, 1946; Hollowday,
(Bryce, 1924; Galliford, 1946; Viaud, 1947; Hal- 1949; Bartos, 1951). Most of these rotifers belong
bach, 1973; Koste, 1978; Schluter, 1984), it has to 3 main genera, namely Rotaria (R. socialis
also been recorded on the carapace of Moina recti- Kellicott, R. magna-calcarata Parsons), Testudi-
rostris and Polyphemus pediculus (Koste, 1978). nella (T. elliptica (Ehr.), T. truncata (Gosse),
The rotifer is attracted towards a Daphnia by the T. caeca (Parsons)) and Embata (E. parasitica
64

(Giglioli), E.laticeps (Murray), E. commensalis glass, the rotifers often expelled epithelial cells
(Western)). These rotifers do not appear to cause ingested from the hosts gills. These observations
any harm to their host and are generally consider- indicated a parasitic association between the
ed to be epizoans, gaining nothing but transpor- species. The type of association between the
tation from their association with Asellus. species mentioned earlier is not known.
The branchial plates and appendages of Gam-
marus pulex are another common site for epizoic Uniramia
rotifers (Gigliolo, 1863; Hudson & Gosse, 1886; Many rotifers live epizoic ally on the aquatic larvae
Plate, 1886; Hood, 1895; Murray, 1906; Varga, of insects, especially those of the dragonfly (Ro-
1931; Bartos, 1951; Koste, 1968, 1978). These are taria rotatoria, Brachionus rubens, B. caudatus
mostly of the genus Embata (E. parasitica (Giglio- Barrois)(Chandra & Kameswara, 1976; Sharma,
Ii), E.laticeps (Murray), E. hamata), although 1979), damselfly (Lepadella ovalis Muller,
Proales daphnicola, Philodina convergens Murray, B. caudatus) (Chandra & Kameswara, 1976;
Dicranophorus hauerianus var. siedleckii Sharma, 1979) and mayfly (Philodina convergens,
Wiszniewski and Encentruin grande (Western) Murray) (Bartos, 1951). Epizoic rotifers have also
have also been found. Again, none of these been found on the cases of caddis larvae (Rotaria
species seems to have adverse effects on the host. tardigrada Ehr., Embata parasitica, Habrotrocha
A variety of rotifer species are commonly found collaris Ehrb.) (Scherren, 1897; Bartos, 1951) and
in the branchial cavities of the freshwater crayfish, attached to the body and legs of the waterboat-
Astacus species (Carlin, 1939; Koste, 1978). man, Corixa sp. (Albertia naidis Bousfield, B. ru-
These include Lepadella astacicola Hauer, bens) (Varga, 1931; Koste, 1978). Little is known
L. branchiola Hauer, L. parasitica Hauer, L. bor- of the relationship between these rotifers and their
ealis Harring, L. Wiszniewski, Cephalodella cras- hosts, but the associations are not thought to be
sipes (Lord) and several varieties of Dicranophorus truly parasitic.
hauerianus Wiszniewski (Carlin, 1939; Koste,
1978). L. borealis, L. astacicola, L. lata and Echinodermata
D. hauerianus have also been found in the gill Zelinkiella synaptae (Zelinka) has been found in
cavity of the crayfish Cambarus affinis (Carlin, great abundance living in the body cavities of the
1939; Wulfert, 1957; Koste, 1978). Little is echinoderms Synapta inhoerens and S. digitata
known of the relationship between these species, (Lankester, 1868 (?); Zelinka, 1888; Cuenot,
but the rotifers may well be true parasites of these 1892). No detailed information of this relation-
Crustacea. ship is given, but Cuenot (1892) suggests that
Rotifers have also been found in the branchial Zelinkiella is commensal on the integument of the
chambers of both freshwater and marine crabs. Echinoderm, and is unlikely to be a true parasite.
Piovanelli (1903) describes two bdelloids, Macro-
trachela cancrophila (Piovanelli) and Anomopus Chordata
telphusae (Piovanelli), from the branchial chamber An extensive search of the literature revealed only
of Telphusa j/uviatilis, while Mane-Garzon and one record of a rotifer which was either parasitic
Montero (1973) record another, Anomopus chas- or epizoic on fish. This was described by
magnathi (Mane-Garzon & Montero), from the Wiszniewski (1946), who found the cold sten-
branchial chamber of Chasmagnathus granulata. othermal species Encentrum kozminskii
In addition, Proales paguri Thane-Fenchel was Wiszniewski on the skin and gills of carp from fish
found on the gills of the hermit crab Pagurus culture ponds. The author describes the species as
bernhardus (L.) (Thane-Fenche1, 1966). In the a true parasite which feeds on the mucus, and
latter case, the rotifers were seen browsing on the probably the epithelium, of its host. He concludes
gills and sucking the epithelium tissue of their that such an infestation could be harmful to the
host. When lightly compressed under a cover- fish, but this has not been confirmed.
 65

Rotifers have rarely been found living in close illoricate, within the Bdelloidea, Monogononta
association with reptiles. However, Magnusson and Seisonidae. The relationships between these
(1985) found the giant (up to 1500 J-tm) rotifer rotifers and their hosts may be specific or non-
Limnias ceratophylli Schrank living on the trunk, specific, obligate or opportunistic. Their method
limbs, tail and jaws of the Amazonian crocodile, of attachment varies - some are attached by the
Melanosuchus niger. The author suggests that this foot, others by the mouthparts. However, littld, is
association is species specific, as the rotifer was known of the relationships between the species
not found on other crocodile species in the area involved. Clearly, this interesting but neglected
(Caiman crocodilus, Paleosuchus trigonatus, P. pal- area of rotifer research warrants further attention.
pebrosus). M. niger did not seem to suffer any ill
effects from the association, indicating that the
rotifer was simply epizoic on the crocodile. How-
Acknowledgements
ever, the author did express the concern that
heavy infestations could reduce the value of the
I wish to thank Dr. A. E. Bailey-Watts for reading
crocodile hide.
and improving the manuscript, Mrs. L. A. Dick-
Giesen (1934) published what appears to be the
son for help in obtaining the literature and Mrs.
only recorded case of a parasitic rotifer in mam-
M. J. Ferguson who kindly typed the manuscript.
mals. He found a considerable number of
bdelloids in urine samples from a seriously ill
woman whose symptoms included sore throat,
aching joints, stomach disorders, exhaustion, References
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Thanet. Science Gossip 1: 173-175. tisches Radertier Sitz. - Ber. K. B5hm. Ges. Wiss. Prag.
Plate, L. H., 1886. Untersuchungen einiger an der Kiemenb- (1882): 390-397.
Hittern des Gammarus pulex lebenden Ektoparasiten. Z. Viaud, G., 1947. Recherches experimentales sur les tropismes
wiss. Zool. 43: 175-241. des Rotiferes. L'oscillorheotropisme des Brachionus rubens
Plate, L. H., 1888. On some ectoparasitic Rotatoria of the Ehrenberg, cause de la fixation de ce Rotifere phoretique
Bay of Naples. Ann. Mag. Nat. Hist., London, ser. 6, 2: sur les Daphnies et autres crustaces d'eau douce. Ann. Sci.
86-112. nat. Zool. 9: 39-62.
Rees, B., 1960. Albertia vermisculus (Rotifer) parasitic in the Williams, J., 1852. On the occurrence of parasitic Rotifera in
earthworm Allolobophora caliginosa. Parasitol. 50: 61-65. Volvox globator. Trans. microsc. Soc. London, 3: 129-131.
Rich, F. & M. A. Pocock 1933. Observations on the genus Wiszniewski, J., 1946. Sur un rotifere, parasite des carpes.
Volvox in Africa. Ann. s. afro Mus. 16: 427-471. Zool. pol. 4: 7-10.
Rousselet, C. F., 1898. Notes on some little-known species of Wollny, R., 1879. Parasitism of Notommata on Vaucheria. J.
Pterodina. J. Quekett microsc. Club, Ser.2, 7: 24-30. r. microsc. Soc. 2: 291.
Rousselet, C. F., 1911. Rotifera (excluding Bdelloidea). Proc. Wulfert, K., 1957. Ein neues Radertiere aus der KiemenhOhle
Royal Irish Acad. 31, No. 51, 10 pp. von Cambarus affinis. Zool. Anz. 158: 26-30.
Rousselet, C. F., 1914. Intelligence in parasitic rotifers. Wulfert, K., 1960. Die Radertiere Saurer Gewasser der
Knowledge 37: 191,270-273. Dubener Heide. II Die Rotatorien des Krebsscherentiim-
Sauer, F., 1978. A rotifera as parasite in Volvox. Mikrokos- pels Bei Winkelmiihle. Arch. Hydrobiol. 56: 311-333.
mos 67: 110-111. Zelinka, c., 1888. Studien iiber Radertiere. II. Raumparasi-
Scherren, H., 1897. Rotifers commensal with Caddis worms. tismus und Anatomie von Discopus synaptae. Z. f. wiss.
Nature 56: 224. Zool. 47.
Schliiter, M., 1984. Kontinuierliche Massenkultur des plank-
tischen Rotators Brachionus rubens. Ber. Kernforschung-
sanlage Julich 1959, 195 pp.
Hydrobiologia 186/187: 69-73, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 69
© 1989 Kluwer Academic Publishers.

Interrelations of rotifers with predatory and herbivorous Cladocera: a

review of Russian works

Lilian K. Matveeva
A.N. Severtsov Institute of Animal Evolutionary Morphology and Ecology, USSR Academy of Sciences,
Leninsky prospekt 33, Moscow 117071, USSR

Key words: rotifers, cladocerans, epibionts, predation, Polyphemus

Abstract

Publications in Russian on the influence of epibiontic rotifers on cladocerans as well as on predator-prey

interrelations between cladocerans and rotifers are reviewed. Proales daphnicola and Brachionus rubens
are common epibionts of Ciadoc era. The biology of these species is described including the choice of host,
feeding, reproduction and impact on the host. Representatives of the families Daphniidae and Moinidae
are most readily colonized. At high densities of epibiontic rotifers, a high percentage of young Cladocera
die. Predators consuming cladocerans (mainly Bosminidae) belong to the family Asplanchnidae. Rotifers
are consumed by the predatory cladocerans Leptodora and Polyphemus. The results offunctional response
experiments by the author with P. pediculus feeding on Synchaeta pectinata, Asplanchna priodonta, colonial
and solitary Conochilus unicornis, and Platyias patulus are given. In the range of prey densities of
100-16001- 1 afunctional response was found in all the rotifers except P. patulus. Colonies of C. unicornis
were not consumed. The highest level of feeding rate saturation was observed in Synchaeta pectinata.
Various adaptations in prey morphology prevented effective predation: coloniality, large size of the prey
and hard lorica with spines.

Introduction these being Proales daphnicola and Brachionus

rubens. P. daphnicola is an obligatory commensal
Interrelationships between rotifers and clado- of Cladocera (Vlastov, 1953a, b). It occurs most
cerans have drawn increasing attention in recent often on Daphnia pulex and D.longispina and,
years. There are several investigations on this much more rarely, on Scapholeberis, Ceriodaphnia
topic in Russian that are poorly known abroad and Polyphemus. Proales feeds, breeds and devel-
because of linguistic difficulties. The aim of this ops on cladocerans. According to Vlastov
paper is to review these works. I also present (1956b), it feeds on the unicellular euglenacean
results of my own experiments on the feeding of alga Colacium vesiculosum common in epibioses of
Polyphemus pediculus on rotifers. c1adocerans and, to a lesser extent, on bacteria.
The rotifers cannot live long or propagate in the
Rotifers as epibionts of Cladocera absence of actively swimming cladocerans. Most
eggs (65.5%) laid by Proales are attached to the
Two species of epibiotic rotifers living on head of cladocerans on their dorsal side. At a
Cladocera are known from the Soviet literature, temperature of 21-22 0 C, eggs developed in
70

20-21 hours. The mortality of Daphnia caused by intensive settling, the biomass of rotifers on an
Proales in nature was not estimated; however, adult daphnid was equal to the weight of the host.
movements of a young Daphnia are disrupted by On young daphnids it exceeded the weight of the
as few as two attached rotifers. host by 5-17 times, rendering the crustacean
B. rubens exploits crustaceans only as a sub- motionless. The impact of rotifers on a crustacean
stratum; it collects its food in mid-water. depended on its size. Large daphnids (larger than
Brachionus firmly adheres to the carapace of 2 mm) even fully covered with rotifers and bearing
crustaceans using an adhesive secretion produced a sleeve, could remain swimming for 32 hours.
by its two pedal glands. This secretion collects Even less burdened juveniles could not swim after
detritus particles, in 3-5 hours becoming as thick 1.5-3 hours; newborn daphnids were rendered
as 135 J-lm. Other Brachionus specimens do not motionless by 3-4 individuals of Brachionus
settle onto the surface covered with detritus. Hav- (Markevich & Rivier, 1978). Therefore, popula-
ing molted, an old crustacean swims freely for tions of Daphnia affected by B. rubens lacked new-
some minutes and then again becomes covered born individuals. Even in large daphnids
with rotifers. Brachionus are most numerous on (2.0-2.5 mm) covered with Brachionus the rate of
Daphnia one hour after molting and grow less movement dropped by 2 times and the character
numerous in 5 hours (Rivier & Markevich, 1978). of movement was modified (fewer jumps, shorter
Selectivity of Brachionus colonizing substrata races, more variable direction). Dead crustaceans
has been studied in nature and under laboratory were rapidly abandoned by rotifers; in 1.5 hours
conditions (Markevich, 1978). In a pond they the rotifer number decreased by 5-7 times.
definitely prefer D. pulex and Moina brachiata. The joint cultivation of Daphnia and Brachionus
The entire surface oftheir shells becomes covered demonstrated that in pond water Daphnia survival
with rotifers. On Simocephalus vetulus and rate, growth and fecundity were lower in the pres-
Scapholeberis mucronata, B. rubens occurred as ence of epibionts (Markevich & Rivier, 1975;
single individuals. In an experiment, rotifers were Rivier & Markevich, 1978). If fresh organic
offered 36 species of Crustacea, plants and material and detritus were added, the situation
inanimate objects. At increasing densities of reversed. The filtering activity of daphnids appar-
Brachionus their selectivity decreased. At a den- ently was not enough to suppress bacterial growth
sity up to 5 x 104 1- 1 they settled only on Daphnia and to prevent deterioration of environmental
and Moina. At over 16 x 104 1- 1 a slime sleeve is conditions.
formed (threads ofrotifers sitting on cladocerans
hung down from cladocerans bodies and were
used for settlement by other B. rubens). At Rotifera as predators of Cladocera
19 x 104 1- 1, rotifers settled on other Cladocera
such as Ceriodaphnia, Polyphemus, Bythotrephes, Most species of the family Asplanchnidae can
Leptodora, Sida, Diaphanosoma, Scapholeberis, consume small Bosminidae. Bosmina remains are
Simocephalus and Bosmina. Copepoda and found in stomachs of A. girodi, A. herricki,
Chydoridae, with the exception of Eurycercus A. sieboldi and A. priodonta (Tribush, 1960,
lamellatus, were not colonized. At higher den- Ghilarov, 1977, Timokhina, 1983). In experi-
sities, rotifers settled also on plants and dead mental conditions, A. priodonta and A. herricki
objects. The intensity of settling on D. pulex (0.4-0.8 mm long) consumed 3-4 individuals of
depended directly on the density of Brachionus in B.longirostris daily. Food assimilation was
the environment and, as it increased from 5 x 104 15-20% and the assimilation efficiency reached
to 43 X 104 1- 1, their quantity on one Daphnia of 34-36% (Sorokin & Mordukhai-Boltovskaya,
2 mm increased from 16 to 301. In one case of a 1962, Monakov & Sorokin, 1972). Adult females
sleeve formation, up to 700 rotifers were recorded of A. herricki digested Bosmina in 6-6.5 hours at
on one Daphnia (Markevich & Rivier, 1975). With 18°C and in about 4 hours at 21-22 °C
 71

(Kutikova, 1970). I have noted up to 4 Bosmina Functional response experiments on Polyphemus

in the stomach of A. herricki simultaneously.
Because most studies of the influence of
Cladocera as predators of Rotifera Polyphemus on rotifers have been qualitative, I
have conducted several quantitative experiments.
Predaceous cladocerans, such as Polyphemus and The efficiency of feeding of invertebrate predators
Leptodora, are known to consume rotifers. In on rotifers is known to be affected by such prey
experimental conditions, young and adult characteristics as coloniality, body size, the pres-
L. kindti readily consumed A. herricki, A. prio- ence of a hard lorica and spines (Williamson,
donta, E. dilatata and, in lesser quantities, 1983, Stemberger & Gilbert, 1987). Therefore, I
Conochilus sp., K. cochlearis, K. quadrata. How- chose several species of rotifers as prey which had
ever rotifers were not a satisfactory food for one or the other of these characters. Females of
Leptodora which lived for only 2-12 days when Polyphemus (0.7-1.1 mm in length and with well
fed a mixture of A. herricki, C. unicorn is and developed but empty brood chambers) were
Volvox instead of 1.5 months when consuming a caught in June-August 1988 in the littoral zone of
better diet (Mordukhai-Boltovskaya, 1960). A Lake Glubokoe (Smirnov, 1986) and transfered
study of the gut contents of 200 specimens of to clean water for 2-5 hours. Then, they were
Leptodora demonstrated that, in the Rybinsk added to 100 ml vessels filled with 50 Jim-filtered
reservoir on May, 8.5% fed on rotifers. In the gut lake water containing one of the following prey
of an adult Leptodora collected in nature, species: Synchaeta pectinata, Platyias patulus, soli-
K. cochlearis was found (Mordukhai-Boltovs- tary or colonial Conochilus unicornis or Aspianchna
kaya, 1958). Under experimental conditions, priodonta. One predatory animal was added to the
juveniles of Leptodora 3.5 mm long consumed first three prey types, 5-7 to the two latter. The
30-40 B. calyciflorus daily at a prey density of rotifers were collected from the littoral zone which
800-13001-1 (L. V. Polishchuk, personal com- they successively dominated during summer. The
munication). experiments lasted 3 hours at 21-24 0 C. During
Rotifers and protozoans are the main food of this period there was no rotifer mortality in the
newborn and adult males of P. pediculus. Adult control vessels which lacked predator. Poly-
females also prefer ciliates and soft-bodied rotifers phemus feeding rate was estimated by the dif-
rather than crustaceans: prefered prey were ference in rotifer numbers at the beginning and the
Asplanchna, Synchaeta, Polyarthra and in some- end of the experiment. Because the sizes of the
what lesser quantities Conochilus colonies polyphemids used in these experiments varied,
(Butorina & Sorokin, 1966; Butorina, 1970). feeding rates were calculated per unit of dry
Abrupt jumps of Filinia interfere with its con- weight (Dumont et ai., 1975).
sumption by predators, female Polyphemus catch The increase of feeding rate as a function of
this rotifer about half as frequently as Polyarthra. prey density was significant for S. pectinata
Among loricate rotifers, they prefer Keratella, (F(5,1O) = 13.495, P = 0.0004), solitary C. uni-
consuming it at prey densities of more than 50 1- 1 cornis (F(5,7) = 7.084, P = 0.0115) and
(Butorina, 1969, 1970). Nevertheless, for egg- A. priodonta (F(2,6) = 10.474, P = 0.0110), but
bearing females, larger than 0.4 mm, rotifers are insignificant in P. patulus (F(3,8) = 0.097,
not a completely suitable food. Culturing on this P = 0.959) (Fig. 1). The functional response
food decreased size and fecundity but did not curve of P. pediculus reached a plateau at a density
modify longevity. The maximum assimilation effi- of about 800 1- 1 for S. pectinata and 300 1- 1 for
ciency of Keratella at an experimental density of C. unicornis. No such saturation was found in
35 x 104 1- 1 was only 3.6%, while that of new- A. priodonta. The low feeding rate for A. priodonta
born Copepoda and Cladocera was 30-45 % as prey (Fig. 1) was probably due to its large size
(Butorina, 1971). preventing easy capture by the predators. For
72

(Bosmina longirostris, mean length 232 Jim) in the

v
0

R/W 0 0 0
range of densities of 50-1000 prey 1- 1 (t = 2.355,
0-0 P < 0.05). Thus, as in other invertebrate preda-
100 0
tors, the successful feeding of P. pediculus was
0
186-337 dependent upon the size of prey, the formation of
0
colonies and the development of a hard exo-
0
87'312
skeleton with spines. Probably in nature this
0 +
predator most efficiently preys upon populations

I- ~
+
376'618
of soft-bodied rotifers of about 300 Jim in length.
50 +
+
+
156'231

,.; ... ••
•- -
•
~ Acknowledgements

0
0
•
•
_

i i
50
. -
--'--w

i i i
•
i
100
-I
150
,
N/l00 ml
This work has been done at the hydrobiological
Station 'Lake Glubokoe'. I am grateful to Dr. C.
Ricci for the arrangements concerning my par-
Fig. I. Functional responses of Polyphemus pediculus for dif-
ticipation in the 5th International Rotifer Sym-
ferent prey. Ordinate: predator feeding rate per unit dry
weight (R/W, prey Jlg- 1 hr- 1 ); abscissa: density of prey (N, posium, and to my husband, Dr. V. Matveev, for
animais/IOO ml). Open circles = Synchaeta pectinata, filled various help throughout this study. I would like to
circles = Asplanchna priodonta, squares = Platyias patulus, thank Dr. P. Starkweather for language revision
crosses = solitary Conochilus unicornis. The averages for two of the manuscript.
major dimensions of body size (Jlm) (n = 20) are also
shown.

References
example, several experiments with smaller
A. priodonta (average dimensions 259 x 446 Jim) Butorina, L. G., 1969. 0 prichinakh obrazovaniya stay u
yielded a 1.5-2 fold increase in the feeding rate of Polyphemus pediculus (L.). Informatsionniy bulleten, Insti-
the predator. The plateau was highest in tut biologii vnutrennikh vod 3: 68-71 (On causes of swarm
formation in Polyphemus pediculus (L.). In Russian).
S. pectinata, intermediate in solitary C. unicornis
Butorina, L. G., 1970. Ob izbirate1nosti pitaniya Polyphemus
and lowest in P. patulus (Fig. 1). The relatively pediculus (L.). Informatsionniy bulleten, Institut biologii
low density feeding rate saturation in solitary vnutrennikh vod 7: 46-50 (On feeding selectivity of Poly-
C. unicornis was probably due to the fact that phemus pediculus (L.). In Russian).
these animals either were not a familiar food for Butorina, L. G., 1971. Intensivnost pitaniya Polyphemus
pediculus (L.) v zavisimosti ot kontsentratsii korma. Infor-
the crustacean or were distasteful because of
matsionniy bulleten, Institut biologii vnutrennikh vod 10:
gelatin remnants. Not a single colony of 35-39 (Feeding intensity of Polyphemus pediculus (L.) in
C. unicornis was consumed by P. pediculus at a relation to food concentration. In Russian).
density of 10-40 colonies per 100 ml. Poor Butorina, L. G. & Yu I. Sorokin, 1966. 0 pitanii Polyphemus
feeding on P. patulus seemed to be due to its hard pediculus (L.). Trudy Instituta biologii vnutrennikh vod
12(15): 170-174 (On feeding of Polyphemus pediculus (L.).
lorica and well developed spines.
In Russian).
The absolute maximum feeding rates of Dumont, H. J., I. Van de Ve1de & S. Dumont, 1975. The dry
P. pediculus were observed at a prey density of weight estimate of biomass in a selection of Cladocera,
1-1.2 x 103 1- 1 and were as high as 7 S. pectinata Copepoda and Rotifera from the plankton, periphyton and
hour - 1 predator - 1, or 4.7 C. unicornis or 1.5 benthos of continental waters. Oecologia 19: 75-97.
Ghilarov, A. M., 1977. Nablyudeniya nad sostavom pishchi
P. patulus. Notwithstanding the high rate of
kolovratok roda Asplanchna. Zoologicheskiy zhurnal 56,
Synchaeta consumption, P. pediculus significantly 12: 1874-1876 (Observations on food composition of
lowered its intensity offeeding on the rotifer in the rotifers of the genus Asplanchna. In Russian).
presence of equal amounts of an alternative prey Kutikova, L. A., 1970. Kolovratki fauny SSSR. Nauka Pub!.,
 73

Leningrad: 744 pp. (Rotifers in the Fauna of the USSR. In Smirnov, N. N., 1986. Lake Glubokoe. Hydrobiologia 141,
Russian). 1/2.
Markevich, G. I., 1978. Ob izbiratelnosti Brachionus rubens Sorokin, Yu. I. & E. D. Mordukhai-Boltovskaya, 1962.
pri zaselenii substrata. Informatsionniy bulleten, Institut Izuchenie pitaniya kolovratok Asplanchna s pomoshchyu
biologii vnutrennikh vod 39: 37-40 (On selective coloni- C 14• Bulleten Instituta biologii vodokhranilishch 12: 17-20
zation of substrata by Brachionus rubens. In Russian). (Feeding of Asplanchna studied using C 14• In Russian).
Markevich, G. I. & I. K. Rivier, 1975. Vliyanie epibiontnykh Sternberger, R. S. & J. J. Gilbert, 1987. Defenses of plank-
bespozvonochnykh na kopepod i kladotser. In: Povedenie tonic rotifers against predators. In W. C. Kerfoot & A. Sih
vodnykh bespozvonochnykh, Borok: 49-52 (The effect of (eds), Predation: Direct and indirect impacts on aquatic
epibiotic invertebrates on copepods and c1adocerans. In communities. Univ. Press of New England, Hanover &
Russian). London: 227-239.
Markevich, G. I. & I. K. Rivier, 1978. Vliyanie Brachionus Timokhina, A. F., 1983. Pitaniye kolovratok rodaAsplanchna
rubens na dvigatelnuyu aktivnost nekotorykh Cladocera. (Ploimida, Asplanchnidae). In G. G. Winberg (ed.),
Informatsionniy bulleten, Institut biologii vnutrennikh Troficheskie svyazi i ikh rol v produktivnosti prirodnykh
vod 39: 45-48 (Influence of Brachionus rubens on motor vodoyemov. Leningrad: 81-84 (Feeding ofrotifers of the
activity of some Cladocera. In Russian). genus Asplanchna. In Russian).
Monakov, A. V. & Yu. I. Sorokin, 1972. Some results on Tribush, T. M., 1960. Nekotorye nablyudeniya nad
investigations on nutrition of water-animals. In Z. Kajak kolovratkami sem. Asplanchnidae Rybinskogo vodokhrani-
& A. Hillbricht-Ilkowska (eds), Productivity problems of lishcha. Bulleten Instituta biologii vodokhranilishch 6:
freshwaters. Warszawa-Krakow: 765-773. 18-19 (Some observations on rotifers of the family
Mordukhai-Boltovskaya, E. D., 1958. Predvaritelnye dannye Asplanchnidae in Rybinsk reservoir. In Russian).
po pitaniyu khishchnykh kladotser Leptodora kindti i Vlastov, B. V., 1953a. Evropeyskie i severo-amerikanskie
Bythotrephes. Doklady AN SSSR 122,4: 723-726 (Prelimi- kolovratki iz sem. Notommatid - sozhiteli Cladocera i ikh
nary data on feeding of predatory c1adocerans Leptodora vidovaya prinadlezhnost. Zoologicheskiy zhurnal 32, 6:
kindti and Bythotrephes. In Russian). 1110-1114 (European and North-American rotifers ofthe
Mordukhai-Boltovskaya, E. D., 1960.0 pitanii khishchnykh N otommatidae family - cohabitants of Cladocera and their
kladotser Leptodora i Bythotrephes. Bulleten Instituta species attribution. In Russian).
biologii vodokhranilishch 6: 21-22 (On the feeding of Vlastov, B. V., 1953b. Vzaimootnosheniya mezhdu Cladocera
predatory c1adocerans Leptodora and Bythotrephes. In i zhivushchimi na nikh kolovratkami iz roda Proales.
Russian). Trudy Vsesoyuznogo hydrobiologicheskogo obshchestva
Rivier, I. K. & G. I. Markevich, 1978. Vliyanie Brachionus 5: 299-317 (Interrelations between Cladocera and rotifers
rubens na biologicheskie pokazateli nekotorykh Cladocera of the genus Proales living on them. In Russian).
pri sovrnestnom obitanii. Informatsionniy bulleten, Institut Williamson, C. E., 1983. Invertebrate predation on plank-
biologii vnutrennikh vod 39: 41-44 (Influence of tonic rotifers. Hydrobiologia 104: 385-396.
Brachionus rubens on biological parameters of some
Cladocera in case of cohabitation. In Russian).
Hydrobiologia 186/187: 75-80, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 75
© 1989 Kluwer Academic Publishers.

Competitive interactions between the rotifer Synchaeta oblonga and the

c1adoceran Scapholeberis kingi Sars

John J. Gilbert
Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire 03755, USA

Key words,' competition, interference, Scapholeberis, Synchaeta, zooplankton

Abstract

Competition experiments showed that the small cladoceran Scapholeberis kingi rapidly excluded the rotifer
Synchaeta oblonga from mixed-species cultures, but was itself unaffected by the presence of S. oblonga.
Short-term experiments testing the effect of S. kingi on the survivorship and reproduction of S. oblonga
showed that the former imposed a high mortality on the latter, even though shared food resources were
abundant. These results indicate that adult S. kingi mechanically interferes with S.oblonga either by
ingesting, or by rejecting in a damaged condition, individuals swept into its branchial chamber. In contrast
to many other small species of cladocerans, and like large species of Daphnia, S. kingi has the potential
to markedly suppress populations of some rotifer species through a combination of interference and
exploitative competition.

Introduction cladoceran genera (Bosmina longirostris, Cerio-

daphnia dubia, Holopedium gibberum,
Considerable direct and circumstantial evidence Diaphanosoma brachyurum) with body lengths less
from field and laboratory studies shows or indi- than 1.2 mm interfered slightly, if at all, with one
cates that rotifers are strongly suppressed by large of the most susceptible rotifer species (K. coch-
Daphnia - those with body lengths (head to base learis) Bums & Gilbert, 1986a, b; Gilbert &
of tails pine) greater than about 1.2 mm (Gilbert, MacIsaac, 1989), and one of these cladocerans
1988a). Such Daphnia can monopolize shared (c. dubia) only interfered with newborn individ-
food resources and also can mechanically inter- uals of this rotifer (Gilbert & MacIsaac, 1989).
fere with (damage or eat) some small or soft- Furthermore, laboratory competition experiments
bodied rotifers swept into their branchial cham- using a variety of different-sized cladoceran
bers (Gilbert, 1985, 1988b; Gilbert & Sternberger, species and three rotifer species (Keratella testudo,
1985; Bums & Gilbert, 1986a, b). K. cochlearis, Synchaeta oblonga) showed that one
In contrast, rotifers seem to be much less sup- or more of these rotifers could coexist for many
pressed by small cladocerans and, in fact, often weeks with, and sometimes competitively sup-
abound with them in natural zooplankton com- press, two small cladoceran species (B. longiro-
munities (Gilbert, 1988a). These cladocerans are stris, Daphnia ambigua) but were rapidly excluded
much less able than large Daphnia to sequester by three large Daphnia species (D. galeata
food resources and to interfere with rotifers. For mendotae, D. pulex, D. magna) (Gilbert &
example, Daphnia and species of four other Sternberger, 1985; MacIsaac & Gilbert, in press).
76

In the present study I examine competitive A short-term experiment was conducted to

interactions between the small, soft-bodied rotifer determine the effect of S. kingi on the survivorship
Synchaeta oblonga and the small cladoceran of S. oblonga in the 2 cm-diameter concavities of
Scapholeberis kingi Sars - probably S. armata glass, 9-spot plates. Each concavity contained
armata (Herrik) according to the revision of 1 ml of a dense Cryptomonas suspension
Dumont and Pensaert (1983). Unlike previous (5-10 x 104 cells ml- 1), 2 randomly-selected
studies with other small cladoceran species, this S. oblonga, and either no or 1 adult S. kingi. Nine
study surprisingly shows that S. kingi rapidly replicate cultures of each treatment were in alter-
excludes S. oblonga from mixed-species cultures nating concavities of each of two 9-spot plates.
and can impose a high mortality rate on S. oblonga Each 9-spot plate was placed in a separate moist
through mechanical interference. chamber. After 16 hours, the numbers of surviv-
ing S. oblonga and S. kingi in the cultures were
determined. The pooled frequencies of living and
Materials and methods dead (missing or eaten) S. oblonga in the cultures
with and without S. kingi were compared with
Synchaeta oblonga and Scapholeberis kingi were Fisher's exact test (Sokal & Rohlf, 1981).
isolated from water bodies within 30 Ian of Another short-term experiment was conducted
Dartmouth College, cloned, and maintained on to determine the effect of S. kingi on the reproduc-
Cryptomonas sp. in glass-tiber-ftltered lake water tive rate of S. oblonga in covered, 20-ml, glass
as described elsewhere (Gilbert, 1985, 1988b). beakers. Each beaker contained 10 ml of a dense
Adult body lengths of female S. oblonga and Cryptomonas suspension (5-10 x 104 cells ml- 1),
S. kingi were about 150 J-lm and 0.4 to 0.6 mm, 5 randomly-selected S. oblonga, and either no or
respectively. All experiments were conducted at 2 adult S. kingi. Six replicate cultures with and
20 DC in a photoperiod (LD 16: 8) with dim light without S. kingi were organized as 6 matched
(",,300 lux). pairs. After 23.3 to 24.2 hours, the numbers of
Competition experiments were conducted in living S. oblonga and S. kingi in each culture were
covered, 50-ml glass beakers containing 40 ml of determined. Six replicate death rates of S. oblonga
a Cryptomonas suspension (104 cells ml- 1) in a due to the presence of S. kingi (ds ) were deter-
9: 1 (v/v) mixture of lake water and the mined as described elsewhere (Gilbert, 1988b)
Cryptomonas growth medium (Stemberger, 1981). from the reproductive rates of the S. oblonga in
Initial population sizes were 20 randomly-selected the 6 pairs of cultures with and without S. kingi.
S. oblonga in the single- and mixed-species cul-
tures and 3 adult (experiment 1) or juvenile
(experiment 2) S. kingi in the single- and mixed- Results
species cultures. Three replicate cultures were set
up for each treatment. All individuals in all popu- In both of the competition experiments, S. oblonga
lations of the two species were counted and trans- was significantly suppressed by S. kingi while
ferred by pipette to fresh Cryptomonas suspension S. kingi was unaffected by S.oblonga (Fig. 1,
every 2 days until one of the species went to Tables 1 and 2). In experiments 1 and 2, the
extinction. Oviposited eggs of S. oblonga were S. kingi excluded S. oblonga from mixed-species
transferred along with the swimming individuals, cultures after 4 and 16 days, respectively. The
but were not counted. Population growth trajec- S. kingi populations in experiment 2 were initiated
tories for each species with and without the other with juveniles and did not begin to noticeably
were analyzed using univariate, repeated- affect the S. oblonga populations until day 6, when
measures ANOVA and Student-Newman-Keuls they contained adults. The S. kingi populations in
(SNK) tests (SAS Institute, 1985) on population- experiment 1 were initiated with adults and mark-
size data from time zero. edly depressed S. oblonga population sizes by day
 77
1000.------------------~
Table 1. Changes in population sizes over time for Synchaeta
oblonga and Scapholeberis king; in 40-ml, single-species and

800
f t
mixed-species cultures. Experiment I. Scapholeberis popu-
lations initiated with gravid adult females. Values are means
of 3 replicates ± 1 SE.

Day Culture

600 Synchaeta Scapholeberis

Single Mixed Single Mixed

species species species species

o 20 ± 0 20 ± 0 3±0 3±0
2 39 ± 7 10 ± 2 20 ± 1 21 ± 1
4 88 ± 23 1± 1 59 ± 4 57 ± 2
200

over time (10.3 vs. 49.1 and 110.6 vs. 410.2 for
experiments 1 and 2, respectively; SNK tests,
P < 0.05) (Table 2). In neither experiment did the
80 presence of S. oblonga significantly affect the
population growth trajectories or the mean popu-
lation sizes of S. kingi (treatment x time effects,
P> 0.05; SNK tests, P> 0.05) (Table 2). Visual

!
60 inspection of the cultures throughout the competi-
.Cr)
'<:0
=: tion experiments showed that Cryptomonas cell
~
~
densities were greatly depleted or negligible before
I:::>
40 renewal in cultures with S. kingi at days 2 and 4
~
t::i in experiment 1 and in all cultures by day 8 and
~
t thereafter in experiment 2.

1!
The results of the short-term survivorship
20
experiment showed that the presence of S. kingi
~ significantly increased the mortality of S. oblonga.
The numbers of surviving S. oblonga with and
01 ~0 c:eI c:eI
4
ceI 0'8
I I
12
,I
16 without S. kingi were 9 and 17 out of 18, respec-
tively (Fisher'S exact test, P = 0.007). Crypto-
Day
monas cells were present in all cultures at the end
Fig. 1. Changes in population sizes over time for Synchaeta of the experiment.
oblonga and Scapholeberis king; in 40-ml, single-species (open
circles) and mixed-species (closed circles) cultures. Experi- The results of the short-term reproductive rate
ment 2. Culture conditions: see Table I. Scapholeberis popu- experiment showed that S. kingi imposed a mean
lations initiated with juvenile females. Values are means of3 death rate (d s ) of 1.2 day- 1 on the S.oblonga
replicates ± 1 SE (drawn when greater than circle). populations (Table 3). The mean reproductive
rate of S. oblonga populations was significantly
lower with S. kingi present ( - 1.1 day - 1) than
2. In both experiments, the population sizes of with it absent (0.08 day - 1 )(Student's t = 4.4 with
S. oblonga with and without S. kingi diverged sig- 10 df, P < 0.01, 2-tailed). Cryptomonas cell den-
nificantly over time (treatment x time effects, sities remained high in all cultures at the end of the
P < 0.001) and had significantly different means experiment.
78

Table 2. Analysis of results of competition experiments with Scapholeberis kingi (S.k.) and Synchaeta oblonga (S.o.) (See Table I
and Fig. I). Effects of each species on the other analyzed with repeated-measures AN OVA (type III model) and
Student-Newman-Keuls (SNK) tests. Significant levels of P > 0.05, < 0.05, <0.01, and <0.001 indicated by NS, *, **, and ***,
respectively.

Treatment Experiment ANOV A (F-value) SNK

test
Treatemnt Replicate Time Treatment
x time

S.k. on S.o. 1 2.3 NS O.4 NS 6.1* 19.0*** *

2 7.1** 1.8NS 86.8*** 65.6*** *
S.o. on S.k. 1 1.1 NS 0.8 NS 251.1 *** ONS NS
2 5.0* 3.2* 74.1 *** 0.9 NS NS

Table 3. Short-term experiment on the effect of Scapholeberis competition from adult S. kingi. The high mor-
kingi on the reproductive and death rates of Synchaeta tality rates imposed on S. oblonga by S. kingi in
oblonga. Experimental conditions: lO-ml cultures initiated
the short-term experiments can only be inter-
with 5 S. oblonga, with and without 2 adult S. kingi females,
and terminated after 0.97-1.01 days; 5-10 x 104 cells ml- 1 preted by some kind of interference, as food was
Cryptomonas sp; 20°C; photoperiod (LD 16: 8, ~300 lux). still present in the cultures at the end of the experi-
Symbols for rates: re and rs (reproductive rates of S. oblonga ments. Similarly, the very rapid declines of the
in control cultures without S. kingi and in cultures with S. oblonga populations in the mixed-species cul-
S. kingi), ds (death rate of S. oblonga due to presence of
tures of the competition experiments when
S. kingi, equals re-rs).
S. kingi adults were present strongly suggest inter-
Replicate Rate ference rather than exploitative competition.
and Exploitative competition alone probably would
statistic re day-l rs day-l ds day-l not have such a pronounced effect, since the
dense populations of S. oblonga in the single-
0.19 -0.23 0.42
2 0.19 - 1.64* 1.83* species cultures were severely food-limited during
3 -0.23 - 1.63 1.40 the last 6 days of experiment 2 yet declined only
4 0 -0.93 0.93 slightly during this period (Fig. 1).
5 0.34 - 0.51 0.85 It is of interest that the results of the two com-
6 0 - 0.59* 1.59*
petition experiments were so similar despite con-
Mean 0.08 -1.09 1.17 siderable differences in initial conditions. The
ISD 0.20 0.62 0.53 Scapholeberis individuals at the start of experi-
ment 1 were adult and rapidly reduced the den-
* Underestimate of rate; all S. oblonga killed or eaten, but 1 sities of the Synchaeta populations from their ini-
individual assumed to be alive at end of experiment so that
tial level (0.5 individual ml- 1). In contrast, the
rs could be calculated from logarithmic formula.
Scapholeberis individuals at the start of experi-
ment 2 were juvenile and permitted the Synchaeta
populations to increase ten-fold (from 0.5 to '" 5
Discussion individuals ml- 1) over a 12-day period before
they became adult and rapidly suppressed the
The combined results of the different experiments rotifers. Thus, the outcome of competition
show than Synchaeta oblonga was rapidly ex- between these two species was the same at rela-
cluded by Scapholeberis kingi from mixed-species tively low and high rotifer densities.
cultures and that this exclusion probably was pri- The mechanism by which S. kingi interferes
marily due to direct, mechanical, interference with S. oblonga is not known. It could be entirely
 79

predation or some combination of predation and 2 mm strongly interfered with K. cochlearls and
damage to rejected individuals. The high mor- also rapidly drove this rotifer to extinction in com-
tality rates attributed to interference in all of the petition experiments (Gilbert 1985; Gilbert &
experiments certainly was due to the very high Sternberger, 1985; Burns & Gilbert, 1986a, b;
population densities of the Scapholeberis (up to MacIsaac & Gilbert, in press). The same was true
'" 1 individual ml- 1 ). Preliminary observations of for S. kingi with S. oblonga in the present study.
encounters between S. kingi and S. oblonga failed Ceriodaphnia dubia, which interfered only with
to detect either type of interference. There appears juvenile K. cochlearis, strongly suppressed this
to be no information on the food niche of any rotifer but did not completely exclude it from
species of Scapholeberls, and so the potential of mixed-species cultures even after 8 weeks
these cladocerans to eat large food items is not (MacIsaac and Gilbert, in press). Bosmina longi-
known. rostris, which did not interfere with K. cochlearis,
Of all the small « 1.2 mm body length) only weakly suppressed both K. cochlearls and
cladoceran species tested thus far, S. kingi S. oblonga and, in turn, was weakly suppressed by
(0.6-0.8 mm adult body length) is the only one to them (MacIsaac & Gilbert, in press). Finally,
markedly interfere with a rotifer. Ceriodaphnia Daphnia ambigua, which interfered slightly or not
dubia (--0.7 mm adult body length) interfered at all with K cochlearls, only weakly suppressed
with juvenile but not older Keratella cochlearis, K. cochlearis and K. testudo, and in one experi-
and Daphnia ambigua ('" 1.0 mm adult body. ment it was excluded by K. cochlearis (Kirk &
length) interfered only to a limited extent, if at all, Gilbert, unpublished; MacIsaac & Gilbert, in
with K. cochlearis (Burns & Gilbert, 1986a, b; press).
Gilbert & MacIsaac, 1989). Bosmina longirostris Cladoceran species that strongly interfere with
( '" 0.4 mm adult body length), Diaphanosoma a rotifer species have not been significantly sup-
brachyurum ('" 0.8 mm adult body length), and pressed by that rotifer species in competition
Holopedium gibberum (0.7-0.8 mm adult body experiments. This was true for the three large
length) did not interfere with K. cochlearls (Gilbert (> 2 mm adult body length) species of Daphnia
& MacIsaac, 1989). (Gilbert, 1985; MacIsaac & Gilbert, in press) and
The susceptibility of S. oblonga to cladoceran also for the S. kingi in the present study.
interference is very similar to that of K. cochlearis. Another cladoceran known to interfere with
This was shown for post-juvenile individuals of rotifers is Moina hutchinsoni. Females of this
these two species in experiments with Daphnia species attain body lengths of 1.4 to 1.6 mm
pulex (Gilbert, 1988b). Juvenile S.oblonga was (Goulden, 1968) and can eat the soft-bodied
more vulnerable to D. pulex than older rotifer Hexarthra (Edmondson, 1974). Exami-
K. coch/earls (Gilbert, 1988b), but the relative nation of the guts of 108 M. hutchinsoni from Soap
susceptibilities of juvenile individuals of the two Lake, Washington over 7 dates in 1974 revealed
rotifer species have not been compared. At any one set of H. fennica trophi in each of 3 individuals
rate, the small cladocerans that did not interfere (N. G. Hairston, Jr., pers. comm.). While these
with K. cochlearls probably would not interfere Moina may not have preyed extensively on the
with S.oblonga, and, conversely, K. cochlearis Hexarthra they may have damaged and killed a
should be as susceptible to interference from high proportion of individuals swept into and then
Scapholeberls kingi as S. oblonga was in the pres- rejected from their branchial chambers. It is note-
ent study. worthy that one field study provides strong corre-
The ability of a cladoceran species to interfere lational evidence for the competitive suppression
with a rotifer species is positively correlated with of rotifers by a much smaller species of Moina,
its ability to competitively exclude that species M. micrura (Gasith & Perry, 1985).
from mixed-species cultures. Three species of
Daphnia with adult body lengths greater than
80

Acknowledgements Gilbert, J.1., 1985. Competition between rotifers and

Daphnia. Ecology 66: 1943-1950.
Gilbert, J. J., 1988a. Suppression of rotifer populations by
This research was supported by National Science Daphnia: a review of the evidence, the mechanisms, and
Foundation research grants BSR-8415024 and the effects on zooplankton community structure. Limnol.
BSR-8717074. I am very grateful to H.J. Oceanogr. 33: 1286-1303.
MacIsaac for performing the ANOVA and for Gilbert, J. J., 1988b. Susceptibilities of ten rotifer species to
interference from Daphnia pulex. Ecology. 69: 1826-1838.
improving the manuscript.
Gilbert,1. 1. & R. S. Sternberger, 1985. Control of Keratella
populations by interference competition from Daphnia.
Limnol. Oceanogr. 30: 180-188.
References Gilbert, J. J. & H. J. MacIsaac, 1989. The susceptibility of
Keratella cochlearls to interference from small c1adocerans.
Burns, C. W. & J. J. Gilbert, 1986a. Effects of daphnid size Freshwat. BioI. 22: in press.
and density on interference between Daphnia and Keratella Goulden, C. E., 1968. The systematics and evolution of the
cochlearis. Limnol. Oceanogr. 31: 848-858. Moinidae. Trans. am. phil. Soc. NS 58(6): 1-98.
Burns, C. W. & J.1. Gilbert, 1986b. Direct observations of MacIsaac, H. 1. & 1. J. Gilbert. Competition between rotifers
the mechanisms of interference between Daphnia and and c1adocerans of different body sizes. Oecologia (Berlin).
Keratella cochlearls. Limnol. Oceanogr. 31: 859-866. In press.
Dumont, H.1. & J. Pensaert, 1983. A revision of the SAS (Statistical Analysis System) Institute, 1985. User's
Scapholeberinae (Crustacea: Cladocera). Hydrobiologia guide: statistics ed. 5. Cary, North Carolina, USA.
100: 3-45. Sokal, R. R. & F. 1. Rohlf, 1981. Biometry. W. H. Freeman
Edmondson, W. T., 1974. Secondary production. Mitt. int. and Co., San Francisco, California, USA.
Ver. Limnol. 20: 229-272. Sternberger, R. S., 1981. A general approach to the culture
Gasith, A. & A. S. Perry, 1985. Use of limnocorrals for of planktonic rotifers. Can. 1. Fish. aquat. Sci. 38: 721-724.
pesticide toxicity studies: effect on zooplankton com-
position and dynamics. Verh. int. Ver. Limnol. 22:
2432-2436.
Hydrobiologia 186/187: 81-102, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 81
© 1989 Kluwer Academic Publishers.

Salinity and temperature influence in rotifer life history characteristics

Maria R. Miracle & Manuel Serra

Dep. de Ecologia, Fac. de C. Biol6gicas, Univ. de Valencia, 46100 Burjassot (Valencia), Spain

Key words: rotifers, rate of population increase, life span, temperature, salinity

Abstract

A review of temperature and salinity effects on rotifer population dynamics is presented together with
original data of these effects for three clones of Brachionus plicatilis. There is a clear relationship between
temperature and the intrinsic rate of increase, r: an increase of temperature - within the natural
environmental range - produces an exponential increase of r, and the slope of the response depends on
the genotype. The effect of salinity is also genetically dependent; the highest r for each clone is observed
at the salinity close to that of its environmental origin. The response of r to temperature is mainly a
consequence of the response of the individual rates of development and reproductive timing. The effect
of temperature on fecundity (number of descendents per individual life time) is negligible when tempera-
ture values are within the normal habitat ranges. On the other hand, salinity seems to affect primarily
fecundity. The interaction salinity-temperature may be important in clones or species living in fluctuating
environments with positive response to the more frequent combinations found in the corresponding
habitats.

Introduction ments on the relationships of temperature and

salinity to the intrinsic rate of population increase,
Extended studies of aquatic populations usually r, which is assumed to be the best measure of
confirm a community typology that matches fitness in non-saturated environments. Rotifers in
environmental heterogeneity. The main factors strongly fluctuating planktonic environments,
determining the community typology are most fre- which have to adapt to regular colonizing events,
quently (1) temperature, (2) salinity and the pa- may frequently be subject to selection for high r.
rameters associated with it, and (3) trophic status The paper is divided into two main sections. In
of the waters. For instance, principal components the first, a brief bibliographic review of the effects
analysis, for data on the species abundance in of temperature and salinity on popUlation dynam-
zooplanctonic communities from a wide range of ics parameters is given. Physiologic effects on
conditions, usually identify, as the first principal individual rotifers will also be summarized to help
components, variables that are highly correlated interpret the effects on populations. In the second,
with the above mentioned factors (e.g., Miracle, original data combining the two factors from a
1974; Armengol, 1978; Miracle et aI., 1987; series of experiments will be discussed. Our in-
Nogrady, 1988). Temperature, salinity and food tention is to evaluate the interaction between tem-
are also the most frequently studied factors in life perature and salinity, i.e., the dependence of the
history analysis. effect of one factor on the level of the other.
This paper reviews results of laboratory experi-
82

Temperature: effects on individuals 1987) or clones (Pourriot & Deluzarches, 1971)

have lower slopes and globally smaller rates than
The direct positive effect of temperature on warm-water adapted ones. Therefore, in many
physiological rates in individual rotifers and on cases, cold-adapted species show faster low-
rotifer populations have been clearly demon- temperature development but much slower high-
strated. Several of these studies deal with the temperature development than warm-adapted
relationship between temperature and duration of species and vice versa. However, other factors
embryonic development. Vinberg & Galkovskaya such as the r-K strategy of the species have to be
(1979), Herzig (1983), Duncan (1983) and considered. The effect of temperature is also de-
Galkovskaya (1987) have summarized data on pendent on its stability. If temperature fluctuates
the subject. The duration of embryonic develop- around a mean value, the egg development time
ment (D) in rotifers is a curvilinear function of deviates from that at the constant mean (Ruttner-
temperature (T), which can be adequately de- Kolisko, 1975, 1978); it is shortened, as would be
scribed by Belehradek's equation liD = aCT - b)C expected, by the effect of oscillating temperature
(a,b,c, fitted parameters) or Arrhenius' equation on the metabolism. Figure 1 presents idealized
liD = ae-b/T(Waltz, 1983) as in other organisms responses to temperature change of activities and
(Herzig, 1983). However other equations have functions discussed in this section.
also been proposed as best fit (i.e., It is often assumed that duration of embryonic
liD = a + bT + cP, Galkovskaya, 1987; development is only dependent upon the tempera-
liD = aebT, Duncan, 1983). Furthermore, when ture (Herzig, 1983). However, relations with other
homogeneous data from one species, studied by parameters have also been described, mainly with
the same author, in a limited range oftemperatures, egg volume (Pourriot & Deluzarches, 1971),
are used, the relationship between liD and Tis which is in tum dependent on temperature
nearly linear (Edmondson, 1965; Pourriot & (Pourriot, 1973a) or on the feeding conditions of
Deluzarches, 1971). The slope of the curve and the mother (YUfera, 1987).
the absolute rate values are dependent on geno- Duration of the pre-reproductive period is also
type. Cold-water adapted species (Galkovskaya, a clear function of temperature. Its variation

O2 (A) act. (BJ 1 tel

cons. D

~\, /
T T T

02 (A) act. (B) r (D)

cons. ,,
,
\

T T
Fig. I. Ideal curves showing relationships between temperature and (A) oxygen consumption, (B) activity (filtration rate),
(C) inverse of development rate, and (D) intrinsic rate of growth.
 83

parallels that of egg development. Thus, under the behaviour of cold vs. warm water species is simi-
same feeding conditions the ratio postembryonic/- lar to the trends described for egg development
embryonic development time remains quite con- time.
stant (Pourriot & Deluzarches, 1971; Ruttner- Galkoskaya (1987) also reports data on O2
Kolisko, 1975). However, when rotifers are sub- consumption in natural popUlations of Brachionus
jected to fluctuating temperature, the above calyciflorus and Hexarthra mira; the same expo-
mentioned ratio could be very different because nential function was found but with smaller slope
there is some evidence (Ruttner-Kolisko, 1978) than the cultivated individuals. Another dif-
that the duration of the pre-reproductive period ference was that, after a maximum, a sharp decline
does not deviate significantly from that obtained occurred when temperatures were too high.
at the corresponding mean temperature. H. mira, usually occurring in lower temperatures
Post-embryonic growth is also highly depend- than B. calyciflorus, also showed lower O2 con-
ent on temperature. Growth in length in rotifers sumption. Other metabolic rates have rarely been
can be described by the Bertalanffy equation studied. Nitrogen excretion rates, investigated for
(Lebedeva & Gerasimova, 1985; Carmona et aI., B. calyciflorus by Galkovskaya et al. (1987),
this volume). Growth is almost totally restricted showed the same exponential function with tem-
to the juvenile period (defined as lasting until the perature as O 2 consumption and a quite similar
laying of the first egg): the growth rate undergoes QlO (= 2.2).
a sharp decrease coincident with the beginning of There is also only fragmentary information on
reproduction and from then on, matter and energy rotifer activities, but from this, it seems that there
are directed towards reproduction instead of exists a clear response to temperature. The
growth. The somatic growth rate in juveniles is a observed response, when a wide range oftempera-
clear function of temperature as well as the dura- tures is tested, could be interpreted either as a
tion of the growth period. Consequently, the cal- sigmoid curve with a decline at unsuitably high
culated QlO for both somatic growth (during the temperatures, or as two sigmoid functions with a
pre-reproductive period) and egg development are plateau, at a specific optimum range of tempera-
similar, ranging approximately from 2 to 3 tures, in between. Tested activities are mainly fil-
(Lebedeva & Gerasimova, 1985, Pourriot & tration and ingestion rates (Hirayama & Ogawa,
Deluzarches, 1971, Duncan, 1983). 1972; Galkovskaya, 1987), and locomotion (Epp
Very few studies exist on the effect of tempera- & Lewis, 1984, Snell et al., 1987), with interpre-
ture on rotifer metabolism, and they have variable tation of the obtained results varying amongst the
results. Pourriot (1973b) and Galkovskaya et al. authors. Rates of beat of vibratile flames and of
(1987) find an exponential increase in oxygen con- output from the contractile vesicles in Asplanchna
sumption with temperature. On the other hand, have been also investigated (Pontin, 1966). Up to
Epp & Lewis (1980), studying a clone of 24 0 C, the relationship between the activity of
B. plicatilis, suggest that there is a plateau in oxy- vibratile flames and temperature is a exponential
gen consumption, within the range of environ- function.
mental temperatures in which the rotifer is most
frequently found. The species studied by Pourriot Temperature: effects on popUlations
(1973b) and Galkovskaya et al. (1987) were
Brachionus calyciflorus and Rinoglena frontalis. The intrinsic rate of increase
QlO was higher for B. calyciflorus (QlO = 2.3-2.4)
and much lower for the cold stenotherm From comparison of the maximum rates of
R.frontalis (QlO = 1.4). Thus, the oxygen con- increase of zooplanktonic organisms (Allan,
sumption for R.frontalis is higher than that of 1976, 1980) it can be inferred that (1) rotifers
B. calyciflorus at low temperatures, but much show amongst the highest r values of all
lower at medium and high temperatures. This zooplankton species, and (2) that the crucial
Table 1. The intrinsec rate of increase (r, days - 1) of some rotifer species growing at different temperature (if not preadapted, the pre-experimental temperature is 00
.j::.
indicated in bracket).
Rotifer Temperature (0C) Reference
species
5 10 15 16 17 19 20 22 24 25 27 30 32 35 36 37 40 43

Brachionus
plicatilis
[I] 0.16 0.38 0.62 King &
[2] 0.12 0.32 0.50 Miracle,
[3] 0.30 0.36 0.44 1980
plicatilis
[4] 0.12 0.39 0.43 0.65 0.74 0.91 0.49 Hirayama &
Kusano,
1972
plicatilis
[5] 0.5 0.7 .l...lQ 0.68 0.4 0.5 Snell, 1986
plicatilis
[6] <0 <0 0.54 0.74 0.98 1.J2 1.07 <0 Pascual &
YUfera,
1983
plicatilis
[7] 0.13 0.25 YUfera &
Pascual,
1980
plicatilis
[8] 0.12 0.53 Nagata, 1985
plicatilis
[9] 0.36 0.57 0.96 Ruttner-
Kolisko,
1972
Brachionus
calyciflorus
[10] 0.23 2.18 0.38 [28°C] Galkovskaya,
0.34 [35°C] 1983
calyciflorus
[11] 1.56 2.83 2.18 Galkovskaya,
[12] 2.18 2.95 1.94 1987
[13] 1.2 1.7
calyciflorus
[14] 0.34 0.48 0.82 Halbach,
0.60 [Tvariable] 1973
calyciflorus
[15] <0 0.1 <0 <0 In: Stark-
[16] 0.2 0.6 1.7 0.9 weather,
[17] 0.3 0.7 1.9 2.5 1987
[18] 0.4 2.2 1.9 3.7
Brachionus
dimidiatus
[19] 0.53 0.70 1.04 Pourriot &
[20] 0.38 0.43 Rougier,
[21] 0.46 0.74 1.22 1975
[22] 0.47 0.52
[23] 0.30 0.42
Euchlanis
dilatata
[24] 0.81 1.78 King, 1972
[25] 0.68 2.10
[26] 0.61 2.13
Asplanchna
sieboldi
[27] 0.60 0.66 1.08 [Saccatte morphotype] Gilbert, 1976
[28] 0.72 0.85 1.22 [Campanulate morphotype]
Brachionus
angularis
[29] -0.005 0.073 0.103 0.352 0.154 Walz,1987
Keratella
cochlearis
[29] 0.004 0.034 0.098 0.082 0.076 Walz,1983

Food conditions and other observations:

[1]-[3] Chlorella (10 6 cell/ml); the three sets of values corresponds to three clones. Clone [3] coming from cold waters than clones [I] and [2].
[4] Chlorella (2.27 x 106 cell/ml).
[5] Chlorella vulgaris (10 JLgCjml).
[6] Nannochloris oculata (45-60 x 106 cell/ml).
[7] Nannochloris (6 x 106 cell/ml) plus Saccharomyces cerevisiae (0.35 gJl per day).
[8] Chlorella saccharophila (1-2 x 106 cell/mI).
[9] Dunaliella sp.
[10] Chlorella (5 x 106 cell/ml).
[1l]-[13] Chlorella: [Il] [12] [l3]
5 x 106 50 X 106 240 X 106 cell/ml.
In [12] the experimental conditions were different that in [10] and [II].
[14] Kirchneriella lunaris (10 6 cell/ml).
[15]-[18] Euglena gracilis, original data of B. Skrdla and P. L. Starkweather:
[15] [16] [17] [18]
0.1 1.0 10 100 JLg dry weight/ml.
[19] Dunaliella salina.
[20] Diogenes sp.
[21] Synechococcus cedrorum.
[22] Synechocystis sp.
[23] Saccharomyces cerevisiae.
[24]-[26] Chlamidomonas reinhardti (0.5 x 106 cell/ml); the different values are average r from groups of clones collected at different times of the year:
[24] [25] [26]
May I June 5 June 12
[27] Paramecium aurelia (in excess); the different values are average r from groups of replicates.
[28] Asplanchna brightwelli (in excess); the different values are average r from groups of replicates.
[29] Stichococcus bacillarius (75 x 106 cell/mI).

00
V'I
86

advantage of one species against another, under as a result of the decomposition of the surplus
a particular set of conditions, may lie in how r food (Stemberger & Gilbert, 1985; Galkovskaya,
responds to anyone of the changing conditions. 1987).
Moreover, r summarizes all life tables parameters, From Table 1, three kinds of response can be
because it combines survival, fecundity and the discriminated. The first 5 species may be con-
timing of development and reproduction. For this sidered warm water adapted; they show their
reason, we will center the review mainly on this highest or maximum r values at temperatures over
parameter, and secondarily on life span as a 27 a C. These maxima range most frequently from
measure of the stability or replacement time of the 1 to 2. Exceptional r values of 2.8-2.9 have also
generations. been found for clones with a high food supply,
Table 1 compiles literature data on intrinsic acclimatized to high temperatures (37 a C).
rates of increase measured at different tempera- Keratella coch/earls can be considered as a cold
tures. Because of the important influence of other water species, the studied clone has a very low
factors on r, mainly genotypes and food quantity maximum r «0.1) localized at 15 ac, which is
and quality, we have only included data sets from the result of a low fecundity and slow develop-
experiments conducted at different temperatures ment, thus, its longer life insures the maintenance
by the same author, so that all other conditions of its population. The studied clone of Brachionus
would remain the same. In addition, it should also angularis has an intermediate position; it reaches
be considered that the different authors estimate a rather low maximum r (0.35) at 20 ac.
r values by different methods; they may be ob- In the first group of species, the averaged rela-
tained from individual, batch or continuous cul- tive increase of r, when the temperature rises from
tures, and by means of different mathematical 20 to 25 ac, is around 10% per degree of tem-
approaches. perature increase, although it varies from 2 to
Very few species of rotifers have been studied 35%, which correspond to ratios r25 oelr2o 0e
to determine the relationship between r and tem- from 1.1 to 2.6 (mean around 1.5). Using the QIO
perature under laboratory controlled conditions. concept for r,
Brachionus plicatilis and B. calyciflorus are the InQIO = 10 (lnr2 -lnr1 )/(T2 - T 1 ),
most studied. Different clones of these species we obtain values from 1.2 to 6.7 (mean around
have maxima of r at different temperatures. This 2.3). Slightly smaller values of these parameters
may be attributed to (l) selection at the environ- were found for increases from 25 to 30 a C. In
mental temperatures of origin, and (2) the inter- most cases within clones, the relationship
action of food with temperature. This interaction between r and temperature is almost linear from
is a complicated balance between the effect of 20 to 30 ac.
temperature on filtering rates, low when the tem- On the other hand, the studied strain of
perature is low, and the accelerated consumption Keratella cochlearis showed much higher relative
of food for growth and reproduction with increas- increases: around 120% from 5 to 10 ac and
ing temperatures. Thus, at low temperatures, if the 36% from 10 to 15 ac, with corresponding ratios
food level is low, the reduction of food intake rate rIO oelr5 °e = 7 (QIO = 50) and r15 oel r1o 0e = 2.8
may not be compensated for by the corresponding (QIO = 8). Brachionus angularls again had an inter-
decrease of the metabolic rate. On the other hand, mediate position.
at high temperatures the accelerated metabolism Although data are very fragmentary, it can be
could only be satisfied if there were high quantities seen that if, for comparison, we plot (Fig. 2) the
of food in the medium. This is clearly illustrated three types of response of r with temperature,
by the data between the interaction of food den- standardizing the data of each clone to their cor-
sity and temperature given by Starkweather responding maximum r value, rmax' and their
(l987, Table 1). However, it has also been optimum temperature (i.e., temperature at which
observed, especially at high temperatures, that an r is maximum, TrmaJ, a family of similar curves is
exaggerated excess of food could be detrimental obtained. They are composite, highly asymmetric,
 87

r
1.0 B. plicatilis D-

·t ····
. . ...
. .... '.'.
'.'.
'.
•
l: 1
2
3
A

rmax
0 ;5
. .. . .. ' D-

• D-
~.. ... ... .. .. ~ .D- • ". D-

..
••
0.0

B
1.0
L-
~.
S. angu/aris 4

0.5 ••
........ ••
0.0 .. ,

C
5
1.0
K. coch/earis
...0 .............. t
... '0 .............. ..
0

0.5
.d

o·
0.0

-15 -10 -5 0 5 10T-Tr max

0
1.5
rmax .1
d-1 6- 2
1.0 .3

0.5
•4
05
0.0

10 15 20 25 30 35 T,OC

Fig. 2. A, B, and C. Relative r values (rlrmax on the ordinate) as a function of the distance from the optimum temperature
(T - Tr=aJ on the abcissa. D. Maximum r(rmax ) and its corresponding temperature (Tr=aJfor the indicated rotifers. (1: Hirayama
& Kusano, 1980; 2: Snell, 1986; 3: Pascual & YUfera, 1983; 4: Walz, 1987; 5: Walz, 1983).
 00
00
Table 2. Duration oflife (in days) of some rotifers growing at different temperatures (LT 50, age for a survival probability of 0.5).

Rotifer Temperature (OC) Reference

species
5 10 14 15 18 19 20 22 24 25 26 27 28 30 32 35 37 38 40

Brachionus
plicatilis
[1] 6.2 6.2 6.8 King &
[2] 6.7 6.0 6.0 Miracle,
[3] 16.5 13.5 11.0 1980
plicatilis
[4] 16.2 19.2 8.0 2.9 [LT 50] Hirayama &
Kusano,
1972
plicatilis
[5] 9.9 6.4 3.3 Shell, 1986
plicatilis
[8] 11.5 26.5 Nagata, 1985
plicatilis
[9] 15 10 7 Ruttner-
Kolisko,
1972
Brachionus
calyciflorus
[12] 2.61 2.20 1.5 Galkovskaya,
[13] 2.72 1.75 1.16 1987
calyciflorus
[14] 17.2 11 5 [LT50] Halbach,
1973
Brachionus
dimidiatus
[19] 9.5 7.7 3.5 Pourriot &
[21] 9 8.9 3.6 Rougier,
1975
Euchlanis
dilatata
[24] 3.3 2.5 King, 1972
[25] 3.1 3.2
[26] 3.4 3.76
Asplanchna
sieboldi
[27] 6.1 4 2.5 [Saccatte morphotype] Gilbert, 1976
[28] 6.9 5.26 2.9 [Campanulate morphotype]
Asplanchna
brightwelli
[30] 4.7 3.2 2.6 Snell &
King, 1977
Brachionus
angularis
[29] 19.4 14.9 4.1 4.5 2.8 [LT 50] Walz, 1987
Keratella
cochlearis
[29] 27.0 21.8 14.9 8.8 6.3 Walz, 1983
Hexarthra
fernica
[31] 12.5 6.3 4 Ruttner-
Kolisko,
1975
Phi/odina
roseola
[32] 35 47 (42) 15 5 3 Lebedeva &
Gerasimova,
1987

Food conditions and other observations:

From [I] to [29], see the footnote of Table I.
[30] Paramecium aurelia.
[31] Chlorella vulgaris (10 6 cell/ml).
[32] Chlorella vulgaris (II x 10 6 cell/ml); the value in parentheses was included by the authors from Ricci, 1976.

00
'-0
90

curves divided by a sharp maximum. The left benthic rotifers (especially bdelloids: Ricci, 1978;
slope is higher than the right slope in the cold- Lebedeva & Gerasimova, 1987, but also Lecane
adapted species and the opposite occurs in warm- tenuiseta, Hummon & Bevelhymer, 1979) have
adapted species. Below this maximum, r is an much longer life spans than planktonic rotifers, as
exponential function of temperature, but in a expected by their occupation of more stable
homogeneous experiment and for a specific range environments. However, there are exceptions,
of temperature this function could be almost such as Euchlanis dilatata which showed the
linear. This response to temperature reflects shortest of all the reviewed life spans. Apart from
metabolic, embryonic and post-embryonic devel- the littoral-benthic species, the mean life span
opment rates, and could be fitted to Belehradek's varies, most frequently, from 3 to 20 days, and the
or Arrhenius' equation. The second part of the ratio between life spans for a 5 0 C increase is
curve is most probably a disequilibrium between most frequently from 1.2 to 2, which indicates
reproduction and survival; reproduction at a too diminutions between 4 and 20% (averages 1.6,
early stage, as a consequence of high temperature, 11%).
could diminish further survival (Snell & King, Within a species, clones show longer life spans
1977). This is most dramatic when the set of according to size and their adaptation to colder
environmental conditions is not quite suitable for temperatures (King & Miracle, 1981). Also the life
the rotifer at the experimental temperature. span of cold-adapted Keratella cochlearis is some-
In most cases, in the first group of warm-water what longer than that of the other studied plank-
species, net reproduction, R o , varies little tonic species.
(Hirayama & Ogawa, 1972; King & Miracle, Life span decreases with a rise of temperature,
1980; Pourriot & Rougier, 1975; Halbach, 1970; the relationship again being similar to the duration
Ruttner-Kolisko, 1972), within a certain tempera- of embryonic and post-embryonic development,
ture range. The effect of temperature on r is intervals between egg depositions, etc. Life span
mainly due to its effect on metabolism and devel- matches the corresponding acceleration of repro-
opment if the rotifer remains under a suitable set ductive effort (descendants per female per day)
of conditions. Ro is only low at the most extreme with temperature, and an inverse relationship
temperatures. exists between the two responses. As Snell &
For species in colder temperatures, Ro is also King (1977) pointed out, reproduction decreases
variable (Walz, 1983, 1987) and has an important the probability offuture survival. However, in this
effect on r. In this case, values of Ro are low owing case the physiological basis for an inverse re-
to the fact that reproduction starts very late, and lationship between reproduction and survival is
then the juvenile period constitutes a high per- the acceleration of development, the organisms
centage of the mean life span (approx. 50% at low more or less attain their potential number of de-
temperatures versus approx. 30% at the optimum scendents during their life time, but over a longer
temperature). Under these conditions the repro- or shorter period with different timings.
ductive period is short. This delay in initial repro-
duction reduces r by increasing the generation
time. At very high temperatures, gross repro- Salinity effects on individuals
duction may not vary, but low survival decreases
Ro as we mentioned before. At extreme values of Total dissolved salts and relative specific ionic
temperature gross reproduction diminishes also. concentrations are important factors condition-
ing rotifer distribution (Ruttner-Kolisko, 1971;
Miracle et al., 1987). Several rotifer genera have
The life span halobiont species living in a very wide range of
salinities. Brachionus, Hexathra, Notholca, Syn-
Table 2 compiles all the data found on mean life chaeta are the most relevant. Moreover a number
span at different temperatures. Most littoral- of essentially freshwater forms can also be salt
 91

tolerant to a certain extent. Nevertheless, studies osmoregulates. It seems however that both
on salinity effects on rotifer individuals or popu- (Asplanchna and Notommata) are incapable of
lations are extremely scarce. hypoosmotic regulation. These rotifers, as well as
Osmotic regulation has seldom been investi- other rotifers, may behave as a group of 'essen-
gated. However, there is good evidence to suggest tially freshwater species that are salt tolerant' as
that highly evolved rotifers can regulate the salt described in Bayly (l972). These animals are
concentration of the pseudocoelomic fluid by hyperosmotic regulators until the salinity of the
means of their flame cells and contractile vesicle. external medium more or less coincides with that
This has been confirmed for Asplachna (Pontin, of the body fluids (Fig. 3, A). At this point, many
1964, 1966; Braun et al., 1966) in which larger species reach their upper limit of tolerance, which
species or forms have a higher number of flame may be widened by acclimatization through
bulbs and in which the vibratile flames show an increases of the isosmotic point. Other species of
activity inversely related to the concentration of this group which are somewhat more salt tolerant,
the medium. In addition, when the animal is behave as osmoconformers for salinities over
placed in a more dilute medium, the pro- isosmoticy. Many rotifer species could be placed
tonephridium responds by sodium conservation, in this group although little or no information
increased water excretion and decreased total exists on this subject.
solute excretion. Kabai & Gilbert (1978) relate the Of the typical halobiont species which can
osmoregulatory capacity of these viviparous tolerate a wide range of salinities, only Brachionus
organisms to the independence of the embryos plicatilis has been studied. According to Epp &
from the external environment, supported by the Winston (l977), B. plicatilis showed a very close
negligible effects that very severe osmotic de- correspondence between internal and external
creases (1O-100 fold dilutions of the normal osmolarity within the range 41 to 957 mOsmoll- 1
medium) had on the response of body wall- with slight but consistent trend towards hyper-
outgrowth to tocopherol, in A. sieboldi. At ex- osmolarity, which was more pronounced as the
treme dilutions, A. sieboldi fecundity was drasti- osmotic pressure of the external medium
cally reduced. There is also evidence, from elec- decreased. This could be represented as Fig. 3
tron microscopy studies of protonephridia (B). B. plicatilis, together with H.fennica and
(Clement, 1968), that Notommata copeus also H. jenkinae were cited by Bayly (l972) as belong-

-0
c:
A
. ...
B
....
.'

-"
0

...ca
-
c:
Q)
(,)
;:,
LL
>a
c:
0
0
"m
0

Concentration of Medium
Fig. 3. Ideal relationships expected in rotifers among those proposed for other invertebrates (Bayly, 1972). (A) Hyperosmotic
regulation till isosmotic point with different acclimations in freshwater species; (B) euryhaIine halobiont species.
92

ing to the group of 'halobiont but entirely non- the external concentration, in osmo-conformers.
marine forms which osmoconform'. He considers They relate this to the assumption of Potts &
that osmotic regulation of extracellular fluids is Perry (1964) that osmo-conformers respond to
not necessary for successful colonization of saline any change in the accustomed osmotic concen-
waters. All the body cells of the animal may be tration by reducing their metabolism. This may be
able to tolerate or regulate their osmolarity instead true for concentrations well below the optimum,
of having a group of specialized cells of a specific such as those described above, when the body
organ regulating the internal fluids. Copepods of cells are beyond their upper limit of tolerance or
these groups, according to Bayly (1972), are capa- regulation. Epp & Winston (1978) also found that
ble of slight hyperosmotic regulation, which can activity and O 2 consumption were less reduced in
be improved with previous acclimatization his- previously acclimatized rotifers, especially at the
tory, but they are incapable of any hyposmotic higher of the above mentioned concentrations.
regulation.
Little or no information exists on osmoregu-
lation in others rotifers, although genera such as Salinity effects on populations
Synchaeta and Notholca are subdivided in an array
of species occupying almost all types of waters, The direct effects of salinity on life history traits
including estuarine and marine environments. are strongly dependent on the species and geno-
There is some evidence that metabolic and type. Raising rotifers at two salinities, 0.2 and
activity responses to salinity correspond to an 0.4 g 1- 1, Lansing (1942) found that the mean life
optimum curve with a more or less extensive span of Rotifer vulgaris (= Rotaria rotatoria)
plateau. Both the shape and position of the maxi- decreases with salinity, without any significant
mum with respect to the salinity axis, is strongly variation in fecundity, while Proales sp. shows a
dependent on genotype. Each species or clone has small life span increase, but a reduction in the
a particular optimum curve and no general trend number of eggs laid per day. According to
of a direct or inverse response to salinity has been Aranovich & Spektorova (1974), Brachionus
found. Work in rotifers on activity response to calyciflorus decreases its survival as the salinity
salinity is restricted to B. plicatilis. An optimum rises from 2 to 109 1- 1, the negative slope being
curve can be extrapolated from the data of progressively steeped. In addition, fecundity de-
Hirayama & Ogawa (1972) on filtration rates, creased with salinity, but levelled off at the highest
with a maximum occurring at moderate salinities salinities assayed. Thus the decrease in growth
(= 12 g 1- 1). Data on oxygen consumption is rate when salinity increases above the optimum,
very fragmentary but Ruttner-Kolisko (1972) is due first to a decrease in fecundity and second
found rates to be low at salinities of 1 g 1- 1 com- to a decrease in survival.
pared to a maximum rate at 17 g 1- 1 and a rate Aranovich & Spektorova (1974) also observed
only slightly below the maximum at 35 g 1- 1. Epp that survival was higher if gradual acclimatization
& Winston (1978) studied rates from 10 to was performed. Such acclimatization seems
100 mOsmol, corresponding approximately to important in the response of rotifers to salinity.
0.35-3.5 g 1- 1 salinity, which are too low for the This has also been pointed out before in relation
normal behavior of B. plicatilis. These authors to osmoregulation, metabolism and activity.
found that rotifers, preadapted to higher salinities Table 3 compiles all the data that we could find
(487 mOsmol), showed a reduction in O 2 con- on parthenogenetic rates of increase in rotifers, at
sumption and activity when transferred to the different salinities. These studies are limited to
above mentioned low salinities, which was more two species of Brachionus, B. plicatilis and
pronounced the lower the salinity. These results B. dimidiatus. The halobiont species B. plicatilis
were interpreted by the authors as being a con- grows in a very broad range of salinities, but data
sequence of tissue hydration and the dilution of on r, for most B. plicatilis clones, can be inter-
active ions and enzymes, which follows the fall of preted as an optimum curve with a maximum or
Table 3. The intrinsec rate of increase (r, days - I )of the indicated species growing at different salinities.
Rotifer S alinity (gil) Reference
species
o 1 2 3 4 5 7 9 10 13 16 17 19 20 21 23 24 25 27 29 30 35 36 38 40 42 44 48 50

Brachionus
plicatilis
acclimatization

3d 0.64 0.82 0.91 0.59 0.42 0.32 0.21 0.34 0.13 Lubzens et al.
8d 0.54 0.34 0.89 0.40 0.42 0.33 0.18 0.01 0.03 1985 [1]
14 d 0.30 0.36 0.80 0.68 0.48 0.42 0.21 0.08 0.05
16 h 0.70 0.57 0.40
without <0 <0 0.95 0.92 0.70 0.35 Pascual & YUfera,
with <0 MQ 0.85 0.75 0.78 0.70 1983 [2]

food level,
x 106 cells/ml

0.1 0.05 <0 <0 <0 Lubzens,

0.5 0.44 0.45 0.44 <0 1981 [3]
1.0 0.66 0.76 0.43 0.35
8.0 0.95 0.81 0.64 0.50

water type

sea water 0.46 0.62 0.98 0.72 0.56 Ruttner-Kolisko,

NaHC0 3 solution 0.88 0.72 0.62 =0 <0 1972 [4]

0.83 0.940.84 0.69 0.5 0.4 from Ito, 1966 [5]

0.85 0.67 0.69 0.66 0.77 0.9* Snell, 1986 [6]

Salinity (gJI)

0.5 1.1 2.2 4.5 9

Brachionus
dimidiatus
0.429 0.417 0.419 0.424 0.417 Pourriot & Rougier,
1975 [7]
Food, temperature conditions, and other observations:
[1] Chlorella stigmatophora (3 x 106 cell/mI); 25°C; pre-experimental salinity, 40 gJI; 16th acclimatization, values were calculated in mass cultures.
[2] Nannochloris oculata (40-60 x 106 cell/ml); pre-experimental salinity, 36 gil; 24°C; growth was not observed at 60 and 80 gil.
[3] Chlorella stigmatophora; 17-23 dc.
[4] Dunaliella sp.
[5] Chlamydomonas sp.; 1.2-2.9 x 106 ceil/mi.
[6] Chlorella vulgaris (10 JlgCfml); 25°C; high sexual reproduction at 20 gJI; *: delayed growth.
[7] Synechococcus cedrorum; 20°C.
\0
W
 94

a plateau located at moderate salinities between 1985). The reproductive response curve of sexual
10-20 g 1-1. Some clones originally from females to an environmental gradient is more
southern latitudes and high salinity show rather constrained and peaked than that of asexual
high rates at high salinities (Pascual & YUfera, females, with maxima located at the optimal con-
1983; Snell, 1986; however, in Snell, 1986, rwas ditions which are probably the same for both
estimated from batch culture growth curves, amictic and mictic growth. However, the sexual .
measured at different time intervals and with a or asexual reproductive responses to temperature ,
high percentage of mixis in the intermediate salini- are very different, because sexual reproduction is
ties ). much diminished at extreme temperatures, and
Increased food increases r and broadens the male rates of increase do not follow the same
range of tolerance, but, in contrast to the response patterns as those of amictic females. By contrast,
to temperature, the maximum is maintained at in a salinity gradient, the maximal sexual repro-
about the same position on the salinity axis ductive effort is more or less coincident with the
(Table 3, Lubzens, 1981). optimum salinity for parthenogenetic growth.
It has been proposed that measures of fitness
of rotifer populations are better based on the pro-
duction of sexual resting eggs, rather than on Interaction between temperature and salinity
parthenogenetic growth rates. Several studies
have been carried out to determine the importance In order to establish the combined effect of tem-
of sexual reproduction in relation to temperature perature and salinity on the intrinsic rate of
(Hino & Hirano, 1984; Snell, 1985) and salinity increase, r, we followed individual cultures of
(Ito, 1960; Lubzens et al., 1980; Lubzens, 1981; three clones of Brachionus plicatilis (CU, sPa,
Hino & Hirano, 1988; Snell, 1985; Lubzens et al., and FCA; Serra & Miracle, 1983, 1985, 1987;

-
.
CU .... SPO FCA
.....I

"
............ 1.5 1
1
f#-.
16 ............
................
......
6.'
--.----
6I ---_
-., /
/
It-- - -- - - ---t

...... ............ , 6 /
I ...... ~
1.0 11 " 1
I 6
1
0.5 I•. ······-t·· ·· ···. .... .
6.:
o
ll. . ... ......• <::> o ".

0.0
9 12 24 9 12 24 9 12 24

Salinity (g/I)

Fig. 4. r values of three clones of Brachionus plicatilis growing at the indicated salinities and at 20°C (dotted lines), 25°C (solid
lines), and 30 °C (dashed lines). Closed symbols represent the values calculated for amictic females; open symbols correspond
to the total number offemales (mictic plus amictic). The triangles indicate which clone shows a maximum r at each temperature-
salinity combination.
 95

Serra, 1987), at three salinities (9, 12, and we have plotted together our results on the
24 g 1- 1; diluted sea water), and at three tempera- response of r to temperature and those calculated
tures (20, 25, and 30 ° C). The r values were calcu- from data of King & Miracle (1981), where also
lated from daily survival and fecundity schedules a bigger clone (SP) was compared with two
from sets of 50 newborn females grown at each of smaller and more thermophilic clones.
the nine experimental conditions and observed The effect of salinity on the intrinsic rate of
until their death. They were derived from parents increase is less pronounced than that of tempera-
coming from log-phase cultures pre-adapted at ture. It is conditioned mainly by clone, but has a
the corresponding experimental combinations. high interaction with temperature. Thus, for clone
The cultures were grown at light and the medium CU the highest value of r has been found at
containing 0.5 x 106 cell ml- 1 of Tetraselmis sp. middle salinity, while SPO shows a decrease of r
was renewed daily. as the salinity rises. The FCA clone shows a trend
Two types of r values were calculated. The first to maintain high r values at high salinity, most
type considered only the amictic females - dis- notably when temperature is high.
regarding the mictic ones - and assumed that all Figure 4 indicates which clone would hypotheti-
their daughters were amictic females. Thus, it esti- cally be dominant in each condition (triangle vs.
mates the intrinsic rate of increase, assuming the circles). According to these results, CU would
absence of mixis. The second type included all have advantage in relatively cold and low salinity
individuals either mictic or amictic. Both r values waters, SPO in warm waters with relatively low
were computed using an integrated solution of the salinities, and FCA in warm waters with high
Lotka's integral equation, assuming a negative salinities. FCA shows also a good response to
exponential interpolation between the observed conditions where salinity and temperature are
survival schedules (Serra, 1987). positively correlated.
Figure 4 shows the effects of genotype, tem- This behaviour of clones is quite closely
perature and salinity on r. The r values calculated adjusted to the one expected if they were adapted
from the subset of amictic females have a clearer to their habitats. Thus, the origin of FCA is a
pattern in relation to these factors; because of coastal marsh with highly variable salinity, reach-
this, we will focus on it. The main feature is an ing more than 35 g 1- 1. We obtained this clone
increase of r as the temperature rises, showing from the Institute of Fisheries Research of
most frequently a linear relationship for our data Castellon, where it was cultured in sea water and
points. These r values, within a temperature, at approximately 26 ° C. SPO was isolated by us
showed great variability due to clone and salinity, from a relatively stable coastal lagoon, in where
but they never overlapped with those at other salinity was 13 g 1- 1 and temperature, 26.6 ° C.
temperatures. The slope ofthe r-temperature rela- Finally, CU was isolated from a endorreic
tionship varies for the different clones. Clone CU, athalasic lagoon, which, at the time of collection,
bigger in size and, according to its origin, adapted had a salinity of 25 g 1- 1 and a temperature of
to lower temperatures shows a much smaller 17°C.
slope than the other two clones (Fig. 5). The same The response of r to some combinations of
comments made before for the duration of temperature and salinity suggests a synergestic
embryonic development (in different species, effect between these parameters, which has been
Pourriot& Rouger, 1975, or clones, Amren, 1964) explored studying their additive interaction. The
or oxygen consumption (Pourriot, 1973b) could interaction (rT . S ' Fig. 6) has been calculated as
be applied here. At low temperature, cold-adapted the difference between the observed value of r, at
genotypes have relatively higher r than more each condition combination, and the expected
thermophilic ones, while the latter have relatively value of r assuming an additive effect of salinity
higher rates at higher temperatures. The same and temperature. The expected r is calculated as
results are obtained in other studies when dif- the addition of the following three terms:
ferent B. plicatilis clones are compared. In Fig. 5 (1) grand mean for the clone, (2) the deviation to
96

this grand mean of the averaged r for the specific perature and salinity is more complex. This clone
temperature, and (3) the deviation from this grand shows a relative advantage at low and middle
mean of the averaged r for the specific salinity. levels of both salinity and temperature, and if only
The results are shown in Fig. 6. these conditions are considered, the pattern of
The most interesting response is that of FCA. CU is similar to that of FCA; the r is above the
It shows a strong interaction between the studied expected value at both low-low and middle-
factors. At the high salinity-high temperature, at middle combinations. Finally, clone SPO pres-
middle salinity-middle temperature, and at low ents a 'smooth relief, pointing out a poor response
salinity-low temperature FCA has a higher r than to specific combinations of temperature and
would be expected without interaction. This can salinity.
be related to the fact that a positive correlation To investigate the underlying causes of the
between temperature and salinity is frequent in obtained variation of r, we have explored the rela-
the natural waters of Mediterranean region, where tionships between r and other life history traits,
in summer high temperature and dryness causes being particularly easy for global - age inde-
an increase of water salinity. Thus, FCA, the pendent -life history traits. An approximation of
clone isolated from a variable marsh, seems to be r could be obtained from the net reproductive rate,
adapted to the fluctuations of natural conditions. R o, and the cohort generation time, Tc ' i. e.,
For clones CU, the interaction between tem- rc = In (Ro)/Tc. In turn, Ro can be decomposed in

1.S
slope
r vs. T

1.0

O.S

SP

0.0..1-1....,Sr"0--.....---2'"T0-0- - - r - - -2'S-0--.....----3'T"0-0

LENGTH, JIm
Fig. 5. The slope of the relationship between r and temperature as a function of the mean body length of four Brachionus plicatilis
strains, measured at two fixed temperatures (data for LA, Me, and SP strains were obtained from King & Miracle, 1980).
 97

CU

-0.1

24
Salinity (gil>

SPO

----- - -- -- --- - --- -

0.1

0.0

-0.1

12
Salinity (g/l)

FCA

Salinity (g//)

Fig. 6. Salinity-temperature interaction, rT . S (days -\, see text) for three clones of Brachionus plicatilis cultured at the indicated
conditions. The bars represent that part of r which is due to interaction, and are solid until the interaction equals zero and empty
when it is positive.
98

CU o •
2.0

1.5
-+.
~. .
......
sPo
FCA o II
•

• •
•••• "11 At)

1.0 ifo.
'.
'11
:.h.
".
0.5 0& 0 0
'. 0
·0·· ............................ c ...

t:.
0.0 +--__._--~-.,....-....,..-.....,--~-~-__._--~-...,.._
o 1 2 3 4 5 6 7 8 9 10

Tc (d)
Fig. 7. Relationship between r and the cohort generation time, Tc (days), in Brachionus plicatilis.

2.0 ...
•• CU o •
... •...
SPO
1.5
FCA o II •
.
•
'
'.
• A
t)

1.0
• II
'.11
..
~
.
'.
'. '. f)
il··· .. ....
'" " .
...•.. 0 0
0.5 A •.... ~ ..•..
o · .. ···0 ...
o c

5 10 15
eo (d)

Fig. 8. Relationship between r and the mean life expectancy, eo, in Brachionus plicatilis. The dotted line has been calculated as
= K, K being the average of the ri . eOi for the 27 experiments, that is K = 6.3. This equation that assumes the potential
r' eo
population growth throughout the life ofan individual is the same for all conditions, i.e., assumes that the intrinsic rate of increase
relative to lifespan is constant.
 99

the product between the duration oflife, measured Our results show that temperature not only
as the mean expectancy oflife, eo, and the average affects the r values, but also the relationships
number of daughters per day: Fd = Ro/eo' between r and the other demographic parameters
As expected, rand Te (Fig. 7) are related by an analyzed. In contrast, genotype and salinity do
asynthotic inverse curve. The values are mainly not cause special relationships between the
arranged according to temperature, and there are studied traits.
not clear distinctions between values belonging to Two important conclusions can be deduced
different clones or salinities. Looking at the slope from our results: (1) At changing temperatures
of this relationship, it is observed that the covaria- the rate of growth and life span or generation time
tion between rand Te is high at 25 0 C, lower at are adaptatively and physiologically adjusted, so
30 0 C, and nearly zero at 20 0 C. the number of descendants per female (R o)
The mean expectancy of life, eo, presents a remains constant. If we plot (Fig. 8) the relation
pattern similar to Tc in its relationship with r between re and Te by inferring rc from the poten-
(Fig. 8). This similarity could be expected as a tial R o (the maximum R o found in our results; i.e.,
consequence of the relationship between Tc and 24) by re = In24/Te , we obtain a line which
the duration of life. matches almost exactly that of expected r assum-
The relationship between r and Fd is also ing constancy in the r relative to life span (dotted
strongly dependent on temperature (Fig. 9), and line in Fig. 8). (2) Both salinity and genotype have
within the same temperature, a linear correlation important effects on fecundity, which can be seen
between r and Fd has been found. When a in the relationship between r and Fd at different
functional regression analysis between these temperatures. On the other hand, their effect on
parameters was performed, lines with similar life span is much less apparent.
slopes were obtained.

20°C
2.0 CU 0
t:.
•
r SPO
(d -1)
1.5
FCA 0 II
•
- - r =0.49 Fd + 0.17
l.c.c.'0.73 lit
1.0

r=0.33 Fd + 0.20
.rp. . ~. I.c.c.: 0.95 **
0.5
O ... 0- .... &
..... r =0.37 Fd + 0.05
I.c.c.: 0.92 .*
O.O~----~--~----~--~----~----~--~----~-----
o 1 2 3 4
Fd (offspring' ~-l'd -1)
Fig. 9. Relationship between r and the reproductive effort, Fd , average offspring per female and day of life} in Brachionus
plicatilis. The relationship is plotted for each temperature, indicating the corresponding regression equation with its significance
(I.c.c., linear correlation coefficient).
100

General considerations can be observed in other invertebrates (Fava

et al., 1983). The genotype is adapted to an
On the basis oflaboratory studies we observe that optimum salinity in which r is maximum. Then, r
temperature has a direct effect on r. This influence decreases as salinity conditions move away from
is mainly derived from the response direct of de- this optimum. The decrease of r is due first to a
velopmental rates or metabolic activities to in- decrease of fertility and, if salinity moves still
creasing temperature, and the resultant r values farther from the optimum, a decrease of survival.
reach a maximum. Beyond this point, r sharply In the response of rotifer rate of increase, the
decreases (Fig. 2). The temperature of the maxi- interaction of temperature-salinity is important.
mum is the upper limit for the normal functioning In our studies of Bracltionus plicatilis clones from
of the animal in the particular set of remaining habitats with greatly flu~tuating temperature and
conditions. The slope of the exponential curve salinity, a positive interaction or beneficial effect
and the temperature at which the maximum oc- is found for the low/low and high/high tem-
curs depend on genotype (Fig. 2), and cold- or perature/salinity combinations within the normal
warm-adapted genotypes are clearly differentiat- limits of those factors in the habitats. Kinne in
ed. Cold-adapted species or clones have in several papers (reviewed in Kinne, 1970;
general, smaller slopes than warm-adapted ones Alderdice, 1972) found similar results for other
(Fig. 2). This confirms the general observation invertebrates and fishes. However, this is not
that cold-adapted species have relatively higher general and Dorgelo (1976) compiles other benefi-
rates of increase at lower temperatures than cial combinations for survival in crustaceans,
warm-adapted ones, and the latter have relatively such as high/low temperature/salinity combina-
higher rates at higher temperatures. It is interest- tions in the tropics. The responses of species or
ing to note also that within species cold-adapted ecotypes seem to match their habitats of origin. In
genotypes have a larger size (Fig. 5). This trend is temperate climates a positive correlation between
coincident with the direct effect of temperature on temperature and salinity is usually found during
the size of genotypically identical individuals, the annual cycle. Temperature-salinity positive
which have a bigger size when reared at low tem- interactions in B. plicatilis ecotypes correspond to
peratures (Serra & Miracle, 1987). Slope and the expected annual covariation of those factors
temperature of the maximum are also more or less in their habitats. In the tropics, where temperature
influenced by environmental factors. Food in par- is more constant and salinity varies with the wet-
ticular determines the temperature limits between dry seasons (corresponding to summer and win-
which the response of r is exponential (Table 1). ter) other factors may be more important. Also in
The effect of temperature on net fecundity, Ro, coastal brackish water environments the inter-
is negligible within normal temperature limits. On action between continental and marine waters
the other hand, life span shows an inverse rela- could determine different salinity-temperature
tionship with temperature, as expected by the relationships.
acceleration of development rates. Thus, tem-
perature (within suitable values for the normal Acknowledgments
functioning of the organisms) influences the tim-
ing, but not fertility (i.e., the number of des- We thank 1. Serrano for her technical assistance
cendents per individual), and an increase of r in laboratory. We also thank Dr. C. Dawson for
results from the acceleration of development. The her language advice in preparing the manuscript.
decrease of r when temperature values move
beyond suitable limits is due first to a decrease in
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© 1989 Kluwer Academic Publishers.

Empirical evidence for a complex diurnal movement in Hexarthra

bulgarica from an oligotrophic high mountain lake (La Caldera, Spain)1

P. Carrillo, L. Cruz-Pizarro & R. Morales-Baquero

Dpto. Biologia Animal, Ecologia y Genetica. Fac. Ciencias. Universidad de Granada, 18701 Granada,
Spain

Key words: Hexarthra bulgarica, diurnal vertical movement, horizontal distribution

Abstract

A detailed 24 hour sampling program has been carried out at 26 depths of 6 stations located along the
two main transects of lake La Caldera. The resultant data has allowed us to derme for H. bulgarica a
general daily trend of movement which couples a typical nocturnal vertical migration with an 'horizontal'
one that is particularly conspicuous at dawn and dusk when the population seemed to moved toward
or away from the shore, respectively.
These results confIrm our previous data and suggest that light is responsible for these complex
movements.

Introduction attempts have been made to obtain fIeld data

coupling vertical and horizontal movements for
Data on rotifer spatial distribution and movement an entire day.
of populations come, basically, from two main
approaches. Diurnal vertical migration is a com-
Material and methods
mon phenomenon, but is variable and far less
important than in crustaceans in terms of ampli- Selected site and species
tude and velocity of migration. Horizontal dis-
tribution also varies and the resultant patchiness La Caldera is an oligotrophic high mountain lake
may reflect shore avoidance processes. in southern Spain. It is a typical 'winter-kill' lake
Although the underlying mechanisms involved of glacial origin which remains frozen for 8 to 9
in both phenomena as well as their adaptive value months a year. Martinez (1975, 1980) gives data
still remain uncertain, light has an important role on its morphometric and physico-chemical char-
as a releasing (and directing) stimulus (Ringel- acteristics and the papers from Martinez (1977),
berg, 1980; Preissler, 1977a, b). Our previous Cruz-Pizarro (1978, 1981), Morales (1985, 1988)
data (Cruz-Pizarro, 1978, 1981) suggestthat these and Sanchez-Castillo et al. (1988) consider dif-
movements are important, thus we have measured ferent aspects of its plankton composition and
a 24-hour cycle of movement for H. bulgarica in ecology. Of particular note is its small size, the
lake La Caldera. As far as we know, no previous absence of littoral rooted vegetation and of visible
inlet and outlets, and the lack of a true thermo-
1 Research supported by CAICYT Project No. 3069/83. cline and oxycline.
104

N
+
o 50m

Fig. 1. Bathymetric map of lake La Caldera and location of sampling stations.

Hexarthra bulgarica, is the dominant rotifer in 4 % formaldehyde. Counts were made with an
species in the lake, both in number of individuals inverted microscope at 100 x magnification. Sur-
and in biomass. Data from Cruz-Pizarro (1978, face light measurements (Luxmeter) were
1981) reveal a defmite nocturnal vertical migra- recorded every two hours at the B station.
tion and suggest a simultaneous horizontal move-
ment of the population.
Results

Sampling and counting Data obtained from all stations and depths for the
daily cycle are shown in Fig. 2. At 15 hours, the
To entend and explain these previous observa- first sampling carried out, individuals are located,
tions, we carried out in August 1986, during the predominantly, at the deepest layers of stations C
peak of population density for the species, a 24 h and D and, to a lesser extent, at B and E also. The
sampling program which included different relative low densities are noteworthy.
sampling stations and depths along our two main Results from the 18 hour sampling suggest an
transects of the lake (Fig. 1). offshore movement has taken place giving rise to
The samples were taken from two small boats the great differences observed between pelagic
working simultaneously (starting from different and littoral catches. This pointed movement
points) so that each sampling series required less seems to continue such that at 21 hour most
than 25 minutes. A double Van Dorn sampler of individuals are found close to the eastern shore
8 I capacity was used. Plankton was removed which remains longer in the light.
with a 45 J1.m mesh net and immediately preserved The situation at midnight reveals a change in
 105

15H 3H
15 15

3 3

18H 30 8H

15 15

3
3
M

0
....
...
'e

-
0
C)

30
21H
30
10H

15 15

3 3

45 45

24H 12 H
30

15 15

B E B E

Fig. 2. Population density along the daily cycle at the different sampling stations and depths.
106

DUSK DAWN DUSK DAWN

A A ®
D
D

C
C

B B

E
E

F
F

D D

C C

B B

E E

F F

I i i
24
, , ,
10
i
12
15 18 21 3 8 15 3 10 12
HOURS

Fig. 3. Diel changes in density distribution along the sampling points at four selected depths (I: surface; 2: one-third maximum
depth; 3: two-third maximum depth; 4: bottom). Shading intensity related to population abundance (orgjm3 ) . Dashed lines
indicate secchi disk visibility (m) at midday on B station.

the direction of the movement and the population mum depth and bottom (irrespective of each par-
is located mainly at the surface layers of the ticular station depth) to emphasize positions of
limnetic zone. An increase in both sample density the population peak densities during the daily
and heterogeneity of vertical distribution are note- cycle.
worthy. At the 8 hour sampling an apparent Time variations in the pattern of vertical and
movement occurs into the F station and the popu- horizontal aggregation of individuals have been
lation has a more homogeneous distribution in quantified by difference of the 'patchiness index',
space. At 10 hour,just after dawn, individuals are P.I. = X/X, following George & Edwards (1976).
concentrated at the southern shore near the Figure 4a shows P.I. values at the different sam-
bottom. This situation continues until midday pling points during the daily cycle and fig. 4b
when, again, differences between pelagic and lit- shows the corresponding P.I.' s of the four selected
toral catches are quite evident. 'horizontal' planes.
In Fig. 3, we have selected four depths: sur- Deviations from a random distribution are the
face; one-third maximum depth; two-thirds maxi- lowest during nighttime, particularly near the
 107

o 15
* 18
.21 1/1 ...
.. 24 :l

ell
•
A 8
0
3 .r.
ell {< 10
Q) 2.0
r:::: 012
..c
...ca
U

Q.

ca
1.5 0
......
U

Q)

>
1.0

A 0 C B E F
Samp l ing points

®
ell 3.0
CII
Q)
r::::
..c 2 .5 Bottom
...cau
Q.
2D
%max. depth
...
ca
r::::
1.5
0 Surface
N
...0 % max · depth
:I: to

15 18 21 24 3 8 10 12

HOURS

Fig. 4. Time variations of vertical (A) and horizontal (B) patchiness index (see text) at the different sampling stations and selected
depths.
108

bottom, whereas the greatest patchiness measures References

were obtained around dusk and dawn in the upper
and deepest layers, respectively. The extremely Cruz-Pizarro, L., 1978. Comparative vertical zonation and
diurnal migration among Crustacea and Rotifera in the
high aggregation value of the population at 21
small high mountain lake La Caldera (Granada, Spain).
hour at maximum depths is noteworthy. Verh. int. Ver. Limno!. 20: 1026-1032.
Cruz-Pizarro, L., 1981. Estudio de la comunidad zoo-
planct6nica de un lago de alta montana (La Caldera,
Discussion Sierra Nevada, Granada). Ph. D. Thesis. Univ. Granada.
186 pp.
George, D. G. & R W. Edwards, 1976. The effect of wind on
Approving some limitations inherent in the lack of the distribution of chlorophyll a and crustacean plankton
sample replicates, these results depict a general in a shallow eutrophic reservoir. 1. appJ: Eco!. 13: 667-690.
daily trend of movement for H. bulgarica. The Martinez, R, 1975. First report on the limnology ofthe alpine
vertical component is a typical nocturnal migra- lake La Caldera, in the Penibetic Mountains (Sierra
tion with an upward movement after dusk and a Nevada, Granada, Spain). Verh. int. Ver. Limno!. 19:
1133-1139.
downward one just before dawn gives rise, during Martinez, R, 1977. Phytoplankton species, biomass and
nighttime, to the highest values of vertical P.I. at diversity in lake La Caldera (Sierra Nevada, Granada,
pelagic stations. This seems to be coupled with an Spain). Acta hydrobio!. 19: 95-107.
'horizontal' migration (highest values of horizon- Martinez, R, 1980. Seasonal variations of phytoplankton
tal P.I.) that is, likewise, particularly conspicuous biomass and photosynthesis in the high mountain lake La
Caldera (Sierra Nevada, Spain). Development in Hydro-
at dawn and dusk. biology 3: 111-119.
Light changes should be able to explain the Morales-Baquero, R, 1985. Estudio de las comunidades de
adaptive value of these movements as, particu- Rotiferos Monogonontes de las lagunas de alta montana
larly for vertical migration, other major proposed de Sierra Nevada. Ph. D. Thesis. Univ. Granada. 296 pp.
hypothesis (energetic and demographic, predator- Morales-Baquero, R, 1988. Body size variability of
Euchlanis dilatata Ehrenberg in high mountain lakes of
avoidance, increased chance for mating and gen- Sierra Nevada (Spain). Arch. Hydrobio!. 112: 597-609.
etic exchange) are unlikely to hold for this, basi- Preissler, K., 1977a. Do Rotifers show 'Avoidance of the
cally parthenogenetic species, in a virtually iso- shore'? Oecologia (Berl.). 27: 253-260.
thermic environment where visually orienting Preissler, K., 1977b. Horizontal distribution and 'avoidance
planktivorous are absent. of shore' by rotifers. Arch. Hydrobiol. Beih. 8: 43-46.
Ringelberg, J., 1980. Introductory remarks: causal and teleo-
Further information is needed on light intensity logical aspect of diurnal vertical migration. In W. C.
and angular distribution in the shore region of the Kerfoot (ed.), Evolution and Ecology of zooplankton Com-
lake. Subsequent studies should also consider munities. The University Press of New England, Hanover
advective factors (wind-induced water move- (N.H.); Lond.: 65-68.
ments) in patch formation and evaluate down- Sanchez-Castillo, P., L. Cruz-Pizarro & P. Carrillo, 1989.
Caracterizaci6n del fitoplancton de las lagunas de alta
ward movements over a slopping bottom.
montana de Sierra Nevada (Granada, Espana) en relaci6n
con las caracteristicas fisico-quimicas del medio.
Limnetica 5: 37-50.
Hydrobiologia 186/187: 109-117, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 109
© 1989 Kluwer Academic Publishers.

Morphological variation in Kellicottia longispina

W. T. Edmondson & Arni H. Litt

Department of Zoology, NJ-15, University of Washington, Seattle, Washington 98195 U.S.A.

Key words: zooplankton, Rotifera, biogeography, arctic limnology

Abstract

The lengths of the body, the posterior spine and the three longest anterior spines were measured for 25
specimens of Kellicottia longispina from each of the eight lakes distributed from Imikpuk at Point Barrow,
Alaska (latitude 71 ° 15') to Lake Washington (latitude 47° 38'). Collections were available for more than
two dates from six of the lakes. Temperature ranged from 1.2° to 18°C. Mean lengths and ratios were
examined in relation to latitude and temperature. Each population differed from the others in some aspect
of absolute size, variability, or shape as expressed by the ratios of the dimensions. The population from
Point Barrow is similar but not identical to Olofsson's var. heterospina.

Introduction species described as Ertemia may have been

rhizopod shells into which a rotifer had crept (see
Kellicottia longispina is a widespread, common Hudson & Gosse).
planktonic rotifer that can become very abundant. Although K. longispina is easily recognized,
It is strikingly different in appearance from all even causal examination shows that there are dif-
other rotifers, even from its only congener, ferences in size and shape among different popu-
K. bostoniensis. Interestingly, both species were lations. A form from arctic Norway is so different
described from North America. Kellicottia bos- that it was given the varietal name heterospina by
toniensis was confined to that continent until Olofsson (1917) (Fig. I-c). A similar form was
about 1943 when it was first noticed in Sweden relatively abundant in samples collected by the
(Arnemo et al., 1968). It subsequently spread to late G.W. Comita from Imikpuk (Freshwater
Finland (Dr. Pertti Eloranta, personal communi- Lake) near Point Barrow, Alaska in 1951. Its
cation). occurrence in this small arctic lake raised
Kellicottia /ongispina was found in 'Niagara questions about its distribution in the Arctic and
water' near Buffalo, N.Y. and assigned to Anuraea the occurrence of forms resembling the ones in
by Kellicott in 1879 (Fig. I-a). Following Hudson temperate regions. Therefore we examined col-
& Gosse (1886) it was known as Notholca until lections from other lakes in northern Alaska and
1938 when Ahlstrom created Kellicottia to recog- material from five lakes at lower latitudes (Fig. 2,
nize its unusual structure. The species was noticed Table 1). We measured major anatomical struc-
in England and Europe soon after its discovery in tures in an attempt to define objectively what
North America. Hood's Ertemias tetrathrix (1888) made the populations look so different (Fig. I-d,
probably was this species (Fig. I-b). Some Fig. 3, Table 2). We also examined correlations
110

c
a b c d

Fig. 1. Four drawings of Kellicottia !ongispina. a. Anuraea !ongispina, from original drawing by Kellicott (1879). b. Ertemias
tetrathrix from Hood (1888). c.K. !ongispina var. heterospina after Olofsson (1917). d. Diagram indicating symbols used throughout
the text.

among the various dimensions and between mor- Edmondson, 1955). For reasons not understood,
phometric features and the location and environ- Kellicottia was extremely scarce in that year. In
mental conditions of the lakes. addition to our own material from Bare Lake,
Hall Lake and Lake Washington, we were kindly
provided with material from Great Slave by the
Materials and methods late Donald S. Rawson and from Lake Pend
Oreille by Raymond A. Stross.
Collections from two arctic lakes in addition to The three arctic lakes are small tundra ponds.
Imikpuk were made by Comita in 1951. Imikpuk Great Slave Lake is very large and oligotrophic
was studied more in detail in 1952 (Comita, 1956; (Rawson, 1956). Bare Lake is small and oligo-
 111

,,
,,
, ,,
"
""
-- -- --
..........
..........

GREAT SLAVE L~

•PEND OREILLE LAKE

Fig. 2. Location map.

trophic, but was fertilized on June 11, 1952 Measurements were made of the three longest
(Nelson & Edmondson, 1955). Lake Pend Oreille anterior spines (A,D,E), the posterior spine (C),
is large and mesotrophic. Hall Lake is small and and the length (B) and width (W) of the body
slightly dystrophic. Lake Washington is moder- (Fig. I-d). After a preliminary graphical analysis
ately large and mesotrophic (Edmondson, 1963). of the dimensions and their ratios to each other,
Twenty-five specimens from each collection we selected dimensions or proportions that
were measured under light compression. showed distinct relation to location or strong cor-
112

Table 1. Data on collections used for measurements of Kellicottia /ongispina.

Name Date Latitude Temperature

°C

1 lmikpuk 09 August 1951 71 ° 20' 12.5

2Imikpuk 18 August 1951 6.6
3 Imikpuk 06 September 1951 4.2
4Imikpuk 18 September 1951 1.2
5Ikroik 01 August 1951 71 ° 20' 13.0
6 Ikroik 30 August 1951 5.6
7 Paddle Lake 22 August 1951 71 ° 15' 9.2
8 Great Slave Lake 03 July 1949 6ZO 00' 4.0
9 Great Slave Lake 11 August 1949 12.0
10 Great Slave Lake 22 August 1947 8.8
11 Great Slave Lake 21 September 1947 7.0
12 Bare Lake 03 June 1952 58° 00' 8.0
13 Bare Lake 17 June 1952 11.1
14 Bare Lake 25 June 1952 14.5
15 Pend Oreille Lake 05 April 1953 48° 25' 4.3
16 Pend Oreille Lake 20 June 1953 13.8
17 Pend Oreille Lake 06 July 1953 18.2
18 Pend Oreille Lake 26 October 1953 12.0
19 Hall Lake 02 January 1952 47° 50' 4.4
20 Hall Lake 24 April 1952 11.5
21 Hall Lake 07 July 1952 14.2
22 Lake Washington 23 April 1980 47° 38' 9.8
23 Lake Washington 27 May 1981 14.2
24 Lake Washington 04 June 1980 13.6
25 Lake Washington 09 October 1981 13.7

relations with other features for further statistical the animals varied greatly among the lakes, with
analysis by standard tests (Table 2). We examined the Imikpuk animals being outstanding (Fig. 4-T).
length-frequency plots for all of the sets of Populations from some of the other lakes show
measurem~nts. more variation with temperature than those from
In the following section a selection ofthe results Imikpuk. There is a clear tendency for the total
is shown in graphical form, with a brief summary, length (T) to decrease in lakes to the south. Great
to substitute for lengthy verbal descriptions. Slave and Paddle are out of line, but their means
have large standard deviations and the range of
Great Slave overlaps that of all the lakes to the
Results south of it. Rotifers are larger at higher tempera-
ture in three of the lakes and smaller in two, but
The clearest generalization that we can make is only the greatest differences in size are significant.
that the population of each lake was different The variation among lakes is more easily seen
from all the others in at least one feature. Some in plots of the range of all the samples for each
of the features varied among samples taken from lake (Fig. 4). Of all the features, the length of spine
a lake at different times. We looked to see whether A shows the clearest geographical pattern
the variation was related to time or temperature, (Fig. 4-A). The population of Ikroik is out of line
as might be expected of cyclomorphosis. and shows few similarities to that ofthe other two
The following comments call attention to facts arctic lakes.
shown by Fig. 4 and Table 2. The total length of Differences in the proportions of spines and
 113

IMIKPUK IKROIK PADDLE LAKE BARE LAKE

,",

~~---------------

I
'...
h.;" t· 4, \
f ~}
....
!

GREAT SLAVE LAKE PEND OREILLE LAKE HALL LAKE LAKE WASHINGTON

Fig. 3. Photograph of representative individuals from each lake.

body determine the different appearances of the is consistent with the shorter posterior spine.
population. The way the total length is shared by Within its range of variation in each lake, A : C is
the spines and body is shown by ratios A : T, not correlated with body size. The ratio A : B has
B : T and C : T. The ratio of A : C relates the two a clear relation to latitude as did the absolute size
most conspicuous features. In four of the eight of spine A, but the ranges of the ratio are propor~
lakes values of A: C vary from 1.0 to 1.4. In tionately much larger. Body length does not vary
Imikpuk they are nearly equal, varying from 1.0 to as much among the lakes as the spines, and its
1.1, but in Great Salve the ratio is 1.7 to 1.9, as variations have less effect on appearance.
Table 2. Upper. Mean values and ratios of some ofthe features measured. See Fig. I-d for symbols. T is totallength (A + B + C).
W is width of body at widest point. Note: The ratios are means of ratios of individual measurements; the values may be slightly
different from the ratios of the tabulated mean lengths. Lower. Standard deviations of the means, n = 25. All measurements are
in micrometers. Smallest unit of measure for lakes 1 to 7 was 121-1, for lake 8 was 41-1.

OBS Lake TAB C D E W A:T B:T C:T A:B A:C A:D D:E C:B W:B

Imikpuk 964 427 134 402 170 133 64 0.44 0.14 0.42 3.20 1.06 2.51 1.29 3.04 0.47
2 Imikpuk 954 433 127 394 176 144 60 0.45 0.13 0.41 3.40 1.10 2.46 1.23 3.09 0.48
J Imikpuk 971 444 140 ~M 179 144 64 0.46 0.15 0.40 3.17 1.15 2.48 1.25 2.73 0.45
4 Imikpuk 940 422 143 374 169 143 65 0.45 0.15 0.40 3.00 1.15 2.57 1.22 2.63 0.45
5 Ikroik 749 347 125 277 133 88 67 0.46 0.17 0.37 2.80 1.25 2.66 1.53 2.23 0.54
6 Ikroik 712 343 122 246 150 100 64 0.48 0.17 0.35 2.80 1.40 2.30 1.53 2.01 0.52
7 Paddle 815 364 124 328 150 113 61 0.45 0.15 0.40 2.96 1.13 2.60 1.34 2.66 0.50
8 Great Slave 673 336 120 217 137 89 55 0.50 0.18 0.32 2.82 1.58 2.47 1.57 1.81 0.46
9 Great Slave 724 346 124 254 173 143 56 0.48 0.17 0.35 2.82 1.41 2.00 1.22 2.07 0.46
10 Great Slave 667 338 125 204 172 133 59 0.51 0.19 0.30 2.73 1.72 1.98 1.30 1.64 0.47
11 Great Slave 622 329 120 173 166 130 59 0.53 0.19 0.28 2.75 1.95 1.99 1.29 1.44 0.49
12 Bare 718 342 127 248 112 48 71 0.48 0.18 0.35 2.72 1.38 3.12 2.39 1.97 0.56
13 Bare 763 350 134 278 138 70 66 0.48 0.17 0.35 2.90 1.38 2.98 2.29 2.11 0.53
14 Bare 719 347 120 252 118 53 64 0.46 0.18 0.36 2.61 1.26 2.55 2.02 2.07 0.49
15 Pend Oreille 706 331 114 260 162 118 62 0.47 0.16 0.37 2.92 1.27 2.06 1.39 2.30 0.55
16 Pend Oreille 686 317 127 242 170 132 64 0.46 0.19 0.35 2.49 1.31 1.86 1.30 1.90 0.50
17 Pend Oreille 655 301 122 232 168 140 60 0.46 0.19 0.35 2.47 1.31 1.81 1.20 1.89 0.49
18 Pend Oreille 668 302 131 235 167 133 60 0.45 0.20 0.35 2.32 1.29 1.82 1.25 1.80 0.46
19 Hall 638 298 108 233 137 97 54 0.47 0.17 0.36 2.76 1.28 2.19 1.42 2.15 0.50
20 Hall 661 308 115 238 150 108 54 0.47 0.17 0.36 2.69 1.30 2.06 1.41 2.07 0.47
21 Hall 594 278 118 198 145 122 54 0.47 0.20 0.33 2.36 1.41 1.92 1.19 1.68 0.46
22 Washington 653 290 122 242 160 115 60 0.44 0.19 0.37 2.38 1.20 1.83 1.38 1.98 0.50
23 Washington 640 296 126 219 166 131 63 0.46 0.20 0.34 2.36 1.35 1.79 1.27 1.75 0.50
24 Washington 612 274 122 216 157 126 55 0.45 0.20 0.35 2.25 1.27 1.75 1.25 1.77 0.45
25 Washington 622 262 123 237 157 127 57 0.42 0.20 0.38 2.13 1.12 1.68 1.23 1.92 0.46

OBS Lake TAB C D E W A:T B:T C:T A:B A:C A:D D:E C:B W:B

1 Imikpuk 37 12 8 25 10 12 5 0.011 0.007 0.013 0.173 0.062 0.127 0.140 0.209 0.051
2 Imikpuk 39 18 8 22 9 10 2 0.012 0.007 0.002 0.225 0.056 0.125 0.102 0.195 0.039
3 Imikpuk 41 22 5 28 9 9 6 0.016 0.008 0.017 0.220 0.086 0.147 0.089 0.213 0.040
4 Imikpuk 58 32 10 35 20 21 5 0.023 0.011 0.022 0.282 0.119 0.429 0.261 0.267 0.035
5 Ikroik 41 21 8 19 21 14 6 0.010 0.011 0.010 0.234 0.052 0.386 0.201 0.193 0.051
6 Ikroik 31 21 8 20 13 15 5 0.020 0.014 0.G18 0.301 0.117 0.206 0.167 0.211 0.058
7 Paddle 87 36 9 54 19 21 4 0.023 0.014 0.030 0.267 0.137 0.294 0.118 0.421 0.053
8 Great Slave 69 35 8 40 17 10 7 0.025 0.021 0.031 0.350 0.221 0.296 0.243 0.352 0.061
9 Great Slave 81 30 7 58 11 12 5 0.029 0.022 0.043 0.310 0.248 0.158 0.075 0.470 0.050
10 Great Slave 84 41 13 49 20 19 3 0.034 0.022 0.040 0.370 0.316 0.234 0.137 0.360 0.004
11 Great Slave 47 25 8 31 18 13 4 0.026 0.G18 0.032 0.282 0.292 0.167 0.091 0.270 0.045
12 Bare 16 12 11 II 13 8 3 0.157 0.G15 0.012 0.308 0.078 0.370 0.410 0.208 0.055
13 Bare 29 13 7 17 12 10 4 0.011 0.010 0.014 0.199 0.077 0.313 0.573 0.186 0.047
14 Bare 23 10 7 18 14 13 4 0.013 0.009 0.G15 0.166 0.087 0.269 0.304 0.176 0.033
15 Pend Oreille 36 19 7 20 IS 12 4 0.016 0.010 0.017 0.233 0.097 0.164 0.104 0.218 0.051
16 Pend Oreille 51 25 9 25 14 11 4 0.018 0.012 0.016 0.204 0.105 0.111 0.103 0.153 0.056
17 Pend Oreille 28 15 7 19 10 13 4 0.016 0.013 0.G18 0.199 0.106 0.131 0.089 0.191 0.040
18 Pend Oreille 41 21 8 22 10 9 2 0.013 0.016 0.016 0.202 0.078 0.136 0.075 0.192 0.030
19 Hall 17 10 5 14 15 13 5 0.G15 0.008 0.G15 0.173 0.089 0.210 0.107 0.144 0.039
20 Hall 30 15 8 15 8 11 5 0.009 0.010 0.013 0.168 0.066 0.127 0.133 0.175 0.052
21 Hall 24 14 6 16 II 12 5 0.015 0.012 0.G18 0.166 0.109 0.118 0.093 0.178 0.050
22 Washington 38 26 5 17 20 11 4 0.022 0.009 0.019 0.195 0.113 0.273 0.193 0.137 0.037
23 Washington 28 19 4 12 12 7 3 0.016 0.008 0.013 0.141 0.092 0.088 0.087 0.096 0.024
24 Washington 34 22 8 17 14 13 4 0.017 0.G15 0.G18 0.225 0.100 0.141 0.099 0.200 0.038
25 Washington 38 18 3 27 14 12 4 0.G18 0.014 0.023 0.168 0.111 0.141 0.081 0.227 0.038
 1000,---------------____________________,
P T 3 .1 A:D 2 .5 D:E
...------------1
.3

..-- ~"""""",5
~~~
~...~..~
...........
~8 • ••••••••• 6
•
; •
7
550~ITI---.----~--~--,_--_.--~--_,----r_--._~ 1.6 -'-----,----,---_r---,----,----,_--,----,-.J 1.0 -'---.---,----r---"'T----,----,...---.----,-.J
o 10 18C)c

475,-----------------------------------, 1 .9 -r---------~-----------,

~ A 3 .4 A:B A:C

•
••
•

250~--,_--_r--_.----._--_r--~----,_--~
2.0 ...L.._-._---.__.-_--.__.-_...,.__,-_-.-' 1.0 -'-----.---,----r---.,.---.----,,----,----..-'

0.53~r------------------ . .---------------------, 0.21 , _ - -- - - - - - - - - - - - - - - - - ,

A:T B:T 0 .41 C:T

0 .42 -'---,----.---.,.---,----,,----,_--,----r-.J 0 . 12 -'1- - ,,..---.---,--,----.--,--...,.--,,; 0 .27 ~

2 3 4 5 8 7 8 2 3 4 5 8 8 2 3 4 5 8 7

Fig. 4. Mean lengths of all features measured and a selection of ratios for all lakes, arranged by latitude and lake number. See Fig. I-d for symbols. Large dots represoent ......
......
overlapping measurements. Upper left. Total length (T) of all samples plotted against temperature. Numbers on curves represent lakes: I-Imikpuk, 2-Ikroik, 3-Padodle, Vl

4-Great Slave, 5-Bare, 6-Pend Oreille, 7-Hall, 8-Washington. Other panels show the individual values arranged in a vertical series for each lake. Lake are numbered
at the bottom, arranged in order of latitude.
116

Two general patterns emerge, one a trend from dates in some of the lakes, but most of the dif-
north to south, the other humped or depressed in ferences are small and statistically insignificant
the middle because of the special features of the (t-test, (J. = 0.05). We examined the samples that
Great Slave population. showed the biggest differences by plotting length
Another feature that catches the eye is the posi- against frequency. The relative frequency of dif-
tion of the body along the total length, determined ferent stages is affected by changes in birth and
by the length of the two spines A and C. Great death rates. An increase in mean value can be
Slave specimens have the body further back on caused either by an increase in the frequency of
the axis because the C spine is shorter than in the largest size or by a decrease in the frequency
other lakes (Fig. 4-C: T). One's subjective of the smallest, or both (Fig. 5). However, we may
reaction to the appearance of the animals is be seeing simply the replacement of a cohort of
affected by the shape of the body as well as by the large animals by a cohort of different size.
proportions of the spines. The shape of the body Any time the lengths of spines of a rotifer show
varies considerably, as measured by the ratio systematic changes, we must consider the possi-
W : B, but not with a very clear distributional bility of predation and chemomorphosis, as with
pattern (see Table 2). The appearance of the body Braehionus and Asp/anehna. As far as we know
is affected by the fact that in some, as in Imikpuk, each of these lakes has a tactile invertebrate
the maximum width of the body is well behind the predator. In addition to the predatory Limno-
front of the lorica, while in others it is at the front ea/anus, Imikpuk had two species of Anostraca
end. Our measurement system did not include which are capable of swallowing the big
that aspect of shape. Kellicottia.
We looked for evidence of cyclomorphosis or
other form changes in graphs of dimensions
plotted against temperature and time. Some of the Conclusions
dimensions and ratios change between sampling
The Imikpuk form of Kellieottia /ongispina is not
identical with Olofsson's var. heterospina which
was characterized on the basis oflarge size, ratios
A: D > 3 and D: E > 1.5. The Imikpuk form
agrees on the ftrst point, although it is 10% larger,
but ratio A: D is only 2.5 and D: E is 1.25. The
other two arctic popUlations agree even less with
°
50% the description of heterospina. It appears that the
Kellieottia fauna of the Arctic is diverse morpho-
logically (see also Turner, 1987).
The question remains whether Kellieottia exhi-
o bits cyclomorphosis or chemomorphosis. Our
samples show that some populations vary con-
50%
siderably over time, but the samples were not
taken frequently enough to define the time course
of changes. These problems are well worth
pursuing in lakes that reliably produce large, varia-
o ble populations. The study of chemomorphosis
600 700 800 would have two components. First, a fteld study
Fig. 5. Length-frequency histograms of total length of to identify lakes with different tactile predators.
K. iongispina in Bare Lake. Above - 03 June 1952, Middle - Then one would do laboratory experiments.
17 June 1952, Below - 25 June 1952. These will be difficult because Kellieottia is hard
 117

to culture, largely because it sicks to the surface References

of the water so easily. Possibly some success
could be achieved by tethering individuals by the Arnemo, R., B. Berzins, B. Gronberg & I. Mellgren, 1968.
posterior spine to the bottom of a culture dish on Dispersal in Swedish waters of Kellicottia bostoniensis
(Rousselet) (Rotatoria). Oikos 19: 351-358.
a microblob of petroleum jelly. We know that the Comita, G. W., 1956. A study of a calanoid copepod popula-
spines are tightly curled around the body in the tion in an arctic lake. Ecology 37: 576-595.
egg and rapidly straighten after hatching, but Edmondson, W. T., 1955. Seasonal life history of Daphnia in
nothing seems to be known about the subsequent an arctic lake. Ecology 36: 439-455.
Edmondson, W. T., 1963. Pacific Coast and Great Basin.
rate and pattern of growth. It is surprising that an
pp.371-392 in D. G. Frey, ed. Limnology in North
animal as conspicuous as Kellicottia /ongispina is America. University of Wisconsin Press.
not better known. Hood, J., 1888. Chats about rotifers (Ertemias tetrathrix).
Hardwickes' Science Gossip. 24: 27-28.
Hudson, C. T & P. H. Gosse, 1886. The Rotifera; or wheel-
animalcules. Longmans, Green & Co. London.
Acknowledgements
Kellicott, D. S., 1879. A newrotifer. Am. 1. Microsc. Pop. Sci.
4: 19-20.
The work at Point Barrow was supported by the Nelson, P. R. & W. T. Edmondson, 1955. Limnological
United States Office of Naval Research. Prepa- effects of fertilizing Bare Lake, Alaska. U.S. Fish and
ration of this paper was made possible by the Wildlife Service, Fish. Bull. 102: 413-436.
Olofsson, 0., 1917. Siisswasser-Entomostraken und Rota-
Andrew W. Mellon Foundation.
torien von der Murmankuste und aus den nordlichsten
Norwegen. Zoo!. Bidrag Uppsala, 5: 259-294.
Pejler, B., 1977. On the global distribution of the family
Brachionidae (Rotatoria). Arch. Hydrobio!./Supp!' 53. 2:
255-306.
Rawson, D. S., 1956. The net plankton of Great Slave Lake.
1. Fish Res. Bd Can. 13: 53-127.
Turner, P. N., 1987. Some rotifers from Alaska, U.S.A. with
a note to researchers. Microscopy 35: 541-548.
Hydrobiologia 186/187: 119-128, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 119
© 1989 Kluwer Academic Publishers.

Morphological structure and functional patterns of Keratella cochlearis

(Gosse) populations in stratified lakes

Galina A. Galkovskaya & Inessa F. Mityanina

Institute of Zoology, Byelorussian Academy of Sciences, Academycheskaya 27, Minsk 220072, USSR

Key words: Keratella cochlearis, diurnal vertical distribution, morphological structure, population and
energetic patterns

Abstract

Vertical distribution of the rotifer Keratella cochlearis in stratified water columns of mesotrophic and
eutrophic lakes during summer stagnation has been studied. Coexisting morphs K. cochlearis hispida
(Lauterborn, 1898), K. c. tecta (Gosse, 1851) and K. c. cochlearis (Gosse, 1851) inhabit different layers
in the water column and are vertically subdivided. The distribution ofmorph abundance and reproductive
potential indicate that substitution of morphs within the vertical water column may be due to trophic
conditions. The maximum population productivity is observed at the epi-metalimnion border. The
maximum density zone lies below the zone of the highest productivity. The principle of 'sliced functioning'
is used to explain the adaptive significance of the morphological structure of populations under hetero-
geneous environmental conditions.

Introduction mental spatial heterogeneity. These data are

necessary to understand how these populations
The rotifer Keratella cochlearis, like many other form functional units.
species of planktonic rotifers, is characterized by The purpose of this investigation is to study
high spatial and temporal heterogeneity of species population functioning of K. coch/earls in strati-
populations. This is especially true in stratified fied lakes with consideration of morphological
lakes. Variability in morphotype, lorica length, structure and spatial distribution.
and posterior spine length occurs seasonally as
well as in response to lake trophy (Carlin, 1943;
Gallagher, 1957; Green, 1981; Hofmann, 1980, Methods
1983, 1987; Hillbricht-Ilkowska, 1972). In these
studies, the short-spined morph of K. cochlearls is Zooplankton samples were collected in June-July
generally found in the epilimnion and the long- 1985 and July, 1987 from oligo-mesotrophic Lake
spined morph is found in the hypolimnion. How- South Volos, Byelorussia (area 1.2 km, max.
ever, there are no data on eco-physiological or depth 40.4 m (Yakushko, 1971» and in June 1986
production parameters of K. cochlearis popu- from eutrophic Lake Mikolajki, Poland (area
lations pertaining specifically to affects of environ- 4.6 km, max. depth 27.8 m (Hillbricht-Ilkowska
120

et aI., 1971)). Samples were collected from three For calculating population and energetic
stations of Lake South Volos in the vertical water parameters, morphometric measurements, em-
column from surface to bottom in 5 m increments. bryonic development rates, and oxygen consump-
In addition, daily collections were made every 3 tion rates were calculated for each population.
hours using the same increments at the station Wet weight of rotifers was calculated relative to
with the maximum depth. In June 1986 and July volume, dry mass was taken as 10% of wet
1987, samples were taken from the stations with weight, and the caloric value of the dry weight was
the maximum depth at 1 m intervals from both 4.7 cal mg- 1.
lakes at midday and midnight. Rotifer and chloro- Birth rate (b) was calculated according the for-
phyll samples were taken simultaneously. mula (Paloheimo, 1974):
Water samples were taken with a 2 1bathometer
and fixed with 40 %formalin, so as to attain a 2 % b = IjDe 'In (1 + EjN),
concentration. Zooplankton samples (11) were
sedimented for 3-4 days, then the overlying liquid where De = duration of embryogenesis, day - 1 ;
was decanted. Each sample was diluted to 100 ml E = number of parthenogenetic eggs, eggs 1- 1 ;
and a 20 ml aliquot from each was completely N = density of parthenogenetic females, ind 1- 1.
counted. The EjN ration was named the egg ratio or rela-

sca1e I
Tim e .) 25°0

0\
12.00 18.00 24.00 3.00 0.00 9.00 12.DO

1\
5 1I\
10

15

0...
Q)

35

40

Fig. 1. Diurnal vertical distribution of Keratella cochlearis in S. Volos Lake, June 24-25, 1985. Distribution ofrelative density
(density in each layer relative to total density at all layers in %). Black circles indicate mean depth of the population 'center'
site.
 121

tive population fecundity. Potential production evenly distributed and formed not more than 15 %
(P) was determined as a product of (e b - 1) and of the total density of K. cachlearls in the water
the density or biomass was expressed as ind 1- 1 column. More than 60% of the total number of
day - 1 or j - 1 day - 1. The net growth efficiency K. cachlearis were found between the surface and
was calculated as the production divided by the 10 m layer. The absence of vertical migration of
sum of production and respiration. K. cachlearis and in other planktonic rotifers in S.
Chlorophyl I concentration in Lake Mikolajki Volos Lake has also been reported by
was determined by Dr. Irena Kufel (Hydrobio- Galkovskaya (1984).
logical station, Institute of Ecology PAN). All Examination of population vertical density
other data were obtained by the authors. profiles of K. cachlearls at three stations (Fig. 2),
and at the same station on different days (Fig. 3),
shows the stability of the vertical distribution of
Vertical distribution of populations of K. coch- this species in both space and time. The surface
learis density is low (50 ind 1- 1 ), density then increases
up to 150-200 ind 1- 1 at a depth of 10 m, and
In Lake S. Volos, June 1985, diurnal vertical decreases abruptly at depths of 15 m and greater.
migration was not observed (Fig. 1). At the sur- Increased sampling intensity of the vertical
face, the density was minimal throughout the day water column up to 1 m shows a clearer picture of
and was generally not more than 10 ind 1- 1. The the vertical profile of the K. cachlearls population
maximum density was observed at 5 m. In the at midday and midnight (Fig. 4). The absence of
20-40 m layer, the number of individuals was vertical migration of K. cachlearis was observed in

Numoer· r'
50 100 150 50 100 150 50 100 150 200

10

'S

20

---
~
25
=
..J
:l.. 30
l.J
:.J
j5

'to
sf 1 st. 2 st5
Fig. 2. Vertical distribution of the Keratella cochlearis population (1 - N, ind 1-1) in water column of S. VOIDS Lake at three
stations. July 1985.
 ......
tv
tv

20 40 60 80 2n 40 60 so 20 1;0 60 80 E/N

40 80 120 80 40 80 120
N
/\\ ~'
5

10~_
15l /- ~ ~_~"o ""-7"0
~ ~ .,:.
7' ~
-/

E 20 k!l if' 9"''''' """ J)

::-: // .4.y
1/ ! # } hlspLda'
P
---t: #,,7
--t-... 25W 0
CL
<lJ }2 cockewu.'$
= 30

LtO

Fig. 3. Variations in density (N, ind 1- 1) and relative fecundity (EjN, %) for Keratella cochlearis hispida (1) and Keratella cochlearis cochlearis (2) in water column
of S. Volos Lake, July 1985.
 123

Numger /.-1
, J

20 60 100 20 60 fOO
E\I~ I
;:fP 0'n
I
if
P

10 ~'r
=~
~
15
- / - - --- i l1M'u~
2 ~~

1
t 20
"

>
-C
+'
0.....

<>J
25
0
D

30

35 -

ItO

Fig. 4. Vertical distribution of densities (N, ind I-I) for Keratella cochlearis hispida (1) and Keratella cochlearls cochleans (2) in
water column of S. Volos Lake at midday (a) and midnight (b). July, 17 (24.00) - 18 (12.00), 1986.
124

eutrophic lake Mikolajki as well (Fig. 5). In both A positive correlation between the density of
lakes, smaller morphs - tecta in Lake Mikolajki K. cochlearis and chlorophyl I concentration was
and hispida in Lake S. Volos - prevail in the found in both lakes (Fig. 6), thus indicating a
epilimnion while the relative density of the larger potential trophic factor contributing to the spatial
morph - each/eans - increases with depth. distribution of morphs of this species.

Numof'/',)
f rf
(.

800

-_--0

'5

10
f
'\

.s:
.+J - - - - j Ud;o..
a....
2 ~
<!J
D

15

Q) m·\clJo.,~
1 ~) m.ld~

Fig. 5. Vertical distribution of densities (N, ind I - 1) for Keratella cochlearis tecta (1) and Keratella coch/earls cochlearis (2) in water
column of Mikolajskie Lake at midday (a) and midnight (b). June, 24 (12.00) - 25 (24.00), 1986.
 125

o - j S. VOl05
X :: 2 Hikolcyshe

8
en tV =.3,9/19 + fJ,68211/z Cft.- x
7
-,
~
6
1"= fJ,'l8

5
i. ..;,

4 o x
'4D
~ x
3
§ 0 o 00
s;:
2 - 00

~ f -

0
~3 -2

Fig. 6. Relationship between density of Keratella cochlearis (N, ind 1-1) and chlorophyll concentration (Ch, p,g 1- 1) based on
data from concurrent sampling of water column layers in Lakes S. Volos (1) and Mikolajskie (2).

Some population and energetic characteristics of A different pattern was observed in

K. cochlearis K. c. cochlearis. The density of this morph
increased gradually with depth. Maximum E/N
In July, 1985, the popUlation size (N) and theE/N values were observed below the maximial density
ratio for two morphs of K. cochlearis in the verti- zone. The maximum occurred in the zone of
cal water column at the same station were 10-15 m.
observed continuously for several days (Fig. 3). The minimum egg ratio and maximum density
Maximum density of K. c. hispida (140 ind 1- 1) of the species popUlation were observed at the
was observed in the 5-10 m layer. K. c. cochlearis meta-hypolimnion border. The apparent density
had a maximum density of 60 ind 1- 1 in the effect is probably a result of trophic limitation. It
10-15 m layer. This morph was not found in the is apparent that the density of species popUlation
0-5 m layer. is determined by K. c. hispida except under trophi-
The vertical profile of the parameters N and cally limiting conditions where it is unable to
E /N from the surface to the bottom showed a reproduce itself and is gradually supplanted by
maximal E/N value for K. c. hispida above or in the larger morph K. c. cochlearis.
the layer with maximal density. In lower layers Embryonic development rates (l/De) in Lake S.
there was an abrupt decrease in the value of this Volos were obtained for rotifers selected from
parameter. The sudden decrease of the E/N ratio water layers with temperatures of6, 12, and 18°C
along with the increased density of K. c. hispida both under natural conditions (Galkovskaya &
allows us to suggest that there may be trophic Mityanina, 1986) and under abruptly changed
deficiencies under these conditions. temperature conditions as well (for example, ani-
126

Tablel. Empryonic development rate (lID e, day-I) and Table 2. Variations in relative populational fecundity (EIN,
embryonic growth intensity (WIDe, 10 - 5 mg wet mass %), embryonic development rate (l IDe' day - I) and birth
day-I) for Keratella cochlearis. S. Volos Lake, June 1984. rate (b, day-I) for two Keratella cochlearis morphs in the
water column of S. Volos Lake. July 1985.
Temperature Experimental temperature, °C
of adapta- Depth, m Keratella cochlearis Keratella cochlearis
tion, °C 18 12 6 hispida cochlearis

IIDe WIDe IIDe WIDe IIDe WIDe EIN IIDe b EIN 1IDe b

18 0.96 3.08 0.66 2.12 0.50 1.60 0 22 1.02 0.202

12 0.75 3.29 0.63 2.77 0.36 1.58 5 24 1.08 0.232 33 0.88 0.250
6 0.63 3.48 0.38 2.10 0.29 1.60 10 6 0.95 0.055 24 0.62 0.134
15 21 0.39 0.075
20 37 0.36 0.112
mals collected from 6 °e water temperature were
tested for embryonic development times at 12 ° e
and 18 °e, etc.). The volume-specific egg wet U sing the empirical curve of the relation
weight was also determined during these experi- between W/ De and temperature, we reconstructed
ments. Using these data we introduce the parame- the expected values oft/De for K. each/earls from
ter WIDe the embryonic development intensity, the values of the egg mass and the temperature of
and consider it in relation to temperature of accli- the corresponding layer. The birth rate for each
mation. The value of this parameter was found to morph and for the whole species was calculated
be independent of the temperature of acclimation based on these values of 1/De' Despite great dif-
(Table 1). When comparing the values of 1/De' the ferences in the rate of embryonic development
effect of preliminary temperature adaptation is between the morphs, the birth rate of both morphs
due to egg mass differences. Using the values of was equal. Maximum birth rates were observed in
W/ De' the Q10 is 1. 7 at temperatures ranging from the epilimnion at a depth of 5 m (Table 2). The
6 °e to 18°C. maximum daily population production in the

Table 3. Some energetic characteristics of the Keratella cochlearis population (1 - K. cochlearis hispida, 2 - K. cochlearis
cochlearis) from S. Volos Lake. July 20, 1987.

Depth Tem- N, ind·l- I EIN B,1O- 2 J'I- 1 P,1O- 3 R, 10- 2

pera- J .1- 1 J .1- 1
ture 2 2 2 day-I day-I
°C

0 20.6 33 0.27 0.40 0.96 4.94

5 19.3 139 0.29 2.06 5.50 19.45
10 8.1 106 61 0.02 0.21 1.27 1.10 0.95 8.06
15 5.7 55 0.38 0.10 0.08 1.59
20 5.0 19 0.16 0.34 0.12 0.46
25 4.9 6 0 0.11 0 0.15
30 4.8 8 3 0.25 0.33 0.10 0.05 0.09 0.27
35 4.8 1 7 0 0 0.01 0.13 0 0.19
40 4.6 6 8 0 0 0.07 0.14 0 0.34

Mean values for the lake: B = 1.51 10 - 2 J '1- I.

P= O.l241O- 2 J·I- I ·day-l.
R= 0.3191O- 2 J·I- I ·day-l.
PIB = 0.08.
K2 = 0.28.
 127

Table 4. Some energetic characteristics of the Keratella cochlearis population (1 - Keratella cochlearis tecta, 2 - Keratella cochlearis
cochlearis) from Mikolajskie Lake. June 24-25, 1986.

Depth Tem- N, ind .1- 1 EIN B,IO- 2 J'I- I P,IO- 3 R,IO- 2

pera- j·I-I j·1- I
ture, 2 2 2 day-I day-I
DC

0 17.0 653 375 0.23 0.15 3.76 2.22 1.10 10.06

5 15.1 171 300 0.28 0.16 0.99 2.58 0.54 4.65
10 11.0 35 55 0.06 0 0.61 0.47 0.02 0.60
15 7.5 46 76 0.26 0 0.21 0.65 0.03 0.62
19 6.8 32 66 0.28 0 0.18 0.57 0.02 0.47

Mean values for the lake: B = 3.09 10 - 2J '1- 1.

P = 0.42 10 - 2 J '1- 1 • day - 1.

PIB = 0.135.
K2 = 0.10.

water column was observed at 5 m as well. Poten- hypolimnion. In particular, it becomes apparent
tial production decreases abruptly at depths of while examining the affects of temperature adap-
10m and greater despite stable, and even increas- tation on embryonic development.
ing, relative fecundities. Cases of simultaneous presence of several
Figure 6 shows that the rotifer density is closely morphs of the rotifer K. eaehlearls in lakes are
correlated with chlorophyl 1 concentration. This known (Green, 1981; Hillbricht-Ilkowska, 1972;
may explain why there is a change of morphs Hofmann, 1987). Pejler (1980) explained mor-
between layers; i.e., the production of each morph phological variability of K. eaehlearis within one
is likely to be dependent upon trophic conditions. water body as a purely physiological adaptation.
The population production rate (P/B) of As a basis for his explanation, he used data on the
K. eaehlearls is higher in the eutrophic lake in absence of sexual reproduction in K. eachlearls
comparison with the mesotrophic lake (0.135 and during one year and some successive years
0.08 respectively), the net growth efficiency of the (Carlin, 1943; Ruttner-Kolisko, 1949) and hence
populations, on the contrary, decreases abruptly found no reason for applying King's (1977)
in the eutrophic lake in comparison with the hypothesis of genetic population interruption to
mesotrophic lake (0.10 and 0.28 respectively) K. eachlearis. Protein polymophism in natural
(Tables 3 and 4). rotifer populations has been demonstrated by
King (1977); i.e., population genetic heterogeneity
has been shown. Only selection characteristics
Discussion can be discussed until it is proven that mutation
accumulation takes place during parthenogenetic
There were no active vertical diurnal movements reproduction within a season.
of K. eaehlearis populations at least between Presently there are not enough data to explain
the epi-, meta-, and hypolimnion in Lake S. Volos morphological variability as a type of morpho-
or Mikolajki during the summer stagnation physiological adaptation occurring within the
period. limits of the genotype, nor to characterize morphs
The absence of diurnal vertical migration and from the point of total genetic interruption as
the relative temporal stability of the population genotypes adapted to different lake conditions.
vertical profile suggest eco-physiological patterns Further investigations in this area are necessary.
of processes taking place in the epi-, meta-, and If morphs are the phenotypic realization of
128

genotypes, strict division of regions inhabited by Galkovskaya, G. A, 1984. Spatial distribution of planktonic
individual morphs may be facilitated in stratified rotifers and their trophic relations in lake ecosystems. V
kn. Limnologia gornykh vodoemov. Yerevan: 54 (In
lakes. The absence of shifting condition gradients, Russian).
i.e., correspondence of the definite part of popu1a- Galkovskaya, G. A. & I. F. Mityanina, 1986. Birth rate char-
tion to epi-, meta-, hypoliminion can promote acteristics of natural population of the rotifer Keratella
selection even during successive parthenogenetic cochlearis (Gosse). DAN BSSR 30: 568-570 (In Russian).
generations. The relative spatio-temporal isola- Gallagher, J. J., 1957. Cyclomorphosis in the rotifer Keratella
cochlearis (Gosse). Trans. am. micro Soc. 76: 197-203.
tion of population components may create a Green, J., 1981. Altitude and seasonal polymorphism of
'sliced functioning' of population as a whole. Keratella cochlearis (Rotifera) in lakes of Auvergne, Cen-
Discussing K. cochlearls popu1ations of deep- tral France. BioI. J. limnean Soc. 16: 55-61.
water mesotrophic and eutrophic lakes in terms of Hillbricht-Ilkowska, A, 1972. Morphological variation of the
a heterogeneous medium, we fmd the realization Keratella cochlearis (Gosse) (Rotatoria) in several
Masurian lakes of different trophic level. Pol. Arch. Hydro-
of optimal population productivity may be due to bioI. 19: 253-264.
the formation of morphotypic-distinct functional Hillbricht-Ilkowska, A., E. Pieczynska & E. Pieczynski,
subunits. 1971. Studies on the productivity of several Mazurian
High correlations between rotifer densities and lakes and on the effect of fish on lake biocenosis.
chlorophyll concentration permits one to consider Wiadomosci ekologiczne 17: 124-146.
Hofmann, W., 1980. On morphological variation in Keratella
spatial morphotypic shifting as a component of a cochlearis population from Holstein lakes (Northern
popu1ation's adaptive ability in response to varia- Germany). Hydrobiologia 73: 255-258.
bility in trophic conditions. Hofmann, W., 1983. On temporal variation in the rotifer
Keratella cochlearis (Gosse): the question of ' Lauterborn-
cycles'. Hydrobiologia 101: 247-254.
Acknowledgements Hofmann, W., 1987. Population dynamics of hypolimnetic
rotifers in the Pluss-see (North Germany). Hydrobiologia
147: 197-201.
The authors are grateful to Dr. I. Kufel who has King, C. E., 1977. Genetics of reproduction, variation and
provided unpublished data. Also to Dr. J. adaptation in rotifers. Arch. Hydrobiol. 8: 187-201.
Ejsmont-Karabin and Dr. A. Karabin for their King, C. E., 1980. The genetic structure of zooplankton popu-
help in sampling Mikolajskie Lake and Dr. V. lations. In W. C. Kerfoot (ed.), Evolution and Ecology of
Vezhnovets for his help in sampling Lake S. Zooplankton Communities. Special Sumposium 3. Univ.
Press New England: 315-329.
Volos. Kutikova, L. A., 1970. Rotifer fauna of the USSR. Fauna
USSR 104. Akad. Nauk. SSR, Leningrad, 744 pp. (In
Russian).
References Paloheimo, J. E., 1974. Calculation of instantaneous birth
rate. Limnol. Oceanogr. 19: 692-694.
Bosch, F. V. D. & J. Ringelberg, 1985. Seasonal succession Pejler, B., 1980. Variation in the genus Keratella. Hydro-
and population dynamics of Keratella cochlearis (Gosse) biologia 73: 207-213.
and Kellicottia longispina (Kellicott) in lake Maarsseveen 1 Ruttner-Kolisko, A, 1949. Zum Formwechsel und Art-
(Netherlands). Arch. Hydrobiol. 103: 273-290. problem von Anuraea aculeata (Keratella quadrata). Hydro-
Carlin, B., 1943. Die planktonrotatorien des Motalastrom. biologia I: 425-468.
Lund: Carl Bloms Bortryckeri, 255 pp. Snell, T., 1977. Clonal selection: Competition among clones.
Djokosetiyanto, D. & N. Lair, 1983. Strategies d'utilisation Arch. Hydrobiol. 8: 202-204.
des ressources dans un lac meso-oligotrophe: Migration Yakushko, O. F., 1971. Byelorussian Poozerje. Historical
verticales. Ann. Stat. bioI. Besse- en Chandess. N 17: development and present situation of lakes of Northern
65-91. Byelorussia. Minsk, 336 pp. (In Russian).
Hydrobiologia 186/187: 129-136, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 129
© 1989 Kluwer Academic Publishers.

The development of Hexarthra spp. in a shallow alkaline lake

Alois Herzig & Walter Koste

Biologische Station Neusiedler See, A-7142 Illmitz, Austria and Ludwig-Brill-StrafJe 5, D-4570
Quakenbrilck, FRG

Key words: Hexarthra, salinity, alkalinity, wind action, temperature

Abstract

In Neusiedler See, a shallow alkaline lake with fluctuating water level and salinity, four species of
Hexarthra occur: H. mira, H. fennica, H. jenkinae (occasional) and H. polyodonta. The analysis of long-
term data reveals a general phenological pattern which does not change from year to year. They fIrst occur
in May, develop a maximum in June/July, sometimes a second one in August/September and disappear
in October. But the species succession is different in the various years, occasionally only one species
(H. mira or H. polyodonta) being present. There is a fairly consistent relation between the chemical
conditions and the prevalent species; an increase in salinity favours the development of H. polyodonta.
Low temperature and wind generated suspended particles have a negative influence on the development
of the Hexarthra populations. Smaller populations of Hexarthra are in a relation to the occurrence of
Leptodora indicating predation pressure of the latter species. In N eusiedler See the Hexarthra populations
seem to be controlled to a great extent by abiotic factors, but predation by Leptodora and most probably
by young fIsh seems to play an important role too.

Introduction and Hexarthra polyodonta (Hauer, 1957). In

earlier papers on the plankton of this lake H. mira,
Morphological and ecological characteristics of H. fennica and H. intermedia Wiszniewski 1929
the genus Hexarthra Schmarda 1854 make these are mentioned (Varga, 1926, 1934; Zakovsek,
rotifers challenging objects for fIeld and labora- 1961). One to three species may be detected within
tory investigations. These animals have complex a year and all of them develop throughout the
and subtle appendages which produce a skipping warmer season (May-October). During summer
motion. They inhabit the pelagic zone offresh and 8-61 % of the rotifer plankton may consist of
inland saline waters as well as the sea. Some of Hexarthra specimens.
the species tolerate a wide range of salinities. A The present paper gives a description of the
preference for high temperatures is obvious phenology of these species over the years and a
(Ruttner-Kolisko, 1974; Koste, 1978). discussion of the possible influences of abiotic
For the past 20 years (1968-1988) four species factors (e.g. salinity, temperature, wind) and
have occurred in Neusiedler See: Hexarthra mira biological interactions (e.g. predator-prey) on the
(Hudson, 1871), Hexarthra fennica (Levander, population development.
1892), Hexarthrajenkinae (De Beauchamp, 1932)
130

Materials and methods Methods

The lake In 1968 and 1969, samples were taken with a

Ruttner bottle at 8-16 stations at three different
Neusiedler See is a shallow (z = 1.3 m) well- depths; in 1970-1974 integrated samples were
mixed lake which is characterized chemically by taken with horizontal tows using an Isaacs-Kidd
its high alkalinity (7.5-14.6 meq 1-1) and a con- high-speed sampler. The mesh size of the filters
ductivity of 1150-2800 J-lS (18°C). The surface used was 50 J-lm. Since 1975 samples have been
area (321 km2) is divided into two different habi- collected with vertical net hauls (30 J-lm mesh
tat types. The open water zone (143 km2) is char- size). The sample interval has varied between
acterized by a high concentration of suspended 6-15 days in summer, 2 weeks in spring and
solids which are stirred up from the bottom by autumn and 3-5 weeks during winter. In order to
wind action. The marginal areas are densely obtain a reasonable estimate of the population in
covered by Phragmites communis (178 km2) and the lake 8-20 separate stations have been sampled
have a clear water colored by humic substances. and from these counted samples a mean for the
The low altitude (115 m a.s.l.), the influence ofthe whole lake was calculated (for details see Herzig,
Pannonian climate, large surface area, shallow 1974, 1979).
water and continuous mixing result in high water Data on chemical analysis is taken from
temperatures during the summer months (maxi- Neuhuber (1971), Ruttner-Kolisko (1971), Berger
mum > 28 ° C) and extremely low ones & Neuhuber (1979), L6ffier & Newrkla (1985)
throughout winter. Since 1970 the lake has and unpublished reports of the Biological Station
become increasingly eutrophic. A detailed de- Neusiedler See.
scription of the physiography and ecological
aspects may be found in Loffier (1979).
Results and discussion

Taxonomy In the quantitative plankton samples from

Neusiedler See, Hexarthra spp. consistently
The identification of the various species was done appear in the summer plankton (Zakovsek, 1961;
on preserved material only and was based on the Herzig, 1979; Ruttner-Kolisko pers. comm.).
trophi of the mastax. Hexarthra is characterized During 1951-1979 the highest relative contribu-
by a mastax with malleoramate trophi and the tion of species in this genus to the rotifer com-
number of teeth on the uncus is used in taxonomy. munity varied between 8 and 61 % and the sum-
According to Ruttner-Kolisko (1974) the various mer mean relative abundance varied between 5
morphological characteristics (e.g. size, relation and 18%. The abundance of Hexarthra spp. for
of the ventral arm/body length, number of bristles, several years is shown in Fig. 2. In general the fIrst
number of teeth on the uncus) by which the specimens appear in May (rarely in April) at
species of this genus are distinguished show con- water temperatures of 8-10 ° C; subsequently an
siderable variation in different environmental exponential growth phase takes place (T ° C:
conditions; there seems to be a tendency towards 17-20) and maximum abundance is reached in
an increase in the number of teeth on the uncus June/July or August when water temperatures
as salinity increases. Nevertheless, according to exceed 21°C; Hexarthra quickly disappears in
Koste (1978), the four species can be identified on October, as soon as water temperature drops
the basis of the number of teeth and display clear below 15°C.
ecological differences. Drawings of the trophi of Both qualitative and quantitative changes in
the various species are given in Fig. 1. seasonal abundance are seen in Fig. 2. The popu-
lations of 1968-1970 are dominated by H. mira
 131

Fig. 1. Trophi of Hexarthrafennica (A), Hexarthra mira (B),

Hexarthra polyodonta polyodonta (C), Hexarthra polyodonta
soaplakeiensis (D); ventral view of Hexarthra polyodonta (E);
drawings by W. Koste.
132

Fig. 2. Seasonal changes of Hexarthra abundance. Dominant species are indicated by capital letters. M-Hexarthra mira, F-
Hexarthrafennica, P-Hexarthra polyodonta. 1977, 1978, 1979: Vertical bars - abundance of Leptodora kindti. (abundance per m3 ).
 133

and H. fennica. Within this period H. jenkinae and Table 1. Mean values (± SD) of electrical conductance and
H. polyodonta were found only once (beginning of alkalinity for the period of occurrence of the various species.
June 1969 and beginning of July 1968 respec-
Electrical Alkalinity
tively). In 1976 H. polyodonta occurred together conductance (JlS) meq'I- 1
with H. mira. In 1977 H. mira was the dominant
species, but H. polyodonta was present in low Hexarthra mira 1327 ± 225 8.48 ± 0.9
numbers. Since 1978 H. polyodonta has frequently Hexarthra jennica 1228 ± 87 8.18 ± 0.72
Hexarthra polyodonta 1846 ± 220 10.29 ± 0.95
been the only Hexarthra species found in the sum-
mer rotifer plankton.
The various Hexarthra populations hatch from Table 2. Mean values (± SD) of electrical conductance and
resting eggs. Resting egg production was observed alkalinity for various years.
in Neusiedler See from late August until the dis-
Years Electrical Alkalinity
appearance of the animals. In addition a con- (meq- 1-1)
conductance (JlS)
tinuous input of Hexarthra resting eggs may occur
from the various shallow lakes ('Seewinkel- 1968/69/70 1269 (± 103) 8.20 (± 0.63)
lacken') or gravel pits east of Neusiedler See. 1975(76 1836 (± 172) 10.19 (± 0.41)
Some of these water bodies chemically resemble 1977/78/79 1867 (± 232) 10.35 (± 0.73)
Neusiedler See, others are more alkaline, and in
some chlorides and/or sulphates prevail. Accord-
ingly H. jenkinae and H. polyodonta may be conductance and alkalinity were observed
observed in lakes containing soda; H. fennica (Table 2). The relation between alkalies (A) and
occurs in water bodies with a smaller amount of alkaline earth compounds (E) (E-A (meq 1- 1):
soda and in waters characterized by chlorides and - 1.22 - + 1.63) and pH (7,8-9.0) remained rela-
sulphates (L5ffier, 1957, 1959; Ruttner-Kolisko, tively stable (Berger & Neuhuber, 1979;
1971). H. mira is frequently found in waters oflow Brossmann in L5ffier & Newrkla, 1985). The
salinity and in gravel pits (Herzig, unpUblished). higher the values of electrical conductance and
The majority of these shallow lakes dry up com- alkalinity, the more likely is the prevalence of
pletely during summer or autumn and resting eggs H. polyodonta (Fig. 3). An analysis of variance
of various Hexarthra populations may be intro- (one factor ANOVA for repeated measures)
duced by wind into Neusiedler See. Passive trans- reveals clear differences (significant at 95 %)
fer of the eggs may also take place by water fowl between H. polyodonta and H. fennica and
which frequently migrate between these smaller H. mira. No significant differences exist between
water bodies and N eusiedler See. H. fennica and H. mira. This is consistent with the
Variation in the occurrence of these species results of Hutchinson (1932) who was not able to
may reflect variation in the number of viable rest- detect any great contrast in the occurrence of the
ing eggs. Alternatively, the chemical conditions of two species. This overlap in occurrence seems to
Neusiedler See may not suit all species and/or happen as long as the salt content remains rather
populations. According to Ruttner-Kolisko low « 2 g 1- 1). At higher concentrations, pre-
(1971) H.jenkinae, which was found only once, dominantly in chloride dominated inland waters,
mainly inhabits waters with a higher salt content only H. fennica will occur (Ruttner-Kolisko, 1971,
(> 5 g 1- 1) and higher alkalinity (> 50 meq 1- 1). 1974).
These values are far above the levels reached in The results of the investigation so far suggest
N eusiedler See. that the increase in salinity did favour the devel-
Nevertheless there is a fairly consistent relation opment of H. polyodonta. None the less more
between chemical conditions and the prevalent comparable field studies and especially labora-
species (Fig. 3, Table 1). Between 1968 and 1979 tory experiments are needed to get a better know-
moderate, but significant, increases in electrical ledge about how the various Hexarthra species
134

;' .,

10 17 I<

",
./
.....
/'
/'
/' \
/' \
/' \
/'
/' \
\
--- --- \
\
\
\
\
\
\
\

10 11 12
ALKALINITY , ... q. I·11

Fig. 3. Occurrence of Hexarthra spp. in relation to temperature, electrical conductance and alkalinity in Neusiedlersee.

cope with a changing chemical environment. several days of calm weather (mean wind speed
Besides the alteration in species composition well < 12 km h - 1) lead to rapid population growth
expressed quantitative changes took place within (daily growth rate > 0.2). But even at these tem-
the years 1968-1979 resulting from the impact of peratures high wind speed (mean > 18 km h - 1,
abiotic factors and biological interactions. Irre- maximum > 40 km h - 1) produces suspended
spective of the species, the population develop- inorganic particles which mechanically interfere
ment of Hexarthra is mainly influenced by tem- with the animals (cf. Diaphanosoma brachyurum,
perature and wind action. A correlation analysis Herzig, 1975, 1979) and may have a negative
revealed highly significant positive correlations effect on the population development (e.g. Fig. 2,
between abundance and temperature (r = 0.575), August 1970). Frequently strong summer winds
as well as between population growth rate and are associated with a rapid temperature decline
mean temperature for the period considered which may last for about one to two days. This
(r = 0.579). The correlation between population combination of abiotic factors has a detrimental
growth rate and mean wind speed was highly effect on the Hexarthra populations (e.g. Fig. 2,
significant and negative (r = - 0.455). June and July 1968, August 1969).
In summer, temperatures above 22 °C and As mentioned earlier, hatching of resting eggs
 135

is followed by a period of exponential growth (e.g. development data derived for H. jennica, Ruttner-
Fig. 2, 1968, 1976). This exponential growth Kolisko, 1975) gave values of 0.2-0.42 for the
phase can be delayed (e.g. Fig. 2, 1969, 1970). As summer months and lead to the suggestion that,
temperature and food conditions (Dokulil, 1984) without interference by Leptodora, Hexarthra
were comparable for all years, but heavy wInds could have develcped II much larger population.
prevailed (mean wind speed> 20 km h - I) during There is little evidence from the literature that
these periods in 1969 and 1970, the growth delay Leptodora is preying upon rotifers; only two
may again be attributed to mechanical inter- references can be found in the review paper on
ference by suspended particles. 'Invertebrate predators of planktonic rotifers' by
These results are consistent with the idea that Williamson (1983). Recently Edmondson & Litt
low temperature and high amounts of wind- (1987) demonstrated a relation between Cono-
generated suspended particles can negatively chi/us hippocrepis and Leptodora kindti; in three
influence the development of Hexarthra popu- years the decrease in Conochilus came during the
lations. Nevertheless a multiple regression analy- peak of Leptodora. No further information on
sis based on the data from the summer months feeding behavior or feeding rates is available but
(y = population growth rate; Xl = mean tempera- at least the authors could identify trophi of
ture; X 2 = mean wind speed for the same period) Conochilus in the Leptodora stomach.
demonstrates that only 25.4 % of the total So far we have neither information on stomach
variance is explained by these two factors. One contents nor experimental work that demonstrate
therefore has to search for an explanation of the this predator-prey relationship. But it seems likely
74.6% residual variance. that the size (up to 400 11m), the reasonable soft
There is evidence from long-term analysis of body, and the skipping movement of Hexarthra
the phytoplankton that the food conditions species may make them suitable prey items for
improved throughout the years 1968-1979 Leptodora kindti.
(Dokulil, 1979, 1984) resulting in an increase of
the zooplankton (Herzig, 1980). The increased
abundance of zooplankton favored the develop-
ment of planktivorous fish. The current idea is References
that planktivorous fish and the still unknown
Berger, F. & F. Neuhuber, 1979. The hydrochemical problem.
quantities of fish fry and fish that are less than one In H. Lamer (ed.), Neusiedler See. The limnology of a
year old (mainly cyprinids) to some extent control shallow lake in central Europe. Dr. W. Junk bv Pub!', The
the zooplankton populations (especially during Hague, Boston, London. Monogr. BioI. 37: 89-99.
the summer months). So far we do not know Dokulil, M., 1979. Seasonal pattern of phytoplankton. In H.
Lamer (ed.), Neusiedler See. The limnology of a shallow
whether these fishes have an impact on the rotifer
lake in central Europe. Dr. W. Junk bv Pub!., The Hague,
populations and in particular on Hexarthra spp. Boston, London. Monogr. Bio!. 37: 203-234.
In any case, the Hexarthra populations show Dokulil, M., 1984. Assessment of components controlling
quite a variation in their quantitative appearance; phytoplankton photosynthesis and bacterioplankton pro-
peak values range from 151- I to more than duction in a shallow, alkaline, turbid lake (Neusiedler See,
Austria). Int. Revue ges. Hydrobio!. 69: 679-727.
601- I (see Fig. 2). But in 1978 and 1979 the
Edmondson, W. T. & A. H. Litt, 1987. Conochilus in Lake
maximum numbers remain below 10 or even Washington. Hydrobiologia 147: 157-162.
below 5 animals per litre. These much smaller Herzig, A., 1974. Some population characteristics of plank-
populations are in a fairly consistent relation to tonic crustaceans in Neusiedler See. Oecologia 15:
the occurrence of Leptodora kindti which was 127-141.
Herzig, A., 1975. Der Neusiedler See - charakteristische
detected for the first time in mid June 1977 and
Eigenschaften und deren Auswirkungen auf das
since then has become more and more abundant Zooplankton. Verh. Ges. Okologie, Wien 1975: 189-196.
(Fig. 2, 1977, 1978, 1989). Preliminary estimates Herzig, A., 1979. The zooplankton of the open lake. In H.
of birth rates of Hexarthra (applying embryonic Lamer (ed.), Neusiedler See. The limnology of a shallow
136

lake in central Europe. Dr. W. Junk bv Pub!., The Hague, punktuellen Nahr5toffeintrages auf die Eutrot'hierung von
Boston, London. Monogr. Bio!. 37: 281-335. Seen. Teil 2: Neusiedler See, Attersee, Universitatsverlag
Herzig, A., 1980. Effects of food, predation and competition Wagner, Innsbruck, Veroff. Ost. MaB Progr. 8: 121 pp.
in the plankton community of a shallow lake (Neusiedler Neuhuber, F., 1971. Ein Beitrag zum Chemismus des
See, Austria). In M. Dokulil, H. Metz & D. Jewson (eds), Neusiedler Sees. Sitz. ber. Ost. Akad. Wiss., mathem.-
Shallow lakes. Contribution to their limnology. Dr. W. naturwiss. K!., Abt. I, 179: 225-298.
Junk bv Pub!., The Hague, Boston, London. Develop- Ruttner-Kolisko, A, 1971. Rotatorien als Indikatoren fUr
ments in Hydrobiology 3: 45-51. den Chemismus von Binnensalzgewassern. Sitz. ber. Ost.
Herzig, A, 1987. The analysis of planktonic rotifer popu- Akad. Wiss., mathem.-naturwiss. K!., Abt. I, 179:
lations: A plea for long-term investigations. Hydrobiologia 283-298.
147: 163-180. Ruttner-Kolisko, A, 1974. Plankton Rotifers. Biology and
Hutchinson, G. E., G. E. Pickford & J. F. M. Schuurman, Taxonomy. E. Schweizerbart'sche Verlagsbuchhandlung,
1932. A contribution to the hydrobiology of pans and other Stuttgart. Die Binnengewasser XXVI/I, Supp!., 146 pp.
inland waters of South-Africa. Arch. Hydrobio!. 24: 1-154. Ruttner-Kolisko, A., 1975. The influence of fluctuating tem-
Koste, W., 1978. Rotatoria. Die Radertiere Mitte1europas. perature on plankton rotifers. A graphical model based on
Monogononta. Gebruder Borntraeger, Berlin, Stuttgart, life data of Hexarthrafennica from Neusiedler See, Austria.
Bd. 1,673 pp. Symp. Bio!. Hung. 15: 197-204.
L{imer, H., 1957. Vergleichende limnologische Untersuchun- Varga, L., 1926. Die Rotatorien des FerW (Neusiedler See).
gen and den Gewassern des Seewinkels (Burgenland). Arch. Balat. I: 181-225.
Verh. Zoo.-Bot. Ges. Wien, 97: 27-52. Varga, L., 1934. Neue Beitrage zur Kenntnis der Rotatorien-
Lamer, H., 1959. Zur Limnologie, Entomostraken- und fauna des Neusiedler Sees. Allatani Kozlemenyek 31:
Rotatorienfauna des Seewinke1gebietes (Burgenland, 139-150.
Osterreich). Sitz. ber. Ost. Akad. Wiss., mathem.-natur- Williamson, C. E., 1983. Invertebrate predation on plank-
wiss. K!., Abt. I, 168: 315-362. tonic rotifers. Hydrobiologia 104: 385-396.
Lamer, H. (ed.), 1979. Neusiedler See. The limnology of a Zakovsek. G., 1961. Jahreszyklische Untersuchungen am
shallow lake in central Europe. Dr. W. Junk bv Pub!., The Zooplankton des Neusiedler Sees. Wiss. Arb. Bgld. 27:
Hague, Boston, London. Monogr. Bio!. 37: 543 pp. 1-85.
Lamer, H. & P. Newrkla, 1985. Der EinfluB des diffusen und
Hydrobiologia 186/187: 137-144, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 137
© 1989 Kluwer Academic Publishers.

On choice of substrate and habitat in brachionid rotifers

Birger Pejler & Bruno BerziJ;ls

Institute of Limnology, P.O. Box 557, 751 22 Uppsala, Sweden

Key words: rotifers, Brachionidae, ecology, substrate, habitat

Abstract

Information on the distribution of28 rotifers of the family Brachionidae from diverse waters in south and
central Sweden was analyzed to reveal their relationships to substrate and habitat. Some brachionids are
preferably planktic, others periphytic and/or benthic. Some non-planktic habitats are utilized more than
others, but there is no evidence of a chemical attraction from any substrate. Instead, some substrates seem
to be avoided, possibly depending on a poorer flora of periphytic algae. Besides substrate type, the
following factors are found to be important for creating separate ecological niches in the brachionid
family: temperature, oxygen content, trophic degree, chemical environment, food choice and sensitivity
to predation. It is possible to delineate separate ecological niches for all brachionid rotifers, implying that
Hutchinson's ideas about the 'plankton paradox' are contradicted. Some species are specialists, other
are generalists, the latter being characterized by a great morphological variation. The species are adapted
in different ways to their preferential habitats, as regards foot, egg-carrying, protrusions and other lorical
structures etc. Longer spines, for instance, are generally found in more transparent water, being a
supposed protection against visual predators.

Introduction and central Sweden. His data include information

on several abiotic parameters (see BerziJ;ls &
Most ecological studies of rotifers have been on Pejler 1987 and in press) and substrate, as well as
planktic forms. Good studies have also been the occurrence of different microzoans (planktic,
made on sessile rotifers (e.g. Edmondson 1944, periphytic and benthic). The material has now
1945; Wallace, 1980; Francez, 1984), as well as been computerized and will be published in
on psammic ones (e.g. Ruttner-Kolisko, 1956; several steps. Data on substrate exist for pro-
Tzschaschel, 1983). As regards some other non- tozoans and rotifers and they will be reported
planktic environments, important information separately for different taxonomic groups. In the
may be obtained in, e.g., Meuche (1939), Wulfert present paper only one rotifer family is discussed,
(1951), Donner (1964, 1970 and 1972), Martin the Brachionidae.
(1976 and 1977), Bateman & Davis (1980), and Bruno BerziJ;ls is solely responsible for the col-
Francez & Devaux (1985). However, a com- lection of data and the taxonomic identification.
prehensive study comparing major substrates has Both authors participated in the computerization.
never been made before. This has now been possi- Soon after the computer treatment was finished
ble by utilizing the large amount of data collected Bruno BerziJ;ls unfortunately died (Pejler, 1987),
by Bruno BerziJ;ls during the years 1945-1982 and thus Birger Pejler wrote the present paper (as
from various types of water bodies in southern well as the others).
138

Material and methods Results

The entire study comprises about 350 lakes, 50 As in the other papers dealing with the present
ponds, 20 pools, 150 running water localities, and material, a drastic data reduction was made. The
15 mires. However, in most cases only planktic entire data set is contained in computer sheets
samples were investigated, together with physical and tapes, archived at the Limnological Institutes
and chemical analyses. This is reflected in in Lund and Uppsala.
Table 1, where the number of samples from dif- The reduction was made in different respects:
ferent substrates is denoted. Most material 1) Only substrates from which at least 50 samples
derives from southern Sweden, especially the were taken are included in Table 1. Chara con-
provinces ofSmaIand and Skane (= Scania). The stitutes an exception, depending on its richness of
waters in this part of Sweden exhibit a very broad species. 2) Abundance is not reported in Table 1,
range of varying character, from decidedly oligo- only occurrence. 3) In the computer sheets, lakes,
trophic to hypertrophic, from oligohumic to poly- ponds, running waters and mires are separated,
humic environments, etc. but in Table 1 they are united. However, in the
Zooplankton material was collected with a discussion the additional information obtained
water sampler (l or 5 1). The samples were filtered from the computer sheets will be considered and
through a 20 J-lm mesh net, and the concentrates sometimes referred to in the text.
were fixed with formalin and counted. In most In the table the planktic environments are
cases net samples were collected as well; they mentioned first (also the mire hollows might be
were studied alive, focusing on species which are regarded as an open-water environment in minia-
difficult to identify in a fixed condition. ture). Then the benthic substrates are brought up,
Periphyton was collected in the following way: followed by submersed vegetation, floating-leaved
moist macrophyte material of at least 0.51 was plants and helophytes. An artificial substrate,
squeezed into a bottle, dipped again and cotton, is given at the end of the series. The cotton
squeezed. This was repeated several times. The was dipped into the water for some days and then
sample was investigated alive. Subsamples were removed for investigation.
sucked up several times and analyzed microscopi- The figure' l' in the table means that the species
cally until no additional species were encountered. has been found in 1-10% of the samples collected
Densities were estimated on a scale from 1 to 10, from the substrate in question, as against 10-50%
where the Figure 1 designated stray individuals, 6 for figure '2' and > 50% for figure '3'. The latter
a common occurrence and 10 abundant occur- figure was obtained only in plankton and only for
rence. Periphytic and benthic species were seldom one brachionid, Keratella c. cochiearis, perhaps
found in high abundance. By investigating the the most common freshwater metazoan in the
epifauna of stones and logs an area of world. Interpretation of results will focus on rank
100-200 cm2 was scraped off. The material was 2- and 3-occurrences, but some attention will also
sucked up into a bottle directly from the knife, be paid to the patterns of distribution on various
being kept at the substrate (air was slowly let out substrates.
from the bottle, causing an influx of water and As regards the sum of species, the planktic
scraped-off material). environment dominates strongly over other 'sub-
For acquiring benthos a small pot was slowly strates'. This may partly depend on the very high
moved over the bottom, whereby a layer of bottom number of plankton samples taken, but certainly
material, some millimeters thick, was removed. also upon a generally higher tendency to a free-
An area of c. 100-200 cm2 was sampled. The swimming life in brachionids than in most other
material was analysed alive, 20-30 sub samples rotifer families. The following species were usually
per sample. The estimation scale from 1 to 10 was found in the plankton of lakes and/or running
used in this case as well. waters: Brachionus anguiaris, Kellicottia iongispina,
Table 1. Occurrence of brachionid rotifers on different substrates. Only substrates with at least 50 samples are included (exception: Chara). The figure '1' means
that the species was found in 1-10% of the samples from the respective substrate, while '2' corresponds to 10-50% and '3' to >50%.

"a '"
:a
~ ~
s ~
~ ~
..c ] ;; c
0 ~ ..c ~
~.D C
" "c ~
Co .;.
~ ..c ~ :s
~c
~ OJ) .D ~
S "
0 ~ ~ z c ~
~ 0 § .; :s '"
gj s 6 ~
~
0 "
c -'" ] .§ .9 .;. a
~ e? c '0 § .~
'" ~ a oS
"
c0 '§ ~ .D 0 ~ ~ ~ 0 ~ ~ .~
~
:;;; ..c
~ "0 ... 5 ..c
0 :3 c
'"
..c
"'"
Co >< ~ '5'" gj gj
i:? ..c
c
c ><
~ ~ ~ 'g ~'".D .~
~
..c ..c s til a'" ""Co ·s g
.5 " s'" ;;'"
:§" " '" ..c 0 0 Co
f'"
Co '0 cr " "c Co
'"
'~" ~
~ ~ ~
i:C'" ::E ~
:Ef-< ~ ~ ti: U ....l ::E ....l
'" Z ~ ~ U 6' ~
1 f-< U
'" '" '" ~ '"" '"
Number of samples 4838 50 215 336 273 53 59 176 27 122 61 94 194 78 60 77 125 136 67 161 234 72 58 74 67 52 130 97 50

Anuraeopsis f/Ssa (Gosse)

Brachionus a. angularis (Gosse)
Brachionus budapestinensis (Daday)
Brachionus calyciflorus (Pallas)
Brachionus I. leydigi (Cohn)
Brachionus patulus (Mull.)
Brachionus q. quadridentatus (Hermann)
Brachionus q. brevispinus (Ehrbg) 2
Brachionus u. urceo/aris (L.) 2
Brachionus u. rubens (Ehrbg) 1
Kellicottia longispina (Kellicott) 2
Keratella c. cochlearis (Gosse) 2 2
Keratella c. hispida (Lauterborn)
Keratella c. macracantha (Lauterborn)
Keratella c. robusta (Lauterbom) 2
Iferatella c. tecta (Gosse)
Keratella hiemalis (Carlin)
Keratella i. irregularis (Lauterborn)
Keratella i. wartmanni (Asper and Heuscher)
Keratella paludosa (Lucks) 2
Keratella q. quadrata (Mull.)
Keratella serrulata (Ehrbg) 2 2
Keratella ticinensis (Callerio)
Keratella valga (Ehrbg)
Notholca acuminata (Ehrbg)
Notholca caudata Carlin
Notholca squamula (Miill.)
Platyias quadricornis (Ehrbg)

Sum of species 22 4 2 4 2 6 2 4 2 0 6 6 4 9 3 2 10 10

......
W
\,0
140

Keratella cochlearls (with its different forms), waters; e.g., isoetids (Lobelia, Isoetes) prefer oligo-
K. hiemalis, K. irregularis, K. quadrata and No- trophic lakes, Fontinalis running waters and
tholca caudata. In contrast, a pronounced Sphaerotilus strongly polluted environments.
heleoplanktic (pond-preferring) occurrence was Thus an inclination of a rotifer for a certain sub-
found for Anuraeopsis jissa, Brachionus patulus, strate often also reflects its preference for the
B. quadridentatus, Keratella serrulata, K. ticinen- habitat as a whole.
sis, K. valga, Notholca squamula and Platyias
quadricornis.
The most typical cold-water eulimnoplankters, Discussion
viz. Keratella cochlearls macracantha, K. hiemalis
and Notholca caudata, were not found in any Relation to substrate and habitat
benthic or periphytic habitat, probably because
no vegetation was investigated during winter. On There is no reason to suspect a chemical con-
the other hand, brachionids tolerating warm water finement to a certain substrate for any brachionid
often appeared in non-planktic habitats as well, similar to what has been found for certain sessile
but then preferably in environments containing rotifers (Wallace, 1980). The high occupation fre-
large areas of open water (floating-leaved vege- quency of cotton taken in concert with the absence
tation, etc.), in which the rotifers may have been of brachionids on some of the available plants,
swimming freely among the plants. (The method instead suggests an avoidance of certain natural
used makes it impossible to distinguish between substrates. It is impossible to decide if such an
different degrees of connection to the plants or avoidance depends on chemical repellants or on
bottom substrates.) an indirect influence. The nature of the periphytic
When studying the different sums of species for algal growth is probably of importance, but was
different substrates, the total number of samples not investigated in the present connection. It may
from the respective substrates has to be con- be supposed that the physical structure of a sub-
sidered. However, even if doing so, it is rather strate plays a role, and this could be one reason
evident that some substrates and habitats are for the attractiveness of the cotton.
more attractive than others. Sphaerotilus, for Based on results presented earlier it is evident
instance, is avoided by all brachionids, as well as that different species prefer different habitats, e.g.
by the members of most other rotifer families. lakes vs. ponds or mire hollows. However,
This is in contrast to the case for ciliates, where species showing a very similar pattern in Table 1
some species even occur in> 50% of the samples may be found to differ strongly when subjected to
(to be published in future papers). Only Keratella more detailed examination. Thus, in the south-
serrulata was found together with Utricularia, Swedish mire Akhultsmyren, Keratella paludosa
exclusively in most samples from mires. The low and K. serrulata were very clearly segregated, the
preference for this substrate by brachionids is former almost exclusively occurring in stands of
surprising with regard to its attractiveness for Carex rostrata and among Sphagnum apiculatum,
many other rotifers (Wallace, 1980; Pejler & the latter close to sediments and, especially, in the
Berzil)s, in press). In general, the benthic sub- free water of mire hollows and soligenous fen
strates do not seem to be very popular among soaks ('drag' in Swedish) as well as among
brachionids, and for the periphytic habitats the Utricularia (Berzil)s, unpublished). On the whole,
power of attraction varies strongly, even between K. serrulata tends more to a planktic life, e.g. also
macrophytes living in about the same way, as occurring in the zooplankton of acidified lakes
between Phragmites and Typha. The high occupa- (Morling & Pejler in press). Hauer (1935) desig-
tion frequency of the artificial substrate, cotton, is nates K. paludosa as a typical bog species
remarkable. (,Leitform im engeren Sinne'), while K. serrulata
Different macrophytes prefer different types of is considered to have a broader ecological range.
 141

Other niche-creating factors occur at rather low pH -values, especially Keratella

serrulata but also K. ticinensis and K. valga
Reasoning from the competitive exclusion prin- (BerziJ;ls & Pejler, 1987). K. paludosa surely
ciple, each species should have its own ecological belongs to this category as well, but it did not
niche, and the question arises as to which forces reach the '50-sample block' needed for inclusion
are important for this diversification. It is evident in the diagrams of the referred paper. Another
from the above that the choices of substrate and brachionid, Brachionus urceolaris sericus
macrohabitat are far from sufficient for this expla- (Rousselet), is a specialist on very acid conditions
nation, and this might be the proper place for a (Morling & Pejler, in press) but it was not included
discussion of other important factors. in the present material because of the lack of such
The relation to temperature divides the brachio- localities in southern and central Sweden. Humic
nids into separate groups. Keratella hiemalis and material may have a varying toxicity depending on
all Notholca species are cold stenothermal; the the pH-level (e.g., Petersen & Persson, 1987).
other species are eurythermal or warm steno- From Pejler & BerziJ;ls (in press d) it appears that
thermal (Pejler & BerziJ;ls, in press a). Similarly, the following brachionids have an unusually good
some species evidently can tolerate lower levels of ability to withstand a high concentration ofhumo-
oxygen concentration, whereby especially Keratella limnic substances; Keratella serrulata, K. ticinen-
ticinensis, K. valga, K. hiemalis, Platyias quadri- sis, K. valga, Brachionus patulus, B. quadridentatus
corn is, Brachionus patulus and Anuraeopsis fissa and Anuraeopsis fissa.
ought to be mentioned (Pejler & BerziJ;ls, in press For co-occurring species a differential choice of
b). For both temperature and oxygen the spatial food may help to prevent competition. Thus, all
and temporal segregation of the separate species Notholca species seem to prefer diatoms, in con-
is much more pronounced within the same locali- trast to all other brachionids (Thane-Fenchel,
ties than when comparing different waters as 1968; Pourriot, 1977; May, 1980a, b; Laxhuber&
done in the regional material presented here Hartmann, 1988). There are also differences
(cf. BerziJ;ts & Pejler, 1989). between other genera and species although per-
Also with respect to trophic degree, there are haps not equally clear-cut (Pourriot, 1977; Pejler,
obvious differences existing between different 1977; Starkweather, 1980; Bogdan et aI., 1980).
species. Thus, Brachionus angularis, B. calyci- When offered the same food, Keratella cochlearis
jlorus, Keratella irregularis and Anuraeopsis fissa and Brachionus angularis were influenced in dif-
may be regarded as indicators of eutrophy, ferent ways, the former behaving as a K -strategist,
Kellicottia longispina rather the reverse (Pejler, the latter as a r-strategist (Walz, 1983). Finally, a
1965, 1983; Pejler & BerziJ;ls, in press c). In some varying sensitivity to predation may be decisive for
cases different forms of the same species are good the environment where a species will be able to
indicators of different trophic degree; K. coch- live (Williamson, 1983).
learis macracantha is found only in oligotrophic
lakes during summer (but during winter in
eutrophic lakes as well), K. eachlearis tecta only Concluding remarks
occurring during summer in eutrophic environ-
ments (Pejler, 1962b). As regards K. quadrata, the In conclusion, it seems possible with present
results are contradictory (Pejler, 1965, p.478; knowledge to delineate separate ecological niches
Pejler & BerziJ;ls, in press c). Probably different for all brachionid rotifers. This also agrees with
ecological races occur, the oligotrophic form results and opinions put forward by Makarewicz
belonging to the 'frenzeli-group' (Carlin, 1943). & Likens (1975) and Miracle (1977). Con-
A different tolerance to deviating chemical sequently, our results contradict ideas of a
environments certainly also plays an important 'plankton paradox' suggested by Hutchinson
niche-creating role. Thus, some brachionids can (1961) and supported by Ghilarov (1984). In fact,
142

these ideas are also contradicted by more recent Egg-carrying is another phenomenon con-
results on Daphnia eueullata and D. galeata, two sidered to constitute an adaptation for a planktic
coexisting species used by Ghilarov in support of life. According to the general formulations by
his position. These two species show a pro- Koste (1978) and Sudzuki (1957), all brachionids
nounced difference in their vertical distribution in possess this habit irrespective of whether they are
the Polish lakes studied by Pijanowska & planktic or not. This is in contrast to the condition
Dawidowicz (1987) and are therefore likely to in some other rotifer families, where only true
have a different vulnerability to predation. planktic species (and not even all of these) show
Some species, like Keratella paludosa and this characteristic. In the genus Notholca, eggs are
Notholea eaudata, seem to be relatively restricted carried for a very short time (cf. Pejler, 1962a).
in their ecology, while others are very euryecious. Another feature typical of planktic organisms is
The most extreme example of the latter category the possession of varying types of protrusions
is Keratella eochlearis, which shows an enormous (spines, etc.). These have been interpreted as
morphological variation, and is composed of means of increasing the buoyancy. However, in
several forms (Pejler, 1980). The morphological brachionids their variation in relation to viscosity
variation is certainly underlaid by genetical and is the reverse of what is expected, the longest
physiological differences which may explain why extensions being found in cold water, when visco-
different forms exhibit different ecological require- sity is highest (Pejler, 1962b). In addition, the
ments. spines are much longer in oligotrophic than in
eutrophic lakes during summer at the same tem-
perature (op. cit.). Thus the most long-spined
Adaptations to different environments forms, Kellicottia longispina, Keratella cochlearis
macracantha and f. frenzeli of K. quadrata, are
The brachionids show a smooth transition from a found in cold water and/or oligotrophic waters,
non-planktic via a semiplanktic and heleoplanktic while the short-spined Braehionus angularis and
to a eulimnoplanktic way of life. One adaptation Keratella cochlearis tecta are typical of eutrophic
of true eulimnoplankters, discussed by de waters during summer (Pejler & Berzi1).s in press
Beauchamp (1909), is the disappearance of the c). Long-spined forms are characteristic of trans-
foot. However, a foot is also lacking in some parent water and may provide protection against
species dwelling in very small open waters, like visual predators (cf. Nilsson & Pejler, 1973,
mire hollows, and in close connection to plants pp.69-70). Evidently, non-visual predators like
and sediments. Thus, all Keratella species are cyclopoids and Asplanehna also avoid the long-
devoid of a foot, even the semi- or non-planktic spined forms as suggested by Stemberger &
K. paludosa, K. serrulata, K. ticinensis and K. val- Evans (1984) and by Stemberger & Gilbert (1984,
gao Certainly these species must differ from 1987).
coexisting forms of Braehionus and Platyias which In addition, soft-bodied forms are more readily
may be fixed to a substrate when feeding. On the ingested than loricated rotifers (Anderson, 1970;
other hand, some species with feet are able to Gilbert & Williamson, 1978; Stemberger, 1982;
invade the pelagic zone. B. angularis may become Williamson, 1983). It is a striking fact that the
planktic during blooms of cyanobacteria ('blue- species with the longest spines, Kellicottia
green algae'), when they use these phytoplankters longispina, has a weaker lorica than most Keratella
as a substrate, as observed by Wesenberg-Lund forms. These two adaptive traits might be con-
(1930, p. 119). B. urceolaris rubens, on the other ceived to compensate for each other. Forms living
hand, is mainly found on Daphnia and other in acid waters often have an unusually thick lorica
cladocerans (Koste, 1978). The possession or with diverse structures. Thus, the extremely acido-
lack of an adhesive organ surely contributes to biontic form Braehionus ureeolaris serieus is dis-
determining the ecological niche of a species. tinguished from B. u. urceolaris through its
 143

strongly sculptured lorica (Koste 1978), and the Bateman, L. & c. Davis, 1980. The Rotifera of hummock-
hollow formations in a poor (mesotrophic) fen in
same is characteristic of Keratella paludosa
Newfoundland. Int. Revue Hydrobiol. 65: 127-153.
(Hauer, 1935; Wulfert, 1956) and K. serrulata Beauchamp, P. M. de, 1909. Recherches sur les Ro@:res: les
(Hendelberg et al., 1979), as well as in several formations tegumentaires et l'appareil digestif. Arch. Zool.
mire-living rotifers of other families. This might be expo gen., ser. 4, 10: 1-410.
explained by the fact that fishes are more or less BerziJ;ls, B. & B. Pejler, 1987. Rotifer occurrence in relation
to pH. Hydrobiologia 147: 107-116.
lacking in such waters, whereby their role as
Bogdan, K., J. Gilbert & P. Starkweather, 1980. In situ
predators is taken over by invertebrates (e.g., clearance rates of planktonic rotifers. Hydrobiologia 73:
Nyberg, 1984). These invertebrate predators gen- 73-77.
erally prey upon smaller animals than fishes do, Carlin, B., 1943. Die Planktonrotatorien des Motalastrom.
and consequently rotifers need a stronger lorica as Zur Taxonomie und Okologie der Planktonrotatorien.
Medd. Lunds Univ. limnol. Instn. 5: 1-256.
a protection. Possibly a thick lorica may also
Donner, J., 1964. Die Rotatorien-Synusien submerser
serve as a protection against plants feeding on Makrophyten der Donau bei Wien und mehrerer
small animals, and it ought to be pointed out that Alpenbiiche. Arch. Hydrobiol. Suppl. 27: 227-324.
K. serrulata is the only brachionid found among Donner, J., 1970. Die Radertierbestande submerser Moose
Utricularia, where it occurs frequently (see der Salzach und anderer Wasser-Biotope des Fluss-
gebietes. Ibid. 36: 109-254.
Table 1). A pustulation is often found, not only in
Donner, J., 1972. Die Radertierbestande submerser Moose
mire-living rotifers, as part of an interspecific or und weiterer Merotope im Bereich der Stauraume der
infraspecific variation. Attempts at explaining its Donau an der deutsch-osterreichischen Landesgrenze.
adaptive value (which it certainly has) are almost Ibid. 44: 49-114.
never found in the literature. Only Ruttner- Edmondson, W. T., 1944. Ecological studies of sessile
Rotatoria. Part I. Factors affecting distribution. Ecol.
Kolisko (1972, p. 173) links the occurrence of
Monogr. 14: 31-66.
Keratella cochlearis hispida (a pustulated form- Edmondson, W. T., 1945. Ecological studies of sessile Rota-
series) with the turbulence of the water; these toria. Part II. Dynamics of populations and social struc-
rotifers being said to occur mainly in shallow and tures. Ibid. 15: 141-172.
small waters (an observation which agrees fairly Francez, A.-J., 1984. Rotiferes sessiles observes en
Auvergne. Cahiers Naturalistes, Bull. nat. paris., N.S. 40:
well with the authors' experience from south and
73-80.
central Sweden). However, with regard to the fact Francez, A.-J. & J. Devaux, 1985. Repartition des rotiferes
that biotic relations have been increasingly found dans deux lacs-tourbieres du Massif Central (France).
to be important in ecology during recent decades, Hydrobiologia 128: 265-276.
it may be suspected that these structures function Ghilarov, A. M., 1984. The paradox of the plankton recon-
as protections against predators as well. sidered; or, why do species coexist? Oikos 43: 46-52.
Gilbert, J. & c. Williamson, 1978. Predator-prey behavior
and its effect on rotifer survival and associations of
Mesocyclops edax, Asplanchna girodi, Polyarthra vul-
Acknowledgements garis, and Keratella cochlearis. Oecologia 37: 13-22.
Hauer, J., 1935. Rotatorien aus dem Schluchseemoor und
seiner Umgebung. Verh. naturw. Ver. Karlsruhe 29:
Data analysis and subsequent routine work was
47-130.
supported by the National Environmental Protec- Hendelberg, M., G. Morling & B. Pejler, 1979. The ultra-
tion Board of Sweden. Among the persons structure of the lorica of the rotifer Keratella serrulata
engaged in this treatment we especially wish to (Ehrbg). Zoon 7: 49-54.
mention Jan Bertilsson. Hutchinson, G. E., 1961. The paradox of the plankton. Am.
Nat. 95: 137-145.
Koste, W., 1978. Rotatoria. Die Radertiere Mitteleuropas.
References Ein Bestimmungswerk begr. von Max Voigt. Uberordning
Monogononta. Vol. 1-2. 673 pp. + 234 pI.
Laxhuber, R. & u. Hartmann, 1988. The influence oftem-
Anderson, R. S., 1970. Predator-prey relationships and
perature on the life cycle of the cold-stenothermal rotifer
predation rates for crustacean zooplankters from some
Notholca caudata Carlin. Verh. int. Ver. Limnol. 23:
lakes in western Canada. Ca. J. Zool. 48: 1229-1240.
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144

Makarewicz, J. & G. Likens, 1975. Niche analysis of a logical effects of humic materials under acidified condi-
zooplankton community. Science 190: 1000-1003. tions. Sci. Tot. Envir. 62: 387-398.
Martin, L. V., 1976. Rotifers in the Sphagnum pools on Pijanowska, J. & P. Dawidowicz, 1987. The lack of vertical
Thursley Common. Microscopy 33: 90-93. migration in Daphnia: the effect of homogenously dis-
Martin, L. V., 1977. Rotifers in the Sphagnum pools on tributed food. Hydrobiologia 148: 175-181.
Thursley Common. Part 2. Ibid. 33: 236-241. Pourriot, R, 1977. Food and feeding habits of Rot ifera. Arch.
May, L., 1980a. Studies on the grazing rate of Notholca Hydrobiol. Beih. 8: 243-260.
squamula Muller on Asterionella formosa Hass. at dif- Ruttner-Kolisko, A., 1956. Der Lebensraum des Limnopsam-
ferent temperatures. Hydrobiologia 73: 79-81. mals. Verh. dtsch. zool. Ges. Hamburg 1956: 421-427.
May, L., 1980b. On the ecology ofNotholca squamula Muller Ruttner-Kolisko, A., 1972. Rotatoria. Binnengew1lsser 26:
in Loch Leven, Kinross, Scotland. Ibid. 73: 177-180. 99-234.
Meuche, A., 1939. Die FaunaimAlgenbewuchs. Nach Unter- Stemberger, R, 1982. Mechanisms controlling selection and
suchungen im Litoral ostholsteinischer Seen. Arch. Hydro- rates of predation on rotifers in Cyclops bicuspidatus
bioI. 34: 349-520. thomasi. Ph. D. Thesis, Univ. Mich. 95 pp.
Miracle, M. R, 1977. Migration, patchiness, and distribution Stemberger, R & M. Evans, 1984. Rotifer seasonal suc-
in time and space of planktonic rotifers. Arch. Hydrobiol. cession and copepod predation in Lake Michigan. 1. Great
Beih. 8: 19-37. Lakes Res. 10: 417-428.
Morling, G. & B. Pejler, in press. Acidification and zoo- Sternberger, R & 1. Gilbert, 1984. Spine development in the
plankton development in some west-Swedish lakes rotifer Keratella cochlearis: induction by cyclopoid cope-
1966-1983. Limnologica 21. pods and Asplanchna. Freshwat. BioI. 14: 639-647.
Nilsson, N.-A. & B. Pejler, 1973. On the relation between fish Stemberger, R & J. Gilbert, 1987. Defenses of planktonic
fauna and zooplankton composition in North Swedish rotifers against predators. In: W. C. Kerfoot and A. Sih
lakes. Rep. Inst. Freshw. Res. Drottningholm 53: 51-77. (eds.). Predation. Direct and indirect impacts on aquatic
Nyberg, P., 1984. Impact of Chaoborus predation on plank- communities. 386 pp.
tonic crustacean communities in some acidified and limed Sudzuki, M., 1957. Studies on the egg-carrying types in
forest lakes in Sweden. Ibid. 61: 154-166. Rotifera. II. Genera Brachionus and Keratella. Zool.
Pejler, B., 1962a. Morphological studies on the genera Mag., Tokyo 66: 11-20. (Japanese, with English summary.)
Notholca, Kellicottia and Keratella (Rotatoria). Zool. Thane-Fenchel, A., 1968. Distribution and ecology of non-
Bidr. Uppsala 33: 295-309. planktonic brackish-water rotifers from Scandinavian
Pejler, B., 1962b. On the variation of the rotifer Keratella waters. Ophelia 5: 273-297.
cochlearis (Gosse). Ibid. 35: 1-17. Tzschaschel, G., 1983. Seasonal abundance of psammon
Pejler, B., 1965. Regional-ecological studies of Swedish rotifers. Hydrobiologia 104: 275-278.
fresh-water zooplankton. Ibid. 36: 407-515. Wallace, R, 1980. Ecology of sessile rotifers. Hydrobiologia
Pejler, B., 1977. On the global distribution of the family 73: 181-193.
Brachionidae (Rotatoria). Arch. Hydrobiol. Suppl. 53: Walz, N., 1983. Continuous culture of the pelagic rotifers
255-306. Keratella cochlearis and Brachionus angularis. Arch.
Pejler, B., 1980. Variation in the genus Keratella. Hydro- Hydrobiol. 98: 70-92.
biologia 73: 207-213. Wesenberg-Lund, C., 1930. Contributions to the biology of
Pejler, B., 1987. Bruno BerziJ;ls in memoriam. 1909-1985. the Rotifera. II. The periodicity and sexual periods. K
Ibid. 147: 1-2. danske vidensk. Selsk., nat.-math. Afd., Raekke 9, Bd 2,
Pejler, B. & B. BerziJ;ls, 1989. Rotifer occurrence in no. 1. 230 pp.
relation to temperature. Ibid. 175: 223-231. Williamson, c., 1983. Invertebrate predation on planktonic
Pejler, B. & B. BerziJ;ls, 1989. Rotifer occurrence III rotifers. Hydrobiologia 104: 385-396.
relation to oxygen content. Ibid. 183: 165-172 Wulfert, K, 1951. Das Naturschutzgebiet auf dem Glatzer
Pejler, B. & B. Berzilfs, 1989. Rotifer occurrence and Schneeberg. Die R1ldertiere des Naturschutzgebietes.
trophic degree. Ibid. 172: 171-180 Arch. Hydrobiol. 44: 441-471.
Pejler, B. & B. BerziJ;ls, 1989. Rotifer occurrence in Wulfert, K, 1956. Die R1ldertiere des Teufelssees bei
relation to water colour. Ibid. 184: 23-28 Friedrichshagen. Ibid. 51: 457-495.
Petersen, R & u. Persson, 1987. Comparison of the bio-
Hydrobiologia 186/187: 145-152, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 145
© 1989 Kluwer Academic Publishers.

Temporal analysis of clonal structure in a moss bdelloid population

Claudia Ricci 1, Manuela Pagani 2 & Anna Maria Bolzern 2

1 Dipartimento di Biologia Animale, via Accademia Albertina 17, Torino, Italy; 2 Dipartimento di Biologia,
via Celoria 26, Milano, Italy

Key words: electrophoresis, isozymes, moss population, bdelloid rotifers

Abstract

Clonal structure of a population of Macrotrachela quadricornifera (Rotifera, Bdelloidea) from a terrestrial

moss in Northern Italy, was investigated over a 16 month period. Every month, 40-60 specimens of
M. quadricornifera were collected from about 0.1 m2 of moss. The individual animals were homogenized
and their isozyme phenotypes analyzed by electrophoresis on vertical polyacrylamide gel. One enzyme,
phosphoglucose isomerase (PGI), was used as a marker to distinguish the different clones present in the
sample. A few clones were established from the rotifers sampled and patterns of esterases (J.. and /3, and
malic enzyme were studied. Nine electrophoretic patterns for PGI were seen. One was dominant, a second
was almost always present, but in lesser amounts. The remainder were present occasionally.
There seemed to be no seasonal replacement of the clones and the composition of the population
appeared to be unaffected by variations in temperature. Relative humidity seemed to be the more
important factor in regulating the number of electromorphs of the rotifer population.

Introduction tors affecting the clonal dynamics of partheno-

genetic zooplankton (Hebert, 1977; Korpe1ainen,
Much attention has been paid to the seasonal 1986).
structure of parthenogenetic populations of Lakes and ponds may be considered predicta-
cladocerans and rotifers in freshwater plankton. ble environments that vary through the year
Here, several clones coexist in the same environ- without dramatic changes. The situation faced by
ment, with temporal changes in their relative fre- animals inhabiting terrestrial mosses, where con-
quencies (e.g. Hebert, 1977 ; Young, 1979; King & ditions change dramatically and rapidly, is dif-
Zhao, 1987; Hebert et al., 1988). In contrast to ferent.
the competitive exclusion principle, the co- The present study concerns the clonal ecology
existence of clones with partially or totally over- of a bdelloid rotifer, Macrotrachela quadricorni-
lapping niches can be maintained by changes in jera, in a terrestrial moss. Bdelloid rotifers repro-
the environment which they inhabit. It is generally duce through obligatory apomictic the1ytoky and,
assumed that different clones compete for the although they are aquatic animals, they can
same resources with different competitive abilities inhabit water bodies and terrestrial mosses
and that varying the conditions of the environ- because they can respond to desiccation by enter-
ment may shift the selective advantage from one ing an anhydrobiotic state. When anhydrobiotic,
clone to another over time. Changes in environ- any bdelloid can be a propagule, able to colonize
ment are commonly recognized as the major fac- a new environment and give rise to a clonal popu-
146

lation: its success in establishing such a popula- examined under a stereo-microscope and the
tion will depend on its ability to survive the rotifers collected with a Pasteur pipette. Each
anhydrobiotic state and on its growth rate. It rotifer was identified under the light microscope.
seems reasonable to assume that competition M. quadricornifera was easily recognized by the
among different clones in the same habitat will two appendages on its foot, dorsal to the spurs,
affect the clonal composition of a population and by its mastax.
throughout the different seasons. Macrotrachela quadricornifera Milne 1886 is a
At present, there is no experimental evidence cosmopolitan species-complex which lives in
that any species of bdelloid rotifers is composed ponds, lakes and terrestrial mosses. Despite the
of different clones, but most of parthenogenetic many varieties described (Donner, 1965), the dif-
morpho species investigated so far, have been ferences in morphology of the specimens con-
found to be composed of several clones (Parker, cerned in this study were considered unimportant
1979). Moreover, a recent investigation of electro- and the animals were classified as M. quadricorni-
phoretic patterns of six enzymes from 5 strains of fera sensu strictu.
the same morpho species M. quadricornifera (un- To obtain a total of about 50 specimens, more
published) has revealed that the different strains moss than usual had to be collected on some
can be distinguished by their enzymatic patterns. occasion. When the number of rotifers were exces-
Therefore, as is will known for other partheno- sive, some animals were isolated and cultured
genetic species (e.g. Snell, 1979; King, 1980; under laboratory conditions. The animals were
Hebert & Crease, 1983; Lynch, 1984), bdelloid fed on a mixture of algae (Chlamydomonas
species also consist of many different clones. reinhardti) and bacteria (E. coli), sometimes with
yeast (Saccharomyces cerevisiae) added.
Shortly after identification, the animals were
Material and methods processed for electrophoretic investigation.
Extracts for electrophoresis were prepared by
Specimens of Macrotrachela quadricornifera were homogenizing each bdelloid in 5 ,Ill sample buffer
collected from moss on a wall near Milan, (0.05 M Tris HCI, pH 8, + 10% sucrose + 0.02%
Northern Italy, between February 1987 and June bromophenol). The entire extract was loaded into
1988. Each moss sample was approximately a sample slot. Electrophoretic separation was
0.1 m2 • Samples were taken every 45 days from performed in 0.8 mm thick 7.5% polyacrylamide
February to October 1987 and monthly from gel with 4.5 % of N,N' -methylenebisacrylamide,
November 1987 to June 1988. On some occasion using a mini slab vertical gel apparatus (Scientific
no M. quadricornifera were found in the moss. Idea, Corvallis, U. S.A.) in a continuous system of
The moss had not been disturbed, for instance 0.082 M Tris-borate buffer, pH 8.5. Since the size
by pesticide treatments. No attempt was made to of the rotifers enabled analysis of a single locus
classify it botanically since the presence of system only, phosphoglucose isomerase, PGI
bdelloid species is not thought to be related to (E.C. 5.3.1.9) was used to distinguish electro-
moss species (Dobers, 1915; Bartos, 1951). morphs. Previous experience with bdelloid elec-
The moss was brought to the laboratory and trophoresis (unpublished) showed that PGI has
the rotifers were extracted as soon as possible, good staining resolution and is polymorphic in
usually on the day of sampling. For extraction, the this species, therefore, although insufficient, it is
moss was hydrated, suspended in a beaker with effective for detecting at least part of the varia-
some water, immersed into an ice bath and heated bility of the population. Two additional enzymes,
from above with a ordinary incandescent lamp. malic enzyme (ME) and esterases (Est (J. and jJ),
Within a couple of hours, rotifers and other moss were tested in the clonal populations when
inhabitants moved downward to escape the light possible. The staining technique of Harris &
and heat. The contents of the beaker were Hopkinson (1976) was used for the three
 147

enzymes, with minor modifications (unpub- clonal diversity in each sample. All metereological
lished). Since the data were mostly for a single parameters were calculated over 10, 15 and 20
locus, only PGI electromorphs are considered days prior to sampling.
and are listed according to speed of migration
through the gel, with 1 being slowest and 6 being
fastest. Results
Physical parameters of the sampling site, such
as rain, relative humidity and average daily air 550 morphologically classified individuals of
temperature, were are also related to the changes M. quadricornifera were screened, and 9 electro-
in clone diversity over the investigation period. phoretic patterns for the enzyme PGI were dis-
Rain, humidity and temperature were obtained tinguished (Fig. 1). The term 'clone' is used as a
from the Brera Metereological Station (Milan). synonym for electromorph, and does not neces-
The Shannon index, the Evenness or, simply, sarily refer to a single lineage.
the number of electromorphs were used to express The distribution of the clones throughout the

1/2 2/2 2/4 1/3 3/3 3/4 4/4 5/5 5/6 M2

mm

- CJ
5

-
4

- 10

-
15
5

6
20
Fig. 1. Patterns for phosphoglucose isomerase (PGI) of the population of M. quadricornifera. M2 is a laboratory clonal population
used as marker. Migration distances are expressed in mm (right axis). Banding patterns are numbered (1-6) according to speed
of migration through the gel.
148

1.5';

1/2 4.7%

Fig. 2. Percentages of the electromorphs throughout the study period. See text for explanation.

n.,. 4
c
1
0
n
0:-
~
3

1987 1988

Fig. 3. Temporal variation of the number of electromorphs from each collection.

 149

Table 1. Number of M. quadricomifera, Shannon index, number and percentage of the electromorphs from each collection.

Electromorphe
Sample N Shannon
date M.q. index 1/2 2/2 2/4 1/3 3/3 3/4 4/4 5/5 5/6

N % N % N % N % N % N % N % N % N %
Feb '87 41 0.60 2.5 2 5 2.5 37 90
Apr '87 62 0.11 2 61 98
May'87 0
Jun '87 0
Jul '87 42 1.35 21 50 4 10 17 40
Sep '87 74 0.20 1.4 1.4 72 97.2
Nov'87 59 1.20 1.7 32 54.2 25 42.4 1.7
Dec '87 74 0.34 I 1.4 3 4 70 94.6
Jan '88 43 0.84 1 2.4 8 18.6 34 79
Feb '88 0
Mar '88 0
Apr '88 0
May'88 57 0.21 2 3.5 55 96.5
Jun '88 98 1.61 19 19.4 2 2 3 3 10 10.2 62 63.4

investigation period was partitioned as shown in decreased to less than 50 % on some occasions.
Fig. 2: a very common electromorph (4/4) repre- The minor clones occurred sporadically. There
sented approximately 79% of all rotifers, a second was no evidence of seasonal succession of any
(3/3) about 14%, a third (1/2) approximately 5%, clone, as the dominant clone was present
and a fourth (3/4) 1%. Up to 5 less frequent throughout the year and no replacement of the
electromorphs, occurring as less than 0.5%, are others was apparent.
grouped into a single zone containing electro- Temporal variation of clones was analysed in
morphs 2/4 (0.2%), 5/5 (0.2%), 5/6 (0.2%), 2/2 relation to physical parameters. Relative humidity
(0.4%) and 1/3 (0.5%). (r = 0.94, P ~0.001), calculated as the average
Clonal diversity, the number of clones recorded value over 10 days prior to sampling (Fig. 4) was
on each sampling occasion, is shown in Fig. 3. No most closely related to the number of clones. No
bdelloid rotifers were extracted from moss relationship was detected between number of
samples taken in Mayor June 1987. Both of these clones and mean temperature over 20 (r = - 0.20,
collections were made after very heavy rain, and P> 0.5) or 10 days prior to sampling (r = - 0.22,
a washing out of the moss by the rain was P> 0.5). The same was true of rainfall (average
suspected. The ground below the wall was over 10 days before sampling) and number of
sampled without success. In February, March clones (r=0.52, P>O.I). In addition, we
and April 1988 the weather was exceptionally dry attempted to relate rainfall in the 5 days prior to
and again the extraction was unsuccessful. How- sampling to the number of clones, suspecting a
ever, the most frequent clone (electromorph 4/4) washing effect on the moss population, but no
was collected before and after both climatic dis- relationship was found (r = 0.25, P> 0.1).
turbances, suggesting continuity of occurrence in In the cultivation of clones established from
the bdelloid population. single animals, we obtained contrasting results.
Table 1 lists the numbers and percentages of Some lineages reproduced promptly when fed
the electromorphs present on the sampling dates. E. coli and Chlamydomonas, while others did not.
Clone 4/4 was almost always dominant, but The latter increased after addition of S. cerevisiae
 150

n
3

c
1
0
n
E~
::: 2

"
1

relative humidity (%)

Fig, 4, Correlation between number of electromorphs and relative humidity of the air, calculated as the average value over 10
days prior to sampling. r = 0.94, P ~ 0.001.

to the food mixture, These clones, when analyzed would have been to rear animals in order to estab-
for PGI enzymatic patterns, were the 4/4 electro- lish clonal cultures in which to screen more loci.
morpho The other loci investigated in laboratory However, we thought it more important to get
clones confirmed the results ofPGI staining: PGI information about the whole population of a
electromorphs gave similar ME and EST electro- single species, also, maintenance of clones estab-
morphs, but no composite electromorph could be lished from collected animals is often difficult.
identified because of the paucity of the cultured One could easily miss a number of clones only
clones. because of difficulties of culturing under labora-
tory conditions.
The amount of polymorphism at the PGI locus
Discussion in M. quadricornifera is very high, with as many as
6 alleles arranged into 9 different 'genotypes'. If
The most serious limitation of this study is the more loci had been examined, the number would
analysis of a single locus system, used as a simple probably have increased further, implying that
marker. This approach underestimates the num- this popUlation consists of many distinct clones,
ber of clones, since each electrophoretic clone Moreover, the sample size is not always large
probably contains several genotypes due to dif- enough to obtain a satisfactory number of
ferentiation at other loci. An alternative approach bdelloids. A greater variability might have been
 151

detected if a larger number of animals had been Lynch, 1984; Rossi & Menozzi, in press). How-
studied. ever, variations in temperature in terrestrial
The results suggest high stability of clonal mosses are so dramatic and sudden that no
structure among the rotifers investigated. One specialization is possible. In fact, there is not even
clone seemed to be dominant throughout the adaptation to summer or winter seasons.
entire period, winter and summer, sharing its Competition among clones leading to the exclu-
leadership with a second common electromorph sion of all but a few genotypes in planktonic
(3/3). The sporadic electromorphs may represent rotifers (Snell, 1977; King, 1980) has been
unsuccessful colonization events in the moss. hypothesized, while King & Zhao (1987), study-
However, bearing in mind how unstable and ing a population of Brachionus plicatilis, found that
unpredictable terrestrial moss is and how easily a several clones 'may coexist over an extensive
bdelloid rotifer should in theory colonize a habitat, period', supporting King's (1977) model of inc om-
the population stability was unexpected. plete genetic discontinuity. All authors (e.g.
Temperature and water content are the major Loaring & Hebert, 1981; Hebert & Crease, 1980;
variables affecting the life of the moss inhabitants. Weider et aI., 1987) agree that clonal diversity in
In any terrestrial moss, rapid fluctuations in tem- natural habitats is maintained by environmental
perature occur daily. A field study of the popula- disturbance and temporal heterogeneity, and that
tion dynamics of moss tardigrades showed that clones are not ecological analogues. Their dif-
humidity and rainfall ten to twenty days prior to ferences in competitive ability and intrinsic rates
sampling affect the popUlation (Morgan, 1977). of increase, coupled with habitat heterogeneity,
Here, the water content has not been investigated, shape the clonal composition of the environment.
but relative humidity of the air might also be Our results, if reliable, are discordant with this
related to it. When humidity increased, a larger idea, since we found a single dominant clone in a
number of electromorphs were detected suggest- very unstable habitat. However, because the ter-
ing that bdelloid colonization takes place when restrial moss dries out completely from time to
environmental conditions offer more resources. time, the ability to withstand anhydrobiosis may
On the other hand, when the water content is too be the determining factor for successful main-
low to sustain life, rotifers survive as anhydro- tenance of a clone. No studies of differences in
bionts. However, the new colonization is ineffec- anhydrobiotic capability among clones have been
tive and the few well established clones continue performed, but this may be worth future investi-
to dominate the habitat. Clonal competition can- gation. Moreover, the founder effect (Hebert &
not be ruled out, of course, since fluctuations in Moran, 1980) may be important in preventing
electromorph frequencies are found. However, other genotypes from colonizing the same habitat.
the clonal frequencies do not vary greatly and In conclusion, the results of this far from
clonal replacement does not appear to take place. exhaustive study show unexpected dynamics of
The differences in food requirements of the clo- clones of a strictly parthenogenetic animal in an
nal populations under laboratory conditions sug- unstable and unpredictable environment. The
gest that at least some clones may use different dominance of single clone in the moss through the
food resources in nature. This supports the study period can be attributed to the founder
hypothesis that different clones can co-occur effect, competitive superiority and anhydrobiotic
because they occupy separate niches (Loaring & viability, all of which are important factors. Only
Hebert, 1981). further investigation, involving field and labora-
Generally, the results of this study are in con- tory studies, can reveal which biological capabili-
trast to the situation in parthenogenetic inverte- ties of such a parthenogenetic population account
brates living in plankton, where seasonal clonal for its unusual dynamics.
succession is commonly observed in response to
changing temperature (Carvalho & Crisp, 1987;
152

Acknowledgements natural populations of Daphnia carinata King. Heredity

45: 313-321.
King, C. E., 1977. Genetics of reproduction, variation, and
We particularly thank Giancarlo Fava and Paolo adaptation in rotifers. Arch. Hydrobiol. Ergeb. Limno!. 8:
Bisol. Very helpful suggestions on the extracting 187-201.
procedure were given by Umberto Parenti. Linda King, C. E., 1980. The genetic structure of zooplankton
May read and provided excellent comments on populations, p. 315-328. In: W. C. Kerfoot (ed.), Evolution
and ecology of zooplankton communities. Univ. Press
earlier draft ofthe manuscript. The study has been
New England, Hanover.
supported by grant CT 84.01201.11 from CNR King, C. E. & Y. Zhao, 1987. Coexistence of rotifer
and by MPI 40% fund to C.R. (Brachionus plicatilis) clones in Soda Lake, Nevada.
Hydrobiologia 147: 57-64.
Korpelainen, H., 1986. Competition between clones: an
experimental study in a natural population of Daphnia
References magna. Hereditas 105: 29-35.
Loaring, J. M. & P. D. N. Hebert, 1981. Ecological dif-
Bartos, E., 1951. The Czechoslovak Rotatoria of the Order ferences among clones of Daphnia pulex Leydig. Oecologia
Bdelloidea. Vestnik Cs. Zool. spol. 15: 241-500. 51: 162-168.
Carvalho, G. R., 1987. The clonal ecology of Daphnia magna Lynch, M., 1984. The genetic structure of a cyclical
(Crustacea: Cladocera).1. Temporal changes in the clonal parthenogen. Evolution 38: 186-20l
structure of a natural population. J. animo Ecol. 56: Morgan, c.1., 1977. Population dynamics of two species of
453-468. Tardigrada, Macrobiotus hufelandii (Schultze) and
Dobers, E., 1915. Uber die Biologie der Bdelloidea. Int. Echiniscus (Echiniscus) testudo (Doyere), in roof moss
Revue ges. Hydrobiol. Hydro!. Supp!. zu Bd. 7: 1-128. from Swansea. J. animo Eco!. 46: 263-279.
Donner, J., 1965. Ordnung Bdelloidea. Akademie Verlag. 297 Parker, E. D., Jr., 1979. Ecological implications of clonal
pp. diversity in parthenogenetic morphospecies. Am. Zoo!. 19:
Harris, H. & D. A. Hopkinson, 1976. Handbook of enzyme 753-762.
electrophoresis in human genetics. North-Holland, Rossi, V. & Menozzi P. The clonal ecology of Heterocypris
Amsterdam, Neth. incongruens (Crustacea: Ostracoda). Ecology, in press.
Hebert, P. D. N., 1977. Niche overlap among species in the Snell, T. W., 1977. Clonal selection: competition among
Daphnia carinata complex. J. animo Eco!. 46: 399-409. clones. Arch. Hydrobiol. Ergeb. Limnol. 8: 202-204.
Hebert, P. D. N. & T. J. Crease, 1980. Clonal coexistence in Snell, T. W., 1979. Interspecific competition and population
Daphnia pulex (Leydig): another planktonic paradox. structure in rotifers. Ecology 60: 494-502.
Science 207: 1363-1365. Weider, L. J., M. J. Beaton & P. D. N. Hebert, 1987. Clonal
Hebert, P. D. N. & T. J. Crease, 1983. Clonal diversity in diversity in High-Artic populations of Daphnia pulex, a
populations of Daphnia pulex reproducing by obligate polyploid apomictic complex. Evolution 41: 1335-1346.
parthenogenesis. Heredity 51: 353-369. Young, J. P. W., 1979. Enzyme polymorphism and cyclic
Hebert, P. D. N. & c. Moran, 1981. Enzyme variability in parthenogenesis in Daphnia magna. I. Selection and clonal
Hydrobiologia 186/187: 153-156, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 153
© 1989 Kluwer Academic Publishers.

Horizontal distribution of the plankton rotifers Keratella cochlearis

(Bory de St Vincent) and Polyarthra vulgaris (Carlin) in a small
eutrophic lake

A.P. Saunders-Davies
33 Park Road, Esher, Surrey KT10 8NP, UK

Key words: horizontal distribution Keratella cochlearis, Polyarthra vulgaris, lux

Abstract

The plankton rotifers Keratella cochlearis and Polyarthra vulgaris were sampled at 10 cm below the surface
at different distances from two dissimilar shores and in the centre of a small eutrophic lake. Light and
depth were measured at each sampling point. In each case the numbers of rotifers per liter increased with
distance from the shore. There was a significant correlation between the numbers for the two species for
the two shores, but none in the centre. In the case of one shore there was a strong correlation between
rotifer numbers and supra-surface ambient light.

Introduction the water was uniformly turbid with a Secchi

depth of 10-20 cm. The South West (SW) shore
The vertical distribution of plankton rotifers has shelves much more gently and a number of the
been well-studied (George, 1970; Stewart & surrounding trees of alder (Alnus) and willow
George, 1987), but there have been fewer investi- (Salix) have shed branches which have re-rooted
gations of the horizontal distribution. Plankton and formed long 'gullies' or inlets. The height of
rotifers are known to be strongly phototactic. This the fallen branches is about 1-2 m.
paper describes the variation of rotifer numbers
with distance from two shores and relates this to
the mean supra-surface light levels. Methods

Traverses were made on 6 occasions of a gully on

Site studied the SW shore together with similar traverses of
the opposite NE shore sampling at 2.5 m and 5 m
Brooklands lake is a small (6 hectare) eutrophic intervals. Sampling was carried out 10 cm below
lake with an average depth of 1 m, and a maxi- the surface with closing bottles of 125 ml capacity.
mum of about 1.5 m. It is approximately Three samples were taken at each site. The
L-shaped. Part of the North East (NE) shore is a samples were then decanted into specimen tubes
re-entrant curve exposed to the prevailing West (4 x 15 cm), ftxed with a 10% formalin solution
and South West wind. It is comparatively free of and allowed to sediment for a minimum of 24 hr.
macrophytes. It supports a vigorous phyto- and The supernatant was carefully siphoned otT leav-
zooplankton community. At the times of sampling ing the bottom 4 ml undisturbed. These 4 ml were
154

then vigorously stirred and transferred to a during the 2 months of the study. Therefore the
perspex chamber and the numbers of rotifers were numbers in each sample were expressed as a per-
counted. centage of the total numbers for each traverse.
The light was measured with a Gossen Lunasix This avoids any weighting by occasions when the
meter fitted with an integrating cone which total numbers were high. The figures are shown in
recorded the light over a 180 0 hemisphere. Days the Table together with mean light levels, depth
on which the sky was overcast were chosen to and mean numbers liter - 1 for each site. The num-
minimise the effects of direct sunlight. bers per sample increased with progressive dis-
The temperature was measured at each sam- tance from the shore. In the case of the NE shore
pling point and found not to vary by more than both species show a significant correlation against
0.2 0 C between sites, except on one occasion light levels, with values for the coefficient of corre-
when the temperature in the centre was 2.1 0 C lation of 0.96 for Keratella, and 0.97 for Poly-
higher than in the shaded areas. arthra. This is significant at the 0.2 % level for
Polyarthra and at the 1% level for Keratella. The
slopes of the regression lines (and intercepts) were
Results remarkably similar, with values of 26. 98 %per log
lux for Polyarthra and 25.05% for Keratella. As
My major interest was the horizontal distribution might be expected from the foregoing, the correla-
of the rotifers. However the relative numbers of tion between the two species was also significant,
each species varied considerably, and the num- in fact at the 0.2 % level.
bers per liter varied by as much as a factor of 10 In the case of the SW shore there was also a

Table. Rotifer numbers as a percentage of the total numbers for the traverse, together with distances from the shores, mean
log lux, standard deviation, mean numbers per litre and depth. Percentages shown represent the mean of 6 traverses.

Table lAo Rotifer numbers, average light values and depth for the SW shore

Distance ~ (m) 2.5 5 10 15 20 25 30 35

Depth (m) 0.84 0.90 1.00 1.05 1.40 1.40 1.37
Log mean lux 8.51 9.20 9.41 9.70 9.83 9.93 9.89 9.89
Polyarthra % total 7.58 4.21 10.90 16.11 45.78 43.84
Std dev for 6 runs 3.83 3.13 6.69 8.92 9.86 6.50
Mean no/liter 57 27 55 142 389 296

K. cochlearis % total 3.36 6.05 5.88 12.32 34.83 37.57

Std dev for 6 runs 1.57 6.12 4.11 9.15 9.83 8.03
Mean no/liter 488 510 749 1481 5245 6190

Table lB. Rotifer numbers, average light values and depth for the NE shore

Distance ~ (m) 2.5 5 10 15 20 25

Depth (m) 0.90 1.20 1.40 1.38 1.50 1.48
Log mean lux 9.29 9.55 9.72 9.88 9.93 9.93
Polyarthra % total 7.84 10.71 18.51 26.63 26.60 21.19
Sd for 6 runs 6.74 2.96 7.49 8.49 5.83 6.63
Mean no/liter 1,058 1,520 3,153 4,154 4,132 3,240

K. cochlearis % total 9.29 12.20 18.27 24.71 25.55 21.72

Std for 6 runs 7.80 6.75 3.18 5.84 4.09 6.03
Mean no/liter 2,595 4,934 6,536 9,784 6,970 7,880
 155

30

28 .

26
o
+
24
+
o
22

Ij)
20
cr
w 18 .
III
2
:J 16
z
.-J 14 .
<l:
f-
0 12 +
1-
~ o
10
+
8 0

4·

0
9.2 9.4 9.6 9.8 10.0

LOG MEAN LUX

o P.vulgoris + K.cochleoris
Fig. 1. Numbers of rotifers plotted as a percentage of the total for the traverse for the NE shore as a function of mean light level
in log lux. Points represent the mean of 6 traverses. (Cross = K. cochlearis, square = P. vulgaris). Regression line shown is for
both species.

marked rise in numbers per liter as the distance Discussion

increased. These were plotted in a similar way to
those of the NE shore (Fig. 1). The correlation The profiles of the relative distribution as a
between the two species was again highly signifi- function of distance from the shore indicates that
cant with a correlation coefficient of 0.99. In this planktonic rotifers follow the pattern established
case, however there was no significant correlation for pelagic crustaceans (Burckhardt, 1910;
between light levels and the rotifer numbers. For Ruttner, 1974). Similar profiles for plankton
both species there was a slow increase during the rotifers were described by Preissler (1977),
length of the gully and then a sharp rise when the although the species present in his samples were
entrance to the gully was cleared. different.
In the centre of the lake the light levels were The strong and significant correlation, coupled
constant over the length of the traverses. In the with the known phototaxic response of the plank-
data from 6 traverses of 20 m each there was no ton rotifers concerned, argues for a causal relation
correlation between the species, and the distribu- between light levels and rotifer numbers for the
tion appeared to be completely random and inde- distribution off the NE shore. The slight drop in
pendent for each species. numbers for the last (25 m) point at first sight
appears anomalous, particularly since the point
on the graph represents the mean of 6 samples
156

and thus is not a single aberrant data point. The References

swimming speeds of K. cochlearis over a range of
temperatures averages 0.5 mID sec - 1 with a maxi- Burckhardt, G., 1910. Hypothesen und Beobachtungen uber
mum of 0.8 mm sec - 1, while Polyarthra averaged die Bedcutung der vertikalen Planktowanderung. Int.
Revue H ydrobiol. 3: 156-172.
0.6 mm - 1 with a maximum of 0.9 mm sec - 1. George, M. G., 1970. Diurnal Migration in three species of
(Saunders-Davies - in preparation). A surface rotifers in Sunfish Lake, Ontario. Limnol. Oceanogr. 15:
wind will generate currents at the surface of 1-3 % 218-223.
of the wind speed (Harvey, 1974). A wind speed Harvey, 1. G., 1976. Atmosphere and Ocean. Artemis.
of less than 0.2 ms - 1 is described as 'calm' Pedgely, D. E., 1978. A Course in Elementary Meteorology.
HMSO.
(Pedgley, 1978) but this could generate a surface Preissler, K., 1977. Do Rotifers Show 'Avoidance of the
water current of 2 mm sec - 1. It would seem that Shore'? Oecologia (Berl.) 27: 253-260.
in all but the most sheltered areas, water move- Ruttner, F., 1974. Fundamentals of Limnology. 150 pp.
ments are an important factor in determining the Stewart, L. J. & M. G. George, 1987. Environmental factors
distribution of rotifers in the horizontal plane. influencing the vertical migration of planktonic rotifers in
a hypereutrophic tarn. Hydrobiologia 147: 203-208.
There will also be convection currents generated
by insolation. The problems of measuring small
water currents has been commented on by many
writers, including Harvey (1976).
Hydrobiologia 186/187: 157-161, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 157
© 1989 Kluwer Academic Publishers.

Oviposition behavior of the littoral rotifer Euchlanis di/atata

Elizabeth J . Walsh
Department of Zoology, Oregon State University, Corvallis, OR, 97331-2914, USA

Key words: oviposition behavior, Euchlanis dilatata

Abstract

Several components of egg-laying behavior in 3 clones of E. dilatata were investigated. Using nearest-
neighbor analyses it was shown that individual animals lay their eggs randomly within culture dishes in
the absence of an artificial substrate while eggs laid in the presence of a substrate are clumped in 2 of
the 3 clones studied. Animals preferentially oviposited on artificial substrates containing eggs when given
a choice between artificial substrates with and without eggs of conspecifics. When given a choice of
natural substrates, animals consistently laid more eggs on the plant species from which they were
collected. This preference persisted for 2 generations after animals had been removed from plants. In
addition, observations of neonate hatching behavior demonstrated that neonates often spend relatively
long times in the immediate vicinity of hatching.

Introduction
Littoral rotifers are ideal study organisms for
Oviposition behavior can directly influence female investigations of oviposition behavior because
fitness by influencing offspring reproductive suc- they reproduce asexually and live in complex
cess (Thorpe, 1945; Chew & Robbins, 1984). Via habitats offering a wide variety of potential egg
(1986) has shown in leafminers that the choice of deposition sites. During asexual reproduction,
egg deposition site affects pupal weight of off- cloning generates a large number of genotypically
spring. Butterflies have genotypic preferences for identical individuals useful for a variety of be-
oviposition site both between and within host havioral tests.
species which can affect larval performance This research investigates two questions relat-
(Rausher, 1979; 1984). Learning may playa role ing to oviposition behavior in the rotifer Euchlanis
in determining butterfly egg-laying sites depending dilatata. First, are there genetically-based differ-
upon other conflicting constraints such as time ences in rotifer egg-laying behavior? The pattern
(Papaj, 1986; Papaj & Rausher, 1987). In Droso- of egg deposition in the presence and absence of
phila, Del Solar (1968) has shown that egg depo- artificial substrates is compared among three
sition pattern is genetically determined. Theoreti- clones. Secondly, what factors influence ovipo-
cally, the pattern of egg deposition may also sition site selection? Whether cues from egg/egg-
influence competitive interactions (Atkinson & shells of clonemates are used in determining egg-
Shorrocks, 1984) and subsequent sympatric spec- laying site is tested and substrate preferences are
iation (Rice, 1984). Egg deposition pattern, choice investigated. Additionally, neonate hatching be-
of deposition site and the ecological and evol- havior was monitored because it may also play a
utionary consequences of these behaviors have role in site recognition and subsequent habitat
not been explored in rotifers. preferences in adults.
158

Study organism Table 1. Egg deposition behavior in the presence and ab-
sence of artificial substrates by E. dilatata~ R is the index of
aggregation, where R = 1 represents a random distribution;
Euchlanis dilatata is a freshwater rotifer commonly R < 1, clumped; and R > 1, superdispersed. n represents the
found in the littoral zone of nutrient-rich ponds number of individuals tested from each clone. The number of
and lakes. This rotifer attaches it eggs directly to eggs laid per individual varied between 3-7.
littoral vegetation (Edmondson, 1960). In labora-
Artificial substrata
tory mass cultures, eggs tend to be laid in clumps
(King, 1967) but variables potentially affecting Present Absent
this behavior have not been studied. Because this
rotifer reproduces by cyclical parthenogenesis Clone n X(R) Range(R) n X(R) Range(R)
(King, 1967), individual females isolated from
natural populations produce genetically identical DL 21 0.396 0.089-0.949 13 1.098 0.313-4.75
QP 7 0.446 0.200-0.773 8 0.831 0.552-1.25
daughters (King, 1977). The original female and SC 28 1.050 0.155-1.940 9 2.563 0.259-6.05
her daughters constitute a clone.
Clones used in laboratory experiments were
derived from collections from three geographically
isolated, natural populations of E. dilatata (R) is calculated by measuring the distance
(DL = Devil's Lake, Lincoln City, OR; between individual eggs. When R = 1 the egg
QP = Quarry Pond, East Corvallis, Oregon; and distribution is considered random, when R < 1
SC = Soap Creek Pond, North Corvallis, Ore- the distribution is clumped and when R > 1 the
gon). Animals were maintained in a temperature pattern is said to be superdispersed. For 2 of the
controlled chamber (25 DC) with a 161: 8d pho- 3 clones tested, animals laid their eggs in clumps
toperiod. Rotifers were cultured in Pourriot-Gil- in the presence of artificial substrates and ran-
bert medium (Gilbert, 1968) at pH 7.5 and fed domly in the absence of artificial substrates
Chlamydomonas reinhartii (Univ. Texas culture (Table 1). Females of the third clone, SC, deposit-
collection 89) grown on Bristol's agar (Starr, ed its eggs randomly both in wells containing
1978). Clones used in experiment 1-4 described artificial substrates and in those without them.
below were kept under these conditions for at
least 10 generations prior to experimentation. Experiment 2
To determine potential cues used in choosing an
egg deposition site, females were given a choice
Methods and results between artificial substrates with and without the
eggs of clone-mated attached. Two replicates of
Experiment 1 sixty ovigerous females from clone SC were plac-
The pattern of egg deposition in the presence or ed into glass culture dishes (5.0 cm diameter)
absence of an artificial substrate (nylon monofIla- containing six 1-cm pieces of monofIlament line:
ment line) was compared among the 3 clones. three lines contained 15-25 eggs and three were
Females of each clone were individually placed in blank (i.e., without eggs). The number of ad-
1 cm diameter culture wells. Half of the wells ditional eggs deposited was recorded after 12 h.
contained a l-cm piece of monofIlament, half Animals laid significantly more eggs on those
were empty. The pattern of egg deposition was lines containing eggs than blanks (ANOVA, F
recorded daily, using a grid system. Neonates test, p < 0.001; Fig. 1).
were removed upon hatching.
Nearest neighbor analyses (Vandermeer, 1981) Experiment 3
were performed to determine whether eggs were An experiment was conducted to ascertain
laid randomly, clumped or superdispersed within whether morphological cues (i.e., the shape of the
culture wells. For this test, an index of aggregation eggs) or chemical cues from the eggs are used in
 159

40.----------------------------. The number of eggs laid on monofIlament de-

*** pended upon the presence or absence of eggs and
en
Ol
Ol
30 eggshells (ANOVA, F test,p < 0.001). More eggs
(l)
were laid in the presence of intact eggs of clone-
0
c 20
mates than on either blank monofIlament or mo-
0
~ nofIlament containing eggshells (Newman Keuls
"0
multiple comparison test, p < 0.05) (Fig. 2).
"0
0 10 -
"'" Experiments 4 and 5
0 rl The role of substrate preference in determining
blank eggs egg-laying site was further investigated in two
Fig. 1. Egg deposition behavior of clone SC on artificial
experiments. Females were offered a choice
substrates with and without eggs of clonemates. Bars repre- among 3 natural substrates. Individuals of Euch-
sent the number of additional eggs laid per monofilament lanis dilatata were isolated from a mass culture of
line. *p < 0.05; **p < 0.01; ***p < 0.001. millfoil (Myriophyllum exalbescens) collected from
Devil's Lake. Fifty females were placed into each
of 5 replicate 100 ml beakers containing 0.25 g
(wet weight) of millfoil, Elodea, and coontail (Cer-
choosing an egg deposition site by E. dilatata. atophyllum) in 50 ml millipore fIltered lake water.
Animals were given the choice of laying eggs on Before use, all plants were rinsed thoroughly in
a blank monofIlament, one with eggshells attach- fIltered lake water and inspected for eggs. No eggs
ed, or one with intact eggs. The procedure de- were found on any plant. Plants were inspected
scribed in experiment 4 was followed with these for eggs and eggshells after 54 h.
modifications:
(1) 150 instead of 60 females of clone SC were For experiment 5, these modifications were made:
placed individually into the experimental cul- (1) all animals were obtained from a single clone
ture dishes. collected from millfoil.
(2) monofIlament lines containing approximately (2) half ofthe replicates contained animals grown
80 eggshells were included along with those in the presence of millfoil until the time of
containing eggs and blanks. experimentation and half contained animals
which had been removed from millfoil for 2
generations. Six beakers were set-up: 3
70 containing females grown in association with
millfoil and 3 which had been grown in culture
60
en dishes for 2 generations.
Ol
Ol 50 *
(l)

0 40 Animals preferentially laid eggs on millfoil when

c
0 given a choice among millfoil, Elodea and
~
30
"0 coontail. A one-tailed t-test based on the numeri-
"0 20 cal difference in the number of eggs laid on sub-
0
10 *** strate pairs (millfoil versus Elodea and millfoil
"'" versus coontail) was conducted. The overall
0
probability that animals prefer millfoil over both
blank shells eggs
coontail and Elodea is 0.9945 (I-tailed t-test).
Fig. 2. Egg deposition behavior of clone SC on artificial
substrates with eggs, with eggshells, or without either. Bars
This preference for millfoil persisted even after
represent the number of additional eggs laid per monofila- animals have been removed from the plant for 2
ment line. *p < 0.05; **p < 0.01; ***p < 0.001. generations (Table 2).
160

Table 2. Egg-laying pattern in the presence of three natural Discussion

substrates of a single clone. dM _E represents the mean differ-
ence in the number of eggs laid on millfoil versus Elodea. dM . c
represents the mean difference in the number of eggs laid on
Oviposition behavior varied among the three
millfoil versus coontail. Significance levels for a one-tailed clones of E. dilatata tested. Choice of egg depo-
t-test are given for animals which were raised in contact with sition site was affected by the presence of other
the natal substrate and those which had been removed from eggs, eggshells, and substrate-type. Individuals of
it for 2 generations.
some E. dilatata clones lay their eggs in clumps in
Millfoil vs Elodea Millfoil vs coontail
the presence of artificial substrates. This clumping
pattern has also been observed in several other
dM _E P dM _C p monogonont species (Mytilina, Monostyla, E. tri-
quetra, E. calypida, pers. obs.; Proales decipiens,
Experiment 1 0.4252 *** 0.4830 *** Noyes, 1922) and in one unidentified bdelloid
Experiment 2
Raised w/millfoil 0.5093 0.4839
rotifer (pers. obs.). Potential benefits of laying
* **
Raised w/o millfoil 0.6047 * 0.6195 * eggs in clumps for rotifers include exploitation of
favorable physical conditions for embryonic de-
velopment and subsequent resource utilization by
neonates and avoidance of interclonal competi-
Experiment 6 tion. A clumped distribution of eggs may also
Neonate hatching behavior was compared among have consequences for the genetic structure of
the 3 clones. One cm pieces of monofilament line populations, particularly if genotype-specific
were placed into mass cultures of DL, QP, and clumps are formed and habitat preferences exist.
SC. Eggs were deposited onto the monofilament The results of experiment 3 demonstrate that
line. The line was then removed and placed into these clones lay their eggs preferentially in the
a small culture well. I recorded the amount of time vicinity of other eggs and eggshells. This is also
individual neonates from the three clones spent in supported by observation of egg aggregations in
contact with the eggshell from which it hatched laboratory mass cultures (King, 1967; pers. obs.).
and nearby eggs as well as observations of posture E. dilatata also chooses to deposit eggs on the
and movement following hatching. plant species from which they were collected. It
Neonate behavior was similar between individ- should be noted that in extensive collections of
uals of clones DL and QP. Each clone DL indiv- individuals of these three plant species, E. dilatata
idual (n = 10) remained in contact with the mo- is most often found on millfoil (unpubl. data).
nofilament line for longer than 20 minutes. Simil- This plant preference may have a genetic compon-
arly, individuals from clone QP (n = 8) spent an ent because it persists over two generations with
average of 24.0 ± 11.7 min in the immediate vi- no chemical or physical contact with any plant
cinity of hatching. Neonates from these clones speCles.
slowly extended their coronas, bent over and then Neonate hatching behavior was observed to
made coronal contact with their eggshell and determine whether recognition of the egg site was
nearby eggs for the durations given above. In possible. In two of the three clones animals ap-
contrast, neonates of clone SC (n = 12) spent peared to be imprinting on chemical and/or me-
very little time (X = 0.97 ± 0.81 min) in contact chanical cues present at the site of hatching. The
with the monofilament line. They typically extend- coronal area of E. dilatata contains most of the
ed their coronas and immediately swam free of the sensory structures of this animal (Stoszberg,
monofilament line. 1932). Neonates of clones DL and QP
maintained coronal contact with their eggshells
and nearby eggs for extended periods and did not
appear to be feeding during this time. This be-
havior may be similar to that described during
 161

oviposition in Notommata copeus (Clement et aI., Clement, P., E. Wurdak, & J. Amsellem, 1983. Behavior and
1983). These data suggest that some form of im- ultrastructure of sensory organs in rotifers. Hydrobiologia
104: 83-129.
printing may occur. However, neonates from Del Solar, E., 1968. Selection for and against gregariousness
clone SC did not display this behavior indicating in the choice of oviposition sites by Drosophila pseudoobscu-
that there is variability in this behavior among ra. Genetics 58: 275-282.
clones. A genetic basis for this is supported by the Edmondson, W. T. 1960. Reproductive rates of rotifers in
results of the egg-laying pattern experiment (1) natural populations. Mem. 1st. ital. Idrobiol. 12: 21-77.
Garcia-Dorado, A., 1987. Polymorphism from environmental
reported here as well as significant differences in heterogeneity. Some features of genetically induced niche
body size, embryonic development time, and life preference. Theor. Pop. BioI. 32: 66-75.
history traits among these clones (unpubl. data). Gilbert, J. J., 1968. Dietary control of sexuality in the rotifer
While the data presented here are intriguing, the Asplanchna brightwelli Gosse. Physiol. Zool. 41: 14-43.
effect of oviposition behavior on the genetic struc- King, C. E., 1967. Food, age, and the dynamics oflaboratory
population of rotifers. Ecology 48: 111-128.
ture of natural populations of rotifers is still un- King, C. E. & T. W. Snell, 1977. Sexual recombination in
known. In an attempt to assess these effects, rotifers. Heredity. 39: 357-360.
research in progress explores genotypic asso- Noyes, B. 1922. Experimental studies on the life history of a
ciations between E. dilatata clones and the plants rotifer reproducing parthenogenetically (Proales dicipiens).
with which they are associated. 1. expo Zool. 35: 225-255.
Papaj, D. R, 1986. Interpopulation differences in host prefer-
ence and the evolution oflearning in the butterfly, Battus
philenor. Evolution 40: 518-530.
Acknowledgements Rausher, M. D., 1979. Egg recognition: Its advantage to a
butterfly. Anim. Behav. 27: 1034-1040.
I thank Charles E. King, Annette M. O1son & Rausher, M. D., 1984. The evolution of habitat preference. II.
Evolutionary genetic stability under soft selection. Theor.
Deanna Olson for improving the manuscript. Eric
Pop. BioI. 31: 116-139.
Atkinson, Michelle Schroeder and Chip Rutledge Rausher, M. D. & D. R Papaj, 1987. Genetic differences and
provided field and laboratory assistance. This phenotypic plasticity as causes of variation in oviposition
research was supported by grants from ZORF preference in Battus philenor. Oecologia. 74: 24-30.
and Sigma Xi. Travel funds were provided by Rice, W. R, 1984. Disruptive selection on habitat preference
and the evolution of reproductive isolation: A simulation
Dan & Judy Jensen and Jim & Sally Nelson. study. Evolution 38: 1251-1260.
Starr, R c., 1978. The culture collection of algae at the
University ofTexas at Austin. 1. Phycol. Suppl. 14: 47-100.
References Stoszberg, K., 1932. Euchlanis. Brachionus and Rhinoglena. Z.
wiss. Zool. 142.
Atkinson, W. D. & B. Shorrocks, 1984. Aggregation oflarval Thorpe, W. H., 1945. The evolutionary significance of habitat
Diptera over discreet and ephemeral breeding sites: the selection. 1. animo Ecol. 14: 67-70.
implications for coexistence. Am. Nat. 124: 336-351. Via, S., 1986. Genetic covariance between oviposition prefer-
Chew, F. S. & R K. Robbins, 1984. Egg-laying in butterflies. ence and larval performance in an insect herbivore. Evolu-
Symposia of the Royal Entomological Society of London, tion 40: 778-785.
No. 11. The Biology of Butterflies. R. I. Vane-Wright and Vandermeer, J. H., 1981. Elementary Mathematical Ecology.
P. R Ackeny (eds.) pp. 65-79. Academic Press, London. Wiley, New York. 294 pp.
Clement, P., 1987. Movements in rotifers: correlations of
ultrastructure and behavior. Hydrobiologia 147: 339-359.
Hydrobiologia 186/187: 163-165, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 163
© 1989 Kluwer Academic Publishers.

Developmental times of Synchaeta oh/onga eggs from the Danube

(Austria)

Wolfgang Zoufal
Schweglerstr. 42/4, A-JJ50 Wien, Austria

Abstract

The relationship between temperature and development time of Synchaeta oblonga eggs from the Danube
was investigated. Growth rates were also measured in populations established from the Danube.

Introduction of the river-rotifer populations it was necessary to

perform laboratory experiments.
The Altenw6rth run-of-river station, which is lo-
cated at stream km 1979.8 at the Austrian part of
the Danube, was the subject of a 'Man and Material and methods
Biosphere Ecosystem Study' from July 1985 to
March 1988. Zooplankton samples were taken at All experiments were performed at the actual tem-
monthly intervals. Compared with literature data perature of the Danube at the day of collection
from this part of the river (Schallgruber, 1944; (kept constant at ± 0.1 cC). For the enclosure
Naidenow & Saiz, 1975; Naidenow, 1985) the experiments river-water was collected with a 50
observed zooplankton densities were much liter Schindler plankton trap at stream km 2000
higher, showing maxima in summer of 100,000, and placed in 3 1plastic bottles and kept for up to
130,000 and 230,000 m - 3 in the years 1985, 1987 6 days in dark without adding food. Daily one of
and 1986 respectively. Synchaeta (mainly Syn- the bottles was filtered and the animals were
chaeta oblonga) was the most abundant genus preserved with 4 % formalin. A 30 Jlffi nylon
followed by Keratella and Brachionus. netting was chosen to ensure that neither small
Field data from a river, even with short rotifers nor rotifer eggs were lost. Even when
sampling-intervals, can never be interpreted as counting at 100 x magnification it was impossible
the result of local factors alone because they are, to distinguish species for all genera.
at least in part, the result of input from other Low zooplankton densities and the small
environments. Information about the growth of volume of the bottles allowed reliable calculation
populations may be obtained by comparing of population growth only for Synchaeta oblonga.
samples from different localities in the river. Population growth (r) and birth (b) rates were
Although the backwater-area has a length of 27 calculated as follows:
kilometers even at low discharge no significant
increase of densities towards the dam could be
observed. Populations do not have enough time in
the backwater-area (retention time about 10-15
where: N = number of individuals and t = time;
hours) to show an increase in numbers or
biomass. To get an idea about the dynamic aspect
164

where: E = eggs per female and D = egg-develop- 1.0

ment time.
Development time of eggs from individuals
collected in the river was determined. The animals
were kept in 10 ml dishes. All eggs were isolated
after laying and checked until hatching at 2-hour
intervals.

Results

As no relationship between temperature and

embryonic development time for Synchaeta 50 100 150
oblonga was available from the literature, it was TlHE (HOURS)
necessary to determine one from my own data. A
1.0
Belehnidek equation was fitted by the method of
least squares and is presented in Fig. 1. The
derived parameters are: a = 16.0932, b = - 1.9
and c = 1.0796.
Birth rates, calculated for the enclosures, are
constant throughout the experiments, reaching
values of about 0.4-0.8 (Fig. 2a). The population
growth rates are in a range of - 0.8 to 1.2
(Fig. 2b). No significant correlation between the
maximum growth rates and temperature was
found.
-0.5 L...-_ _ _ _- - ' -_ _ _ _ _!........l_ _ _ _--'--

50 100 150
TlHE (HOURS J

Fig. 2. Birth and growth rates of Synchaeta oblonga in the

enclosure experiments at different temperatures: 0: 8.6 °C;
e: 9.8 °C; 0: 11.4 °C; . : 14.8 0c.

Discussion

The embryonic development time of Synchaeta

oblonga is very short at low temperatures and can
be compared with the cold-stenothermous
Rhinoglenafertoeensis (Herzig, 1983). As the tem-
perature of the Danube exceeds 20 0 C only on
some days in summer but drops below than 4 0 C
for several weeks during winter, this short
5 10 15 20 development time can be regarded as an adap-
TEHPERATURE (0C) tation to the temperature conditions in the river.
Fig. 1. Relationship between embryonic development time Although no food was added during the experi-
of Synchaeta oblonga and temperature. ments the rotifers showed high rates of increase.
 165

Presumably, bacteria from the Danube was the References

food source used by the S. oblonga.
Herzig, A., 1983. Comparative studies on the relationship
between temperature and duration of embryonic develop-
ment of rotifers. Hydrobiologia 104: 237-246.
Acknowledgements Koste, W., 1978. Die Radertiere Mitteleuropas. Gebr.
Borntager, Berlin, Stuttgart, 673 pp.
My sincere thanks are due to Univ. Doz. Dr. A. Naidenow, W., 1985. Die Auswirkungen der Wasserbauli-
Herzig for his constructive criticism and help im- chen MaBnahmen und der Belastung auf das Plankton und
das Benthos der Donau. Verlag der Bulgarischen
proving this manuscript. I am grateful to the
Akademie der Wissenschaften: 72-102.
Austrian Bundesministerium filr Wissenschaft Naidenow, W. & D. Saiz, 1975. Der EinfluB des regulierten
und Forschung and the Donaukraft (Donaukraft- Abflusses auf das Plankton der Oberen Donau. In: Be-
werke AG) for financial assistance. This study richte der 18. Tagung der lAD. Regensburg 1975, I. Teil:
was done in the scope of the 'Ecosystem study 239-243.
Schallgruber, F., 1944. Das Plankton des Donaustromes bei
Donaustau Altenworth'.
Wien in qualitativer und quantitativer Hinsicht. Arch.
Hydrobiol. 39: 665-689.
Hydrobiologia 186/187: 167-179, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 167
© 1989 Kluwer Academic Publishers.

Community structure and coexistence of the rotifers of an artificial

crater lake

Geert Bogaert & Henri J. Dumont

Laboratorium voor ecologie der dieren, zoogeograjie & natuurbehoud, State University of Ghent, K.L.
Ledeganckstraat 35, B-9000 Ghent, Belgium

Key words: rotifers, succession, migration, abundance, distribution

Abstract

The community structure and dynamics of the rotifers of an abandoned and inundated limestone quarry
are described. Zooplankton standing crop is low, but the community is rich in species. Ploesoma hudsoni
and Filinia hofmanni are rare or unknown in a perimeter of several hundreds of kilometers around the
lake. This illustrates that (passive?) dispersal across large stretches ofland even occurs in rare species.
Temporal and spatial distribution, together with specific diurnal vertical movements in different seasons
facilitate the coexistence of numerous species in the lake. This complex ecology contrasts with the small
size of the initial propagules that helped colonise the lake.

Introduction 40 years can be considered a short period for

populating such an isolated lake. The objectives
Many studies deal with the dynamics of rotifers in of the present paper were to identify the species
a variety of lake types. In Belgium, numerous assemblage living in the lake and to see how they
studies of stagnant waters focus on shallow artifi- compare to similar but natural lakes elsewhere.
cial basins. These are nearly all under direct Also, because the lake's populations can be
human influence and have recently undergone expected to have originated from a limited num-
eutrophication. The present paper initiates the ber of small initial propagules, each with little or
study of fairly unknown limnological sites in the no genetic variation, this narrow genetic basis
country: the inundated quarries in the south. might well be expressed in an unusual ecology.
Their depth and trophic status, their remarkable
plankton fauna and the unknown way in which
they were colonised, prompted the given research. Site description
In the site studied, quarrying ceased around
1948 and the cratershaped cavity was allowed to The study site is a 47 meter deep, artificial lake
fill with groundwater. Filling-up presumably took (Lake Delwart), a former limestone quarry near
not more than a year. The quarry and the resulting Doornik, Province of Henegouwen, Belgium
lake is not connected to any surface water, and (50° 35' 30" N, 3 0 24' E). The lake is mesotrophic
thus has no in- or outlet. Even though no informa- (Table 1). The shores are steep and a littoral zone
tion on the colonisation of the lake is available, is negligible: there is no Phragmitetum and no
 168

Table 1. The morphometric characteristics of Lake Delwart. Temperature, pH, oxygen concentration and con-
ductivity were also measured.
Area 4.8ha
Diurnal vertical migration was studied over
Length 295m
Breadth 250m 24 hours in winter (20-21/12/1987) and in sum-
Mean breadth 163 m mer (4-5/1/1988). Samples were taken every
Maximum depth 47m 3 hours at 2.5 meter intervals in the upper
Relative depth 19% 20 meters and in deeper waters every 10 (winter)
Shore line 880m
or 5 meters (summer). Mean residence depth of a
Shore line development 1.13
Altitude above sea level +35m population was calculated as
Bathymetry complex

submerged vegetation. Fish were introduced with N; concentration of individuals at depth i,

=
artificially and the ichthyofauna is dominated by d; = depth of ith sample.
perch (Perea jluviatiiis) and carp (Cyprinus carpio).

Results
Methods
Abiotic environment
The plankton was sampled biweekly between
October 12, 1987 and October 8, 1988, at the The lake is slightly alkaline with pH values
deepest point of the lake. Duplicate samples were between 6.32 and 8.70. It was warm and
collected at 5 meter intervals, using a 2 liter monomictic during the study period, with a winter
Friedinger bottle (October-November 1987) with low of 5.3 °C and a summer high of 21.5 0c.
a 35 Jlm screen or a 35 liter plankton sampler Oxygen stratification was clinograde in autumn,
(December 1987-0ctober 1988) with a 50 Jlm nearly uniform during turn-over, positive hetero-
screen. Plankton was preserved in 4 % form- grade in spring, and clinograde again in autumn.
aldehyde and counted in the laboratory using a At the deepest point (45 meter), the water was
dissecting microscope at 45 x magnification. permanently anoxic, due to a sapropelium layer

1987 1988
24/10 21/11 21/12 H,/1 13/2 12/3 9/4 7/5 4/6 2/7 30/7 26/8 24/9
12/10 7/11 5/12 2/1 30/1 27/2 26/3 23/4 21/5 18/6 16/7 12/8 10/9 8/10 N.l0 L-l

~==--------------------=~~~==~==~=~==--
ASPLANCHNA PRIODONTA
J20
BRACHIONUS CALYCIFLORUS ~ Jl0

FILINIA HOFMANNI
c==============___ -======_ J 20

c=:::::
~
____~> ~J30
KELLICOTTIA LONGISPINA

~
KERATELLA COCHLEARIS
J250
KERATELLA QUADRATA

PLOESOMA HUDSONI
c=:::::::= J10
- - - - - - - - - - - - ho
POLYARTHRA SPP.
<===---------------=====~>=--~=::::JJ J250
POMPHOLYX SULCATA
-------------------~<::>~--===----------~ J50
SYNCIIAETA s p p .
J50
TRICHOTRIA POCILLUM
ho
GASTROPUS HYPTOPUS
J 250

Fig. 1. Succession of the rotifers in Lake Delwart during the study period.
 169

900
Vl
~
fJ.1 800
f-i
H
..:1
0
,...,

~
fJ.1
p.
~
fJ.1
~

5z
OCT NOV DEC JAN FEB MAR APR MAY J UN JUL AUG SEP OCT

Fig. 2. Seasonal abundance of cope pods (a), nauplii and copepodites (b), cladocerans (c), rotifers (d) and sum of these categories
(e) in the lake in 1987-1988. Average over the water column, in numbers per 10 liters.

on the bottom. Conductivity values ranged Ridder, 1973). Its nearest known occurrence is in
between 560 and 840 Jl S cm - 1. France, at 250 kilometers from Lake Delwart (De
Ridder, Pers. Com.).

Community composition
Seasonal occurrence related to abiotic conditions
The pelagic zooplankton community is domi-
nated by rotifers: 18 species were found, including Many rotifer species are perennial (Fig. 1): the
a few incursive littoral forms. The number and dominant Keratella cochlearis, with a broad maxi-
species composition is comparable to that of mum from spring to autumn; Keratella quadrata,
many natural deep lakes in the temperate region with maxima in spring and autumn; Polyarthra
(Table 2). Two taxa among this assemblage stand spp., with dominance in autumn and summer.
out: the Filinia species present shows most of the Filinia hofmanni and Kellicottia longispina also
morphological characteristics of Filinia hofmanni, occur during the whole year, the former species
as described by Koste (1980) and Schaber & with a broad maximum from winter to late spring,
Schrimpf (1984). Its ecology follows that reported the latter abundant mostly in winter. Brachionus
by these authors: the species is found at tempera- calyciflorus is absent only during a short period in
tures from 5 to 10 0 C and at oxygen contents from autumn and reaches a weak spring maximum.
1 to 14 mg 1- 1. The highest abundance is at lower There is a marked seasonal separation of the inci-
oxygen concentrations near the anoxic layer in the dence of the raptorial forms: Ploesoma hudsoni
hypolimnion. The species has been reported from (summer and autumn), Asplanchna priodonta
Norway, Austria and Germany. Ploesoma hudsoni (perennial, with highest densities late in autumn
has only been reported twice from Belgium (De and late in spring) and Synchaeta pectinata (late
 ......
25 ~ , , , ' I , , , , I , , , , I I , , , , I , , -.l
25 ~ , , , • I , , , , I ' , , , I ' , ' 25 ~ , ' , , I I I 0
°2 mg . I - 1 ASPI.ANClINA PRTonONTA BRACHIONUS CALYC1FLORUS
~ ~ FXl-oXNXA HOFMANN X ~
0 0
~

20-1
20 20

00
1 f 1 f
15--1 I-
0
~
o. ~ 0 0
0
: • ~.O
10-1 0
"j
10 0 l "1 10-
S
O
t-

0 .
5-1 I- 5~ t- 5-1 II
o·
0

.rJM
0-, tt
0 I
, I I
, I i o I i .. i I
, I i i I 0 I, I i i i ,---' , I
0 5 10 15 20
t 10 15 20 0 5 10 15 20
0 5

25~ , j , • , , , , , ,
25 j , ,, , I I , , ii, , , , I , I , I I i I , I , I , , I
KELLICOTTIA LONGISP:NA 25 KERATELLA
GAS'InOPUS HYPTOPUS ~ t i, COCHLEAR:!:S
••
20-1 20 20-
0

~ ••
15-1 15 15 t-
o 0
Q 0
0 0
0 0 0 0 0
o· • . •
0 0 q,
o.
• ••
10 0 0 10 •
10-1 I-
~ ~ ~

0 •
.0
5-1 I- 5-j I-
~ @ ~ 5~ ••
0

0 i i i
, I I
, i 0 i i
, I
I
,I , I O~ I f
0 5 10 15 20 5 10 15 20 0 5 10 15 20
0
 mg.I-1 25
. . •
25 25
KERATELLA QUADRATA PLOESOMA HUDSON X POLYARTHRA SPP.
0
°2
20-\ .. 20 20 + t
0
0

+•
0 •
15-\ r- 15-1 15 .. f-
..• tJ
.. 0
.. •
0
0 10 ..
'] ':1 5
[
.0 r J G
00 0
<b'
01 i i i I I i
0 i i i I i i i 0 i I I i 2'0
I I
I. • I I I • . . 0 5 10
• 15 to (C
0 5 10 15 20 0 5 10 15 20

25 ~ • i ' i I
25
POMPHOLYX SULCATA
SYNCHAETA SPP

20-
2°l
j
15-1
.. N.10 1-1 +100 -200
0

0
0
0
l
0 , x200-500
o 0
~ "~
10
-<10
10
1 ~ () 10 -20 e500-1000
5-1 5i
.' t r 020-50 e1000-2000
o 1 • • I • • I ••• 0 I • 0 0 • I 01··· i ~ •• i 'l~i '1~'" i' ·1tO(OCh:J50-100
0 5 10 15 20 • >2000
Fig. 3. Specific incidence as a function of temperature and concentration of dissolved oxygen. .....
-...l
.....
172

Table 2. Pelagic species composition of Lake Delwart. Gastropus hyptopus is a winter species. The lit-
toral Trichotria pocillum occurs in the plankton
Summer/autumn species
only in late spring, but in relatively high densities,
Cyclops vicinus vicinus
Thermocyclops oithonoides while Pompholyx sulcata is perennial with a maxi-
Diaphanosoma brachyurum mum In summer.
Ceriodaphnia quadrangula The peak of the rotifer community taken as a
Ploesoma hudsoni whole, is separated in time from that of copepod
Chydorus sphaericus
nauplii and small cladocerans (Fig. 2).
Diacyclops bicuspidatus
Data on temperature and concentration of dis-
Winter species solved oxygen were used to determine some
Cyclops strenuus strenuus
Eudiaptomus gracilis
specific preferences (Fig. 3). For each sampling
Gastropus hyptopus date, the combinations of the maximum abun-
Kellicottia longispina dance depth were plotted per species, sum-
Late winter/spring species marizing their preferential incidence as a function
Filinia hofmanni of these factors.
Pompholyx sulcata
Synchaeta spp.
Perennial and rare species Depth distribution
Brachionus calyciflorus
Keratella quadrata Depth segregation of species was marked during
Euchlanis dilatata
Trichotia pocillum stratification periods (Fig. 4). Under turn-over
Asplanchna priodonta conditions rotifers were more homogeneously dis-
Polyarthra spp. tributed over the well oxygenated part of the water
Stephanoceros fimbriatus column, although some species maintained a
Macrocyclops albidus
depth preference: Synchaeta spp., Gastropus
Daphnia longispina
Alone quadrangularis hyptopus and Filinia hofmanni remained in the
Lecane sp. upper layers. In spring, when surface layers warm
up, most species reached maximum density.
Coexistence during this period is probably due to
winter-early spring). Ploesoma hudsoni coincides a high phytoplankton production. Later, when a
with Ceratium, the preferred food of the species 'clear water phase' in the epilimnion occurred,
(Ruttner-Kolisko, 1974). species segregation developed again: Polyarthra

Table 3. Maximum speed of ascent and descent (meters per hour) for the rotifer species in Lake Delwart during the winter (a)
and summer (b), as calculated from the mean residence depth of a population. Period when maximum speed was reached is given
between brackets. Migration type: N = nocturnal, R = reverse, S = stationary, T = twilight.

A Maximum speed Migration type

Ascent Descent

Asplanchna priodonta 0.80 (21.00-24.00) -1.60 (12.00-15.00) R

Brachionus calyciflorus 0.50 (21.00-24.00) - 0.43 (6.00- 9.00) S
Filinia hofmanni 0.93 (21.00-24.00) -0.67 (3.00- 6.00) N
Gastropus hyptopus 0.53 (21.00-24.00) -0.27 (3.00- 6.00) N
Kellicottia /ongispina 0.57 (21.00-24.00) -0.63 (3.00- 6.00) N
Keratella cochlearis 0.60 (21.00-24.00) - 0.90 (3.00- 6.00) N
Polyarthra spp. 1.23 (21.00-24.00) - 0.43 (18.00-21.00) N
Pompholyx sulcata 0.93 (18.00-21.00) - 0.57 (15.00-18.00) R
 -»t. N.I0 1- 1 AS PLANCHNA PRl:ODONTA ..II!. N.IO 1- 1
~N.IO C l KERATELLA COCHLEARXS GASTROPUS HYPTOPUS

-I f: -I I
"-\T ,
••
-I t , -II
I I I
"
,. 'I -, r I
7/11 16/1 <S
18/6 I 1 ~/IU
12/10 16/1 2/7 16/1 9/4
...m.N.IO 1- t -I
BRACHrONUS CALYCYVLORUS J:IL N.l0 1 KERATELLA QUADRATA
..lIlII.N.IO 1- 1 SYNCHAETA spp.

" -I -\ "-II -I I
!. 11 I
: ,:,
I
Y -I
i I
.. I
" 16/1 18/6 12/10 16/1 2/7
7/11 -I
5/12 12/3 . 4/6
...m.N.IO Cl FXL~NIA HOFMANNY .JU.N.IO 1- 1 POLYAHTIIRA SPP ..

"
.,. .,.
-II
:11
" 12/10 16/1 2/7 10 16/1
2/7
JOt, N.I0 1- 1 KELLICOTTl:A LONC:t:SP:t:NA .JIll. N.IO 1- 1 POMPHOLYX SULCATA

" -\ "-II -I
>0 "10

.",

'"
7/11 16/1 18';6 12/10 16/1 4/6
Fig. 4. Representative depth profiles of rotifer species in different seasons. Depth scale in meters. Vertical bar indicates zone with temperature gradient. Arrow points
at oxyc1ine.
.......
-...l
W
174

0 0
2
1

10 10
0
E
p
T :20 20
H

(m)
30 B C 30 A

40 40

45 5,....._""",,,,--,1,0 DC 7 9 45 0%
~----~----~----~----~

0
2

10
\
\
0
E
P
T 20
H

(m) C
30 B D
I
I night - -
I
I day-
I
40 I
I

45 10, 15, 20, DC 7 S ? o, I 10, 2,0 mg.I-1

Fig. 5. Environmental conditions during the 24 hour sampling in winter (1) and summer (2). a: under water illumination in %
of that of the subsurface (average of day), b: temperature curve (average of 24 hours), c: pH curve (day), d: oxygen curve.
 L1L./. L..1L./.
ASPLANCHNA PRIODONTA KELLICDTTXA LONGISPINA

,. ,

KERATELLA COCHLEARXS

)0

1.0
'0
1.5
45
FTLINIA HOFMANNI
'POLVARTHRA SPP.
o

10
10

1m, 1-10
Iml1-20

-30
JO

'0
'0

GASTROPUS HYPTOPUS
POMPHOLYX SULCATA

-10

Iml '20

JO

5
12.00 15.00 18.00 21.00 24.00 3.00 6 .. 0 0 9.00
12.00 15.00 18.00 21.00 24.00 3.00 6.00 9.00
hour I
hour'

Fig. 6. Migration panels for rotifer species during winter. Depth scale in meters, time in local hours. The line connects the mean depth of the population at different ......
-..l
sampling hours. Vl
176

L.lL%
ASPLANCHNA PRIODONTA

30

(ml 20

3D

5
12.0015.0018.0021.0024.00 3.00 6.00 9.0012.00

hour
Fig. 7. Migration panels for rotifer species during summer. Depth scale in meters, time in local hours. The line connects the mean
depth of the population at different sampling hours.
 177

B Maximum speed Migration type

Ascent Descent

Asplanchna priodonta 0.77 (3.00- 6.00) - 0.23 (21.00-24.00) S? or R?

Filinia hofmanni 1.77 (15.00-18.00) - 2.23 (21.00-24.00) T
Kellicottia longispina 0.53 (3.00- 6.00) - 1.10 (12.00-15.00) N
Keratella cochlearis 0.87 (3.00- 6.00) - 0.87 (12.00-15.00) R?
Polyarthra spp. 0.73 (24.00- 100) - 0.87 (9.00-12.00) R?

spp. and Keratella cochlearis remained in the Discussion

epilimnion where they were slightly segregated.
Most rotifer species accumulated near the The diversity of the rotifer community indicates
thermocline, where bacteria, algae and other food that relatively isolated lakes can get populated by
particles concentrate. Filinia hofmanni and a fairly 'complete' plankton zoocenosis in a
Keratella quadrata sank to the hypolimnion as remarkably short period of time. As stressed
soon as spatial overlap in the upper layers earlier, Ploesoma hudsoni is an epilimnetic species,
increased. Filinia hofmanni is a rare, largely hypolimnetic
species. Further, their nearest known occurrence
is about 250 kilometers away from Lake Delwart.
Diurnal vertical migration While this illustrates the power of passive dis-
persal in rotifers, the mechanism through which
Winter sampling (20-21/12/1987) this has occurred remains undocumented.
The weather was cloudy, with rain. Sunset was at Environmental factors in a lake change in time
16.39 and sunrise at 8.43. A bloom of blue-green and space. The way in which species relate to
algae occurred in the upper 20 meters. Oxygen them in terms of abundance and depth distribu-
stratification was present with the oxycline tion, is determined by their specific ecological
between 20 and 25 meters (Fig. 5a). tolerance, competitive interactions, the occur-
The rotifers migrated over small distances rence of food (Pourriot, 1965) and predation
(Fig. 6, Table 3), but depth segregation was (Stemberger & Evans, 1984). Separate density
diminished compared to the biweekly sampling. peaks among the carnivorous species in Lake
Copepods and cladocerans showed nocturnal Delwart suggest an avoidance of competition
migration behaviour. through seasonal succession. Likewise, maxima
of the herbivorous species are also segregated
Summer sampling (4-5/7/1988) across the year. Cooccurrence with the filter-
There was cloudy weather and rain. Sunset was feeding nauplii and small cladocerans is reduced
at 21.58, sunrise at 5.36. Turbidity in the epilim- by a succession of their density peaks during
nion was slight, with a bloom of bluegreen algae bloom periods. Given these temporal segre-
occurring at a depth of approximately 7 meters gations, differences in food selection (e.g. con-
(Fig.5b). sumption of food particles from different size-
Different rotifers were located in and under the class; Edmondson, 1965; Hutchinson, 1967;
bloom layer and were segregated from species Peyler, 1957; Dumont, 1977; Starkweather,
occurring nearer the surface. Data on migration 1980), which can allow coexistence of even closely
type and speed are given in Fig. 7 and in Table 3. related species, probably do not play the most
Copepods and cladocerans were concentrated in important role in the present study.
the bloom layer during the day and migrated Apart from temporal segregation, overlap
upward at night. between species is reduced by differing depth-
178

distributions: the marked segregations, seen on a a role for biotic interactions in organizing the
small scale during the 24 hour sampling and the lake's community. The positioning and migration
sinking of species like Keratella quadrata and of rotifers is consistent with a response to preda-
Filinia hoJmanni to the hypolimnion when den- tion pressure from copepods and competition
sities in the upper layers increase, illustrate this with cladocerans and other rotifer species, i.e. it
point well. The slight segregation between Poly- is a spatial and temporal adaptive strategy which
arthra spp. and Keratella cochlearis is probably facilitates co-occurrence of numerous species
due to light preferences (Peyler, 1957). Many (Miracle, 1974, 1977; Makarewicz & Likens,
species are abundant over a broad range of tem- 1975; Lane, 1975).
perature and oxygen concentrations, but the It follows that the ecology of the rotifer species
absolute maximum of each species occurs at of Lake Delwart is complex and comparable to
specific combinations of these factors. Filinia hof that of other, older lakes. This indicates that even
manni behaves as a cold-stenotherm (5-10 0q, in a lake as young as Delwart, the (narrow?) gene
as does Brachionus calyciflorus to a lesser degree, pools of the populations present acquired suf-
while Ploesoma hudsoni is warm-stenotherm. ficient variation to successfully respond to chang-
Gastropus hyptopus, Asplanchna priodonta, Ploeso- ing abiotic and biotic conditions.
ma hudsoni and Synchaeta spp. have specific oxy-
gen preferences. The data also illustrate the ability
of rotifers to tolerate low oxygen concentrations, Acknowledgements
as described by different authors (Larsson, 1971;
Peyler, 1957; Elliot, 1977; Pourriot, 1977; We thank Guido Goos for practical help and Dr
Hofmann, 1977); Keratella quadrata, Kellicottia M. De Ridder for help in species identification
longispina and Brachionus calyciflorus survive in and for providing unpublished data.
nearly anoxic layers.
The diurnal vertical movements of rotifers in
the lake are also noteworthy. Migrations are pres- References
ent but not pronounced in winter, while they are
more marked in summer. A relation with the Cushing, D. H., 1951. The vertical migration of planktonic
homogeneous conditions during turn-over in win- Crustacea. BioI. Rev. 26: 158-192.
De Ridder, M., 1973. Atlas provisoire des Rotiferes de
ter and with the higher light levels, oxygen, tem- Belgique. J. Leclercq & c. Gaspar, eds, Gembloux.
perature stratifications and food concentration in Dumont, H. J., 1977. Biotic factors in the population
the trophogenic layer in summer seems obvious. dynamics of rotifers. Arch. Hydrobiol. Beih. 8: 98-122.
Light regulation also seems important, as in many Elliot, J. E., 1977. Seasonal changes in the abundance and
other planktonic organisms (Cushing, 1951 ; distribution of planktonic rotifers in Grasmere (English
Lake District). Freshwat. BioI. 7: 147-166.
McNaught & Hasler, 1964; Hutchinson, 1967). George, M. G. & c. H. Fernando, 1970. Diurnal migration in
Comparing winter and summer, we recorded sig- three species ofRotifers in Sunfish Lake, Ontario. Limnol.
nificant variation in migration patterns. Stewart Oceanogr. 15: 218-223.
& George (1987) studied migration of rotifers in Hofmann, W., 1977. The influence of abiotic environmental
different months and found a similar variation. factors on population dynamics in planktonic rotifers.
Arch. Hydrobiol. Beih. 8: 77-83.
The speed of ascent and descent, as calculated Hutchinson, G. E., 1967. A treatise on limnology. Vol. 2:
from D on different sampling hours is high. It is Introduction to Lake biology and the Limnoplankton.
higher than in the (shallower) lakes studied by Wiley, New York. 1155 pp.
George & Fernando (1970) and by Stewart & Koste, W., 1980. Uber zwei Plankton-Radertiertaxa Filinia
George (1987). It is highest in summer, when australiensis n.sp. und Filinia hofmanni n.sp., mit Bemer-
kungen zur Taxonomy der longiseta-terminalis-Gruppe.
migration amplitudes are larger. Genus Filinia Bory de St. Vincent, 1824, Familie Filiniidae
The numerous cases of spatial segregation Bartos 1959, (Uberordnung Monogononta). Arch. Hydro-
within the vertical migration event again suggests bioI. 90: 230-256.
 179

Lane, P. A., 1975. The dynamics of aquatic ecosystems: a Pourriot, R., 1965. Recherches sur l'ecologie des Rotiferes.
comparative study of the structure and dynamics of four Vie Milieu Suppl. 21. 224 pp.
zooplankton communities. Ecol. Monogr. 45: 307-336. Ruttner-Kolisko, A., 1974. Plankton rotifers: Biology and
Larsson, P., 1971. Vertical distribution of planktonic rotifers Taxonomy. Binnengewasser Suppl. 26. 146 pp.
in a meromictic lake; Blankvatn near Oslo, Norway. Schaber, P. & A. Schrimpf, 1984. On morphology and
Norway. J. Zool. 19: 47-75. ecology of the Filinia-terminalis-longiseta-group (Rotatoria)
Makarewicz, J. C. & G. E. Likens, 1975. Niche analysis of a in Bavarian and Tyrolean lakes. Arch. Hydrobiol. 101:
zooplankton community. Science 190: 1000-1003. 247-257.
Mc Naught, D. C. & A. D. Hasler, 1964. Rate of movement Starkweather, P. L., 1980. Aspects of the feeding behavior
of populations of Daphnia in relation to changes in light and trophic ecology of suspension-feeding rotifers. Hydro-
intensity. J. Fish Res. Bd Can. 21: 291-316. biologia 73: 63-72.
Miracle, M. R., 1974. Niche structure in freshwater Stemberger, R. S. & M. S. Evans, 1984. Rotifer seasonal
zooplankton: a principal components approach. Ecology succession and copepod predation in Lake Michigan. J.
55: 1306-1317. Great Lakes Res. 10: 417-428.
Miracle, M. R., 1977. Migration, patchiness, and distribution Stewart, L. G. & D. G. George, 1987. Environmental factors
in time and space of planktonic rotifers. Arch. Hydrobiol. influencing the vertical migration of planktonic rotifers in
Beih. 8: 19-37. a hypereutrophic tarn. Hydrobiologia 147: 203-208.
Pejler, B., 1957. Taxonomical and ecological studies on
planktonic Rotatoria from northern Swedish Lapland.
Kgl. Svenska Vetenskapskad. Handl. 4: 1-68.
Hydrobiologia 186/187: 181-189, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 181
© 1989 Kluwer Academic Publishers.

The rotifer communities of acid-stressed lakes of Maine

Michael T. Brett
Institute of Limnology, Uppsala University, Uppsala, Sweden

Key words: rotifer communities, acidification, Maine

Abstract

The structure of the rotifer community in relation to lake pH, trophic status, the type of planktivore
assemblage and the crustacean community was assessed in a survey of 23 lakes ranging in pH from 4.4
to 7.3, and in a study of two lakes - one acidic, the other circumneutral - during two summers. In both
investigations the number of rotifer species encountered per sample was strongly reduced with pH.
Although the reason for this is not clear acid-stress, the ultraoligotrophic nature of the acidic lakes, and
competitive interactions with crustacean zooplankters may all have played a role. More importantly the
ecological significance of this relationship is not known. The rotifer Keratella taurocephala was a principle
species in the most acidic lakes, while several common rotifers were notably absent from these lakes.
Although rotifer abundance was correlated with lake pH, the results of this study indicate that rotifer
abundance is not a result oflake pH per se, but oflake trophic status and interactions with the crustacean
community.

Introduction MacIssac et al., 1987; Pinel-Alloul et al., 1987;

Schaffner, 1989).
A considerable amount of research has been Considerable confusion exists as to the rela-
carried out on acidification influences on tionship between rotifer abundance or biomass
zooplankton communities, however, the vast and lake pH. Yan & Geiling (1985) reported
majority of this work has focused on the extensive rotifer communities, 15 to 51 % of total
crustacean component of the zooplankton (Brett, zooplankton biomass, in two acidic metal con-
1989b). Of those studies dealing with rotifers the taminated lakes relative to six circumneutral
general fmdings include reductions in the number lakes, where rotifers comprised no more than 2 %
of species encountered per sample with pH of the total biomass. During the experimental
(Almer et a/. , 1974; Roff & Kwiatkowski, 1977; acidification oflake 223 the importance of rotifers,
Hobrek & Raddum, 1980; Brezonik et al., 1984; as portion of total zooplankton biomass, in-
Chengalath et al., 1984; Siegfried et a/., 1984, creased slightly as pH was reduced from 6.8 to 5.0
1988; Yan & Geiling, 1985; Carter etal., 1986; (Schindler et a/., 1985). In apparent contradiction
MacIssac et a/., 1987), and dominance of the to these results Roff & Kwiatkowski (1977),
communities of acidic temperate eastern North- Carter et al. (l986), and MacIssac et al. (1987)
ern American lakes by Keratella taurocephala have all reported reduced rotifer abundance with
(Chengalath et al., 1984; Siegfried et a/. , 1984, pH. However, Pinel-Alloul et a/. (l987) and
1988; Yan & Geiling, 1985, Carter et a/., 1986; Siegfried et a/. (1987) observed that even across
182

broad lake acidity gradients rotifer community selection was biased to include a broad range of
abundance was more closely related to factors pH values, from 4.45 to 7.20, with relatively little
associated with lake trophic status, i.e. chloro- variation in lake morphology. These lakes were
phyll a and total phosphorus, than to lake pH. small, oligo- to mesohumic, with soft water, and
This study examined the relationship between were generally oligotrophic, see Table 1. Five of
lake pH, trophic status, the planktivore com- the lakes, the most acidic, were fishless and
munity and the planktic crustacean and rotifer contained dense populations of limnetic hemi-
communities in a survey of 23 lakes across a pH pterans - especially Buenoa macro tibialis, while
gradient, and in a seasonal comparison of two four contained allopatric populations of brook
lakes (one acidic and the other circumneutral) char (Salvelinus fontinalis), and the remaining 14
during two summers. contained cyprinid fish, mainly golden shiners
(Notomigonus crysoleucas) (Brett, 1989a).
Each of the lakes was sampled once at its
Methods deepest point for zooplankton and water chemis-
try in July or August of 1984. Zooplankton sam-
The 23 lakes investigated were all forest lakes pling was carried out using a 63 f.1.m Wisconsin net
located in isolated regions of Maine, northeastern drawn at a rate of 0.5 m sec - 1 from 1 to 2 m
United States; i.e. these lakes had little or no above the sediment to the surface, except for Mud
human development in their watersheds. Lake Pond which was sampled from 5 m above the

Table 1. Limnological characteristics ofthe 23 lakes surveyed, all values reported as averages for paired duplicate surface (0.5 m)
and bottom (I m above sediment surface) samples taken on the same dates as the zooplankton samples.

Lake Elevation Surface Maximum pH Alkalinity Conduc- Color Total Planktivore

(m) area depth (JLeq/l) tivity (Pt units) P type
(ha) (m) (JLs/cm) (JLg/I) classification

Unnamed P. (WC) 73 2.8 5.2 4.45 -29.1 18 35 7 Insect

Dead P. 48 4.9 10.8 4.50 - 30.4 14 30 6 Insect
Kerosene P. 67 2.8 5.2 4.50 - 19.1 17 45 7 Insect
MudP. 97 2.2 14.0 4.60 -26.5 25 20 2 Insect
Salmon P. (WC) 42 4.5 6.7 4.85 -12.3 14 35 6 Insect
Lily L. 72 13.0 7.3 5.15 - 1.8 17 40 6 Cyprinid
Ledge P. 948 2.4 7.3 5.45 8.9 17 50 5 Salmonid
Anderson P. 65 5.0 6.0 5.45 2.8 16 5 5 Cyprinid
Eddy P. 793 3.6 6.7 5.65 10.8 16 10 6 Salmonid
Garrock P. 145 0.8 6.0 5.75 29.2 17 60 31 Cyprinid
Grenell P. 388 2.4 5.5 5.80 86.2 18 70 9 Salmonid
Hosea Pug L. 88 23.5 10.7 5.85 13.4 16 35 8 Cyprinid
Round P. 279 2.4 9.5 5.85 88.4 25 45 13 Cyprinid
Salmon P. (HC) 85 4.1 10.7 5.90 55.8 25 15 11 Cyprinid
Unnamed P. (PC) 380 6.8 2.0 5.90 43.0 16 85 27 Cyprinid
Secret P. 385 5.7 10.4 6.00 88.2 22 35 12 Cyprinid
Possum P. 80 12.1 5.5 6.10 7.9 14 35 8 Cyprinid
Harvey P. 291 4.1 2.4 6.15 54.8 26 10 11 Cyprinid
Daicey P. 329 15.4 7.9 6.15 6.28 18 10 8 Cyprinid
Trout L. 274 2.0 3.0 6.20 50.7 22 25 11 Cyprinid
Upper Basin P. 745 14.2 9.0 6.30 14.8 14 5 1 Salmonid
Grindstone P. 394 2.4 9.1 6.40 195.9 31 20 9 Cyprinid
Carry P. 900 6.9 3.7 7.20 534.4 63 20 13 Cyprinid
 183

sediment. These samples were preserved in for- cells until at least 400 individuals were counted.
malin. The entire sample was scanned for rarer species.
Two of the lakes included in the survey, Mud The whole of each sample was counted for
Pond and Salmon Pond (HC), were chosen for crustacean zooplankters. In order to generate
more extensive investigations because they repre- biomass values for the lake survey the first 20
sented extremes in the lake types included in the individuals of each crustacean group, generally
lake survey - Mud Pond was acidic and fishless, genus, were measured and their lengths recorded.
while Salmon Pond (HC) was circumneutral with Then length weight regressions (McCaully, 1984)
fish - and because they were morphologically were used to calculated the average weight for the
similar and located approximately 100 m apart, group or species. When possible, equations for
see Table 2. These lakes were sampled on four the same species, or most common species of a
dates in 1983 and six dates in 1984 at biweekly genus, from the same region were used. Alterna-
intervals during the mid-summer period. The tively the same species from another region or a
sampling was carried out at eight stations in the similar species regression was used. Rotifer
littoral zone using a vertical tube sampler (1 m). weights were adapted from Makarewicz & Likens
Samples were filtered through a 64 /lm filter (1979). Species identifications were based on the
apparatus and preserved in Lugol's solution. The taxonomic works of Edmondson (1959), Ruttner-
results for the two-lake comparison are presented Kolisko (1974), Chengalath & Mulamoothil
as pooled data for all stations on each date. (1975), Kiefer & Fryer (1978), Pennak (1978) and
For both the lake survey and the two-lake com- Sternberger (1979).
parison, rotifers and nauplii were counted by In order to rank the lakes surveyed according
decanting known volumes of the samples, sub- to their presumed predation pressure, the lakes
sampling with a Henson-Stemple pipette, and were first divided into three categories based on
enumerating with 1 ml Sedgwick-Rafter counting the type of planktivore present, and then ranked
within these groupings according to the capture
rate of the dominant planktivore in that lake
Table 2. Selected physical, biological, and chemical charac- (Brett, 1989a). Thus the lake dominated by limn-
teristics of Mud Pond and Salmon Pond (HC). All chemical etic insects with the lowest capture rate, during
data expressed as J.Leq L - 1, except as otherwise noted. sweep net sampling, received the lowest rank
while the cyprinid-fish dominated lake with the
Mund Pond Salmon Pond
(HC)
highest capture rate received the highest ranking.

Surface area (ha) 1.6 2.4

Maximum depth (m) 14 11 Results
Elevation (m.a.s.l.) 97 85

Fish absent present

In the lake survey 29 rotifer species were collected
Notonectidis abundant rare and identified, of which only 12 were found in
Chaoborus rare present 40% or more of the lakes. Of these Keratella
eoehlearis, Kellieottia bostoniensis, Kellieottia
pH 4.6 6.2 longispina, Polyarthra remata, Triehoeerea multi-
Alkalinity -26.5 55.8
Ca 28.3 72.4 erinis, Asplanehna priodonta and Collotheea muta-
Al 37.5 3.6 Mis were greatly reduced or absent from those
S04 104 52 lakes below pH 5.0. In fact these seven species
Total phosphorus (ppb) 2 11 had a 73 % rate of occurrence in the lakes above
pH 5.0., but only a 17% rate in lakes below this
Color (Pt units), Na, K, similar between the lakes
Mg, Fe, Zn, Mn, N0 3 , CI
pH. In contrast the rotifer Keratella tauroeephala
was found in every lake investigated (Table 3).
184

Table3. The rotifer community structure of the lakes surveyed, expressed as individuals per liter. Those species listed indi-
vidually occurred in ~ 40 % of the lakes surveyed, while those occurring in fewer lakes are listed all together under other rotifers.

NRS RA Kt Kc Kb Kl Pv Pr Cn Cu Tm Tc Ap Cm oth

Unnamed P. (WC) 5 2 2 <1

Dead P. 7 6 1 4 <1 <1 1
Kerosene P. 7 19 5 11 1 1
MudP. 5 8 7 <1 + <1
Salmon P. (WC) 5 132 <1 131 +
Lily L. 7 138 5 17 6 27 67 15 +
Ledge P. 9 132 33 <1 5 + + 93 <1 <1
Anderson P. 7 86 25 30 + 8 22 +
Eddy P. 9 17 9 + 2 + 5 + +
Garcock P. 14 1920 2 318 1220 276 + 1 50 11 19 20 + 3
Grenell P. 10 124 6 28 <1 19 + 5 39 4
Hosea Pug L. 14 80 4 27 1 12 28 2 3 2 + +
Round P 14 100 4 44 8 13 31 9 24 2 1 +
Salmon P. (HC) 14 89 10 <1 6 2 31 2 20 13 3 2
Unnamed P. (PC) 8 471 348 43 1 71 + + 6
Secret P. 14 171 1 12 8 87 34 1 + 3 4 + + 24
Possum P. 12 210 18 18 <1 11 106 7 4 35 6 5 +
Harvey P. 14 92 46 10 I 1 5 5 + 23 <1 + 1 +
Daicey P. 9 128 + 3 3 8 11 103 + + +
Trout L. 9 176 3 + 167 5 + + +
Upper Basin P. 5 9 <1 <I 1 <1 8
Grindstone P. 12 250 1 158 20 52 9 4 + 5
Carry P. 8 191 30 2 8 136 12 4 +

NRS = number of rotifer species per sample; RA = rotifer abundance per liter; Kt = Keratella taurocephala; Kc = Keratella
cochlearis including K.c. cochlearis, K.c. tecta and K.c. ssp.; Kb = Kellicottia bostoniensis; KI = Kellicottia longispina;
Pv = Polyarthra vulgaris, including P. dolichoptera from one lake; Pr = Polyarthra remata; Cn = Conochiloides natans:
Cu = Conochilus unicorn us, including C. hippocrepis from one lake; Tm = Trichocerca multicrinis; Tc = Trichocerca cylindrica;
Ap = Asplanchna priodonta; Cm = Collotheca mutablis; oth = other rotifer species, including Lecane lunaris, L. luna, Synchaeta
spp., Ascomorpha ecaudis, Keratella hiemalis, Trichocerca elongata, T. platessa, T. myersi, T. simillis, Polyarthra euryptera, Ploesoma
truncatum, P. hudsoni, Filinia longiseta, and Gastropus styli/er.

Table 4. A correlation matrix for various chemical and biological characteristics of the lakes surveyed. All statistically significant
relationships are shown in bold type.

pH
Total phosphorus 0.474 Total phosphorus
Predation rank 0.589 0.788 Predation rank
Specific conductivity 0.396 0.508 0.304 Specific conductivity
Color - 0.353 0.336 0.174 - 0.114 Color
Rotifer species 0.482 0.704 0.634 0.412 0.148 Rotifer species
Rotifer abundance 0.580 0.669 0.799 0.261 0.357 0.526 Rotifer abundance
Rotifer biomass 0.594 0.550 0.678 0.090 - 0.055 0.548 0.403 Rotifer biomass
Crustacean species 0.169 0.499 0.303 0.326 0.122 0.465 0.054 0.273 Crustacean species
Crustacean abundance 0.145 0.023 0.006 - 0.019 -0.093 - 0.117 0.257 -0.204 - 0.386 Crustacean abundance
Crustacean biomass 0.129 0.229 0.078 0.413 - 0.198 - 0.035 - 0.088 0.031 0.158 - 0.044 Crustacean biomass
Total biomass 0.272 0.389 0.250 0.287 - 0.117 0.293 - 0.041 0.536 0.402 - 0.317 0.718

If r > 0.412 then p < 0.05.

If r > 0.531 then p < 0.01
If r > 0.633 then p < 0.001.
 185

Table 5. Mean zooplankton community parameters when the lakes surveyed are divided into dominant planktivore catagories.
Least-square means were calculated, then the data was ranked and compared for significant differences (P < 0.05).

Class Limnetic insect Salmonid Cyprinid Significant differences

n= 5 4 14

pH 4.6 5.8 6.0 LI*SL; LI*CY

Total phosphorus 1 5 5 12 LI*CY; SL*CY
Rotifer species 5.9 8.3 11.2 LI*CY
Rotifer abundance 2 13.6 48.9 263.2 LI*CY; SL*CY
Rotifer biomass 1 0.9 4.5 21.9 LI*CY; SL*CY
Total biomass 1 66.2 72.6 115.4
RA/CA 3 0.6 3.3 5.9 LI*CY

Zooplankton community biomass proportional composition

Rotifers 1 6 19 LI*CY; SL*CY
Nauplii 5 4 18 LI*CY; SL*CY
Bosmina 9 2 3
Diaphanosoma 8 <1 2 LI*CY; LI*SL
Holopedium 29 32 6 LI*CY; SL*CY
Daphnia <I 17 17 LI*CY
Diaptomus 45 32 24
Cyclopida 3 2 II LI*CY; SL*CY

1. in Jl g per liter.
2. in individuals per liter.
3. RA/CA = rotifer abundance/crustacean abundance.

Table 4 is a correlation matrix for various chemi- less evasive cladocerans Holopedium gibberum
cal and biological characteristics of the lakes and Diaphanosoma birgei were more important in
surveyed. Notably this matrix shows a very high the limnetic insect dominated lakes.
amount of interrelatedness between the predictor In the two-lake study 16 rotifer species were
variables pH, total phosphorus, and predation identified from the acidic lake, and 21 species
rank. These variables, in turn, predicted nearly were observed in samples from the circumneutral
identical variation in the rotifer community lake. The acidic lake averaged 4.5 ± 2.2 (± sd)
parameters - rotifer species, rotifer abundance, species per sample while the circumneutral lake
and rotifer biomass. averaged 10.4 ± 2.2. The rotifer community of the
By dividing the lakes into planktivore cate- acidic lake was strongly dominated by Keratella
gories and analyzing for differences in zooplank- taurocephala (98% of individuals observed) while
ton community composition between them, the rotifer community of the circumneutral lake
strong differences emerge. Namely, in both was composed of 3 to 5 species, with Keratella
absolute terms (rotifer abundance and rotifer taurocephala again the dominant (77 % of indi-
biomass) and relative terms (the ratio between viduals observed), but with Keratella cochlearis,
rotifer abundance and crustacean abundance and Kellicottia longispina, Trichocerca cylindrica, Cono-
the rotifer portion of the zooplankton biomass) chilus unicornis, and Collotheca mutabilis periodi-
rotifers were strongly favored by cyprinid preda- cally common.
tion relative to limnetic insect predation. Like- Notably, the crustacean community of the
wise, cyprinid predation was associated with acidic fishless lake was principally composed of
enhanced nauplii and cyclopoid components of the calanoid Diaptomus minutus and the
the zooplankton, whereas the relatively large and cladocerans Diaphanosoma birgei and Polyphemus
186

pediculus, while the crustacean community of the crustacean community of that lake and to the
circumneutral fish-containing lake was again rotifer community of the circumneutral lake,
dominated by D. minutus, but also contained large except for the month of August 1984 when rather
cyclopoid and Bosmina longirostris components, high abundances were observed (Tables 6, 7).
Tables 6 and 7. Rotifer abundance was generally This rise in rotifer abundance coincided with a
reduced in the acidic lake, relative to the marked decline in the cladoceran abundance

Table 6. Zooplankton community structure in Mud Pond, expressed as individuals per liter.

Date
1983 1984

21/6 7/7 21/7 8/8 12/6 28/6 12/7 26/7 9/8 27/8

Copepods 1 6 8 4 1 26 3 3 6 14 o
Nauplii 37 10 5 4 28 <1 <1 <1 <1 <1
Cladocerans 2 7 4 5 7 4 9 2 2 o
Keratella taurocephala <1 9 7 8 6 8 37 229 287
Other rotifers <1 <1 <1 <1 <1 <1 <1 <1 <1
Total rotifer abundance <1 9 7 8 6 1 8 37 229 287
Number of rotifer 3 5 7 3 3 6 6 8 3
species per sample

1. copepods = 96% Diaptomus minutus, 4% Mesocyclops edax.

2. c1adocerans = 58% Diaphanosoma bigrei, 26% Polyphemus pediculus, 16% Bosmina longirostris.

Table 7. Zooplankton community structure in Salmon Pond (HC), expressed as individuals per liter.

Date
1983 1984

21/6 7/7 21/7 8/8 12/6 28/6 12/7 26/7 9/8 27/8

Copepods I 21 10 7 10 24 17 19 8 46 6
Nauplii 13 32 12 13 12 60 12 5
Cladocerans 2 32 9 5 8 11 5 6 9 20 2

Keratella taurocephala 27 36 36 63 49 93 120 50 77 11

Keratella cochlearis 20 2 26 6 8 4 3 12 4
Kellicottia longispina <1 <1 <1 3 5 8 2 1
Trichocerca cylindrica <1 <1 3 <1 1 4 13
Conchilus unicornis <1 10 1
Collotheca mutabilis <1 <1 2 6
Asplanchna priodonta 1 <1 <1 2 3 <1 2
Polyarthra vulgaris <I 4 <1 1 <1
Other rotifers <1 <1 <1 <1 <1 <1 <1 <1 <1 <1
Total rotifer abundance 48 38 38 97 72 109 135 67 103 19
Number of rotifer 8 7 13 12 12 13 8 11 9 II
species per sample

1. copepods = 72% Diaptomus minutus, 18% Tropocyclops prasinus mexicanus, 10% Mesocylops edax.
2. c1adocerans = 92% Bosmina longirostris, 7% Diaphanosoma birgei.
 187

during this period. In samples collected at the Schaffner, 1989) indicates that acid-stress, at
same time, but from the pelagic as apposed to the least in influencing competitive interactions
littoral region of Mud Pond, crustacean between species, might have an important
zooplankters were much more abundant and the influence on rotifer species number. Furthermore
rotifers were greatly reduced (Table 3, 6). that several common species had greatly reduced
rates of occurrence in lakes below pH 5.0 relative
to lakes above this pH is also indicative of stress
Discussion influences. Notably MacIssac et ai. (1986)
reported that when a severely acidic metal-
The most obvious result of this study, for both the contaminated lake partially recovered, pH change
lake survey and the two-lake comparison, was a from 4.0 to 5.0, the rotifer community shifted from
reduction in the number of rotifer species per one dominated almost exclusively by Keratella
sample with pH. This observation has been made taurocephaia and Synchaeta spp., to one domi-
in several other studies (Almer et ai., 1974, RofT nated by these species plus Poiyarthra spp.,
& Kwiatkowski, 1977; Hobrek & Raddum, 1980; Trichocerca similis, and Conociloides natans. A
Brezonik et ai., 1984; Chengalath et aI., 1984; more fundamental problem is the ecological sig-
Siegfried et aI., 1984, 1988; Yan & Geiling, 1985; nificance of the reduced species number in the
Carter et ai., 1986; MacIssac et aI., 1987), how- acidic lakes. Is this observation ecologically
ever to what extent this is a direct result of lake irrelevant - but technically easy to generate, or is
acidity or the otherwise simplified nature of acidic it of real importance to the functioning of acid-
lakes is poorly understood. In the present mate- stressed ecosystems?
rial the number of species per sample was also The abundance of rotifers in the present study
strongly, in fact more strongly, correlated to total was also correlated with pH as has been observed
phosphorus, lake predation rank, and rotifer den- in several other studies (RofT & Kwiatkowski,
sity. It has been suggested (Pejler, 1983) that ultra- 1977; Brezonik et ai., 1984; Carter et ai., 1986,
oligotrophic lakes, such as the most acidic lakes MacIssac et ai., 1987). However, rotifer abun-
of the present study, are generally species dance was more strongly correlated with total
depauperate relative to mesotrophic lakes, such phosphorus. This in agreement with the results of
as several of the circumneutrallakes of the pres- several other studies which found zooplankton
ent study. The strong relationship between rotifer abundance in acid-stressed lakes to be more
species number and abundance may in fact be closely related to lake trophic status, and hence
important, for instance reduced rotifer abundance presumably the availability of edible algae and
might be indicative of conditions unfavorable for bacteria, than to pH per se (Brezonik et ai., 1984;
rotifers as a whole. One could also speculate that Pinel-Alloul et ai., 1987; Kerekes et ai., 1988;
reduced predation pressure on the crustaceans - Siegfried et ai., 1987). This finding is also con-
invertebrate versus fish, could act to increase sistent with several nutrient enrichment studies of
competitive interactions and hence dominance of acidic lakes (Yan et al., 1982; DeCosta et al.,
large cladocerans over rotifers as a group, thereby 1983; Yan & Lafrance, 1984) showing dramatic
reducing 'niche space' for rotifers in the most increases in zooplankton community abundance
acidic lakes. The fact that Keratella taurocephaia or biomass after enrichment, and is consistent
was so strongly dominant in the most acidic of the with the relationship between nutrient levels and
two lakes compared, and that it was important in community standing crop typical for non-acidic
several of the other acidic lakes examined, as well lakes.
as other acidic lakes in eastern North America Analysis of zooplankton community structure
(Chengalath et aI., 1984; Siegfried et ai., 1984, across planktivore type categories suggests that
1988; Yan & Geiling, 1985; Carter et aI., 1986; planktivore type influenced the relative com-
MacIssac et aI., 1987; Pinel-Alloul et aI., 1987; position of the zooplankton. Cyprinid fish preda-
188

tion was associated with increased representation clearly shown the ability of crustaceans to depress
of the rotifer community, nauplii, and cyclopoids rotifer populations (Gilbert, 1985), and have also
- small or highly evasive groups - at the expense shown that intense predation on the crustacean
of the much larger Holopedium gibberum and component of the zooplankton can greatly
Diaphanosoma birgei, both in absolute, and more increase rotifer abundance (Neill, 1984).
importantly relative terms, e.g. as regards portion
of the zooplankton community biomass. This
would indicate that fish predation, which is Acknowledgements
presumably most concentrated on larger clado-
cerans - for reasons of size and evasiveness, I thank T. Haines who provided considerable
liberated resources or reduced competitive inter- assistance during the course of this study.
actions with crustacean zooplankters thereby
benefitting the rotifers. The extreme case of this
can be exemplified by Garcock Pond which had References
an allopatric population of golden shiners, a very
large population relative to lakes of this region, Almer, B., W. Dickson, C. Ekstrom, & E. Hornstrom, 1974.
which reduced the crustacean community to less Effects of acidification on Swedish lakes. Ambio 3: 30-36.
than one individual per liter, despite the fact that Brett, M. T., 1989a. The distribution of limnetic macro-
invertebrates in acidic lakes of Maine: the Role of fish
this lake had the highest total phosphorus concen-
predation. Aque Fennica (in press).
tration of the lakes examined. In this lake the Brett, M. T., 1989b. Zooplankton communities and acidifi-
rotifer abundance was estimated at 2000 individ- cation processes - a review. Wat. Air Soil Pollut. 44:
uals per liter, an abundance 5 times higher than 387-414.
that recorded in any of the other lakes studied. Brezonik, P. L., T. L. Crisman & R L. Schulze, 1984. Plank-
tonic communities in Florida softwater lakes of varying
Interactions between the crustacean and rotifer
pH. Ca. J. Fish. aquat. Sci. 41: 46-56.
communities was also indicated by the results Carter, J. C. H., W. D. Taylor, R Chengalath & D. A.
from the acidic lake of the two-lake comparison, Scruton, 1986. Limnetic zooplankton assemblages in
where the rotifer K. taurocephala increased Atlantic Canada with special reference to acidification. Ca.
dramatically after first the cladocerans and then J. Fish. aquat. Sci. 43: 444-456.
Chengalath, R, W. J. Bruce & D. A. Scruton, 1984. Rotifer
the copepods were greatly reduced in the littoral
and crustacean plankton oflakes in insular Newfoundland.
zone. This pattern was not observed in the pelagic Verh. int. Ver. Limno!. 22: 419-430.
zone of this lake during the same period, presuma- Chengalath, R., & G. Mulamoottil, 1975. Littoral rotifera of
bly because the chief planktivore in this lake was Ontario - genus Trichocerca. Ca. J. Zoo!. 53: 1403-1411.
not particularly abundant in the pelagic relative to DeCosta, J., A. Janicki, G. Shellito & G. Wilcox, 1983. The
effect of phosphorus additions in enclosures on the phyto-
the littoral zone. Other studies of acid-stressed
plankton and zooplankton of an acid lake. Oikos 40:
zooplankton communities have also indicated the 283-294.
importance ofinteractions between the rotifer and Edmondson, W. T., 1959. Freshwater-biology. John Wiley,
crustacean components of the zooplankton, New York.
namely Yan & Geiling (1985) used the reduced Gilbert, J. J. 1985. Competition between rotifers and
Daphnia. Ecology 66: 1943-1950.
crustacean biomass, relative to six circumneutral
Hoblek, A. & G. G. Raddum, 1980. Zooplankton com-
lakes, and hence reduced competition with munities in acidified regions of south Norway. SNSF-
rotifers, to explain the extremely high rotifer project. IR75/80.
biomasses attained in their two acidic lakes. Kerekes, J. J., A. C. Blouin & S. T. Beauchamp, 1988.
While Yan etal., (1982) showed dramatic in- Trophic response to phosphorus in acidic and non acidic
lakes in Nova Scotia, Canada. P. Biro (ed.) Trophic
creases in K. taurocephala abundance of an acidic
relationships in inland waters symposium, Tihany,
lake after predation by Chaoborus had virtually Hungary. Junk Pub!. Co. (in press).
eliminated the crustacean community. Further- Kiefer, F. & G. Fryer, 1978. Das zooplankton def binnen-
more, several studies of non-acidic systems have gewasser. Die binnengewasser. 26 (2).
 189

MacIsaac, H. J., W. Keller, T. C. Hutchinson & N. D. Yan, Schindler, D. W., K. H. Mills, D. F. Malley, D. L. Findlay,
1986. Natural changes in the planktonic rotifera of a small J. A. Shearer, I. J. Davies, M. A. Turner, G.A. Linsey &
acid lake near Sudbury, Ontario following water quality D. R. Cruikshank, 1985. Long-term ecosystem stress: the
improvements. Wat. Air Soil Pollut. 31: 791-797. effects of years of experimental acidification on a small
MacIssac, H. 1., T. C. Hutchinson & W. Keller, 1987. Analy- lake. Science (Washington, DC) 228: 1395-1401.
sis of planktonic rotifer assemblages from Sudbury, Siegfried, C. A., 1. A. Bloomfield & 1. W. Sutherland, 1988.
Ontario, area lakes of varying chemical composition. Ca. The planktonic rotifer community structure in Adirondack
1. Fish. aquat. Sci. 44: 1692-1701. lakes in relation to acidity, trophic status, and related
Makarewicz, 1. C. & G. E. Likens, 1979. Structure and the water quality characteristics. Hydrobiologia (in press).
function of the zooplankton community of Mirror Lake, Siegfried, C. A., J. W. Sutherland & 1. A. Bloomfield, 1987b.
New Hampshire. Eco!. Monogr. 49: 109-127. Analysis of plankton community structure in Adirondack
function of the zooplankton community of Mirror Lake, lakes in relation to acidification. In R. Perry, R. M.
New Hampshire. Eco!. Monogr. 49: 109-127. Harrison, J. N. B. Bell & 1. N. Lester (eds.). Acid Rain:
McCaully, E., 1984. The estimation of the abundance and Scientific and Technical Advances. Selper Ltd., London.
biomass of zooplankton in samples. In Downing,J. A. and 445-450.
Rigler, F. H. (eds.), Secondary productivity in freshwaters. Siegfried, C. A., J. W. Sutherland, S. O. Quinn & J. A.
Blackwell Scientific Publications, Oxford. 228-265. Bloomfield, 1984. Lake acidification and the biology of
Neill, W. E., 1984. Regulation of rotifer densities by Adirondack lakes: I. Rotifer communities. Verh. int. Ver.
crustacean zooplankton in an oligotrophic montane lake in Limno!. 22: 549-558.
British Columbia. Oecologia 61: 175-181. Sternberger, R. S., 1979. A guide to rotifers of the Laurentian
Pejler, B., 1983. Zooplanktic indicators of trophy and their Great Lakes. US EPA, Report EPA 600/4-79-021.
food. Hydrobiologia 101: 111-114. Yan, N. D. & W. Geiling, 1985. Elevated planktonic rotifer
Pennak, R. W., 1978. Fresh-water invertebrates ofthe United biomass in acidified metal-contaminated lakes near
States. John Wiley, New York. Sudbury, Ontario. Hydrobiologia 120: 199-205.
Pinel-Alloul, B., G. Methot & G. Codin-Blumer, 1987. Struc- Yan, N. D. & c. Lafrance, 1984. Responses of acidic and
ture spatiale du zooplancton des lacs du Quebec: relation neutralized lakes near Sudbury, Ontario, to nutrient
avec l'acidite. Nat. can. 114: 295-305. enrichment. In J. Nriagu (ed.) Environmental impact of
Roff, J. C. & R. E. Kwiatkowski, 1977. Zooplankton and smelters. John Wiley and Sons, INC., New York. 457-521.
zoo benthos communities of selected northern Ontario Yan, N. D., C. J. Lafrance & G. G. Hitchin, 1982. Planktonic
lakes of different acidities. Ca. 1. Zoo!. 55: 899-911. fluctuations in a fertilized, acidic lake: the role of inverte-
Ruttner-Kolisko, A., 1974. Plankton rotifers. Biology and brate predators. In R. E. Johnson (ed.) Acid Rain/
taxonomy. Die binnengewasser. 26(1). Fisheries. Proceedings of a symposium on acidic rain and
Schaffner, W. R. 1989. Effects of neutralization an the addi- fishery impacts in northeastern North America. 137-154.
tion of brook trout (Salvelinus fontinalis) on the limnetic
zooplankton communities of two acidic lakes. Ca. 1. Fish.
aquat. Sci. 46: 295-305.
Hydrobiologia 186/187: 191-200, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 191
© 1989 Kluwer Academic Publishers.

Composition and distributional patterns in arctic rotifers

R. Chengalath & W. Koste

National Museum of Natural Sciences, P.O. Box 3443, Station 'D', Ottawa, Ontario, Canada KIP 6P4;
D 4570 Quakenbruck, Ludwig-Brill-Strsse 5, Federal Republic of Germany

Key words: arctic North America, rotifera, distribution, community composition

Abstract

Based on collections of rotifers from 212 localities in arctic North America, the patterns of distribution
and composition are evaluated. An attempt is made to discern the dominant components of the rotifer
community in arctic habitats. One hundred and sixty five species of rotifers are reported, three of which
represent new records for North America. With increasing latitude and decreasing summer temperatures,
a decline in species richness and change in species composition is observed. Some rotifers that previously
were not adequately described are redescribed and illustrated. The significance of the dispersal capacity
of rotifers in the arctic is discussed.

Introduction American arctic, to redescribe and illustrate some

species that previously were not adequately de-
Studies on the rotifers of arctic North America scribed and to assess the significance of dispersal
have been very few (Ahlstrom, 1940, 1943 ; capacity as well as overall distribution patterns.
Chengalath, 1978; Chengalath & Koste, 1987;
Harring, 1921; Hooper, 1947; Moore, 1978.) and
were generally limited to restricted areas. There Materials and methods
are forty-eight publications, mainly dealing with
fish and general zooplankton studies, that list Qualitative sampling of lakes and ponds
rotifers from the North American arctic throughout Alaska (USA), and Yukon Territory,
(Chengalath, 1984). In most of these studies Northwest Territories, and Bathurst Island
rotifers were identified only to the generic level (Canadian arctic archipelago) were carried out
and no morphological or ecological data were during summer field expeditions. A total of 212
provided. Comprehensive biogeographic studies samples have so far been examined and are in-
are necessary for complete systematic and cluded in this study. The Bathurst Island samples
ecological understanding of any animal group. were collected in 1974 and the rest in the summer
This paper is part of an ongoing investigation to of 1986 and 1987. At each of the five arctic regions
examine extensively the occurrence, distribution, (See Map, Table 1,2), from 22 to 84 different
and species diversity of the Rotifera of Canada. aquatic habitats were sampled with a plankton
We will also attempt to discern the dominant net with a 64 f.lm aperture mesh. The waterbodies
components of the rotifer communities in North in the main North American land mass under
 1.0
tv

/
///
~-," .; , /,/'
~.~ ".
/
: .' "'-;( . ,,,, . ,,

•: '.• ./I . "'~"'

........ • NORTHWEST
• • / • I TERRITORES
. '/ •'YUKON •
, / TERR!lO~

<~ ':'" ~~
\'.•.::;,:•.....~ .t
.... ~ .....~ _ .-;". J ...
'~
'", . . . :7"--.... ..._. .
,i --.. ·. ·-r·_·_···-._._-.. ""'r . . . . ..... _ a._
i ; i

I
! j
, •
I
! ! f
/ / ! .'
/ CANADA I /1
\
! I /
\\ j ! /
~ J I •
D Unglaclate<l region \ I 1 ;
-.. ... , I 1 I
......... I I I i

'-"'-" --'~' . . ,. .1 I \
I •
...... - - _. - ... . . ' 'o ' • •• _ . . . . ... ..' ' .

Map: The study area showing all the localities sampled.

 193

Table 1. Distributional records for sub-arctic and arctic rotifers. (SYN = southern Yukon and Northwest Territories,
SA = southern Alaska, NYN = northern Yukon and Northwest Territories, NA = northern Alaska, BI = Bathurst Island,
Canadian arctic archipelago).

Rotifer taxon Region Rotifer taxon Region

SYN SA NYN NA BI SYN SA NYN NA BI

Ascomorpha ecaudis Perty x x x Eothinia elongata x x

A. saltans Bartsch x (Ehrenberg)
Aspelta aper (Harring) x x x Epiphanes senta (Muller) x
A. circinator (Gosse) x x Euchlanis alata Voronkov x
Asplanchna priodonta x x x E. deflexa Gosse x x
Gosse E. dilatata Ehrenberg x x x x x
Asplanchnopus multiceps x E. incisa Carlin x x x x
(Schrank) E. lyra Hudson x x x x
Bipalpus hudsoni (Imhoff) x E. meneta Myers x x x x x
Cephalodella catellina x x E. oropha Gosse x x
(Muller) E. triquetra Ehrenberg x x x x
C. forficata (Ehrenberg) x x Filinia longiseta Ehrenberg x
C. forficula (Ehrenberg) x x F. terminalis (Plate) x x
C. gibba (Ehrenberg) x x x x x Floscualria melicerta x
C. globata (Gosse) x Ehrenberg
C. gracilis (Ehrenberg) x F. ringens Linnaeus x x
C. megalocephala x x Gastropus stylifer Imhoff x x x
(Glasscott ) Kellicottia longispina x x x x
C. sterea (Gosse) x x x (Kellicott )
C. ventripes (Dixon-Nuttall) x x x Keratella cochlearis x x x
Collotheca companulata x (Gosse)
(Dobie) K. faluta Ahlstrom x
C. edentata (Collins) x x x K. hiemalis Carlin x x x
C. libera (Zacharias) x K. quadrata (Muller) x x
C. mutabilis (Hudson) x x K. recurvispina x x x
C. ornata (Ehrenberg) x x x (J agerskiold)
C. pelagic a (Rousselet) x K. serrulata (Ehrenberg) x x x
Colurella adriatica x x Lecane acus (Harring) x x x
Ehrenberg L. bulla (Gosse) x x x x
C. colurus (Ehrenberg) x L. closterocerca (Schmarda) x x x x
C. obtusa (Gosse) x x x x L. cornuta (Muller) x x
C. uncinata (Muller) x x x x L. flexilis (Gosse) x x x x x
C. dossuarius (Hudson) x x x L. furcata (Murray) x x x
C. natans (Seligo) x x L. hamata (Stokes) x x x
Conochilus hippocrepis x x x x L. lauterborni Hauer x
(Schrank) L. levistyla (Olofsson) x x
C. unicornis (Rousselet) x x x x L. ligona (Dunlop) x x x
Dicranophorus epicharis x x L. luna (Muller) x x x x
Harring & Myers L. lunaris (Ehrenberg) x x x x x
D. forcipatus (Muller) x x L. mira (Murray) x x x x
D. lutkeni (Bergendal) x L. perplexa Ahlstrom x x x
D. robustus Harring & x L. pyriformis (Daday) x
Myers L. quadridentata x
D. rostratus (Dixon-Nuttall x (Ehrenberg)
& Freeman) L. rotundata (Olofsson) x
Dipleuchlanis propatula x L. scutata (Harring & x x x
(Gosse) Myers)
Encentrum grande x L. stichaea Harring x x x x
(Western) L. stokesi (Pell) x x x
194

Table 1. (continued). TableJ. (continued).

Rotifer taxon Region Rotifer taxon Region

SYN SA NYN NA BI SYN SA NYN NA BI

L. ungulata (Gosse) x x x P. loniremis Carlin x

Lepadella acuminata x x x x x P. major Burckhardt x x x
(Ehrenberg) P. minor Voigt x x
L. ovalis (Muller) x x x x x P. remata Skorikov x
L. patella (Muller) x x x x x P. vulgaris Carlin x x
L. quadricarinata x x x Proales decipiens x
(Stenroos) (Ehrenberg)
L. rhomboides (Gosse) x P. fallacioca Wulfert x x x
L. triptera Ehrenberg x x x P. theodora (Gosse) x x
Limnias me1icerta Weiss x Ptygura brachiata (Hudson) x
Lophcharis salpina x x x x x P. crystllina (Ehrenberg) x
(Ehrenberg) P. mucicola (Kellicott) x
L. oxysternon (Gosse) x x Resticula melandocus x
Microcodon clavus x x (Gosse)
Ehrenberg Scaridium longicaudum x x x
Monommata aeschyna x (Muller)
Myers Squatinella rostrum x
M. caudata Myers x (Schmarda)
M. dentata Wulfert x Stephanoceros fimbriatus x
M. grandis Tessin x x (Goldfusz)
M. phoxa Myers x Synchaeta grandis x x
Mytilina bicarinata (Perty) x Zacharias
M. mucronata (Muller) x x x x x S. longipes Gosse x x
M. ventralis (Ehrenberg) x x x x x S. oblonga Ehrenberg x x
Notho1ca acuminata x x x S. pectinata Ehrenberg x
(Ehrenberg) S. tremula (Muller) x
N. foliacea (Ehrenberg) x x x Taphrocampa selenura x
N. labis Gosse x Gosse
N. lapponica (Ruttner- x x Testudinella emarginula x x x
Kolisko) (Stenroos)
N. squamula (Muller) x x x x x T. mucronata (Gosse) x
Notommata allantois x T. naumanni Carlin x
Wulfert T. parva (Ternetz) x x x x
C. cerberus (Gosse x x T. patina (Hermann) x x x x
N. copeus Ehrenberg x x x x Tetrasiphon hydrocora x x x
N. cyrtopus (Gosse) x x Ehrenberg
N. doneta Harring & x x Trichocerca bidens (Lucks) x x
Myers T. bicristata (Gosse) x x
N. glyphura Wulfert x x x x T. brachyura (Gosse) x
N. pachyura (Gosse) x x T. cavia (Gosse) x x x
Playias qudrocornis x x T. collaris (Rousse1et) x
(Ehrenberg) T. elongata (Gosse) x x x
Pleurotrocha petromyzon x x x x T. insignis (Herrick) x x
Ehrenberg T. intermedia (Stenroos) x
Ploesoma triacanthum x x x T. longiseta (Schrank) x x x x
(Bergendal) T. porcellus (Gosse) x x x
P. truncatum (Levander) x x x x T. pusilla (Lauterborn) x x
Polyarthra dolichoptera x x x x T. rattus (Muller) x x x x
Idelson T. rosea (Stenroos) x x
P. euryptera Wierzejski x T. rousseleti (Voigt) x
 195

Table 1. (continued). Results and discussion

Rotifer taxon Region
A list of 165 rotifers found during this study is
SYN SA NYN NA BI given in Table 1, including the three species that
are new for North America. Except for the three
T. similis (Wierzejski) x common species of rotifers, Kerateiia cochiearis,
T. tenuoir (Gosse) x x x Keratella quadrata, and Kellicottia iongispina, all
T. tigris (Muller) x
the other 110 species are recorded from Alaska
T. uncinata (Voigt) x
T. weberi (Jennings) x
for the first time. Bdelloid rotifers were present in
Trichotria pocillum (Muller) x x x x most samples but could not be positively identi-
T. tetractis (Ehrenberg) x x x x x fied to species level and are, therefore, not
Wolga spinifera (Western) x included in this study. The total number of species
encountered in the sample varied at each site from
0-22, with the majority of the samples containing
study vary from small, highly coloured, ponds that 8-12 species. Twenty-one samples contained no
have a low content of electrolytes and profuse rotifers at all. The rotifer community in the high
vegetation to alpine lakes surrounded by snow arctic sites of Bathurst Island is composed of only
with clear blue waters and sandy bottom with 21 species (Tables 1,2), and a high percentage of
little or no vegetation. The high arctic site, ponds lacked rotifers entirely. These ponds are
Bathurst Island, is one of the Parry group of characterised by extremely low allochthonous
islands situated in the central part of the Canadian nutrient input and lower conductivity. However,
arctic archipelago (see Map). The island has at a number of low arctic sites in Alaska, Yukon,
rather low elevation and is composed pre- and Northwest Territories, ponds with low con-
dominantly of sedimentary rocks (Blake, 1964). ductivity contained a rich rotifer fauna, indicating
The samples were collected from within a radius that lack of rotifers may be correlated more
of three miles of a field station at 75 0 43' N, strongly with the lack of available nutrients for
98 0 25' W. The ponds sampled in this region were phytoplankton growth.
essentially shallow basins, with a diameter of The size of the species pool gradually increased
30 m, a maximum depth of 25 cm, and with a soft from the central arctic island southwards
muddy bottom. (Table 2), with the most dramatic change in the
assemblage occurring upon moving inland onto
continental North America. The species encoun-
tered in this study are generally widely distributed
inhabitants of ponds and littoral zone of lakes
(Koste, 1978). However, the occurrence of several

Table2. Numbers of rotifer species inhabiting designated regions in North American arctic.

Region N.lat. W.long. No. of Loricate Illoricate

localities rotifers rotifers
sampled

Southern Yukon & NWT (SYN) 60° 00'-66° 33' 112° 12'-142° 08' 84 88 30
Southern Alaska (SA) 61 ° 30' -66° 33' 112° 15'-152° 00' 30 66 23
Northern Yukon & NWT (NYN) 66° 33'-69° 10' 122° 10'-132° 15' 22 63 18
Northern Alaska (NA) 66° 33'-70° 30' 150° 00' -151 ° 10' 38 66 21
Bathurst Island, Canadian 75° 43' 98° 25' 38 19 2
arctic archipelago, (BI)
196

species represents an extension of their known ditches or living as epibionts of Asellus aquaticus
range. Members of the genera Notholca, Mytilina, and Gammarus sp. (Western, 1891; Koste, 1976).
Lepadella, Lecane, and Trichotria were the most Specimens are usually contracted in preserved
widely distributed and present in all five regions samples to about half their normal size and
(Table 1). Distribution pattern for individual seldom reveal the coronal structure or the
species can provide useful qualitative data; how- posterior end showing the toes. The trophi
ever no attempt was made during this study to (Figs. 2-5) however is very characteristic and is
derme the distribution of each species as a seen invariably through the body. Another inter-
function of anyone or combination of limnologi- esting rotifer, Mytilina bicarinata, (Figs. 10-12)
cal parameters. has previously been reported from temporary
Three little known species of rotifers, Encen- waterbodies and from among vegetation in
trum grande, Lecane rotundata, and Mytilina bi- smaller ponds mainly from northern Europe
carinata, were found in samples from the (Olofsson, 1918; Pejler, 1962). Lecane rotundata
Canadian arctic archipelago, and represent new (Fig. 6) was found in two samples from Bathurst
records for North America. Encentrum grande Island and has hitherto been reported only from
(Figs. 1-5) was first recorded from the arctic by coastal waters of Novaja Semlja, Spitzbergen,
Murray (1908) from Spitzbergen living in moss. and Swedish Lapland (ldelson, 1926; Olofsson,
This species is mostly known from Europe where 1918; Pejler, 1962). The lorica of this species is
it is found in the psammon of shallow ponds and almost spherical in outline and could easily be

50pm
100
}Jm

3
Figs. 1-5. Encentrum grande. 1. Ventral view. 2. Trophi. 3&4. Teeth on unci and rami, apical view. 5. Fulcrum, lateral view.
 197

,
··
,
,
,
,,
,
.
,
100,","

. \" ; :
~\, " .. / : ./~
,
,
:,.
:'

100 pm

11 9

Figs. 6-12. 6. Lecane rotundata, ventral view. 7. Lophocharis oxysternon, lateral view. 8. L. oxysternon, dorsal view. 9. L. oxysternon,
ventral view. 10. Mytilina bicarinata, dorsal view. 11. M. bicarinata, lateral view. 12. M. bicarinata, cross-section of lorica.

mistaken for a number of other Lecane species discern all the parts clearly. It is readily recognised
under low magnifications. by its asymmetry, the left dorsal appendages being
Polyarthra minor and Lophocharis oxysternon almost twice as long and a little wider than the
were found fairly commonly in northern Alaskan rest. Also, one of the appendages in the right
ponds. Both these species have been reported ventral bundle is broader than the other two
from various parts of the world (Bartos, 1951; (Fig. 13). All the appendages have clearly visible
Koste, 1978; Ruttner-Kolisko, 1974). Although mid-ribs and serrated edges. The more or less
Polyarthra minor is considered to be a rare acid broadly oval body seldom exceeds a size of
water form, during the present study it was found 90 ].lm. The vitellarium contains four distinct nu-
in alkaline ponds with low conductivity. The mor- clei (Fig. 13). Lophocharis oxysternon is mentioned
phology of this species has long been neglected in the literature as a variable species (Bartos,
mainly because of its very small size and 1959; Gosse, 1851; Koste, 1978; Kutikova, 1970)
transparency which make it very difficult to but the illustrations provided were far from satis-
198

13

100 pm

Fig. 13. Polyarthra minor, ventral view.

factory. The lorica of this animal is oval shaped to life history patterns, favourable environmental
with distinct ridges along the dorsal side conditions, or absence of predators. Several
(Figs. 7-9). It is also distinguished from other morphs of Notholca acuminata and Notholca squa-
members of this genus by its characteristic lateral mula were found in the high arctic localities. Many
VIew. of these forms are also found in the low arctic and
Species of the genera Notholca and Mytilina have been described previously (Chengalath &
predominated the high arctic rotifer fauna, and Koste, 1987). In several samples in the arctic two
the body sizes of the animals found during this or three morphs of a species with varying body
study were much larger than the previously sizes were present. For example, Notholca acumi-
recorded forms. There are references to giant nata and N. squamula ranged from 180 f.lm to
forms of rotifers in the literature (Chengalath & 383 f.lm in body size. The significance of such size
Koste, 1983; Koste & Shiel, 1980). The reasons differences in rotifers is not fully understood.
for the occurrence of such rotifers may be related Water temperatures are usually cited as a factor
 199

for variation in sizes. However, the body size thenogens producing ephippial eggs asexually.
variation among conspecifics cannot be attributed While physiochemical factors may play a role in
to temperature. Patterns of size separation are preventing successful colonisation in high arctic
found to be common in copepod communities regions, dispersal difficulties including failure to
and it has been suggested that such separation produce resting eggs may be the controlling fac-
minimises competition for resources among tor. Too little is known about the production,
species (Cole, 1961; Hutchinson, 1967). A num- morphology and dispersal of rotifer resting eggs
ber of studies on calanoid copepods have for any definite conclusions to be drawn.
indicated that food abundance is the primary fac- Although passive dispersal of resting eggs and
tor determining their body sizes (Deevey, 1960; ephippial eggs via birds and wind has been docu-
Harris & PafTenhoffer, 1976). It has also been mented (Maguire, 1963), the dispersal capability
shown that copepod body sizes increased greatly and opportunity may be responsible for distri-
when population densities were reduced (Hebert, butional patterns among animals. However, dis-
1985). Perhaps these are also contributing factors persal history alone does not restrict species dis-
to the increased body size of arctic rotifers. tributions. Survival in a new habitat involves
Some patterns of rotifer community composi- other factors such as avoidance of predators,
tion in the arctic emerge from this study. The availability of food, completion of the life cycle
waterbodies in the arctic archipelago support a within the short favourable season including the
very depauperate assemblage of species com- production of resting eggs.
pared with habitats located to the south
(Table 1,2). The severity of the physical environ-
Acknowledgements
ment, low temperature and low primary produc-
tion are clearly factors limiting size and composi-
We thank Colin Eades, Chief, Zoology Division,
tion of rotifer community in the high arctic habi-
National Museum of Natural Sciences, for his
tats. In comparison to other regions, the Alaskan
support and encouragement. Thanks are also due
habitats have a high diversity of rotifers that may
to Chang-tai Shih for critically reading the
be attributable to the fact that much of the area
manuscript. The help of Maria Wing in assisting
escaped glaciation during the pleistocene
with the manuscript and that of Brett Purdy, Jerry
(Hopkins, et al., 1982). The possibility exists that
Chemilowski, and Bonnie Smith, all of the
the diversity patterns of rotifers reflect the effects
Department of Biology, University of Calgary, in
of survival in glacial refugia. Most long distance
collecting the samples from Yukon, Northwest
movements of zooplankton is probably mediated
Territories, and Alaska is gratefully acknowl-
through birds and opportunities for such dispersal
edged. Ian Sutherland, Canadian Heritage Infor-
are undoubtedly greater along the north-south
mation Network, Department of Communica-
axis. The high arctic localities in Bathurst Island
tions, collected the samples from Bathurst Island.
have been colonised by several loricate rotifers,
most notably by members of the genera Notholca,
Mytilina, Trichotria, Lecane and Lepadella. How- References
ever, other groups, especially illoricate rotifers,
Ahlstrom, E. H., 1940. A revision of the rotatorian genera
are conspicuous in their scarcity. This may sug- Brachionus and Platyias with descriptions of one new
gest a very slow dispersal of some rotifers from species and two new varieties. Bull. Am. Mus. nat. Hist. 77:
areas in which they survive. Differences in life 148-184.
cycles among various rotifers may be involved in Ahlstrom, E. H., 1943. A revision of the rotatorian genus
Keratella with descriptions of three new species and five
their differential colonisation success in the arctic.
new varieties. Bull. am. Mus. nat. Hist. 80: 411-457.
Most cladocerans require a minimum of two Bartos, E., 1951. Klic k urcovani virniku rodu Palyarthra
generations in order to produce resting eggs, Ehrenberg. Csaop Narodn. musea. Odd. prirodved.
although some species have become obligate par- 118-119: 82-91.
200

Bartos, E., 1959. Virnici- Rotatoria. Fauna CSR, Praha. 15: Hopkins, D. M., J. V. Mathews, Jr., C. E. Schweger & S. B.
1-969. Young (Ed.) 1982. Paleoecology of Beringia. Academic
Blake Jr., W., 1964. Preliminary account of the glacial history Press. New York. NY.
of Bathurst Island, Arctic Archipelago. Geo!. Surv. Can. Hutchinson, G. E., 1967. A treatise on limnology. Vo!. II.
Pub!. 64-30: 1-8. Introduction to lake biology and the limnoplankton. John
Chengalath, R., 1978. A new species of the genus Notholca Wiley & sons, New York.
Gosse, 1886 (Brachionidae: Rotifera), from Great Slave Idelson, M. S., 1926. Zur Erforschung der Rotatorianfauna
Lake, N.w.T. Ca. J. Zoo!. 56: 363-364. der Gewasser auf Nowaja Semlja. Ber. Wissenschaft.
Chengalath, R., 1984. Synopsis Speciorum: Rotifera. Biblio- Meeresinstitut. Moskau. 12: 77-99 (in Russian).
graphia Invertebratorum Aquaticorum Canadensium 3. Koste, W., 1976. Uber die Radertierbestande. Rotatoria der
National Museum ofN atural Sciences, National Museums oberen und mittleren Hase in den Jahren 1966-1969.
of Canada, Ottawa. 102 pp. Osnabrucker Naturwiss. Mitt. 4: 191-263.
Chengalath, R. & W. Koste, 1983. Rotifera from northeas- Koste, W., 1978. Die Radertiere Mitteleuropas. Ober-
tern Quebec, Newfoundland and Labrador, Canada. Hy- ordnung Monogononta. Borntraeger, Stuttgart-Berlin, 2
drobiologia 104: 49-56. Vols., 673 pp., 234 plates.
Chengalath, R. & W. Koste, 1987. Rotifera from northwes- Koste, W. & R. J. Shiel, 1980. Preliminary remarks on the
tern Canada. Hydrobiologia 147: 49-56. characteristics of the rotifer fauna of Australia (Notogea).
Cole, G. A., 1961. Some calanoid copepods from Arizona Hydrobiologia 73: 221-227.
with notes on congeneric occurrences of Diaptomus Kutikova, 1. A., 1970. Rotifer fauna SSSR. Subclass
species. Limno!. Oceanogr. 6: 432-442. Eurotatoria. Nauka, Leningrad. 742 pp. (In Russian).
Deevey, G. B., 1960. Relative effects of temperature and food Maguire, B., 1963. The passive dispersal of small aquatic
on seasonal variations in length of marine copepods in organisms and their colonization of isolated bodies of
some eastern American and western European waters. water. Eco!. Monogr. 33: 161-185.
Bull. Bingham Oceanogr. Collect. 17: 54-85. Moore, J. W., 1978. Composition and structure of zooplank-
Gosse, P. H., 1851. A catalogue of Rotifera found in Britain ton communities in eighteen arctic and subarctic lakes. Int.
with descriptions of five new genera and thirty-two new Revue ges. Hydrobio!. 63: 545-565.
species. Ann. Mag. nat. Hist. 2: 197-203. Murray, J., 1908. Arctic rotifers collected by Dr. W. Bruce.
Harring, H. K., 1921. The Rotifera of the Canadian Arctic Proc. Roy. Philosophical Soc. Edinburgh. 17: 121-127.
expedition, 1913-1918. Rep. Can. Arct. Exped. 1913-1918, Olofsson, 0.,1918. Studien uber dei susswasser fauna Spitz-
8E: 1-23. bergens. Beitr. z. Systematik, Bio!. u. Tiergeogr. der
Harris, R. P. & G. A. Patfenhoffer, 1976. Feeding, growth Crustaceen u. Rotatorien. Zoo!. Bidr. Uppsala 6: 183-648.
and reproduction of the marine copepod Temora longicor- Pejler, B., 1962. On the taxonomy and ecology of benthic and
nis Muller. J. mar. bio!. Ass. U.K. 56: 675-690. periphytic Rotatoria (Lapland). Zoo!. Bidr. Uppsala. 33:
Hebert, P. D. N., 1985. Ecology of the dominant copepod 327-422.
species at a Low Arctic site. Ca. J. Zoo!. 63: 1138-1147. Ruttner-Kolisko, A., 1974. Plankton rotifers, biology and
Hooper, F. F., 1947. Plankton collections from the Yukon taxonomy. Supp!. Die Binnengewasser 26: 1-164.
and Mackenzie river systems. Trans. am. micros. Soc. 66: Western, G., 1891. Notes on rotifers. J. Queckett Micorsc.
74-84. Club ser. 2: 320-322.
Hydrobiologia 186/187: 201-208, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 201
© 1989 Kluwer Academic Publishers.

Abundance and diversity of planktonic rotifers in the Po River

I. Ferrari 1, A. F arabegolP & R. Mazzoni 2

l Istituto di Zoologia, Universita, 44100 Ferrara, Italy; 2Istituto di Ecologia, Universita, 43100 Parma,

Italy

Key words.' Po River, zooplankton, rotifers, diversity

Abstract

Zooplankton samples from the middle reach of the Po River were collected daily from 27 July to 24 August
1988 from a station located near Viadana. Changes in the biocoenosis structure were analyzed in relation
to variations in flow rate. Rotifers accounted for more than 99 % of the total zooplankton (protozoans
excluded) in every sample. The dominant species were Brachionus calyciflorus, Brachionus bennini,
Brachionus budapestinensis and Epiphanes macrourus. Under scanty flow conditions, the taxocoenosis
showed marked stability. An increase in flow rate acts as a disturbance factor leading to a significant
decrease in both total density and dominance.

Introduction its structure under reduced flow conditions and to

reconstitute after disturbance by floods.
Considerable research has been carried out on Studies on zooplankton variability were con-
zooplankton composition and variability in rivers tinued in July-August 1988 with another short-
of Europe and other continents (Williams, 1966; term sampling series at the same station near
Hynes, 1970; Mordukhai-Boltovskoi, 1979; Lair, Viadana. Changes in zooplankton composition
1980; Pourriot et al., 1982; Shiel, 1985; Davies & and density during this period were analyzed in
Walker, 1986; Guisande & Toja, 1988). Plankton relation to river flow variations. Particular atten-
studies of the Po River since the 1970's have tion was paid to trends of the rotifer taxocoenosis
chiefly involved the analysis of zooplankton and results were compared to those obtained in
samples collected with seasonal or monthly fre- 1985.
quency in the delta (Ferrari et aI., 1983) and in the
middle reach of the river (Ferrari & Benassi,
1984). These studies demonstrated that during Materials and methods
the low flow summer periods the biocoenosis is
characterized by very high densities and is domi- Zooplankton samples were collected daily or
nated by rotifers. every other day from 27 July to 24 August 1988
Ferrari et al. (1987) examined a series of from a station near Viadana. Sampling was
samples collected every other day during July- carried out from the surface to a depth of 1 m with
August 1985 from a station near Viadana (Fig. 1). a 151 Patalas trap. Sixty 1of water were taken for
A close relationship was found between each of the 26 samples collected and filtered
zooplankton structure, hydrometric parameters through a 50 Jlm mesh net. Daily data of river flow
and trophic state of the water. There was also a rate recorded at Viadana were kindly supplied by
definite tendency of the biocoenosis to maintain the Ufficio Idrografico del Po, Parma.
202

PADOVA
•

ADRIATIC
SEA

Fig. 1. Po River basin: the Viadana sampling station is indicated.

Table 1. List of zooplankton species found in the Po River at Viadana.

Ciliophora Dinamoeba mirabilis L. luna Bdelloidea unidentif.

Actinobolina radians L. (Monostyla) bulla Rotifera unidentif.
Cinetochilum margaritaceum Heliozoea L. (M.) closterocerca
Codonella cratera Acanthocystis sp. L. (M.) hamata Cladocera
Coleps incurvus Actinosphaerium eichhorni L. (M.) lunaris Bosmina longirostris
Cyclidium glaucoma L. (M.) quadridentata Scapholeberis sp.
Dileptus falciformis Rotifera L. (Monostyla) sp. Moina micrura
Frontonia leucas Epiphanes macrourus Scaridium longicaudum Ilyocryptus agilis
Mesodinium pulex E. brachionus spinosus Monommata sp. Macrothrix sp.
Metacineta sp. Brachionus angularis Cephalodella catellina Pleuroxus sp.
Nassula tumida B. bennini Cephalodella spp. P. aduncus
Opercularia sp. B. budapestinensis Trichocerca capucina Chydorus sphaericus
Paracineta mobiusi B. calyciflorus T. elongata Leydigia leydigi
Phascodolon vorticella B. diversicornis T. pusilla L. acanthocercoides
Plagiocampa rouxi B·falcatus T. similis Alona quadrangularis
Podophrya sp. B. patulus Trichocerca sp. A. rectangula
Stentor polymorphus B. quadridentatus Synchaeta spp. A. costata
Strobilidium sp. B. urceolaris Polyarthra dolich.-vulgaris
Tintinnidium fluviatile Keratella cochlearis P. major-euryptera Copepoda
Tokophrya quadripartita K. eachlearis tecta Asplanchna brightwelli Nauplii
Trachelophyllum pusillum K. quadrata A. priodonta Harpacticoida copepodites
Urocentrum turbo K. tropica Encentrum sp. Cyclopoida copepodites
Uronema nigricans Anuraeopsis [/Ssa Pompholyx sp. Macrocyclops albidus
Vorticella campanula Euchlanis sp. Flosculariacea unifentif. Paracyclops jimbriatus
V. mayeri Mytilina ventralis Testudinella patina Acanthocyclops sp.
Zoothamnium sp. Trichotria pocillum Hexarthra mira Cryptocyclops sp.
Colurella sp. Filinia cornuta-brachiata Thermocyclops crassus
Sarcomastigophora Lepadella sp. F. longiseta
Rhizopoda Lecane elsa F. opoliensis
Arcella sp. L. flexilis Collotheca sp.
 203

Identification and counting was completed for of both the association pattern among species and
rotifers, cladocerans and copepods. Although the degree of similarity among samples.
abundant in a number of samples, protozoans
were not counted. Other taxa, especially including
drift fauna organisms (e.g. nematodes, oligo- Results and discussion
chaetes, chironomid larvae), present with low
abundances in some samples, were not taken into During the ftrst ten days of the sampling period,
account. river flow rate was decreasing. On 27 July it was
The rotifer taxocoenosis was analyzed by com- 526 m 3 • s - 1 and from 2 to 4 August it was
puting the Shannon-Wiener diversity index for 368 m 3 • s - 1. During the next few days, flow rate
each sample. Density data of the rotifer taxa were increased sharply to a maximum of 1150 m 3 • s - 1
processed by Principal Component Analysis on 7 August. It then gradually decreased to
(PCA) in order to obtain a coincise representation 394 m 3 • s - 1 on 20 August. During the ftnal four

1200 Po River flow rate

m3. &-1

900

600

300
Total Rotifer density
2400 ind .1-1

1200

:1 20 i iii

·:1 o i

H' 4

2
Fig. 2. Po River flow rate at Viadana and parameters ofrotifer community structure: total density, number of species (S), percent
incidence of the most abundant species in each sample (0 Brachionus calyciflorus, • Brachionus bennini, D Brachionus
budapestinensis, • Epiphanes macrourus, * Keratella cochlearis tecta) and Shannon-Wiener diversity index (H').
204

days of the sampling period flow rate again During the first days of August 1988 the lowest
increased with a peak of 827 m3 • s - 1 on 24 flow rate at Pontelagoscuro was above
August. 500 m 3 . s - 1, a value much higher than the all-time
Flow rate was, on average, lower than that low (307 m3 . s - 1) recorded at that station.
recorded during the corresponding sampling Water temperature shows fluctuations which
period (23 July-30 August) in 1985 (Ferrari et ai., are to a large extent related to flow rate variations.
1987). This is confrrmed by flow rates recorded at The highest temperature (27.5 0c) was recorded
Pontelagoscuro, a permanent station located in at the end of July, the lowest (22-23 ° C) coincided
the terminal reach of the Po River, near the city with flood phases on 7 and 24 August.
of Ferrara, with recordings dating back to 1915. Settling measurements carried out on

Po R,ver lIow rate

1200 m3 s-'

900

600

Brachionus calyciflorus
1200 ind·I-1

600

0, '" 1'
B. quadridentatus

300

30:j
1------
i i i i I I i -----
o~==~,-~~~~~~;:==~~~~~~~=;=;=;=;=;=;,~,=;,:;-,
I Iii 8. angulsris
ind·I-1

,nd·I-'

, 1' B. bennini

30: j!...-_':::;=~':;I:;,:::;::::;::::;:::;:::;:==::::;:::::;:==~-.--.--,---,--,-~~,---,---,---,---,:::;-i nd . 1-,

8. tJudapestinensis
ind .1-1

~l~==~=;, : ;: : ;: : ;=;: : ;= : :;: : : :;: : :=:;: :~=- '-'- -r-.-. .-.-ro- r" ;: : :;-'
:;:1;:,

30:j~__~~'~I~'~~~~===;:~==~~~~~'-.-r-ro--r~~
30:j
-r--'-"J-U'I~'I~~-ug~~-st~,~~~--r-r-~-r-r-r-r'-'-'-'-'-,-,-,-~

3 4 5 6 7
Fig. 3. Po River flow rate at Viadana and densities of the dominant rotifer species.
 205

zooplankton samples have evidenced that the vol- ceran species appear: Moina micrura, Bosmina
ume of the sandy fraction was greater than that of longirostris, Chydorus sphaericus and Alona sp.
the plankton fraction. In the samples collected Copepods and cladocerans show very low densi-
during the lowest flow phases of July-August ties; in all samples the two taxa together represent
1985, on the contrary, the plankton fraction less than 1%of the total zooplankton (protozoans
prevailed significantly over the sandy one. excluded) while over 99% is made up ofrotifers.
Phytoplankton collected with the 50 Jlm mesh The overall rotifer density was inversely corre-
net were dominated by Fragilaria crotonensis in the lated to river flow (Fig. 2). During the first ten
first samples of the 1988 series. After the flow days it always exceeded 1000 ind . 1- 1 with a peak
peak of 7 August, phytoplankton diversity in- of 2350 ind '1- 1 on 27 July. On 8 August, when
creased, due particularly to the appearance of river flow abruptly increased, density fell to
blue-green algae. 350 ind '1- 1. Total density then gradually rose to
All the rotifer taxa found in 1988 are also pres- a maximum of 2700 ind . 1- 1 on 18 August; at the
ent in 1985, except for some species of Lecane and flow peak on 24 August it dropped to
Trichocerca (Table 1). Copepods are mainly repre- 400 ind . 1- 1.
sented by cyclopoid nauplii and copepodites and There is also a relationship between river flow
by Acanthocyclops robustus adults. Four clado- and Shannon-Wiener index. This index was cal-

n (15.6'l.)
26
•
11
•

23
•
14
24 17 16 15•
21
• • • • ·13
• 18 10
• •
20
22
. .
19
.
12
0
• 25
•

9
•

1 8
• •
4
6 • 7•
•
2
•

5 3
• •
1 I (20.5t.)
0
Fig. 4. Ordination of the 26 samples (see Fig. 2 and Fig. 3) by PCA performed on density data of 41 rotifer taxa.
206

culated for 41 taxa: some of them are represented including Epiphanes macrourus, B. bennini,
by genera, like Cephalodella, Trichocerca, Syn- Brachionus budapestinensis, Polyarthra spp. and
chaeta and Polyarthra. Synchaeta spp., and diversity remains high.
During the first low water phase (from 27 July Density trends of the most important species
to 6 August) the dominant species is Brachionus are shown in Fig. 3. Other species appearing with
calyciflorus, which accounts for about 50% of all high frequency are Keratella cochlearis, Keratella
rotifers. During this phase Brachionus quadriden- cochlearis tecta, Anuraeopsis jissa, Lecane
tatus, Brachionus angularis, Brachionus bennini and (Monostyla) closterocerca, Cephalodella catellina,
Polyarthra spp. also appear, with densities of Trichocerca pusilla, Asplanchna priodonta, As-
more than 200 ind . 1- 1 in at least one sample. On planchna brightwelli and Filinia longiseta. Some of
8 August there is a marked increase in diversity these species reach density maxima during the last
due to an increased number of species, but a week of August, in particular K. cochlearis tecta
greater evenness in distribution of abundances. and F. longiseta with 123 ind . 1- 1 and Trichocerca
Keratella cochlearis tecta becomes the most abun- spp. with 159 ind '1- 1. The predatory species
dant species, comprising 13 %of the total rotifers. A. brightwelli also attains a relatively high density
During the second low water phase B. calyciflorus (60 ind . 1- 1) during this period. The presence of
is still abundant but does not return to its previous males is generally negligible.
relative importance. Several species are abundant PCA was carried out on the densities of 41

1116.4%)

B. qua~ridentatus

B. ~alyci/lorus

POI~arthra spp.

,
Synchaeta spp. K. cochlearis A.lissa
o ,
K. c. tecta C~PhalOde/la spp.
olD
o o
Trichocerca spp. A. br ightwell,
I

B.benn,", o
o
o
B. angularis
o
F.longiseta

B. budapestlnensis
o

E. macrourus
o

----------------r,o------ 1(84.0%)
Fig. 5. Ordination of the 41 rotifer taxa by peA performed on density data for the 26 samples: the most important taxa, in terms
of both abundance and frequency, are indicated.
 207

taxa, the same ones for which the diversity index increase in diversity. This increase is due to the
was computed (Figs 4 and 5). Samples of the fIrst higher number of species (benthic and phyto-
low water phase (No I-No 9) are distinctly sepa- philous forms dislodged by the current from the
rated from those of the second low water phase bottom and vegetation), but especially to the
(No 13-No 24) (Fig. 4). Samples collected at the reduction in Brachionus calyciflorus density. On
two flow peaks in August (No 11 and No 26) are the other hand, in July-August 1985 total rotifer
closely associated with each other and sharply density was very high (up to 6600 ind '1- I).
detached from all the others. B. calyciflorus, with a maximum density of
PCA on rotifer taxa reveals a density gradient 4450 ind '1- I, remained the dominant species dur-
from the most abundant (Brachionus calyciflorus) ing both low water phases, thus contributing to
to the rarest species (Fig. 5). On the second com- the maintenance of low diversity. High produc-
ponent axis, species at their maxima during the tion of Brachionus males, which is likely to be a
fIrst low flow phase (B. calyciflorus and Brachionus density-dependent phenomenon, was also
quadridentatus) are inversely correlated to species observed.
with the highest densities during the s~cond low Diversity was higher in 1988, above all during
water phase, in particular Filinia longiseta, the last two weeks of the sampling period. This
Brachionus budapestinensis and Epiphanes macro- might be attributed to the higher turbidity due to
urus. dredging activities upstream. Turbidity acts as a
disturbing factor promoting diversity. The inter-
mediate disturbance hypothesis (Connell, 1978;
Conclusions Huston, 1979; Colinvaux, 1986) suggests that
moderate physical disturbances promote diver-
1. During the summer low flow phases, the Po sity.
River waters have a trophic structure comparable 4. The two distinct phases observed in the 1988
to that of shallow, highly productive bodies of sampling series, the fIrst characterized by the
water. Microzooplankton are dominant, espe- clear-cut dominance of Brachionus calyciflorus
cially rotifers which attain densities that are and the second by high rotifer diversity, could be
among the highest reported (Hynes, 1970; interpreted as stages of succession (Margalef,
Pourriot et aI., 1982). High densities of the Po 1974). In the fInal phase biotic interactions proba-
planktonic rotifers might be related to the increase bly become important factors controlling
in concentrations of nutrients (nitrogen and phos- zooplankton community. Predatory behavior of
phorus) as documented by Marchetti et al. (1985) Asplanchna brightwelli could also playa signifIcant
over the past 20 years. role in structuring rotifer communities (Gilbert,
2. Under low water conditions zooplankton tend 1980).
to become a structured community, with a stable 5. Further research should shed light on the
composition, abundance and diversity in time and mechanisms which allow the zooplankton com-
space. During the summer, zooplankton com- munity, under different hydrological and trophic
position in the Po Delta branches (Ferrari et al., conditions, to assemble, persist or modify its
1983; Ferrari & Mazzocchi, 1985) is very similar diversity. Parameters of the community structure
to what has been described here. seem to be valid descriptors of river ecosystem
3. Interesting similarities and differences come to dynamics. In this context, studies on plankton
light when data from the 1985 and 1988 series are can make useful contributions to expansion of the
compared. The ranges of water temperature River Continuum Concept (Minshall et aI.,
(22-28 0c) and flow rates are very similar for the 1985).
two years. The floods observed in both August
1985 and August 1988 have a destabilizing effect
on the rotifer community, leading to a sharp
208

Acknowledgements Ferrari, I., R. Mazzoni & A. Solazzi, 1987. II popolamento

zooplanctonico del flume Po nell'estate 1985. Atti VII
Congresso A.I.O.L., Trieste, 1986: 261-266.
The authors wish to thank Mr. L. Cati and Mr. Gilbert, 1. 1., 1980. Feeding in the rotifer Asplanchna:
G. Allodi of the Ufficio Idrografico del Po, behavior, cannibalism, selectivity, prey defenses and
Parma, who provided us with river flow data and impact on rotifer communities. In: Kerfoot, W. C. (Ed.),
Mr. A. Carrieri of the Institute of Zoology, Evolution and ecology of zooplankton communities. Uni-
versity Press of New England: 158-172.
University of Ferrara, who carried out the statisti-
Guisande, C. & 1. Toja, 1988. The dynamics of various
cal elaboration of the rotifer density data. species of the genus Brachionus (Rotatoria) in the
Protozoan species listed in Table 1 were identi- Guadalquivir River. Arch. Hydrobiol. 112: 579-595.
fied by Dr. P. Madoni of the Institute of Ecology, Huston, M., 1979. A general hypothesis of species diversity.
University of Parma. Thanks also go to Mrs. Am. Nat. 113: 81-101.
Hynes, H. B. N., 1970. The ecology of running waters.
Nancy Birch Podini for her help in translating the
Liverpool University Press, 555 pp.
manuscript. Lair, N., 1980. The rotifer fauna ofthe River Loire (France),
at the level of the nuclear power plants. Hydrobiologia 73:
153-160.
Marchetti, R, G. Pacchetti & A. Provini, 1985. Tendenze
References evolutive della qualita delle acque del Po. Nova Thalassia
7: 311-340.
Colinvaux, P., 1986. Ecology. 1. Wiley & Sons, New York, Margalef, R, 1974. Ecologia. Ed. Omega, Barcelona, 951 pp.
725 pp. Minshall, G. W., K. W. Cummins, R. C. Petersen, C. E.
Connell, J. H., 1978. Diversity in tropical rain forests and Cushing, D. A. Bruns, J. R Sedell & R L. Vannote, 1985.
coral reefs. Science 199: 1302-1310. Developments in stream ecosystem theory. Ca. 1. Fish.
Davies, B. R & K. F. Walker (Eds.) 1986. The ecology of Aquat. Sci. 42: 1045-1055.
river systems. Dr W. Junk Pbls, 793 pp. Mordukhai- Boltovskoi, Ph. D. (Ed.), 1978. The river Volga
Ferrari, I., A. Malice, M. G. Mazzocchi, & G. Matteucci, and its life. Dr W. Junk Pbls, The Hague, 473 pp.
1983. Struttura dello zooplancton dulcicolo nei rami termi- Pourriot, R, D. Benest, P. Champ & c. Rougier, 1982.
nali del Po e in una laguna del Delta. Atti V Congresso Influence de quelques facteurs du milieu sur la com-
A.I.O.L., Stresa, 1982: 505-514. position et la dynamique saisonniere du zooplancton de la
Ferrari, I. & G. Benassi, 1984. Ricerche sullo zooplancton Loire. Acta Oecol., Oecol. Gener. 3: 353-371.
estivo nel tratto medio del Fiume Po. In: Moroni, A. (Ed.), Shiel, R 1., 1985. Zooplankton of the Darling River system,
Contributi di ecologia fondamentale. Zara, Parma: 27-41. Australia. Verh. int. Ver. Limnol. 22: 2136-2140.
Ferrari, I. & M. G. Mazzocchi, 1985. Composizione, Williams, L. G., 1966. Dominant planktonic rotifers of major
dinamica e ruolo troflco dello zooplancton nel Delta del waterways of the United States. Limnol. Oceanogr. 11:
Po. Nova Thalassia 7: 187-214. 83-91.
Hydrobioiogia 186/187: 209-214, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotiler Symposium V. 209
© 1989 Kiuwer Academic Publishers.

Rotifer distribution in relation to temperature and oxygen content

Ernst Mikschi
Goethegasse 4, A-2380 Perchtoldsdorf, Austria

Key words: rotifers, distribution, temperature, oxygen content

Abstract

Lunzer Obersee, a small lake located at an altitude of 1100 m above sea level, was investigated from July
1985 to October 1987. The rotifer community consists of7 dominant species, 7 subdominant species and
34 species which occasionally occurred in the plankton. The dominant species show rather different
demands in relation to temperature and oxygen content; e.g.: Filinia hofmanni was found at a wide range
of oxygen concentrations (0.6-13.3 mg 0 2 1- 1 ) and low temperatures (4-6 QC), living in the upper water
layers (1-7 m) during spring and in the deeper, anoxic zone in summer. In contrast, Asplanchna priodonta
was found at rather high oxygen contents (> 9 mg O2 1- 1), and showed a wide range of temperature
tolerance (4-15 QC).
On the basis of field data the temperature and oxygen requirements of several species are described
and discussed.

Introduction covered by quaking bogs. There are five basins. In

the main basin the maximum depth is 15.5 m; the
Temperature and oxygen content are key factors mean depth of the lake is about two meters.
in restricting rotifer occurrence. Physiological and
population parameters are influenced by tempera-
ture, and the population development of rotifers Methods
is limited by the combined effect of oxygen con-
centration and temperature (Herzig, 1987). The From July 1985 to October 1987 samples were
present study is based on field data and describes taken at the main basin from the surface to a
and discusses the requirements of different rotifer depth of 14 m at one meter intervals using a 10 I
species in relation to these important physical Schindler sampler. Samples were concentrated
parameters. by filtering through a 0.034 mm mesh size net.
U sing the Winkler method oxygen content and
temperature were measured simultaneously at
The lake each sampling date. Except during the time of
ice-cover, samples were taken biweekly. In winter
Lunzer Obersee is a small lake situated at an the sampling interval was extended to about one
altitude of 1100 m above sea level. The area of the month.
lake is 0,144 km 2 , 44% of the surface being
210

Temperature and oxygen content species are monocyclic. The seasonal occurrence
and abundance of the different species is shown
After ice thaw, usually at the end of April or in Table 1.
beginning of May, a rapid temperature increase Occurrence and abundance of different species
occurs in the topmost water layers and inhibits in relation to temperature and oxygen content.
complete mixing. The spring turnover reaches a The analysis of the relationship between these
water depth of only about 7 m and the chemocline two parameters and the occurrence of each
becomes established in 9-10 m (Fig. 1). species was restricted to samples containing more
In general the temperature of the lower stratum than 5 indo 1- 1 (Synehaeta peetinata, Asplanehna
(10-14 m) never rose above 5 °C. The tempera- pnodonta and Filinia hofmanni) and 10 indo 1- 1
ture of the middle stratum (5-10 m) always (Polyarthra doliehoptera, Aseomorpha ecaudis and
remained below 10 0 C. Keratella hiemalis), respectively. Because of the
Rapid cooling in October and November high densities of Keratella eochleans, only samples
resulted in a uniform temperature profile (4-5 ° C). containing more then 100 indo1- 1 were used for
The intensity of the autumnal turnover primarily analysis.
depends on the duration of the homothermous F. hofmanni and K. hiemalis were most abun-
period and the wind intensity during this period. dant at low temperatures ( < 7.5 ° C) under a wide
range of oxygen concentrations. Both species
inhabited the upper layers (0-7 m) during spring
Planktonic rotifers and migrated to the deeper, deoxygenated zone in
summer. P. dolichoptera and S. peetinata preferred
The rotifer community consists of 7 dominant waters with a high oxygen content but developed
species, 7 subdominant species and 34 species at a wide range of temperatures (> 8.7 mg 0 2 1- 1,
which occasionally occurred in the plankton but 5.5-16.0 °C for P. doliehoptera; >9.1 mg 0 2 1-1,
were usually confmed to the littoral or benthic 5.1-12.7 °C for S.peetinata). Both species were
zone. restricted to the upper water layers (0-6 m). As the
Only Keratella eoehleans is a perennial and preceding two species, A. pnodonta showed a
forms two maxima. The more significant one wide range of temperatures being tolerated
occurs in summer (July/August); the second one (6.8-15.4 0C) and - apart from a few samples -

1.-----_- ,
is in winter (January/March). All the other high numbers were found at oxygen contents

m
o r~/ f ,I
, .
I \ 12"
.
\ ..' \
\ \ I'
I \
1' \

, I I \ \. " , \
2 8" : - Smg \ ~, , /
,,
!
: ! \ 8", ,
I I \
\' '
I '
4 .... ' - ......... ' 8" , ,-
........ ,I, , ,' I-
6
\, I
"

4" 4"

L _ __ ___ __ .1

JASONDJ FMAMJ JASONDJ FMAMJJAS

1985 1986 1987

Fig. 1. Temperature and oxygen content in Lunzer Obersee during the years 1985-1987. The isopletes of 5 and 1 mg 0 2 1- 1 and
the isotherms of 4, 8 and 12°C are shown. Black bars indicate ice-cover.
Table 1. Seasonal occurrence (-) and abundance ( *),maximal density found during the period of investigation, vertical distribution (stratum in which a species was
most abundant: U = upper stratum (0-5 m), M = middle stratum (6-10 m), L = lower stratum (10-14 m), ranges of temperature and oxygen content within which
the highest densities were attained, and type to which each species belongs (A, B or C) as described in the text.

Dominant species Occurrence and abundance Max. dens. Vert. High densities Type
(ind. 1- 1) dist.
J FMAMJ JASOND DC mg 0 2 1- 1

Keratella cochlearis *******-----------*****----------- 7,690 UM 5.5-13.5 1.0-13.7 C

Keratella hiemalis -****------- 450 ML 4.4- 8.4 1.4-15.0 A
Polyarthra dolichoptera -- * * * *----- 660 U 5.5-16.0 8.7-18.4 B
Ascomorpha ecaudis --* * * *-- 1,590 UM 5.0-16.0 2.4-18.1 C
Synchaeta pectinata ---****----- 110 U 5.1-12.7 9.1-13.7 B
Asplanchna priodonta --*****----- 200 U 6.8-15.4 8.0-14.4 B
Filinia hofmanni ------------***----------- 110 ML 4.0- 7.5 1.0-11.7 A
Subdominant species
Notho1ca squamula 3 UM 5.1-11.9 6.7-13.5
Synchaeta lakowitziana 4 U 2.0-10.2 8.0-15.7
Trichotria tetractis <1 M
Euchlanis dilatata <1 U
Synchaeta tremula 6 UM 6.5-14.3 7.8-18.2
Keratella quadrata <1 UM
Synchaeta longipes 22 U 5.9-15.0 6.9-15.2

tv
.-
.-
212

exceeding 8 mg 0 2 1- 1. This species inhabited the (5.5-13.5 0c) and oxygen contents (1.0-13.7 mg
upper stratum (0-6 m) while the lake was strati- 0 2 1- 1).
fied and was distributed throughout most of the
water column during the autumnal turnover.
A. ecaudis was abundant over a wide range of Discussion
temperatures (5.0-16.0 0c) and oxygen contents
(2.4-18.7mg 0 2 1- 1 ), but high oxygen contents Some results ofthis study on the dominant rotifers
(> 9 mg O 2 1- 1) obviously were preferred. are summarized in Table 1. All species occurring
K. cochlearis - well known as a generalist with in Lunzer Obersee tolerate low temperatures and
respect to many ecological factors - was abun- high oxygen concentrations. Differences between
dant over a wide range of temperatures species become obvious when comparing the
lowest oxygen content and the highest tempera-
ture tolerated. According to their requirements,
19 mg 021-1
three different groups of animals can be dis-
17 tinguished (Fig. 2):
8
15 Type A: Cold stenothermous species tolerating a
0 wide range of oxygen contents (i.e.
13 ®
~ 0 F. hofmanni and K. hiemalis)
11 • F. hofmanni was described by Koste (1980) as
exhibiting the ecologial behavior of F. terminalis .
•
o.
0
9 Miracle & Vicente (1983) found F. hofmanni being
0
0 clearly restricted to the chemocline and Schaber
7
• >50
5 ® >25
0> 5 19 mg 021-1
3 Filinia hofmanni
17
DC
15 ®
9 11 13 15 17 19
00.
19 mg 02 H 13 o®@ 0,

•
o
t;~08
,
17 11
®o0 00 ®o

15 9
••o
7
13
g• • > 1000
o o® 5 ® > 500
11
o > 100
9 3 Keratella cochlearis

7
®
®
·c
• >50 3 5 7 9 11 13 15 17 19
5 ® >25
o o > 5 Fig. 2. Occurrence and abundance in relation to tempera-
ture ( 0c) and oxygen content (mg O2 1-1) for: (a) Filinia
3 ~ynchaeta Rectinata
hofmanni, (b) Syncheata pectinata, (c) Keratella cochlearis.
Numbers mean indo 1- 1, the continuous line includes all the
·c combinations of temperature and oxygen content occurring
11 13 15 17 19 in Lunzer Obersee during the present study.
 213

& Schrimpf (1984) described F. hofmanni migrat- Bosse1mann (1981) reported a downward move-
ing along the 12 0 C isotherm, disappearing at ment of this species similar to that found in
higher temperatures. Herzig (1987) notes that Lunzer Obersee.
cold stenothermous species can be trapped
between the warm epilimnion and the Type B: Species tolerating a wide range of tem-
deoxygenated hypolimnion. Similar observations peratures but requiring high oxygen con-
were made during the present study (Fig.3a). tent (i.e. S. pectinata, P. dolichoptera and
According to other workers (Ruttner-Kolisko, A. priodonta).
1975; Koste, 1978), K. hiemalis must be charac- Ruttner-Kolisko (1974) described S. pectinata
terized as a cold stenothermous species. being a eurythermous species, showing high den-

Filinia hof manni !ZZI= 50% 0=90%

m
o
2

1mg
O2
,
'S·C

14
ASONOJ FMAMJ JASONOJ FMAMJ JAS
1985 1986 1987

c::J=90%

10

12

14
JASONOJ FMAMJJASONOJ FMAMJ JAS
1985 1986 1987

Fig. 3. Vertical distribution of: (a) Filinia hofmanni and (b) Syncheata pectinata during the years 1985-1987. The total number of
individuals in the whole water column was taken as 100%; layers where 50% and 90% of the individuals are concentrated are
indicated.
214

sities at low temperatures. Similar observations Acknowledgements

have been made by Elliott (1977). During the
present study this species always inhabited the I wish to thank Dr. Alois Herzig for his helpful
upper water layers (Fig. 3b). advice and for revision of the manuscript and Dr.
While Ruttner-Kolisko (1975) found P. doli- Walter Koste for his aid in species identification.
choptera in Lunzer Obersee tolerated low oxygen
contents and its distribution pattern was governed
by temperature and food resources, this species References
was always found at high oxygen contents during
the present study. The discrepancy might be due Bosch, F. v.d. & J. Ringelberg, 1985. Seasonal succession
to the marked decrease in abundance of and popUlation dynamics of Keratella cochlearis (EHRB.)
and Kellicottia longispina (KELLICOTT) in Lake
P. dolichoptera within the last ten years (values Maarsseveen I (Netherlands). Arch. Hydrobiol. 103:
found during the present study usually remained 273-290.
below 10-20% of the numbers reported by Bosseimann, S., 1981. Population dynamics and production
Ruttner-Kolisko (1977)). When occurring at rela- of Keratella hiemalis and K. quadrata in Lake Esrom.
tively low numbers, the availability of suitable Arch. Hydrobiol. 90: 427-447.
Carlin, B., 1943. Die Planktonrotatorien des Motalastrom.
food may be sufficient and it may not be necessary Medd. Lunds Univ. Limnol. Inst. 5: 256 pp.
for P. dolichoptera to migrate to the deeper, Elliott, J. I., 1977. Seasonal changes in the abundance and
deoxygenated zone in order to reach new food distribution of planktonic rotifers in Grasmere (English
resources. Lake District). Freshwat. BioI. 7: 147-166.
The requirements with respect to temperature Herzig, A., 1987. The analysis of planktonic rotifer popu-
lations: A plea for long-term investigations. Hydrobiologia
and oxygen content for A. priodonta are similar to 147: 163-180.
those reported by Elliott (1977). In general, the Hofmann, W., 1983. Interactions between Asplanchna and
success of this species in Lunzer Obersee is Keratella cochlearis in the PluBsee (north Germany).
obviously related to the abundance of K. coch- Hydrobiologia 104: 363-365.
learis (Pourriot, 1977; Hofmann, 1983). Koste, W., 1978. Rotatoria. Die Riidertiere Mitteieuropas.
Gebr. Borntraeger, Berlin, Stuttgart, 673 Seiten.
Koste, W., 1980. Uber zwei Plankton-Radertiertaxa Filinia
Type C: Species showing great tolerance in rela- australensis n. sp. und Filinia hofmanni n' sp., mit
tion to temperature and oxygen content Bemerkungen zur Taxonomie der longiseta-terminalis-
(i.e. A. ecaudis and K. cochlearis). Gruppe. Genus Filinia Bory de St. Vincent, 1824, Familie
A. ecaudis obviously preferred high temperatures Filiniidae Bartos 1959 (iiberordnung Monogononta).
Arch. Hydrobiol. 90: 230-256.
and high oxygen concentrations, but during the Miracle, M. R. & E. Vicente, 1983. Vertical distribution and
time of high abundance, high numbers were also rotifer concentrations in the chemocline of meromictic
found at low temperatures and oxygen contents. lakes. Hydrobiologia 104: 259-267.
The appearance of bacteria populations close to Pourriot, R., 1977. Food and feeding habits of Rotifera. Arch.
the chemocline, which can be used as a food Hydrobiol. Beih. 8: 243-260.
Ruttner-Kolisko, A., 1974. Plankton Rotifers; Biology and
supply (Pourriot, 1977), might cause the migra- Taxonomy. Binnengewasser 26, 1, 146 pp.
tion of a considerable number of individuals to the Ruttner-Kolisko, A., 1975. The vertical distribution of plank-
deeper, deoxygenated zone. The eurythermous ton rotifers in a small alpine lake with a sharp oxygen
character of this species was also described by depletion (Lunzer Obersee). Verh. int. Ver. Limnol. 19:
Ruttner-Kolisko (1974). 1286-1294.
Ruttner-Kolisko, A., 1977. Population dynamics of rotifers as
High tolerances have been reported for
related to climatic conditions in Lunzer Obersee and
K. cochlearis by many authors (e.g. Carlin, 1943; Untersee. Arch. Hydrobiol. Beih. 8: 88-83.
Elliott, 1977), but - like the preceding species - it Schaber, P. & A. Schrimpf, 1984. On morphology and
preferred high temperatures and oxygen contents ecology of the Filinia-terminalis-Iongiseta-group (Rota-
and its distribution pattern seemed to be governed toria) in Bavarian and Tyrolean lakes. Arch. Hydrobiol.
101: 247-257.
by food sources (Bosch & Ringelberg, 1985).
Hydrobiologia 186/187: 2:15-221, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Roti/er Symposium V. 215
© 1989 Kluwer Academic Publishers.

Patterns in the composition of the rotifer communities from high

mountain lakes and ponds in Sierra Nevada (Spain) *

R. Morales-Baquero, L. Cruz-Pizarro & P. Carrillo

Departamento de Biologfa Animal, Ecologfa y Genetica, Facultad de Ciencias, Universidad de Granada.
18071 Granada, Spain

Key words: rotifer communities, high mountain lakes

Abstract

On the basis of periodic collections of rotifers from 29 lakes and ponds over 2500 m above sea level in
the Sierra Nevada (Southern Spain), patterns of species richness, distribution and community com-
position were evaluated. Results allow us to distinguish communities which fall into two major lake types.
One is defined by the presence of typically planktonic species as well as lower specific richness whereas
the other includes communities of mainly benthic and periphytic species. Both lake types seem to be
related to small differences in their physical and chemical characteristics. These relationships and the
influence of littoral vegetation are discussed.

Introduction Material and methods

The study of the composition of rotifer com- Figure 1 shows the area of study. Some basic
munities in different lakes in a specific geographi- limnological characteristics of the lakes were
cal area provides valuable information on those described in earlier pUblications (Sanchez-
factors which control species distribution and Castillo, 1986; Morales-Baquero, 1988) and are
grouping (Tonolli & Tonolli, 1951; Pejler, 1965; summarized here. Lakes rarely exceed 100 m in
Miracle, 1978; Green, 1981; Chengalath etal., length or 3 m in depth and have siliceous sub-
1984; Siegfried et al., 1984; Ebert & Balko, 1987). strates. Littoral vegetation is sparse along the
A series of small alpine lakes are found in the shores consisting of Bryophyta and filamentous
highest part of the Sierra Nevada mountains, green algae. There are no aquatic vertebrates and
where the action of glaciers was able to create the lakes are ice-covered from October through
appropriate conditions in this southern mountain May.
range. Morales-Baquero (1987) has previously Samples were collected from 29 lakes located
published a list of the species found in this area. at altitudes ranging from 2800 to 3050 m above
In this paper we will attempt to discern overall sea level. Lakes were sampled from 2 to 6 times
distribution patterns as well as discussing some of (most frequently 4) during the ice free period in
the physical and chemical factors which control 1981. A conical plankton net (45 Jlm mesh) was
them. swept repeatedly through the open water.

*) Research supported by CAICYT Project n° 3069/83

216

~I -@-

,.

o
...
it

«
"G>

Fig. I. Map of the upper region of Sierra Nevada. (UTM coordinates overimposed).
 217

flS cm-1
Samples were preserved in 4 % formaldehyde
60
solution immediately after collection. Environ-
mental variables were measured with a Hydrolab 50
4041 for simultaneous reading of pH, conduc-
tivity, oxygen concentration and temperature. 40
Similarity values between lakes were obtained
from presence-absence species data using the 30

Serensen index. Lakes were then clustered with

.. . . . ..
the UPGMA method of Sneath & Sokal (1973).
This was followed by a polar ordination analysis
20
.
(Poole, 1974) performed by transforming the
10
.. •
same Sorensen values into a disimilarity matrix. 200 600 1000 1400 m
Fig. 2. Relationships between mean conductivity from study
lakes and distance to the nearest crest line.

Results
tivity values were usually low and never exceeded
Sampled lakes were all well oxygenated, with 55.4 J,lS cm - 1. Conductivity generally tended to
more than 5 mg 1- 1 dissolved oxygen. Most lakes rise as the distance between the lake and the
were acidic and varied widely in range of mean nearest crest line (as determined from maps)
temperatures (8.1 c to 22.7 cC). Mean conduc- became greater (Fig. 2).

Table 1. Distribution of rotifer species in 29 lakes of Sierra Nevada (bdelloids and rare species omitted)

Lakes

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 192021 22232425262728 29

Euchlanis dilatata + + + + + + + + + + + + + + + + + + + + + + + + + + +
Hexarthra bulgarica + + + + + + + + + + + + + + + +
Polyarthra dolichoptera + + +
Lepadella patella + + + + + + + + + + + + + + +
Lepadella quinquecostata nevadensis + + +
Lepadella acuminata + +
Notholca squamula + + + + +
Trichocerca relicta + + + + + + + + + + + + + + + + + + + + + + + + + + + +
Trichocerca tenuior + + + + + + + + + + + +
Trichocerca rattus + + + + + +
Trichocerca bicristata + + + + + + + + + + + + +
Trichocerca cavia + +
Trichotria tetractis + + + + + + + + + + +
Colurella obtusa + + + + + + + + + + +
Lecane lunaris + + + + + + +
Lecane flexilis + + + + + + + +
Lecane furcata + + + + + + + + + + + + + +
Lecane c1osterocerca + + + + +
Lecane kluchor + + +
Cephalodella gibba microdactyla + + + + + + + + + +

1: Mosca; 2: Aguas Verdes; 3: Virgen Inferior; 4: Virgen Superior; 5: Gemelas; 6: Siete Lagunas (4); 7: Siete Lagunas (7);
8: Siete Lagunas (5); 9: Siete Lagunas (6); 10: Dilar (1); II: Dilar (2); 12: Siete Lagunas (2); 13: Rio Seco; 14: Laguneto Laguna
Larga; 15: Pei'i6n Negro; 16: Caldera; 17: Majano; 18: Rio Seco Inferior; 19: Lanjar6n; 20: Pei'i6n Co10rao; 21: Lanjar6n (4);
22: Rio Seco Superior; 23: Larga; 24: Dilar (3); 25: Caballo Inferior; 26: Cuadrada; 27: Caballo; 28: Caldereta; 29: Virgen Media.
 30

->.
.tV..
50

E
II!

90
14 11 29 I 22 23 2 3 4 10 5 7 6 12 8 9 13 15 17 16 18 19 20 21 24 26 25 27 28
B B B B BBB BBB B B

Fig. 3. Average Linkage Cluster Analysis (UPGMA) of communities from 29 lakes (numbers like in Table I). Those with
bryophytic vegetation in their shores are labeled with a B.
0.00 0.07 O. I ~ 0.22 0.29 0.36 0.43 0.50 0.57 0.65 0 .72 0.79 0.86

0 .78 0.78

0.71 0.71

0.64 0.64

0.57 0.57

0.50 0.50

0.4J 19 O.~J
Y
0.36 0 .36

0 .29 0 .29

0.21 0.21

0.14 0 .14

0.07 0.07

om O~
0 .00 0.07 0.140.220.290.360.430.500.570.650.72 0.79 0.86
X

Fig. 4. Two dimensional Polar Ordination of rotifer communities in 29 lakes of Sierra Nevada (data from Table I). Isolines of
conductivity (ps cm - I) has been overimposed in this plane according to the mean values of each one of the lakes. (X,Y: first
and second ordination axis, respectively).
 219

Table 1 summarizes the occurrence of the Polar ordination reflects the division between
species of rotifers found. Most species are ben- the two lake groups (Fig. 4). Ordination of the
thic-periphytic. Only 5 can be considered pelagic: lakes on the basis of disimilarity in fauna con-
Hexarthra bulgarica, Polyarthra dolichoptera, sistently revealed differences in the degree of
Notholca squamula, Euchlanis dilatata and Tricho- mineralization. The isolines of conductivity are
cerca bicristata. plotted in the plane defined by the ordination
Compositional trends are illustrated in the model and are based on mean values for each
results of the cluster analysis (Fig. 3). Two main lake. Conductivity rises steadily from right to left.
groups of lakes not necessarily near one another Both number of species per lake and mean
could be distinguished. The first group comprised density of individuals show a clear pattern
9 lakes containing few species. Their rotifer com- (Fig. 5). Those lakes with fewer species and higher
munities were dominated, with one exception, by densities lie in the low conductivity portion of the
H. bulgarica. The second group generally con- graphs. In most of these lakes H. bulgarica alone
tained a greater variety of species, which were is responsible for the majority of organisms; only
mainly benthic-periphytic, as well as more devel- P. dolichoptera reached similar figures in Lanjaron
oped littoral vegetation. This group included all lake.
lakes with bryophytes in the shores (Fig. 3). The remaining parameters failed to show such

0.86

0.78 0.78

0.71 0.71
NUMBER OF SPECIES
• < 9
0 .64

0.57 \b.:
0.64

0.57
•
e
9 - 13

13 <

0.50 0.50

Y
0.43 • 0.43

0.36 0.36
'<t
<-

0.29 0 .29
(j
0.21 0.21

0.14 • 0.14

0.07 0.07

(a)
0.00 0.00
0.00 0.07 0. 14 0.22 0.29 0.36 0.43 0.50 0.57 0.65 0./2 0 .79 0 .86
X

Fig. 5a. See p. 220 for caption.

 220

0.00 0.07 0.14 0.22 0.29 0.36 0.43 0.50 0 .57 0.65 0 .72 0 .79 0 .86

0 .78 0.78

0.71 0.71
ORG. W 3
• < 10 . 000
0 .64 0.64

e
• 10.000 - 70.000

0.57 0.57 70 , 000 <

0 .50 0.50

0.4.3 .43
Y
0.36 0.36

0.29 0 .29

0 .21 0.21

0 .14 0 . 14

0.07 0.07

0.00 (b)
0.00 0.36 0.43 0.50 0.57 0.65 0 .72
X
Fig. 5. Polar ordination showing distribution of number of species (a) and organisms abundance values (b) in surveyed lakes.
(X,Y: as in Fig. 4).

clear patterns of distribution, although most lakes (Miracle, 1978), or other mountain ranges
with higher pH and dissolved oxygen values are (Pennak, 1957; Thomasson, 1956). The lack of
found in the left hand side of the graph. predatory rotifers in the Sierra Nevada mountains
can likewise be taken as an indication of the
simple trophic structure of these lakes (Pejler,
Discussion 1983).
Patterns in the structure of rotifer communities
The amount of energy in the environment and its in the Sierra Nevada as recorded in this study,
distribution among species determines the num- deserve further comment. In earlier studies,
ber and kinds of species that can be maintained changes in pH were related to changes in the
in equilibrium (Brown, 1981). In this sense, the structure of rotifer communities in Newfoundland
features characterizing the communities of roti- (Chengalath et al., 1984) and Adirondack
fers in the Sierra Nevada are typical of alpine Mountain lakes (Siegfried et al., 1984). In the
systems with limited nutrient input. Hence the Sierra Nevada, however, this factor was not
number of species per lake is on the same order clearly related to rotifer assemblages. Water in
as that found in other alpine lakes such as those our lakes was only slightly butTered and pH varied
in the Alps (Tonolli & Tonolli, 1951), the Pyrenees markedly during the thawing period within indi-
 221

vidual lakes (Morales et aI., 1986), making it diffi- Ebert, T. A. & M. L. Balko, 1987. Temporary pools as islands
cult to rely on mean pH values over the entire in space and in time: the biota of vernal pools in San Diego,
Southern California, USA. Arch. Hydrobiol. 110 (I):
sampling period. Another important factor in the 101-123.
overall structure ofrotifer communities is mineral- Gliwicz, Z. M., 1975. Effect of zooplankton grazing on photo-
ization. Margalef et af. (1976) found changes in synthetic activity and composition of phytoplankton. Verh.
the composition of rotifer communities in 102 int. Ver. Limnol. 19: 1490-1497.
Spanish reservoirs to be related with their degree Green, J., 1981. Associations ofrotifers in Australian crater
lakes. J. Zool., Lond. 193: 469-486.
of mineralization. Green (1981) also observed a Margalef, R, D. Planas, J. Armengol, A. Vidal, N. Prat, A.
drop in the diversity of such communities in Guiset, J. Toja & M. Estrada, 1976. Limnologia de los
Australian crater lakes with conductivities rang- embalses espanoles. Ministerio de Obras Publicas.
ing from 550 to 30420 J.lS cm - I. It is nonetheless Madrid. 422 + 85 pp.
surprising that such slight variations in conduc- Miracle, M. R, 1978. Composici6n especifica de las comuni-
dades zooplanct6nicas de 153 lagosde los Pirineos y su
tivity were so closely related to the ordination of interes biogeogrMico. Oecol. Aquat. 3: 167-212.
Sierra Nevada lakes based strictly on rotifer com- Morales-Baquero, R, 1987. Rotifer fauna oflakes and ponds
munities. over 2500 m.a.s.l. in the Sierra Nevada, Spain, with
The rmding that low conductivity lakes yielded description of a new SUbspecies. Hydrobiologia 147:
greater densities of typically planktonic species, 97-101.
Morales-Baquero, R, 1988. Body size variability of Euchlanis
whereas high conductivity lakes contained pre- dilatata HERENBERG in high mountain lakes of Sierra
dominantly benthic and periphytic species, sug- Nevada (Spain). Arch. Hydrobiol. 112 (4): 597-609.
gests that this change is associated with the devel- Morales-Baquero, R, L. Cruz-Pizarro & P. Carrillo, 1986.
opment of littoral vegetation in these small alpine Lagunas de alta montana en Sierra Nevada: algunas
caracteristicas fisicas y quimicas. Actas II Simp. Agua en
lakes. In the former, phytoplankton species are
Andalucia. 1: 413-424.
probably favored by their greater surface to vol- Pejler, B., 1965. Regional-ecological studies ofswedish fresh-
ume ratio and high turnover rate (Gliwicz, 1975), water zooplankton. Zool. Bidr. Upps. 36 (4) 407-515.
which facilitates the development of filterfeeding Pejler, B., 1983. Zooplanktic indicatory of trophy and their
planktonic rotifers. As the difference in altitude food. Hydrobiologia. 101: 111-114.
Pennak, R. W., 1957. Species composition of limnetic
between lakes and the crest line become greater,
zooplankton communities. Limnol. Oceanogr. 2: 222-232.
nutrient input should be more efficiently used in Poole, R W., 1974. An Introduction to Quantitative Ecology.
the littoral zone. This gives rise to a community McGraw-Hill, New York. 532 pp.
of rotifers of more varied habitat. This also could Sanchez-Castillo, P., 1986. Estudio de las comunidades fito-
explain why some Sierra Nevada alpine lakes of planct6nicas de las lagunas de alta montana de Sierra
similar morphometric characteristics were found Nevada. Unpl. Ph. D. Thesis, Universidad de Granada.
Siegfried, C. A., J. W. Sutherland, S. o. Quinn & J. A.
to have markedly different rotifer communities. Bloomfield, 1984. Lake acidification and the biology of
Adirondack lakes: Rotifers communities. Verh. int. Ver.
Limnol. 22: 549-558.
References Sneath, P. H. A. & R R Sokal, 1973. Numerical taxonomy.
W. H. Freeman, San Francisco. 573 pp.
Brown, J. H., 1981. Two decades of homage to Santa Rosalia: Thomasson, K., 1956. Reflections on Artic and Alpine Lakes.
towards a general theory of diversity. Am. Zool. 21: Oikos. 7 (1): 117-143.
877-888. Tonolli, L. & V. Tonolli, 1951. Osservazioni sulla biologia ed
Chengalath, R, J. W. Bruce & D. A. Scruton, 1984. Rotifers ecologia di 170 popolamenti zooplactonici di laghi italiani
and crustacean zooplankton communities of lakes in cen- di alta quota. Me. 1st. ital. Idrobiol. 6: 53-136.
tral Newfoundland, Canada. Verh. int. Ver. Limnol. 22:
419-430.
Hydrobiologia 186/187: 223-228, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 223
© 1989 Kluwer Academic Publishers.

Rotifer associations of some wetlands in Ontario, Canada

Thomas N ogrady
Department oj Biology, Queen's University, Kingston, Ontario, Canada, K7L 3N6

Key words: rotifers, wetlands, marsh, Pleurotrochopsis anebodica, Trichocerca bicuspes

Abstract

The rotifer assemblage of a small marsh-pond and one large marsh/swamp system in the Kingston,
Ontario area was investigated for four years. The remarkably diverse (177 species) rotifer fauna contained
many species considered very rare. Autecological and taxonomic description of some of these species is
presented, and the wider implications of wetland investigations are discussed. The following species are
new for Canada: Collotheca trilobata, Diplois daviesiae, Encentrum tyrphos, Eosphora thoMes, Itura
chamadis, NotommataJasciola, Pleurotrochopsis anebodica, Trichocerca bicuspes, Trochosphaera solstitialis.

Introduction that of Anderson et al. (1977) dealing with an

acidic swamp and the paper of Goddard &
While there is an enormous literature on the McDiffett (1985) describing the rotifer community
synecology, autecology, population dynamics, of an alkaline marsh, underline rather than solve
seasonal distribution and taxonomy of planktonic the numerous problems.
rotifers, relatively little is known about the same The present paper summarizes data from two
aspects of rotifers occupying wetland habitats. marshes that were sampled during 1984-88 about
There are probably numerous reasons for this. every 10-14 days during ice-free periods. It also
Quantitative and even qualitative sampling of describes the habitats, rotifer associations, and
these habitats is fraught with difficulties and prob- the autecology and taxonomy of the two most
lems (Downing & Cyr, 1985); differentiation of noteworthy rotifer species; a future pUblication
wetland niches is difficult; last but not least, the will attempt to evaluate and ordinate the asso-
taxonomy of many species encountered (primarily ciations using multivariate methodology.
the notommatids) is problematic. These con- Long-term investigations of littoral or wetland
siderations are also true oflittoral associations of rotifer associations are even rarer than those of
lakes, macrophyte beds (Pennak, 1966) and even planktonic populations. Thus their species suc-
bogs. Consequently, we cannot even begin to for- cession, composition of assemblages, their niches,
mulate coherent hypotheses of the synecology of and many autecological parameters are poorly
rotifer associations inhabiting shallow water with known. This paucity of data is compounded by
abundant submerged and emergent macrophyte the vastly larger number of species, smaller num-
vegetation, regardless of the prevailing pH (see ber of individuals, the urgent need for taxonomic
Francez & Devaux, 1985). Two relatively recent revision of some genera, and the methodological
publications using modem ecological methods, difficulties associated with littoral biotopes. The
224

Adineta vaga incisa

Anureopsis fissa Filinia c. corn uta
Ascomorpha agilis 60 terminalis
ecaudis Floscularia (tube)
ovalis Gastropus hyptopus
saltans Itura aurita
Asplanchna herricki aurita intermedia
priodonta chamadis
Asplanchnella girodi myersi
10 Asplanchnopus multiceps veridis
Beauchampia crucigera Keratella c. cochlearis
Brachionus bidentata earlinae
calyciflorus 70 serrulata
patulus curvicornis
quadridentatus tecta
Cephalodella apocolea ticinensis
auriculata Lacinularia flosculosa
catellina ismailovensis
delicata Lecane acuminata
20 exigua leontina
forficula luna
gibba ohioensis
tenuior stichaea
ventripes 80 stokesi
Collotheca c. campanulata ungulata
c. gracillipes Lepadella acuminata
o. ornata ovalis
ornata corn uta patella
trilobata pterygoidea
30 Colurella uncinata rhomboides
Conochilus unicornis triptera
Cyrtonia tuba Limnias ceratophylli
Dicranophorus caudatus Lindia truncata
edestes? 90 Lophocharis oxysternon
forcipatus salpina
grandis Macrotrachela quadricornifera
Dipleuchlanis propatula Manfredium eudactylotum
Diplois daviesiae Microcodides chlaena
Dissotrocha aculeata Microcrodon clavus
40 Diurella brachiura Monommata aeschyna
cavia arndti
vernalis longiseta
weberi Monostyla bulla
Dorystoma caudata 100 closterocerca
Encentrum grande corn uta
sutor? decipiens
tyrphos hamata
Enteroplea lacustris lunaris
Eosphora ehrenbergi quadridentata
50 najas Mytil ina m. mucronata
thoa v. ventralis
thoides ventralis
Epiphanes clavulata macracantha
Euchlanis calpidia Notholca acuminata
d. dilatata 110 squamula
deflexa Notommata allantois
triquetra collaris
 225

advantages of long-term investigations and the research will also help to uncover the suspected
obvious need to extend such work to the vast groups of 'equivalent species' that probably
majority of rotifers living in the benthic/littoral occupy the same niches, and their presence or
zone are amply illustrated in the important and absence depends on the 'fIrst come fIrst served'
sophisticated paper of Herzig (1987). The present principle of fortuitious occupancy of an available
paper shows that extensive and frequent sampling niche; thus any member of such group can occupy
can uncover an unexpected faunal diversity, yield an available niche, but to the exclusion of other
species that are 'rare' or present for a very short members of the 'equivalence group'. Recognition
time only, and contributes to the knowledge of the of such groupings may be able to extricate us from
autecology of these species. It seems, however, the seemingly insoluble and frustrating quandary
that even more data and longer observations are of correlating environmental parameters and
needed to be able to attempt the elucidation of physiological properties of species with ecological
populations dynamics and interrelation of assem- phenomena (lvanova, 1987). Thus, beside con-
blages using methods of numerical ecology. Such tributing to the knowledge of wetland rotifer

copeus Sinantherina semibullata

fasciola socialis
tripus Squatinella longispinata
Platyias quadricornis m. mutica
Pleurotrocha petromyzon m. tridentata
Pleurotrochopsis anebodica 150 - rostrum
Ploesoma lenticulare Synchaeta asymmetrica
120 - truncatum lakowitziana proloba
Polyarthra dolichoptera longipes
dolichoptera aptera oblonga
major pectinata
major proloba stylata
remata tremula
remata proloba Taphrocampa annulosa
vulgaris luminosa T estudinella patina
v. vulgaris 160 - patina intermedia
Proales decipiens Trichocerca bicristata
130 - fallaciosa bicuspes
reinhardti elongata
sigmoidea iernis
theodora intermedia
vernecki longiseta
Ptygura furcillata porcellus
p. pilula pusilla
Resticula gelida rattus
nyssa 170 - rattus carinata
plicata similis
140 Rotaria neptunia tenuior
rota tori a weberi
sordida Trichotria pocillum
tardigrada tetractis
Scaridium longicaudum Trochosphaera soltitialis

Fig. 1. Species list of Bayview Marsh and Little Cararaqui Marsh (Kingston, Ontario, Canada) in 1984-1987.

* The subgenera Trichocerca (Diurella) and Lecane (MonostyJa) are shown as genera.
226

assemblages, more basic ecological principles Methods

could be elucidated by prolonged and frequent
sampling in wetlands. Both marshes were sampled every 10-14 days
with few exceptions, and data obtained between
September 1984 and July 1988 are considered in
Description of study areas this paper. Only qualitative samples were taken
using a plankton net of 28 tlm mesh, due to the
Both marshes are in or the immediate vicinity of unavoidable and well known difficulties of sam-
Kingston, Ontario. The fIrst one, Little Cataraqui pling among macrophytes (Downing & Cyr,
Marsh (Kingston Township), is a marsh pond 1985). Open water, sediment, and water squeezed
about 100 x 50 meters in an area of Carex marsh, from submerged macrophytes were investigated;
a few hundred meters from the Little Cataraqui macrophytes were scanned for sessile rotifers.
Creek and across the road from a bay of Lake Living as well as preserved material was exam-
Ontario, but is not in direct contact with either. It ined, and percentage composition of the rotifer
is about 1-2 meters deep, but there is a 0.5-1 meter assemblage counted.
thick loose sediment layer at the bottom. In sum-
mer it is fIlled with submerged macrophytes. Its
physicochemical parameters are very ordinary:
pH 7.2-8.5, conductivity 400-1800 tlmhos/cm, Results and discussion
CaC0 3 /CaHC0 3 type water chemistry, no H 2 S
during ice-free periods, but due to its immediate The rotifer association
vicinity to a major road, it is somewhat polluted.
The second study area, Bayview Marsh, is about The list of rotifers in the two marshes is combined
20 km west of Kingston in Ernestown Township. in Table 1, since there is considerable overlap and
It is a depression area of about 1 by 2 km in no discernible information could be obtained
calcareous till in a non-cultivated open forest- from separating the two lists. However, Little
meadow area and is fed by precipitation, but has Cataraqui Marsh has a lower diversity, and
a small drainage creek. It is in the fInal stages of contained, on occasion, very few rotifers; this was
filling, and some parts are dry land most of the probably due to competition from Bosmina
year. Two areas of the marsh, about 300 meters /ongirostris or Chydorus sphaericus, judging from
apart, were studied: one is a Carex marsh with a the cladoceran population maxima at such occa-
maximum of 30 cm deep water over about 1 m sions.
sediment, that dries out in July or August until It was also most remarkable that the rotifer
fIlled again by fall rains. The other section is a assemblages of the two areas of Bayview Marsh,
swamp containing numerous dead treetrunks and only about 300 m apart, were often, if not always,
only submerged macrophytes. There are small very different qualitatively as well as quantita-
cyprinid fIsh and wild ducks in this part, because tively. This indicates that wetlands, where water
it contains water all year round. The water movement is normally very restricted, must be
chemistry is as unremarkable as the physiogra- sampled at as many locations as feasible.
phy: pH 7.1-7.8, conductivity The list of species is conspicuous for containing
400-700 tlmhos/cm, and occasional faint H 2 S over 170 taxa from an area as small as these two
smell if the thick sediment is disturbed. The free marshes. However, about 30% of all species
water layer can be up to 80 em deep. Bayview occurred only once, even if present in large num-
Marsh is relatively undisturbed apart from occa- bers at that occasion. It was not rare to fInd over
sional hunting. 30 species in a single sample. Varga (1939)
describes a similarly rich rotifer fauna in his paper
on a small wetland off Lake Balaton in Hungary,
 227

Trichotria spp. 58 Lepadella patella 25

Synnchaeta spp. 48 Cephalodella gibba 24
Euchlanis incisa 48 Mytilina spp. 23
Euchlanis dilatata 44 Polyarthra vulgaris 22
Testudinelia patina 41 Monostyla quadricornis 21
Polyarthra remata 39 Lophocharis spp. 20
Trichocerca longiseta 38 Trichocerca rattus carinata 19
Cephalodella auriculata 34 Anureopsis fissa 18
Monostyla lunaris 30 Keratella cochlearis 17
Squatinella mutica 29 Platyias quadricornis 17
Ascomorpha ecaudis 29 Keratella serrulata curvicornis 15
Brachionus patulus 29 Trichocerca bicuspes 12
Scaridium longicaudum 27 Itura viridis 9
Gastropus hyptopus 27 Brachionus calyciflorus 5
Monommata spp. 26
Fig. 2. Percent occurrence of most common rotifer species in all samples.

whileas the paper of Goddard and McDitTett the dorsal side. The stout and flexible body carries
(l983) lists only 30 species in a year. a series of thorns beginning at the neck, thus
Only 16% ofrotifer species (or genera) encoun- resembling Fadeew's description. The number of
tered can be designated as common or dominant, these thorns is hard to establish. There are appar-
and their list is shown in Table 2. Planktonic ently four groups of three thorns ventrally, per-
genera normally considered 'common' are very haps a few smaller thorns as well. At the very end
poorly represented (Keratella, Brachionus). of the trunk there is an especially prominent
curved ventral thorn, and 5 more equidistant
thorns circling the body. In addition there are two
Taxonomic remarks dorsolateral series of fine teeth at the distal end.
The foot has two segments, the first one carries 5
Probably due to the frequent and prolonged or 6 thoms, the second one only 4. The lateral
sampling a large number of rotifer species were thorny lamellae described by Berzins could not be
found that are considered rare. In view of the found. Toes are small, dagger-like.
sporadic sampling of Canadian waters (see The central ganglion is long and carries an
Chengalath, 1984), this is not surprising. Two of eyespot with 4-5 additional red pigment spots.
these species are discussed and illustrated below, The stomach consists of large, round cells, the
because they have been either poorly described or lines shown by Berzins were not seen. The
no figures are available in the modem literature.

~ffl
. ..:" ~

Pleurotrochopsis anebodica Berzins 1973 (Fig. 3).

The single specimen seen differs in many respects
:"~r . '.
. . ........ ..'

.: ': '" ': d

from the figure given by Berzins (1973) as well as
b

vb)
from the figure of Fadeew (as shown in Koste,
1978), and one has to concur with Koste regard-
ing the unsatisfactory nature of these figures. The
animal seen among submerged macrophytes in
Bayview Marsh had a distinctly segmented
notommatid-type body, but not like shown by
Berzins. The head is large and changes shape Fig, 3. Pleurotrochopsis anebodica Berzins 1973. a, from left,
easily; corona extends down to the ventral side. b, head from left, c, end of body and foot dorsally, d. trophi
There is a rigid rhinoceros-like pointed 'hom' on dorsally, e. trophy sideways,
228

References

Anderson, C. B., A. F. Benfield & A. L. Buikema Jr., 1977.

Zooplankton of a swamp water ecosystem. Hydrobiologia
55: 177-185.
Berzins, B., 1973. Pleurotrochopsis anebodica n.g., n.sp.
(Rotatoria) aus Schweden. Hydrobiologia 41: 449-451.
Chengalath, R, 1984. Synopsis Speciorum. Rotifera. Biblio-
graphia Invertebratorum Aquaticorum Canadiensium
Vol. 3. National Museum of Natural Sciences, National
Museums of Canada., Ottawa. pp. 1-102.
Downing, l A. & H. Cyr, 1985. Quantitative estimation of
epiphytic invertebrate populations. Can. l Fish. Aquat.
Sci. 42: 1570-1579.
Fig. 4. Trichocerca bicuspes (Pell, 1890). a. sideways, Francez, A.-J. & J. Devaux, 1985. Repartition des rotiferes
b. ventrally, semiextended, c. trophi. dans deux lac-tourbieres du massif Central (France).
Hydrobiologia 128: 265-276.
Goddard, K. A. & W. F. McDitTett, 1983. Rotifer distribu-
intestine is smooth. The mastax is small, rami tion, abundance and community structure in four habitats
square, manubria straight, spindle-shaped with a of a freshwater marsh. l Freshw. Ecol. 2: 199-211.
small triangular alula in the middle. The fulcrum Harring, H. K., 1921. The Rotatoria ofthe Canadian Arctic
is wide and paddle-shaped in side view. Measure- Expedition, 1913-1918. Rep. Can. Arctic Exp. Vol. 8 Pt E.
p.3-23.
ments: total length 180 jlm, toe 13 jlm, terminal
Hauer, l, 1958. Radertiere aus dem Sumpfe 'Grosse
ventral thorn 5 jlm. Found among submerged Seewiese' bei Kist. Nachr. Naturw. Mus. AschatTenburg
vegetation in September, temperature 17 0 C, pH 60, pp. 1-19.
8.2, conductivity 640 jlS. Herzig, A., 1987. The analysis of plankton rotifer popu-
lations: A plea for long-term investigations. Hydrobiologia
147: 163-180.
Trichocerca bicuspes (Pell, 1490) (Fig. 4) is another
Ivanova, M. B. (1987). Relationship between zooplankton
one of those species that appear mysteriously, not development and environmental conditions in ditTerent
to be seen again for decades. After its description types oflakes in the zone of temperate climate. Int. Revue
by Pell, it was properly figured by Jennings (1904). ges. Hydrobiol. 72: 669-684.
It was listed next by Harring (1921) from the Jennings, H. S., 1904. Rotatoria of the United States II. A
Canadian Arctic, and listed again in Dismal monograph of the Rattulidae. Bull. U.S. Fish Comission
22: 275-352.
Swamp, Virgin a, by Anderson et al. (1977). It is Koste, W., 1978. Rotatoria. Die Radertiere Mitteleuropas
an abundant and regular summer constituent ofthe begriindet von Max Voigt. Borntrager, Berlin.
Bayview Marsh, both in the open water as well Koste, W., R J. Shiel & M. A. Brock, 1983. Rotifera from
as between submerged plants, seen every year. Western Australian wetlands with description of two new
The species conforms to the figure of Jennings, species. Hydrobiologia 104: 9-17.
Pell, A., 1890. Three new rotifers. The Microscope, Detroit,
but is drawn here again. The two big lateral thorns 10: 43-45.
are unique and unmistakable; the keel is striated Pennak, R. W., 1966. Structure of zooplankton populations
and extends almost to the foot. Right manubrium in the littoral macrophyte zone of some Colorado lakes. Tr.
is L-shaped and hooked, the right ramus carries Am. Micr. Soc. 85: 329-349.
two alulae. Measurements: body length Varga, L., 1939. Adatok a Balaton kerekesfereg-faunajanak
ismeretehez. Az Aszofoi nadas oMI kerekesfergei. (Con-
90-110 jlm, toe 120-140 jlm, thorn 9 jlm, trophi
tributions to the rotifer fauna of Lake Balaton. Rotifers of
43-45 jlm. It occurs from May to September, pH the AszofO bay.) Magyar BioI. Kut. Int. Munkai 11:
7.2-7.9, conductivity 400-650 jlS. 316-371.
Hydrobiologia 186/187: 229-234, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 229
© 1989 Kluwer Academic Publishers.

Opportunist rotifers: colonising species of young ponds in Surrey,

England

Rosalind M. Pontin
26 Hermitage Woods Crescent, St. Johns, Woking, Surrey, GU2l 1UE, England

Key words: rotifers, new ponds, colonisers, herbivores, omnivores, reproduction

Abstract

Two young ponds of known age were investigated and their populations ofrotifers and other organisms
compared. The earliest colonisers appeared after a few days and included the rotifers Keratella valga
(Ehrb.) and Brachionus urceolaris Muller, the cladoceran Daphnia obtusa Kurz and cyclopoid copepods.
The rotifers quickly produced males and resting eggs, as well as large numbers of parthenogenetic females.
These herbivorous species were followed by an omnivorous Asplanchna species. These species continued
to flourish in the ponds several years after digging, in spite of the presence of additional species, although
their season of occurrence changed. The greater growth of macrophytes in one pond may account for
the greater variety of species in this community.

Introduction small beds of Nymphaea and a few clumps of Iris

at the margins.
Surrey is a heavily-populated area of Southern Bulhousen Farm Pond was dug out in February
England. Numerous ponds which have silted up 1979 after it had been dry for at least twenty years,
and have been dry for many years have been having been filled with wood waste from a sawmill
re-dug and allowed to fill with water naturally. (Pontin & Pontin, 1980). It varies from 11/2-2 1/2 m
Two of these ponds were compared to find which deep, the water level being highest in winter and
species can fill the roles of opportunist colonisers falling in summer. Its area is about 120 m 2 • Ini-
and whether they persist in the pond as it ages. tially the pH was 5-5.5, but now averages 6.5 and
conductivity is 160 J1 S cm - 1 at 16 0 C. Macro-
phytes include Glyceria, Hottonia and Myriophyl-
The ponds lum, which grow thickly in spring and summer.

Four Horseshoes Pond was dug out in November

1982 after it had been dry for nearly forty years. Methods
It varies from 1/2-1 1/2 m deep; in winter it is often
flooded and in summer it may be nearly dry. Qualitative samples were collected by throwing a
Consequently its area is also variable, but is about plankton net on a rope from the banks. It is possi-
300 m2 • Initially the pH was 5-5.5, but now ble to reach any part of the ponds by this means.
averages 6.5, while the conductivity averages Three horizontal hauls, each of about 5 m and just
80 J1 S cm - 1 at 16 0 C. Macrophytes include two below the surface, were taken in different direc-
230

tions on each sampling occasion. These were structed to represent this stage in the pond's devel-
combined, examined live and also preserved in opment. Algae of different sizes may be taken by
5 % formalin. The net pore size was 50 x 50 f..lm. the herbivores K. valga, B. urceolaris, G. hyptopus,
Quantitative samples were taken just below the D.obtusa and copepods, either by filtering,
surface, using a vessel of standard volume sus- piercing or seizing. The omnivore Asplanchna,
pended on a rope. Three samples were taken on however, takes K. valga as prey as well as the
each occasion, at positions chosen at random. larger algae.
The samples were combined, making a total of
1050 ml, filtered through bolting silk of pore size
70 x 70 f..lm, preserved in 5 % formalin and Quantitative study
counted in total.
Samples were taken weekly during March- A quantitative study was made in 1987 and the
September with extra samples every two or three results are shown in Fig. 1 and 2. It can be seen
days from Four Horseshoes Pond during late July that the herbivores make up the majority of the
and early August. Observations and counts of population in each pond. As well as those present
stomach contents were made on living animals.
Measurements of pH and conductivity were
FOUR HORSESHOES 1987 41/2 YEARS OLD
made in the field with Whatman microsensors.
LARGE ALGAE
pharus
Synura colOnies +---+---+....-4If-+-
Initial colonisers
HERBIVORES
cyclopoid copepods
The first colonisers appeared in Four Horseshoes adults and nauplil

Pond about ten days after digging. They were

Keratella valga (Ehrb.), Daphnia obtusa Kurz and
cyclopoid copepods including nauplii. They were
still present after one month, but in Bulhousen
Farm Pond, which was dug in winter rather than
in late autumn, no animals were found during the
first month.
KerateUa testudo 9 +--+---H.-t-I-+-
Mter two months, males of K. valga had
appeared in Four Horseshoes Pond, but D. obtusa
had disappeared. Brachionus urceolaris Milller,
both females and males, was now in Bulhousen
Farm Pond, as well as cyclopoid copepods.
After three months, only K. valga females and
copepods were in Four Horseshoes Pond, but
Bulhousen Farm Pond also contained both sexes Brachionus urceolaris 9 -+---+----- ..~+-
of K. valga and B. urceolaris and females of
Gastropus hyptopus (Ehrb.).
After four months, Four Horseshoes Pond
remained as it had been the previous month i.e. OMNIVORE
K. valga females and copepods. Bulhousen Farm Asplanchna sleboldi 9 +--+--+---iII.
Pond also had K. valga females plus copepods
APR MAY JUN JUL AUG
and males and females of Asplanchna brightwelli
Gosse. Fig. 1. Four Horseshoes Pond 1987,4 1/2years after digging.
A relatively simple food chain can be con- Changes in log. numbers per litre with time.
 231

BULHOUSEN FARM 1987 e YEARS OLD Asplanchna. The species present in Four Horse-
shoes Pond was very rapacious, eating K. valga,
LARGE ALGAE
copepod nauplii and B. urceolaris. Asplanchna
predation, therefore, had a considerable effect on
HERBIVORES ~ its prey populations (Fig. 1 and 3). As Asplanchna
cyclopoid copepods
adults and nauplll density increased, it became increasingly can-
nibalistic (Fig. 3). Cannibalism, and its large
Q!phm' .bI.g , ~ toothed jaws, lobed nuclei in the vitelline gland
and the presence of humps in the males, indicate
that this was A. sieboldi (Leydig), although no
humped females were seen.
A. brightwelli occurred in Bulhousen Farm
Pond, but was present only in very small numbers.
Both ponds also contain carnivorous planktonic
larvae of the phantom midge Chaoborus. These
may take considerable numbers of prey, but their
effect has not yet been determined.
The 1987 survey showed all the original colonis-
ing species to be present in each pond and K. valga
in very large numbers. Of the additional species,
BrachlOnus urceolarls 9 -+---+--t--4~I---f- K. testudo and P. vulgaris also had very large
maxima in Bulhousen Farm Pond, although they
~y'nchaeta pectlnata 9 +-+-f-M . ...-+- are represented by only small numbers in Four
Horseshoes Pond. The production of resting eggs
~I!phanes brachionus 9 +-+--I-..MI-.-+- in Keratella and other genera precedes the
~p'landma ~9ht.. elh 9~.
OMNIVORE
,,.
J.. 1
.-.1
~
I maxima, beginning almost as soon as females are
seen. By the time maximum numbers of females
APR MAY JUN JUL AUG
are reached, production of both sexual and
asexual eggs per female is already falling (Fig. 4).
Fig. 2. Bulhousen Farm Pond 1987, 8 years after digging. Bulhousen Farm Pond at 8 years old in 1987
Changes in log. numbers per litre with time.
had more species than did Four Horseshoes Pond
at 4 1/2 years. However, Bulhousen Farm Pond
initially, the following appeared in each pond: also had more species in its early days. Apart from
K. testudo (Ehrb.), Polyarthra vulgaris Carlin and age, other differences between the ponds include
Synchaeta pectinata Ehrb., plus in Bulhousen nutrient levels and macrophyte growth. Conduc-
Farm Pond only, Epiphanes brachionus (Ehrb.). tivity values are twice as high on average in
Estimates of their potential food algae have so far Bulhousen Farm Pond, perhaps because of the
been limited to counts ofthe larger species, Phacus debris originally remaining in the bottom or
and the colonial Synura, which may be eaten by floating on the surface or perhaps because of
piercers and suckers such as S. pectinata and run-off from the nearby farm. This in turn would
G. hyptopus. However, an estimate of the numbers promote greater algal and macrophyte growth,
of small-sized algae is also needed. The stomachs thus providing a greater potential food source and
of K. valga and K. testudo contain very small algal more microhabitats.
cells of about 2 Jlm in diameter, while some of Adjustment of a species to a maturing pond
about 10 Jlm diameter were observed in community might involve a change in timing or
S. pectinata. extent of occurrence, as is suggested by Fig. 5,
Each pond has an omnivorous species of which compares occurrence time for species in
232

FOUR HORSESHOES A: ASRlanchna sieboldi

80
2.4" number of eggs/young
per ~ 8.. sieboldi

60

Keratella,....~~
\valg~
40 Pie charts show
the % of ASRlanchna
stomachs containing:
ALGAE
Keratella va 19~
20 B Brachionus vulgaris
Af Anurapo~ fissa
A ASRlanchna si~boldi A
l/)
a::: n cyclopoid nauplii 4-----A
UJ
co - empty .
~ 30/0+------J...--------'-I--4"""------
z Cyclopoid nauplii

20

10
~n
/\n
3010+-----------------------
B Brachionus urceolaris
20 Af AnuraeoRsis fissa

10 /B-B B\
..B_____ B B
Af-Af 'B-B
O~------~~~~~~~-------------------
1 10 20 30 10 20 30 10 20 30
JUNE JULY AUGUST 1987

Fig. 3. Asplanchna diet in Four Horseshoes Pond 1987. Numbers per litre of A. sieboldi and its food organisms compared with
its dietary choice. Pie charts indicate proportion of A. sieboldi stomachs containing different food organisms.
 233

FOUR HORSESHOES
2.0 Keratella val@ total eggs per female c;>
asexual eggs per female 0
resting eggs per female 0-
1.5

1.0

!;? total number of females

o asexual eggs
o"eggs hatching males
400 0 resting eggs

l/l
a::
~ 300
E
=>
z
o
200

100
$
$- M
II
0
1 10 20 30 10 20 30 10 20 30
APRIL MAY JUNE 1987

Fig. 4. Egg production in Keratella valga in Four Horseshoes Pond 1987. Below: numbers per litre K. valga females, asexual,
sexual and male eggs with time. Above: ratio of eggs per female for the same period.

Bulhousen Farm Pond in its first year with 1987. have extended their season, while K. testudo and
K. valga and copepods, for example, appear to B. urceolaris occurred at different times. These
234

BULHOUSEN FARM Species and season of occurence

1979
Cyclopoid copepods ________________________________________________ 1987

_____________ 1979
Brachionus urceolaris ___________ 1987
____________________ 1979
Keratella valg9 ______________________________________ 1987

____ 1979
_ _ 1987
_________________________ 1979
Keratella testudo ______________________ 1987
______________ 1979
Poly'arthra ___________________________________________ 1987

___________________________________ 1979
__________________________ 1987
____________ 1979
___________________ 1987

10 20 30 10 20 30 10 20 30 10 20 30 10 20
APRIL MAY JUNE JULY AUGUST

Fig. 5. Comparison of season of occurrence of species in Bulhousen Farm Pond in 1987 (8 years after digging) with the same
species in 1979 (first year).

changes could be a consequence solely of weather grazers. Changes in season of occurrence of the
conditions, but could also reflect changes in the opportunists may demonstrate their ability to
total community. adapt to new pressures resulting from the increas-
ing complexity of the community.

Conclusions
Acknowledgements
Initial colonisers include species which are com-
mon in temporary ponds and which can multiply I have pleasure in thanking Mrs. Sarah Wroot for
rapidly to take advantage of untapped algal preparing the original poster presentation of this
supplies. These species are capable of quickly work. I am also indebted to Prof. C.T. Lewis and
producing resting eggs. Williams (1987) points Prof. R.P. Dales and members of the Zoology
out that, in temporary waters, the aquatic phase (now Biology) Department of Royal Holloway
may be so short that powers of dispersal and (now R.H.B.N.) College, for their kind hospi-
plasticity of life cycle are more important to tality.
success than competitive ability. In a persistent
water body, however, competitive ability and
avoidance of predation may become more signifi- References
cant. In these two ponds, the colonising species
Pontin, A. J. & R. M. Pontin, 1980. Bullhousen Pond, Bisley,
were still present after some years in spite of the 1979. Surrey Trust for Nat. Conserv. Newsletter 55: 8-9.
arrival of additional species, including omnivores, Williams, D. D., 1987. The Ecology of Temporary Waters.
which followed the populations of filterers and Croom Helm, Beckenham & N. S. Wales, 205 pp.
Hydrobiologia 186/187: 235-238, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 235
© 1989 Kluwer Academic Publishers.

Percentage of rotifers in spring zooplankton in lakes of different trophy

Stanislaw Radwan & Barbara Popiolek

Academy of Agriculture, Department of Zoology and Hydrobiology, Akademicka 13,20-934 Lublin,
Poland

Key words: zooplankton community, rotifers, lake trophy, Poland

Abstract

Studies carried out on 8 lakes in the LC(czna-Wlodawa Lakeland of eastern Poland indicated that the
qualitative and quantitative structure of zooplankton was clearly correlated with the lake trophic state.
In the spring zooplankton oflakes affected by gradual natural eutrophication were dominated by rotifers.
In the zooplankton oflakes strongly affected by human activities, Cladocera dominated. With an increase
in lake trophy there was an increase in the number of species that were indicators of eutrophy and a
decrease in the number of indicators of meso trophy. The total number of species in individual lakes tended
to increase with an increase in trophy.

Introduction Site and methods of studies

Relationships between the qualitative and quanti- Studies have been carried out on 8 lakes in the
tative composition of the zooplankton community LC(czna and Wlodawa lakeland. The area of these
and the trophic state oflakes have been the object lakes ranges from 12 to 85 ha and the maximum
of numerous studies. These studies have focused depth varies from 4,3 to 38,8 m. Most of the basic
on three issues: 1) determination of species com- physical and chemical factors are at a low or
position and abundance of crustacean plankton medium level. Only two factors - electrolytic con-
in lakes of different trophy, 2) the relationship ductivity and calcium have high values in most of
between the trophic state oflakes and the qualita- the investigated lakes. These lakes are affected by
tive and quantitative structure of main zooplank- different forms of human activity. Almost all of
ton groups (Rotifera, Cladocera, Copepoda), them are under the influence of agricultural
3) determination of groups of species that are activities. Five deeper lakes with a low state of
bioindicators oflake trophy. In the present study eutrophication are progressively affected by
special attention has been paid to the role of recreational activities and three lakes are affected
rotifers in the creation of the qualitative and by strong hydrotechnical activities, because they
quantitative structure of zooplankton in lakes of are connected by a system of canals with two
different trophic status. rivers. The investigated lakes represent different
trophic states, namely: mesotrophic, dystrophic-
eutrophic and eutrophic.
236

In spring of 1987 in the pelagial zone of these taxons, whereas in the Lake Conway system only
lakes quantitative planktonic samples were col- 1 to 3 taxons were found (Blancher, 1984). The
lected. A 21 apparatus of Patalas type was used difference may result from the small area of the
in collecting samples (Patalas, 1954). The col- Polish lakes and from migration of benthic -
lected samples were processed according to the periphytonic species from the littoral to the
generally used method (Hillbricht-Ilkowska & pelagial zone (Radwan, 1973; Kowalczyk &
Patalas, 1967). Three major groups of zooplank- Radwan, 1982). In both lake systems rotifers
ton (Rotifera, Cladocera, Copepoda) were sub- dominated the zooplankton community.
jected to detailed microscopical analysis. In most of the lakes in the L~czna-Wlodawa
lakeland undergoing gradual eutrophication the
quantitative structure of spring zooplankton was
Results and discussion similar. Rotifers predominated in these lakes.
Their percentage share ranged from 60 %to 94 %,
The spring zooplankton of the examined lakes and was highest in the zooplankton of strongly
was composed of 33 taxa of Rotifera, 10 taxa of eutrophic lakes. Eight taxa constituted the domi-
Cladocera and 7 taxa of Copepoda. The qualita- nant group in the rotifer community. Both
tive composition of zooplankton in most of lakes eurytopic species and lake trophy indicators pre-
was relatively uniform, because, as a rule, it was dominated among the rotifers. In mesotrophic
represented by 20 to 23 species. There were 28 lakes Conochilus unicomis Rouss. and Kellicottia
species in one shallow and highly eutrophic lake. longispina (Kell.) preferring waters of low trophy
The species composition of zooplankton in the were found among the dominants (Tab. 1, Fig. 1).
L~czna and Wlodawa lakeland was represented However in eutrophic lakes the clear dominants
by many more species than in the Florida lakes of were Keratella cochlearls tecta (Gosse) and
similar trophy. In each of the 8 lakes of the Keratella quadrata (Mull.) - indicators of high
L~czna-Wlodawa lakeland there were 11 to 18 trophic-status (Tab. 1, Fig. 1). A similar species
Table 1. Number of indicators of waters trophy in lakes of different trophism.

Lake Piaseczno Rog6zno Bialskie Rotcze Czarne Krzczen Glebokie

Indicator Sosn. Cyc. Uscim.

EUTROPHIC WATERS:
Bosmina longirostris (O.F.Mull.) + + + + + +
Branchionus angularis Gosse + +
Filinia longiseta (Ehrb.) + + +
Keratella c. tecta (Goose) + + + + + +
Pompholyx sulcata (Huds.) + + + + + +
Trichocerca cylindrica (Imf.) +
Keratella c. hispida (Laut.) + + + + + +
Keratella quadrata (Mull.) + + + + + + + +
Polyarthra euryptera Wierz. + +

Total 3 4 2 3 7 5 8 8

MESOTROPHIC WATERS:
Bosmina coregoni Baird. +
Conochilus unicomis Rouss. + + + + + + +
Gastropus stylifer Imh. + + +
Kellicottia longispina (Kell.) + + + + + + +

Total 4 3 2 3 2 2 2
 237

U
>-
U

t>l
H
:..:
r.:I
H
0
gj. x:
H
:..: u
3 000 til ~ til
::>
....:I
...;
H c.l
a:I H
t>l :..:
Z N
U
0
gj.
bc:t:
til
0 0
Z til zr.:I tj
2000 'N
"0 N
u
8 ~
.. N
e::; e::; 00
...;
N
o• el
0 u .0
Z
N
U II
W
II
1000 til
...;
H II
p..
II
II
E
I I
II
II
RC RC RC RC RC RC RC

.....
--<Il
......
<II
::I
.....
."

.....>
."
....
I': 1000
....0
~
Q)
M
..c
§
Z

RC RC RC RC

lOool ..
l l L
", I
H

RC RC RC RC

!Zl K. cochlearis II c. un icornis 1m P. vulgaris § A. priodonta

§.. K. quadrata ~
II K. c . hispida ..
~ K. c . tecta 00 K. longispina

III B. long irostris illIJ B. coregoni ~ D. cuccullata Q M. leuckarti

0 Others E Epilimnion M Metalimn ion H Hypolimnion

R Rotifera C Crustacea

Fig. 1. Abundance structure of zooplankton in particular thermal zone in lakes of different trophy.
 238

composition of dominants has been found in 3. With an increase in lake trophy there is an
other Polish lakes (Hillbricht-Ilkowska & increase in the number of indicators of
W~glenska, 1970; Radwan, 1976; Karabin, 1983; eutrophy and a decrease in the number of indi-
Radwan et al., 1984) and in other countries cators of mesotrophy.
(Pejler, 1965, 1981; Ferrari & Rossi, 1970;
Blancher, 1984).
The crustacean plankton in the examined lakes References
was scarce and of a lowest abundance in highly
eutrophized lakes. In slightly eutrophic lakes Blancher, E. c., 1984. Zooplankton - trophic state relation-
Cladocera were the predominant crustacean ship in some north and central Florida lakes. Hydro-
plankton, but in highly eutrophic lakes the biologia 109: 251-263.
Brzc:k, G., C. Kowalczyk, W. Lecewicz, S. Radwan, W.
Copepoda were predominant (Fig. 1). In two Wojciechowska & I. Wojciechowski, 1975. Influence of
lakes affected by strong human influences (hydro- abiotic environmental factors on plankton in lakes of dif-
technical and breeding activities) the Cladocera ferent trophy. Pol. Arch. Hydrobiol. 22: 123-139.
were the most abundant zooplankton. Their per- Ferrari, 1. & O. Rossi, 1976. Compositions and seasonal
centage share ranged from 51 % to 54 %. Among succession of the zooplankton in lake Mergozzo (Northern
Italy). Me. 1st. ital. Idrobiol. 33: 85-104.
them small filter-feeding Daphnia and Bosmina Hakkari, 1., 1972. Zooplankton species as indicators of
predominated (Fig. 1). environment. Aqua Fenn. pp.46-54.
The number of indicator species depended on Hillbricht-Ilkowska, A. & K. Patalas, 1967. Methods of esti-
trophic state, with an increase in lake fertility mate of the production and biomass and some problems
there was an increase in the number of indicators of the quantitative zooplankton methodology. Ekol. Pol. B,
13: 139-172 (in Polish).
of eutrophy and a simultaneous decrease in the Hillbricht-Ilkowska, A. & T. Wc:glenska, 1970. Some rela-
number of mesotrophy indicators (Tab. 1). tions between production and zooplankton structure of
The total numbers of spring zooplankton in the two lakes of a varying trophy. Pol. Arch. Hydrobiol. 17:
examined lakes ranged from 576 individuals 1- 1 233-240.
to 2,667 individuals 1- 1. It increased with increas- Karabin, A., 1983. Ecological characteristics of lakes in
north-eastern Poland versus their trophic gradient. VII.
ing lake trophy. This relationship is very clear in Variations in the pelagic zooplankton (Rotatoria and
6 lakes undergoing gradual eutrophication. It was Crustacea) in 42 lakes. Ekol. Pol. 31: 383-409.
the strongest in deep lakes, first of all, in their Kowalczyk, C. & S. Radwan, 1982. Groups of pelagic
meta- and hypolimnion. However, in the epilim- zooplankton in three lakes of different trophy. Acta
nion, irrespective of the lake trophic status, these Hydrobiol. 24: 39-51.
Patalas, K., 1954. Comparative studies on a new type of self
were considerable fluctuations, due to continuous
acting water sampler for plankton and hydrochemical
changes of major environmental factors (Fig. 1). investigations. Ekol. Pol. Ser. A, 2: 231-242 (in Polish,
English summ.).
Pawlowski, L., 1972. Liste supplementaire de Rotiferes
Conclusions trouves dans la riviere Grabia. Bull. Soc. Sci. L6dz 21:
1-10.
Pejler, B., 1965. Regional - ecological studies of Swedish
1. In the meta- and hypolimnion of deep lakes freshwater zooplankton. Zool. Bidr. Upps. 36: 407-515.
with well developed thermal stratification, the Pejler, B., 1981. On the use of zooplankton as environmental
total number of zooplankton increases with an indicators. Some Approaches to Saprobiol. Problems
increase in trophic state. As a rule, the pre- 1981: 9-12.
Radwan, S., 1973. Pelagic rotifers of the Lc:czna and
dominating species are rotifers. This is not true Wlodawa Lake District. Faunistic-ecological studies.
of the epilimnion (Fig. 1). Rozpr. Nauk. Akad. Roln. Lublin 8: 1-57, (in Polish).
2. In the zooplankton of lakes subjected to Radwan, S., 1976. Planktonic rotifers as indicators of lake
gradual eutrophication, rotifers predominate, trophy. Ann. Univ. Mariae Curie-Sklodowska, Sec. C, 31:
227-235.
however, in the zooplankton oflakes subjected
Radwan, S., C. Kowalczyk, W. Kowalik & W. Zwolski, 1984.
to intensive human activite cladocerans from Littoral fauna in two lakes of different trophy (Bialskie and
the genera Daphnia and Bosmina predominate Piaseczno, Eastern Poland). Verh. int. Ver. Limnol. 22:
(Fig. 1). 991-995.
Hydrobiologia 186/187: 239-245, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 239
© 1989 Kluwer Academic Publishers.

Tasmania revisited: rotifer communities and habitat heterogeneity

RJ. Shiell, W. Koste 2 & L.W. Tan 3

1 Murray-Darling Freshwater Research Centre, P.O. Box 921 Albury, N.S. W. 2640, Australia; 2 LudWig
Brill Strasse 5, Quakenbriick D-4570, Federal Republic of Germany; 3 Western Mining Corporation, P.O.
Box 409 Unley, S.A. 5061, Australia

Key words: rotifers, Tasmania, dune lakes, endemism

Abstract

The results offour field surveys for Rotifera in Tasmania are summarized. Most new species and records
in a 1987 survey were from acid waters (pH < 4.0) of dune lakes on the west coast (42 0 S). Marked intra-
and interhabitat differences in rotifer communities of lakes and ponds were demonstrated by cluster
analysis and related to habitat heterogeneity.

Introduction excluded by acidity; 'several taxa, including ...

rotifers ... are rare or absent in acidic dune lakes'.
Our first survey of Tasmanian natural lakes on the Our attention was directed to these lakes by Dr
central plateau (1980) provided 62 new rotifer P.A. Tyler at the University of Tasmania, who
records (one n. sp., Aspelta tilba). In 1984 the had sampled them for phytoplankton.
survey was extended to impoundments and a The 1987 survey resulted in 159 identified taxa,
range of smaller ponds and stock dams, with 62 59 ofthese first records for the island (with Lecane
further taxa (including n. spp. Lecane tasmanien- herzigi, Notommata tyleri, Trichotria buchneri and
sis, Lepadella tyleri and Testudinella unicornuta) T. pseudocurta new) (Koste et al., 1988). Most of
added to the known Rotifera (Koste & Shiel, the new records came from the western plateau
1987a). In 1985, a third survey (100 sites, includ- area (Fig. 1). Some 250 taxa of Rotifera are now
ing all sites samples earlier), added 63 taxa known from the island, and it is likely that only a
(including Brachionus Iyratus tasmaniensis, Cepha- fraction of the rotifer fauna has been encountered.
lodella lindamaya, Lepadella tana and Testudinella We summarize here the results of the four surveys
mucronata tasmaniensis) (Koste & Shiel, 1986). to date, and discuss some of the peculiarities of
To provide seasonal information from the same the Tasmanian limnetic and littoral rotifer com-
100 localities, a fourth survey was made in spring munities relative to those of the nearby mainland.
1987, with the addition of eight new sites in a
series of dune lakes on the west coast of the
island. We had not investigated dune lakes previ- Sampling sites
ously; work on the mainland (e.g. Timms, 1973)
suggested that dune lakes were not productive of All sites described in the earlier reports were
Rotifera. Indeed, in a review of these habitats, resampled in 1987. Habitats ranged from shallow
Timms (1986: 425) noted that rotifers seem to be roadside pools, ponds, turbid stock dams, to large
240

Fig. 1. Tasmania, with nine lakes mentioned in text. Arrows indicate approximate locations of endemic species figured.

impoundments and natural lakes above 1000 m Species

A.S.L. on the central plateau. A number of stream
and river sites also were sampled. For con- Species lists are provided in each of the survey
venience, the sites are grouped here into (a) lakes, reports cited above, considerably extending the
(b) ponds, and (c) streams, however the division known distribution of most taxa recorded. New
is arbitrary in view of the range of habitats within species apparently confined to Tasmania are
each category. shown in Fig. 1. Notably absent from Tasmania
 241

are Brachionus species known from the mainland Table 1. Summary of rotifer recorded from Tasmanian and
(28 taxa) (Koste & Shiel, 1987b). Only Australian mainland habitats. Lakes includes artificial
impoundments; Ponds includes turbid stock dams.
B. angularis was common in Tasmanian waters,
(R. Murray data after Hillman, 1986).
with occasional occurrences of B. calyciflorus,
B. quadridentatus and a local variant of B. lyratus No. of species
(Fig. 1). The most abundant taxa in limnetic
assemblages were species of Keratella, with nine Tasmania R. Murray
(240) (229)
identified and four (K. australis, K. cochlearis,
K. procurva and K. slacki) widely distributed. 1. Lakes 148 126
Also abundant were species of Lecane (28 spp.), 2. Ponds 189 174*
with four taxa common in the limnetic regions of 3. Streams 50 73
ponds and stock dams (L. bulla, L. flexilis, 4. Lakes and ponds 59 85
5. Lakes and streams 3 45
L. hamata and L. lunaris), and an endemic
6. Ponds and streams 10 54
(L. tasmaniensis) occurring widely in acid waters 7. Lakes. ponds and streams 41 39
in the southwest. Trichocerca species (26) were
widespread, but only T. simi/is abundant. * Billabongs
Of 250 taxa recorded, 116 (48 %) were found
only once (or twice from the same locality in dif-
ferent surveys). Such patchiness is generally in the flows slower, and a complex rotifer plankton
accord with Australian mainland surveys (Shiel & is maintained.
Koste, 1986) and with studies reported elsewhere A simple plot of species numbers against
(Dumont, 1983). Approx. 20% (49 spp.) have not sampling frequency (Fig. 2) suggests that more
been reported from the mainland; in most cases rotifer species await discovery in Tasmania. Typi-
these are acidophile species known previously cally, such plots produce an asymptote as
from tropical regions of South America or South repeated surveys produce fewer unknown taxa.
East Asia, e.g. Testudinella ahlstromi, Trichocerca Our sample series has not reached an asymptote,
braziliensis (Koste, 1978). About half the recorded although first records have declined (90%, 47%,
taxa were oflimited distribution ( < 20 % oflocali- 32 %, 28 % for respective surveys). Given that we
ties). Only 10 species (4 %) occurred in > 20 % of have collected from only 100 of > 4000 available
samples (n = 267): K. slacki (38 %), T. similis sites, it is conceivable that more rotifer taxa ulti-
(35 %), K. cochlearis (32%), K. australis/L. flexilis mately will be recorded from Tasmania than pres-
(30 %), L. lunaris (28 %), L. bulla (23 %), ently are known from the mainland (about 600
B. angularis/Polyarthra vulgaris (21 %) and K. pro- species).
curva (20%).
Comparison of species distribution data from
the Tasmanian series with those from a more
intensive sampling effort reported by Hillman
(1986) from the R. Murray catchment on the 300
U)
mainland suggests that the Tasmanian rotifer 4)

fauna is more diverse (Table 1), i.e. the R. Murray ~200

Q.
U)
data represent > 1500 samples from a ...o
300000 km2 area, the Tasmanian data 267 . 100
o
samples from a 68000 km2 area. More species z
occur in Tasmanian standing waters, with a
marked decrease in streams possibly reflecting the 1980 1984 1985 1987
steeper gradients and greater turbulence at Fig. 2. Cumulative species numbers per sampling survey,
Tasmanian sites. R. Murray gradients are gentler, Tasmania 1980-1987.
242

Community composition rotifer assemblages, although not necessarily the

most species. The rotifer assemblages (l987
Using Sokal's (l961) distance measure to com- survey) often sites (mean no.spp. site- 1 = 10.4;
pare species assemblages (number of taxa and mean diversity (H') = 2.31) are compared by
proportion of each) across all sites in all years, the average linkage cluster analysis in Fig. 3a. Rela-
most striking feature of the rotifer assemblages tive proximity of the lakes is shown in Fig. 1.
was the marked dissimilarity in species com- Although communities of the ten lakes were
position between habitats, even within the same ranked of low similarity (0-22% taxa shared) by
habitat category. The whole data set is too large Jaccard coefficient run concurrently with Sokal's
to reproduce here, however two smaller subsets niche distance, clustering grouped lakes in close
are used below as examples. proximity, e.g. Great Lake and Arthur's Lake,
both dominated by Polyarthra vulgaris, and shar-
(a) Lakes ing Gastropus minor and Filinia longiseta from 16
In summary, over the four surveys, the natural and 10 species, respectively. Similarly, Plateau
lakes on the central plateau had the most diverse (l6 spp.) and Augusta (11 spp.) were both domi-

(al Plateau -
I
l Augusta -
Great -
Arthur's -
r--
Gordon -
Pedder w-
I

-
r-
y I Pedder E-
Garcia -
Dobson -
Fenton -

Y
(bl 1977 -
2028-

2050-
2025-
1992 -
~ 1993 -
2004-

-
r-
-1 2014-
2032-
2045-
1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1
Sokal's niche distance
Fig. 3. Average linkage dendrograms (Sakal's niche distance) for (a) ten lake sites and (b) ten ponds/stock dams (indicated by
site #). 1987 survey.
 243

nated by Synchaeta oblonga and shared Euchlanis abundant pond rotifers were species of Polyarthra,
meneta, Lepadella patella and Lecane jlexilis. Lecane, Filinia, Trichocerca and Synchaeta.
Generally, the widely distributed taxa (K. coch- As in the lake communities, there was little
learis, P. vulgaris/dolichoptera, F. pejleri/longiseta similarity in inter-site rotifer species composition
and T. similis) comprised the shared species; the on each survey. Fig. 3b shows clusters produced
remaining taxa were specific to each lake. No two for ten unnamed pond sites (mean no. spp.
lakes had the same combination of temperature, site - 1 == 17.3; mean H' == 3.25) selected from
pH and conductivity (within the range 7-14 ~C widely seplll'llted tlfetls of th@ id:md i.e. 1977,
pH 4.0-7.7,9.0-33.0 J.lS cm - 1), suggesting other 1992, 1993 are north coast, 2004 and 2014 east
differences in chemistry which may account in coast, 2025 near L. Pedder, 2050 near Garcia and
part for the disparity of resident rotifer assem- so on. Most obviously, tighter clustering suggests
blages by their influences on the algal and bac- that the pond rotifer communities are more uni-
terial communities. form than those of the lakes, i.e. more species are
Notably, a net tow from Lake Garcia, a dune shared. Again, the analysis grouped sites in
lake first sampled in the 1987 survey (Fig. 1) closest geographical proximity, e.g. 1992/1993
(17.0 °C, pH 4.3, 98.3 J.lS cm - 1) had 35 rotifer and 2004/2014, with outliers (1977, 2032, 2045)
species, including 8 new records (3 n. spp., with the most isolated sites.
one (Trichocerca) yet to be described) and the The most diverse pond rotifer assemblage
highest diversity (H' == 4.37) yet recorded from (H' == 3.92) and greatest number of species (32)
Tasmania. The assemblage was a mixture of lim- occurred in a small black-water humic roadside
netic and littoral taxa indicative of shallow pool in the dune lakes area (site 2050, near L.
eutrophic waters, including K. procurva (domi- Garcia, Fig. 1). This pool had the lowest pH (3.1)
nant), 5 spp. of Lecane and 8 of Trichocerca. of the eight sites in the dune series (pH range
Seasonal changes in species composition 3.1-4.3). Dominant taxa were: K. javana, Lecane
within each of the lakes were reflected in low tasmaniensis, Synchaeta oblonga, Cephalodella
similarities between surveys, with only 1-3 of the mucronata and Trichotria buchneri. The high
common species present on successive visits. In species diversity at this site accounts for the
the larger lakes, e.g. L. Pedder, there were anomalous grouping with 2028 in Fig. 3b, which
appreciable community differences at opposite was not geographically close, but which also was
ends of the lake on the same day. In L. Pedder in dominated by the acidophile K. javana, and
the 1987 series (cf. Fig.3a), community domi- shared other acidophile taxa (incl. L. jlexilis and
nants at the eastern end of the lake were L. tasmaniensis) of 15 spp. present.
K. cochlearis/ Synchaeta pectinata/ Conochilus dos- The ten pond data set was run against the ten
suarius, while at the western end, 12 km away, lake set to compare the respective assemblages.
dominants were K. australis/F. pejleri/C. dossua- The dendrogram derived (Fig. 4) maintains the
rius with only 20 % of taxa shared. separation of all lakes except Garcia, which clus-
tered with the pond assemblage, sharing five of
the widely distributed taxa with site 1992, and
(b) Ponds supported our impression of a mixed limnetic/
Smaller standing waters commonly had 8-20 littoral assemblage from the shallow margin of
tychoplanktonic species, with dominants one or this lake.
more species of Keratella (the most common in
ponds and stock dams was K. slacki, followed by
K. australis > K. cochlearis > K. procurva > The dune lakes
K. javana, with incidental occurrences of K. qua-
drata, K. valga, K. hispida and K. tropica). Three The unexpectedly high species richness of the
sites had six co-occurring Keratella species. Other dune lake series is clearly in contrast with the
244

Plateau -

-1 Augusta -
GrI~~t

Arthur's-
-
Pedder w-
J
I Pedder E-
Garcia -
r-

r-
-1 1992 -
2004-

2014 -
r-
2032-
- 1993 -
2045-
1977 -

"--
~y 2028-

2050-
2025-
Gordon -
Dobson -
Fenton -

1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1
Sokal's niche distance
Fig. 4. Sokal's niche distance for combined ponds/lakes data sets. L. Garcia separated from lakes, clustered with ponds.

observations of Timms (1986), and may represent Shiel, 1986). Many thousands of years isolation
a real difference between the dune lakes of during periods of climatic change could account
Tasmania and the mainland. The rotifer com- for the high species diversity and predominance of
munities of these acid waters are among the taxa now recorded primarily in the tropics, but
richest yet recorded from Australia. A possible which may have persisted in Tasmanian waters as
explanation is provided by recent geological evi- relict populations from a time when Tasmania's
dence reviewed by De Deckker (1986), which sug- climate, and that of southern Australia, was tropi-
gests that these western Tasmanian waters have cal. Notably, the west coast of Tasmania, directly
a long, undisturbed evolutionary history. in the path of prevailing westerlies, has the highest
At the height of the last glaciation (> 20 000 rainfall in southern Australia (ca. 5 m annually);
years B.P.), only part of Tasmania was covered even shallow pools are permanent.
by ice. It is likely that some of the many lakes,
particularly in coastal areas, remained ice-free.
Geological evidence points to the persistence of Rotifer biogeography in Tasmania
some of these lakes during an extremely arid cli-
matic phase 18000 years B.P., when their waters Dumont's (1983) question 'Do rotifers have a
may have been refugia for aquatic species as the biogeography?' was applied in the global context,
mainland dried (De Deckker, 1986; Koste & with evidence that although most species were
 245

cosmopolitan or widely distributed, some were was at the Botany Department, University of
remarkably restricted. Our Tasmanian surveys Adelaide; thanks to the then Chairman, George
must be regarded as preliminary, however it is Ganf, for access to facilities. Particular thanks to
apparent that Tasmanian rotifers have their own Terry Hillman, MDFRC, for cluster analysis, and
biogeography. to Kim Jenkins for graphics.
If an arc is drawn on Fig. 1 approximately from
Great Lake southwest through Aspelta tUba, east
of Lakes Dobson and Fenton to B.lyratus References
tasmaniensis, the eastern margin of the central
plateau is demarcated. We have named this arc De Deckker, P., 1986. What happened to the Australian
aquatic biota 18000 years ago? In P. De Deckker & W. D.
'Tyler's Line'. It separates two broad rotifer
Williams (eds) Limnology in Australia. CSIRO & Dr W.
assemblages: west of Tyler's Line, predominantly Junk BV, Melbourne and Dordrecht: 487-496.
acidophiles, contains 11 of the 13 endemic taxa Dumont, H. 1., 1983. Biogeography of rotifers. Hydrobiologia
and most of the species which have not been 104: 19-30.
recorded from the mainland. Waters are dark- Hillman, T. 1., 1986. Billabongs. In P. De Deckker & W. D.
Williams (eds) Limnology in Australia. CSIRO & Dr W.
black humic acid type, low in electrolytes, and
Junk BV, Melbourne and Dordrecht: 457-470.
presumably differ markedly in characteristics Koste, W., 1978. Rotatoria. Die Rlidertiere Mitteleuropas.
such as nutrients and phytoplankton, and rotifer Bestimmungswerk begriindet von Max Voigt. 2 vols.
composition from those east of the line. To the Borntraeger, Stuttgart.
east, which is a rainshadow area, waters are less Koste, W. & R 1. Shiel, 1986. New Rotifera (Aschelminthes)
from Tasmania. Trans. r. Soc. S. Aust. 110: 93-109.
humic, some are alkaline, and electrolytes are
Koste, W. & R 1. Shiel, 1987a. Tasmanian Rotifera: affinities
higher, including some saline and hypersaline with the Australian fauna. Hydrobiologia 147: 31-43.
waters on the east coast, i.e. they reflect geological Koste, W. & R 1. Shiel, 1987b. Rotifera from Australian
and climatic differences between the two areas. inland waters. II. Epiphanidae and Brachionidae (Rotifera:
The heterogeneity of rotifer assemblages across Monogononta). Inv. Taxon. 1: 949-1021.
Koste, W., R 1. Shiel & L. W. Tan, 1988. New rotifers
the island appears to be a response to local, even
(Rotifera) from Tasmania. Trans. r. Soc. S. Aust. 112 (in
microhabitat, variations in their requirements. press).
Shiel, R. J. & W. Koste, 1986. Australian Rotifera: ecology
and biogeography. In P. De Deckker & W. D. Williams
(eds) Limnology in Australia. CSIRO & Dr W. Junk BV,
Acknowledgements Melbourne and Dordrecht: 142-150.
Sokal, R R, 1961. Distance as a measure of taxonomic
similarity. Syst. Zoo!. 10: 71-79.
In 1987 survey was funded by the Australian
Timms, B. V., 1973. A limnological survey of the freshwater
Biological Resources Study. We thank Peter Tyler coastal lakes of east Gippsland, Victoria. Aust. J. mar.
for help with field work, and for hospitality and Freshwat. Res. 24: 1-20.
use of facilities at Hobart. Initial MS preparation Timms, B. V., 1986. The coastal dune lakes of eastern
Australia. In P. De Deckker & W. D. Williams (eds)
Limnology in Australia. CSIRO & Dr W. Junk BV,
Melbourne and Dordrecht: 421-432.
Hydrobiologia 186/187: 247-254, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 247
© 1989 Kluwer Academic Publishers.

Occurrence of Rotifera in the field under natural and

intentionally-changed conditions. II. Lake Numasawa

M. Sudzuki, T. Matsumoto 1 & K. Narita 1

Biological Laboratory, Nihon Daigaku-University, Omiya-shi SaUama-ken, Japan; 1 Naisuimen Fisheries
Experimental Station, Inawashiro-cho, Fukushima-ken, Japan

Key words: seasonal-vertical occurrences, power-water circulation, saprobic rotifers

Abstract

Seasonal and vertical occurrences of representative rotifer species were recorded together with such taxa
as Cyanophycea, Phytomastigophorea, Bacillariophycea, Protozoa, Rotifera and Crustacea, from 1982
to 1986 at two sites SI (natural) and S2 (nearby water circulated since 1952). 1) The following species
were observed from S 1: K. hiemalis, C. ovalis, N. labis, L. patella and Anuraeopsis sp., from S2:
B. urceolaris andA. ecaudis, 2) P. hudsoni appeared in 1984 as a successortoP. truncatum, 3) K. longispina
was negatively associated with P. t. vulgaris, 4) Synchaeta spp suddenly appeared at both sites in 1985,
5) K. cochlearis, B. calyciflorus, Proalides sp., Diurella sp., N. labis, B. urceolaris and bdelloids did not
appear until 1986, 6) The following species decreased or disappeared: A. p. herricki and Collotheca sp.
since 1982, C. coenobasis, C. hippocrepis and K. hiemalis since 1984, 7) A complicated relation was
observed between rotifer population density and other plankton. 8) Occurrence of Rotifera seemed to be
affected by circulation of an electric power plant.

Introduction least once per season, except winter, from 0 to a

50 m depth at 7-11 different strata
Seasonal and vertical occurrences of representa- (0,5,10,15,20,30,40 m occasionally 25,35,45 and
tive plankton in a lake, locally called Numasawa- 50 m) at 2 sites of a lake. S 1 is natural lake water
numa, were investigated from 1982 to 1986 as a and S2 is nearby water that has been circulated by
continuation of a 1980-81 project (Sudzuki et a/., an electric power plant. A total of 256 samples
1983). In this paper, a correlation between some was examined.
10 rotifer species and 6 plankton groups is figured Further detail for materials and methods is
and tabulated. given in Sudzuki et al. (1983).

Materials and methods Results and discussion

All the plankton were concentrated by pulling or Results are presented in Figs. 1-5 and Tables 1-2.
towing a Rigosha's plankton net (diameter = We may make the following observations: 1) Poly-
20 cm, mesh size = 47 jlm, messenger = 500 gr) arthra t. vulgaris is considered to be eurythermal
through 4 m of water. Samples were collected, at and perennial, but prefers to concentrate in the
248

Depth
•
•
1.0
•• + •
ft.f!. .· 1
••
••••
• • ••••
0.4 2 .3

•• ••
11 3.4 1.9 3.0
••
1.1 1.1
+ • • ., 10

•
1.0 0.7 4.9 2.7 3.4 3.1 0.9 1.2 2.2 ~ 2.0
IS • • ., + • + • + + • • •• 15
. 0.5 3.4 • +

• • "•
1.5 3.4 0.5 3.0 A 1.0 0.7 0.7
+ •
20 • + •
0.6 •0.7 ~ +
VI 0.8 0.5 3.2 ~ ++ !ft ~8 20

• • ·VIl
1.9
• •2.0 +
~ ft
3.6
25 • • ., + + • + + 25
•2.8 1.5
• + 1.8 Ai. 1.3 0.5 1.6
• ••• + + • + + • + + • + • • + 30

•
+
••
0.7 1.0 0.5 1.5 1.0 0.4 ., + 0.6 0.4 3.0 1.7
+ 3S

••
35
1.6 1.1 1.2
• • • + • + + • + + + 40
0.3 1.2 0.4 0.9 2.7 0.8 U
45

so •
•
2.3
+
+
P . I . lIulgaris
45

so
0.4

.. +
Depth
•• o

•+.
+ + + + +
1.4 0.7

.: +
•
0.5 . 2.0
+ + ••
0.8 0 .8
10
10 • + + +
0.5 0.1
15
•
0.6
+ •
1.0
+ + 15

20 + + + ~~ + + + + 20

• ••
2.0
4.2
25
•
0.7 1.5 1.4 1.5
+ + + 25

30 +
••
2.0 0.6
+ 30

35 + •
0.5
+ + 35

40

45

K . cochlearis P. hudsoni so

SIS2S2S1
4 4 23 26
IV II II II II IIII
'86 '86 '85 '82 '84 '84

•
Depth

••
+

··1
+
•• •
.:
1.1
•3.2 • • +
•
••
+ + • • • +
1.5 1.1 0.8 1.7

•
1.1
\0 + • • \0
• <I
••
1.3 1.2
••
1.6 0.5 38 4.0
+ + + • 15

•
\5
1.4 1.5 1.4 2.7 0.9
10
• + •
0.4
20

•
0.5 1.9
25
•
1.0 +
+ •
0.5
+

35
+ +
+
+ •
1.6 0.4
• •
0.6 + 1.4• •
1.6
+ + + UI

••
2.4 0.8
+

•
+ + •
1.1 0.8
•
+
•
0.6
+ + +

1.6
;(1
•
1.5
+
0.4
•

... :! .... I .... !

fl . IOllgispill(l
.... t .. I ;-. , ... " ... I
:-.. I ~:! :-.. 1 :"'11 ::"'I "' ."1 I ~ 't =-- I ~2~1:-t ') ~I~I:-t2:"1I!"12 .I ~I~:!""I
]'; 1:- :?I '" 15 15 1 17 19 20 30 3 3 I -1 :n 26 2Y :?Y 1; II I I'" l4 ~j!'" I
\1 II I I I I I II 1"1 II 1'1 1'1 II 1\1 II II II \I 1'1 II II II 1\ \ \ \ \ \1
~.i ~:! ...d .)'!, ~ ",6 k:! lotS 3 1'16 Jo!6 '~5 ~I ~I

Fig. 1. Seasonal and vertical occurrences of 4 rotifers in lake, Numasawa during 1982-86. S 1 = natural lake water, S2 = nearby
water circulated by an electric power plant since 1952. 17 IV '86 = 17th April, 1986, Depth = depth in meter, numbers in
circles = individual density per liter (as to diameter of circles see Carlin, 1943), + = individual density less than 0.1.
249
250

10

15
•
10
Depth

.~
• 1.7
+

~
• 0.6•
0.5
+
. •
4.8
+
+
•
+

1.0
+
+
+
+ +
10

15

20
•
1.3 +
+ •
0.6 •
0.8
+
25
•
0.7
+ + •
0.7
+ •
0,5
+ 25

30 + •
0.7
•
0.5
+ + •
0,5
30

35 35

40 + •
0.4
+ 40

45 45

50 F . o/ ngiseta 50

5 1$2SI S I I I 1 2 S I S I S 251
17 11 21 8 IS 15 IS 19 20 30 3 3 4
IV IV V VI \'1 \'1 VI VI 11 11 I'll III III III
'86 '86 '82 'S3 '8-1 . 4 8' 6 '86 '82 ' 5 '2 '83 '83 8' 6

+.
Depth
~ :s 0•" Ps•
+ + + •
0.3
+ + +

•
O.S •
0.6 1.0 0 .3
+
Ps• ~ '-t + + • •
0,5 0.1
+ +
10 + • • 0 .6 • +
ft+ + • • + + + + + + + 10

•
0.8 0.1 2.2 0.2 0.2
IS
• O•.S
0.5 0.6
+ •
0.5 0.1
• + + + + + + + 15

20 + •
1.0
+ + + + •
0.3
+ + 20

• + +
25

30
•
0,5
0.5
+ a,s•
+ +
+
+ •
0.5
..
+

0,1 0.1
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30

35 + •
0.1
+ 35

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•
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45

50 + A . p . herricki 50

I I 2 2 I I I
17 19 20 30 4 23 26 25 II 18 28 28 18
IV VI VI VII III \'I III II IX X X X X XI
'86 '82 '85 '82 '86 '86 '85 '82 '82 '85 '4 '86 '86 . 2

·
.•..
Depth

•
10

IS
o
2.1

•
1.0 •
~
•
0.7

•• •
1.0

• i
1.7
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• • ~
•
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1.7

~7 ~
•
1.9
10

IS

25
+
1.5
•1.7 •
• •
1.3
2,5

2.9 •
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3 ,2
+
+
•
0.8
+
0.5 2,9 0.7

•
0.6
2.1
•
0.6
• 20

25

30
•O.S • •
2,0 0 ,6
+ •
O.S
30

35 + 35

40

45
•
0.6 •
1.2
+ + 40

4;

50
•
0.8
C, hippocrepi
~ I S 2 S I S2 ~I _'I ~ 2 .' 1
17 17 21 '9 2j 2K 2K IK
II' II' I' \'I I -I III ~ ~ \ \ \1
'S1 -83 ' 3 -S-I - -I ' " . I .? . 5 '2 6

Fig. 3. Seasonal and vertical occurrences of 3 rotifers in lake, Numasawa during 1982-86,
 251

Depth
• • o

•••
0.7 2.1

•
10 2.9 •• • ,·•Po
3}
10

~. • •
0.1
•
0.6 1.4 0.5
IS

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1.2 1.2 2.0
20

• 25

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JO
0.9 0.5 2.8 2.1

•
0.6
•
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1.5

•
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Cyanophycea
45

so

2 5 1 5 2 5 2 5 1 5 1 52 S2 5 I
334423262929 28 18
VI 1'1 1'1 1'1 1'1 1'1 1'1 1'1

.
X XI
's?

I_••••
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'83 '83 '86 '86 '85 '82 '84 '84 '82
Depth

10 •• 1.° e_ ~ •~ • •••
0~3 O~ • •
0.8 1.7

• 2.0
It
••
- • ~• 10

••
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1.1 1.3 2.3

•••
15 3.1 • • 2.0
15

•• •
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1.4 1.0 0.1
20 ••
4.6 4.8 3.5 0.5 18
•
0.5 • 20

25 • • o~
• ~ o~
•• •
2.0 1.0 10
.~
0.8
25

JO ••
4.6 2.1 0.6
• •
0.5 1.0 3.7
JO
35

.i• •••
• 3S
40
••
1.8 2.4 • ~ 0.1 •• 0.1 0.6 4.1
1.8

•
3.0
40

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0.4
3.3
••
1.4 3 .5
•
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Phytomastigophorea
4S

so
5 1 5 2 5 151 5 2 25 1 5 2 5 1 5 1 5 2 5 1 5 2 5 2 5 1 5 1 5 2 5 1 5 2 1525251515251
17 17 21 8 8 17 19 20 JO 3 J 4 4 23 26 29 29 25 II 18 18 19 27 28 28 18
IV IV I ' l 'l VI 1'1 \1 VI VI VII 1'1 1'1 1'1 ''I 1'1 1'1 ''I 1'1 IX X X X X X X X XI
'86 '86 '82 '83 '83 ~ ~ ~ • ~ ~ ~ ~ ~ • ~ w w ~ • w w ~ ~ ~ ~ ~
Depth

•
• • • •
•••
0.4 0.6 1.6 1.4

• •
10

IS
•• ¥~
•• ••
18 4.5 3.6 3 .2
2.3 0.9
••
• 1.0
•
0.6 2" la
•••
I
10

15

20

~
20 •
3.1
• • ~• ••••
•••
0.4 0.9
25 •
• • • •• 0.1 •• 25

e- ••
0.5 0 .4
• •••
3.6 0 .5 1.0 0.5 0.2
••• • JO

••
30 ••
11 1.4 0.5 0.5 4.7 2.6 1.6 ~ • 1.9 0.5 0 .6 3 .9 0.1 0.43.4 0.5 2.6 2.2
35 JS

o~• • • • •
••
•• • • • 3.9
2.2 1.9
'0 • •
0.5 0.1 3 .3 2.8 2.4 0.5 0] 0.1 0.4 3.4 •
1.0
40

• 4S

•
2.6 1.0
;0

';1 ,;~ ';1 SI

0.8
,'2 S I 52 5 1 5251
•
1.8
S I S I 52 SI S2 ~I ~2
Bacillarioph ycea

,2 , 1 ~ 1 .' 2 , 1
so

17 17 21 B ~ 15 15 I 17 19 20 3 3 4 4 26 29 29 25 II I 18' 19 27 2R 28 IR
I\' 1\' \. \'I \ '1 \'I \ '1 \'1 \'1 \'1 \ '1 1'1 1'1 1'1 1'1 1'1 1'1 1'1 IX .'( .'( X X X X .\ XI
~ .~ '~2 • 3 g3
" "81 "lU '86 '86 "82 '85 83 ' ; 8' 2 'S< '8' H2 '85 ' , ' , 11.1 ~2 ~ t\6 '~

Fig. 4. Seasonal and vertical occurrences of phytoplankton in lake, Numasawa during 1982-86.
252

.•••••••
epth
• ft • •
•
~II
~
•
0:1
•
0.7 0.1

10
• ••
1.1

3.2 0.5
~1.0 !J~.~. ~ ••
0.7

•
2.2

2,3
0.1
10
2.4 0.7
. 0.5
~
0.4 4.5 2.0 1.9
15 • • • • • • 15

••• • ••
0.6 0.4 1.5 0.1 1.1 0.5
20
•0.5 2.0 0.1 •
0.5 0.1 3.4 •
0.5
20

25 • 25

••
0.5 1.0 1.0 0.5 0,5
JO
• • • • + JO

35
0.5 0.5 0.5 2.0
•
0.6
0.1

0.1
0.5
•
0,5
0.5
.
0.1
40
0.1 0.1
•
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•
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40

45 45
0.1
50 Prolozoa 50

5251515251525152 1$2S2SISIS2SIS25152S251S12S1
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'86 '82 '83 '83 'St 'St '86 '86 ~ ~ ~ ~ • ~ ~ ~ ~ • ~ ~ ~ '2 ~ ~ ~

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20 • •••• 20
0.5 0.5 1,0 3,6 1.9
25
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4.6 0.3 U 0.5 2,0
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i• • • •
35
• 35

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0.3 1.6 1.3 1 .3
02
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40 •••
0.5 4.2 2,5 1.0 •
0.2 3,5 2.3 1.0
'
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Ja.
~ •
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+
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•
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•
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•
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50
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.
SIS 2 $251525152$15152 25251515251525152 2151 5 251
11 17 15 18 11 19 20 30 3 3 4 4 23 26 29 29 25 II 18 I 19 27 28 28 I
IV IV VI VI VI VI VI ~ VI VI VI VI VI VI VI VI IX X X X X X X X XI
'86 '86 ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ '83 '82 '86 '86 'S2

•• ••• ••
·i····i.
Depth

10

15

20
•
1.1 0.2
•
0,8 0.1

~
•• 0
• .•
1.6

O.B ...... ft···

·2.iiii.ielili
• 0.7• 1,0• 1•.2 0.4• 3,0 4.4 •
•
•••
• 4,2
0.7
1.7 3.7

- 0.2 1 .9 2.6 2.5• 0••.4 0.3•• 1.2••0.6••0.8••1.5 1.5 2..5 41•

• • ••
• • • •••••••••••
• • • • • • • • • ••
• • • • • • •• •••••
3.3 2.2 1.4
3.0 3.3

1.2 1.9
4,9 4.4 3.7

2.4 0.7 1.5 D 0,8 1.0

4.3 . 2.9

2.1 1.7
10

15

20

25
0.3
•2,1 :..i! 0.3 0.4 0.6 0.6 0.7 4.8 0.9
•
• • •• ••••4J ••
2,1 0.8 0.5 0.5 2.2 1.8 0.8 0.9 1.0 1.3 2.5 2 .1 0.4

• • • • • • • • • 0.7• ••• • •• • • • • • • • •
1.6 1.7 1.0 0] 1.1 0,4 1.0 1.4 1.8 1.0

0.1 0.1 0.2 3.0 •

JO
• • 30

•• •
3.0 03 0.5 0.9 0.8 0.9 0,8 1.8 1.4 1.8 2.2 0.7 0.5 0,8 1.4 0.4 0.3 0 .2 O,Y 0,3 0.4 0,6 0.7 0.3
35
• • • • 35

40
•
0.2 0 .6
•0,9 0.2
• •
0.4
0.9
•
0.7
•
0,3 0 .5
••• 1.0 0 .6 2.2
0.8 0.8
•
0.8
•0.2 •• • •••••
0 .8 0 ,5 0.4
0.9

0.4 07 0.5 0.4 04

D,S 0]
•
03
40

45
• • • 45

•
0.6 0.4 0.5
Cr u lacea
50
•
0.5
50

S I S2 ~ I SI S251 S2 5 1 525 1 2 S2 51 SI ,2 .' 1 ~2 ., 1 ~2~2..,1..,1..,~..., 1

17 17 21 IS 15 18 11 19 4 • 23 26 29 29 25 II I 18 l ~ 21 2~ 2x !K
1\ 1\' \. \ '1 \ '1 \ '1 \ '1 '1 \ \'1 \ '1 VI VI VI VI VI VI ~ \ \ \\\\\ \1
~ ~ g ~ 83 ~ s.t ~ 8 6 2 '86 '86 85 82 ~ 8-' H2 H; H' \;1"] "2 1>fi><6 "2

Fig. 5. Seasonal and vertical occurrences of zooplankton in lake, Numasawa during 1982-86.
 253

Table 1. List of taxa investigated. high abundance in Numasawa-numa. Never-

theless, P. hudsoni increases year after year. It is
Rotifera regarded as a summer form (Carlin, 1943; Pejler,
Polyarthra trigla dolichoptera. P.t. vulgaris. Synchaeta sp. 1, S.
sp. 2, Ploesoma hudsoni. P. truncatum. Ascomorpha ecaudis.
1957) or oligo-mesotrophic (Maemets, 1983),
Chromogaster ovalis. Kellicottia longispina. Notholca labis. although Sladecek (l973) regarded it as oligo-
Anuraeopsis sp., Keratella cochlearis. K. hiemalis. Brachionus saprobic. 5) Kellicottia longispina is considered to
calyciflorus. B. urceolaris. Lepadella patella. Lecane inermis. be eurythermal and perennial. 6) Varga (l954)
L. nodosa. L. sp., Monostyla quadridentata. Proalides sp.,
mentioned that K. longispina is a glacial relict in
Diurella sp., Filinia longiseta. Asplanchna priodonta herricki.
Conochiloides coenobasis. Conochilus hippocrepis. Collotheca
Lake Balaton and adapted to rising water tem-
sp., Bdelloidea, etc. peratures. However, Larsson (1971) classified it
Cyanophycea as a perennial summer form even though it has not
Gomphosphaerium sp., etc. been detected from tropical waters (Sudzuki,
Mychophycea
1989) nor ponds in temperate regions (Sudzuki,
Bacillariophycea
Asterionella spp., Cyclotella spp., Tabellaria spp., etc. 1964). 7) K. longispina abundance is negatively
Chlorophycea correlated with P. t. vulgaris, a fact which was
Oocystis spp., Chroococcus spp., Dictyosphaerium spp,. observed by Laxhuber (1987). 8) Keratella
Pandorina spp., Eudorina spp., Uroglenopsis spp., Closterium hiemalis is treated as a winter or cold tolerant
spp., etc.
stenothermal species from the hypolimnion
Phytomastigophorea
Peridineae, Dinobryon spp., Mallomonas spp., Ceratium sp., (Carlin, 1943; Pejler, 1961). In contrast, it rarely
etc. is encountered in Numasawa-numa. 9) Cono-
Protozoa chiloides coenobasis is considered to be a warm
Zoomastigophorea, Actinopodea, Centropyxis spp., Difflugia water hypolimnion inhabitant. 10) Ploesoma trun-
spp., Cyphoderia sp., Euglypha spp., Holotricha, Epistylis spp.,
catum is considered to be a summer inhabitant in
Zoothamnium spp., other Spirotricha, etc.
Trematoda (larva), Nematoda, Chaetonotus spp., the hypolimnion. 11) P. hudsoni appears as a
Crustacea successor of P. truncatum, since truncatum was
Ceriodaphnia sp., Daphnia spp., Alona spp., Bosmina spp., common until 1983 but a shift from truncatum to
Bosminopsis sp., Holopedium sp., Polyphemus sp., hudsoni happened in 1984-85. In 1986 hudsoni
Diaphanosoma sp., Copepoda, Nauplii, shrimps became dominant. 12) Filinia longiseta is con-
Insecta
Chironomus spp., Collembola sidered to be the eurythermal-perennial of the
Arachnida hypolimnion. Nearly the same results were
Acarina, Araneae. obtained (Pejler, 1957); George & Fernando,
1969; Larsson, 1971 & Herzig, 1987). The species
studied by Carlin (1943), Pejler (1961), Berner-
epilimnion as already depicted by Carlin (1943), Fankhauser (l983) and Laxhuber (1987)
Sudzuki (l955), George & Fernando (1969), appeared seldom or never in summer.
Larsson (1971), Berner-Fankhauser (1983), 13) Asplanchna p. herricki also is eurythermal and
Herzig (l987) and Laxhuber (1987). 2) Keratella perennial. The same kind of result was reported
cochlearis was not perennial in Numasawa-numa (Berner-Fankhauser, 1983) in contrast to Carlin's
contrary to Carlin (l943), Sudzuki (l955, 1962), fmdings (1943). 14) Conochilus hippocrepis is con-
Pejler (1961), George & Fernando (1969), sidered to be eurythermal and perennial, but con-
Larsson (1971), Berner-Fankhauser (1983), centrates in the hypolimnion. 15) A correlation
Herzig (1987) and Laxhuber (1987). 3) When between the occurrence of Rotifera and other
K. cochlearis was common in 1986, it suggested plankton groups is not always simple. For exam-
certain pollution (beta-oligosaprobity: Sladecek, ple, P. t. vulgaris occurrence is correlated with
1973 or meso-eutrophic: Maemets, 1983, but a Phytomastigophorea but varies inversely with
conflicting case in Neusiedlersee in 1979: Herzig, Bacillariophycea; F. longiseta abundance is nega-
1987) 4) Ploesoma hudsoni has not yet reached tively correlated with Bacillariophycea; A. p. her-
254

Table 2. Seasonal and vertical occurrences of uncommon rotifers. Density = Number of individuals per liter,
Date = day/month/year, Depth = depth in meter, Sl = site 1 (natural lake water), S2 = site 2 (circulated water by an electric
power plant).

Taxa Density Date Depth Site

Synchaeta sp. 1 0.1-27.7 IV-X/84-86 0-45 S1S2

S. sp.2 3.0-11.6 20/IV/85 5-10 S2
Ascomorpha ecaudis 0.1- 0.7 4/VIII/86 10-35 S2
Chromogaster ovalis 0.5- 4.2 28/X/86 0-30 SI
Notholca labis 0.6 17/IV/86 5 SI
Anuraeopsis sp. 0.5 4/VIII/86 15 SI
Collotheca sp. 0.5- 5.6 VIII-X/83-85 0-30 S1S2
Brachionus calycijlorus 0.5- 1.7 17-18/VI/86 0-30 S1S2
B. urceolaris 2.6 17/VI/86 0 S2
Lepadella patella 0.7 28/X/86 0 SI
Lecane nodosa 0.4- 0.6 VI-VIII/83 0-15 S1S2
L. inermis 0.6- 5.3 VI-X/85-86 0-30 S2
L. sp.l 1.0- 5.3 20/VI/85 0-30 S2
Monostyla quadridentata 0.1- 1.0 VI,VIII/85 0-40 S2
Proalides sp. 1.0- 1.1 17-18/VI/86 20 S1S2
Diurella sp. 0.5- 0.6 28/X/86 20-30 S1S2
Bdelloidea 0.2- 4.3 VI-X/86 0-35 SlS2

ricki is inversely related to Phytomastigophorea George, M. G. & c. H. Fernando, 1969. Seasonal distribu-
and so on. tion and vertical migration of planktonic rotifers in two
From Tab. 2 we may make some observations: lakes. Verh. int. Ver. Limnol. 17: 817-829.
Herzig, A., 1987. The analysis of planktonic rotifer popu-
1) The following species have appeared mainly in lations. Hydrobiologia 147: 163-180.
S 1 or natural lake water: Chromogaster ovalis, Larsson, P., 1971. Vertical distribution of planktonic rotifers
Notholca labis, Lepadella patella and Anuraeopsis in a meromictic lake. Norw. J. Zool. 19: 47-75.
sp. 2) The following mainly in S2 or the water Laxhuber, R., 1987. Abundance and distribution of pelagic
circulated by an electric power plant: Brachionus rotifers in a cold, deep oligotrophic lake (Konigssee).
Hydrobiologia 147: 189-196.
urceolaris, Ascomorpha ecaudis, Monostyla quadri- Maemets, A., 1983. Rotifers as indicators of lake types in
dentata, Synchaeta sp.2, Lecane sp.l. 3) The fol- Estonia. Hydrobiologia 104: 357-361.
lowing have a tendency to increase their individ- Pejler, B., 1957. Taxonomical and ecological studies on
ual densities at both sites (S 1,S2): Brachionus planktonic Rotatoria from northern Swedish Lapland. K.
calyciflorus, Synchaeta sp.1, Proalides sp., Diurella svensk Vetensk Akad. Handl. 4: 3-52.
Pejler, B., 1961. The zooplankton of Osbysjon, Djursholm.
sp. and Bdelloidea. 4) The following have a I.Oikos 12: 225-248.
tendency to decrease at both sites: Lecane nodosa, Sladecek, V., 1973. System of Water quality from the bio-
Collotheca sp. 5) From the fact above we may logical point of view. Ergeb. Limnol. 7: 218 p.
consider that the occurrence of Rotifera may be Sudzuki, M., 1955. Life histories of some Japanese rotifers I.
affected by circulation of an electric power plant. Polyarthra trigla. Sci. Rep. Kyoiku Daigaku 118: 41-64.
Sudzuki, M., 1962. Class Rotatoria. In: Uchida Ed. Sys-
tematic Zoology 2: 9-74 (Nakayama-shoten, Tokyo).
Sudzuki, M., 1964. New systematical approach to the
Japanese planktonic Rotatoria. Hydrobiologia 23: 1-124.
References Sudzuki, M., 1983. Occurrence of Rotifera in the field under
natural and conditions. Hydrobiologia 104: 341-347.
Berner-Fankhauser, H., 1983. Abundance, dynamics and Sudzuki, M., 1989. Rotifera from the Oriental region and
succession of planktonic rotifers in Lake Biel, Switzerland. their characteristics. Special Issue for Centennial Anniver-
Hydrobiologia 104: 349-352. sary of Nihon Daigaku 3: 301-343.
Carlin, B., 1943. Die Planktonrotatorien des Motalastrom. Varga, L., 1954. A relict of the ice age in the water fauna of
Medd. Lunds Univ. Limnol. Inst. 5: 1-256. lake Balaton. Annal. BioI. Tihany 22: 227-234.
Hydrobiologia 186/187: 255-278, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 255
© 1989 Kluwer Academic Publishers.

The skeletal muscles of rotifers and their innervation

Pierre Clement 1 & Jacqueline Amsellem 2

1 Laboratoire Ethologie, Equipe Neuro-Ethologie, IASBSE, 2 Laboratoire Histologie Experimentale
UA-CNRS 244, ICBMC, Universite Lyon I, Mtiment 403,69622 Villeurbanne, France

Key words: muscles, innervation, behavior, cytology, ultrastructure, rotifers, smooth muscles, striated
muscles, motor units, phylogeny

Abstract

The skeletal muscles of rotifers are monocellular or occasionally bicellular. They display great diversity
of cytological features correlated to their functional differentiation. The cross-striated fibers of some
retractors are fast contracting and relaxing, with A-band lengths of 0.7 jlm to 1.6 jlm, abundant sarco-
plasmic reticulum and dyads. Other retractors and the circular muscles are tonic fibers (A band> 3 jlm),
stronger (large volume of myoplasm) or with greater endurance (superior volume of mitochondrial
myoplasm). All of these retractor muscles are coupled by gap junctions and are innervated at two
symmetrical points; they constitute two motor units implicated in withdrawal behaviour.
The muscles inserted on the ciliary roots of the cingulum control swimming. They are multi-innervated
and each of them constitute one motor unit. They have characteristics of very fast fibers; the shortest
A-band length is 0.5 jlm in Asplanchna.
All the skeletal muscles of bdelloids are smooth or obliquely striated as are some skeletal muscles of
monogononts. These muscles are well suited for maximum shortening and are either phasic or tonic fibers.
All rotifer skeletal muscles originate from ectoderm and contain thin and thick myofilaments whose
diameters are identical to those of actin and myosin filaments in vertebrate fast muscles or in insect flight
muscles. There are no paramyosinic features in the thick myofilaments. The insertion, innervation,
coupling by gap-junctions and other cytological differentiations of rotifer skeletal muscles are reviewed
and their phylogeny discussed.

Introduction 1929-1932; Hyman, 1951; de Beauchamp, 1965).

Electron microscopy showed that every rotifer
Muscles of rotifers can be grouped into two muscular fiber has a nucleus (Clement, 1977) and
classes. The visceral muscles lie along the diges- that each skeletal muscle is uni- or bicellular.
tive tract and surround other internal organs. The Studying skeletal muscles and their innervation
second class are the skeletal muscles which con- is essential to analyze the behavior of rotifers.
trol body form, movement, and spatial displace- Their small size (generally from 0.1 to 1 mm)
ment. This paper reviews only skeletal muscles; makes an electrophysiological approach difficult
visceral muscles will be treated in another review. and this has not yet been attempted. Never-
The first rotiferologists described skeletal circu- theless, this technique would be of great interest
lar muscles as anucleate (see review by Remane, because each skeletal muscle is made of only one
256

Plate I: Insertions of the skeletal muscles.

Fig. a. Philodina roseola. The oblique arrows show some of the musculo-epithelial desmosomes. The ribbon-like material is
inserted on the desmosomes and goes through the transversally sectioned myofilaments (small arrows). At right, a dense material
 257

or two fibers. The fiber lengths are as much as 2/3 in the monogonont Trichocerca rattus was made
of the body lengths and this review will show that from seriated ultrathin sections of contracted or
each fiber is differentiated and can be observed in relaxed animals (Amsellem & Clement, 1977,
living animals (light microscopy) through their 1981, 1988). This review gives information about
transparent integument. this work which is largely unpublished and on
From the beginning of the century, the trans- unpublished results obtained in our laboratory on
parence of the integument has permitted locali- several other species of rotifers: the bdelloid
zation of the musculature in monogononts Phi/odina roseola and six monogononts: Brachio-
(Remane, 1929-1932) and bdelloids (Brakenhoff, nus urceolaris sericus (female and male) B. calyci-
1937). However, no functional analysis was done florus (collaboration with A. Comillac), B. plica-
because the cyto-histological observations with ti/is (collabortion with A. Luciani) Asplanchna
light microscopy were not precise enough. brightwelli (collaboration with A. Comillac),
Electron microscopy increased our knowledge B. plicatilis (collaboration with E. Wurdak), Rhi-
of skeletal muscles, with two kinds of observa- noglena frontalis, Notommata copeus.
tions: Phylogenetic relationships are also proposed,
1) Precise cytology from serial ultrathin based on ultrastructural analysis ofrotifer skeletal
sections permitted quantification of cytological muscles. Clement (1981) has used several other
parameters which have been correlated with ultrastructural criteria to show that rotifers are
muscular characteristics (10sephson, 1975; very primitive metazoa. Finally, we present new
Hoyle, 1983; Nicaise & Amsellem, 1983). These evidence showing the extraordinary diversity of
include speed and strength of contraction, en- specializations of rotifer skeletal muscles.
durance, maximum shortening and speed of
relaxation.
2) Precise localization of innervations or gap The epithelial origin of the skeletal muscles of
junctions permits defmition of motor units. There rotifers
are very few motor units in rotifers and their
identification is essential for the analysis of Vertebrate cross-striated muscle fibers come from
behavior (Clement et aI., 1983; Clement, 1987). mesoderm whereas most smooth fibers and car-
Ultrastructural observations of rotifer muscles diac muscle cells come from the mesenchyme. A
were first available from analysis of other organs, few smooth muscular cells, like those around the
like the integument (Koehler, 1965, 1966; acini of the salivary glands, come from ectoderm
Clement, 1969, 1977, 1981, 1985, 1987; and are thus called 'myo-epithelial' cells. In
Schramm, 1979). An exhaustive study of muscles sponges myocytes which are the first expression

joins each desmosome to the skeletal lamina densa (L) which is intracytoplasmic in the syncitial integument of the trunk. The
cuticle is external. x 60000.
Fig. h. Trichocerca rattus. Muscle dense bodies are juxtaposed to desmosomial plates (d.p.) where thin myoflimaments are also
inserted. Thick myofilaments are on the left. In the syncitial integument, in front of each dense body, dense material joins the
desmosome to the skeletal intracellular lamina (L) of the trunk. x 32000.
Fig. c. Brachionus plicatilis. Cingulum ciliary roots are inserted on a muscle cell (M) (transversally sectioned): at the level of
the desmosome, the cross-striated ciliary roots insert in a submembranar fibrillar material (arrow). x 21000.
Fig. d. Brachionus calyciflorus. The cingulum cell contains oblique and axial sections of ciliary roots associated with numerous
mitochondria. The muscle (M) inserts on a basal fibrillar lamina (arrow at left) and directly on the basis of the ciliary roots (arrow
at right). x 5000.
Fig. e. Brachionus calyciflorus. Apex of the corona: a desmosome (D) joins the muscle (M) to the microfibrillar subepithelial
layer (thick arrow). Microvilli (m) cross the cuticle (cu). x 39000.
Fig. f Trichocerca rattus. The desmosome (D) joins the muscle to the dense material of a buccal cell. Buccal cilia are on the
left. x 36000.
258

Plate II: Innervation

Fig. a. Brachionus plicatilis. Cross section of a CRM, showing the myop\asm (M) and the sarcoplasmic extension (S) which
penetrates into the brain (at right). P = pseudocoel. x 5800.
 259

of a muscular differentiation are mesenchymatous In rotifers, even in the absence of a real sarco-
cells (Hoyle, 1983; Pavans de Ceccatty, 1974). In plasmic extension, the muscular synapses occur
Cnidaria and Ctenaria, myoepithelial cells are often on the peripheral sarcoplasm near the
juxtaposed to mesoglean muscular cells (e.g. the nucleus when it is peripheral (Clement, 1987).
giant smooth muscular fiber in the ctenophore Nevertheless, numerous neuro-muscular synap-
Beroe: Hernandez-Nicaise & Amsellem, 1980). ses occur near the myoplasm of the skeletal fibers
In rotifers, there is neither mesenchyme nor even in bdellords (Plate II - d).
typical conjunctive tissue (Clement, 1980). All the
skeletal muscle cells come from ectoderm. Con-
trary to nematodes, rotifer muscle cells only have The insertions of the skeletal muscles of rotifers
a muscular function, but there are some excep-
tions to this rule. They can have an epithelial Insertions on the peripheral skeleton
function when adjacent to neurons. For instance,
around the rotifer brain there are bordering Each ending of circular muscles is inserted on the
epithelial cells with associated muscle cells skeletal syncytial integument of the trunk and
(rotifers have no glial cells, Clement 1977, 1980). sometimes at the level of muscle Z material.
More spectacular is a cylindrical muscle in which Longitudinal muscles constitute two groups: one
a part of a nerve is lodged (Plate II -c). This where muscle fibers join the trunk to the articu-
socket cell is a classical specialization of epithelial lations of the foot and toes (Clement, 1987) and
cells around a sensory nerve as in the nematode another were muscle fibers join the anterior rota-
Caenorhabditis elegans (Ward et al., 1975; Ware tory apparatus to the trunk (Plate I - c, d, e).
et al., 1975). Muscle insertions on the skeletal integument
Rotifer muscle fibers have another charac- always implicate the thin fIlaments which termi-
teristic resembling the muscle cells from the body nate in the dense peripheral material of a des-
wall of the nematode Ascaris (Rosenbluth, 1965) mosomial structure. From the epithelial part of
where each cell sends an extension to the the desmosome, a fibrillar dense material goes
peripheral longitudinal nerve cord for innerva- through the epithelium to join the internal sheet of
tion. Paired rotifer central retractor muscles each the intracytoplasmic internal skeletal lamina
send a sarcoplasmic arm inside the brain where (Koehler, 1965; Clement, 1969, 1985). In the
the two symmetric arms contact by a large gap primitive insertions ofbdelloids, punctual desmo-
junction and are innervated by cerebral neurons somes are regularly spaced (Plate I - a; Koehler,
(Plate II, Clement, 1987; Amsellem & Clement, 1966; Schramm, 1978; Clement, 1981, 1985) and
1988). Similar muscle extensions with the Z dense material (but is it Z dense material
neuromuscular synapses have been observed in or others structural proteins?) corresponding with
Platyhelminths (Chien & Koopowitz, 1972) in the the desmosomes penetrates deeply into the
polychaete Syllis, the onychophore Peripatus, a muscle cell (Plate I - a). In monogononts, punc-
starfish and in the prochordate Amphioxus tual desmosomes are grouped in desmosomial
(Hoyle, 1983). plates (Plate I - b). Such juxtapositions have

Fig. h. Trichocerca rattus. Innervation (arrow) at the end of the sarcoplasmic extension of a CRM, inside the brain. x 66400.
Fig. c. Philodina roseola. A cross section of a cylindrical muscle (M) which is a socket cell for a nerve (ne = a transversal section
of neurite). Part of the gastric gland at left. x 13 300.
Fig. d. Philodina roseola. Multi-innervation (arrows) of a skeletal muscle (M = myoplasm, s = sarcoplasm) by two nerve
endings (nel and ne2). In the muscle fiber sections surrounding the nerves, note thin (t) and thick (T) myofilaments, dense bodies
(d.b.), sarcoplasmic reticulum (sr) and some mitochondria (mi). x 33200.
Fig. e. Philodina roseola. Detail of Fig. d., showing the neuromuscular synapse. Note the numerous synaptic vesicles, the
characteristic dense line intercellular (thick arrow) and the post-synaptic submembranar material (little arrows). x 66400.
260

E
(a)

Plate III: Fig. a, b, c. Axial sections of the retractors of Trichocerca rattus.

Fig. a. Relaxed LRM (lateral retractor muscle): A band (A) and I band (I) and lined Z bodies in the myoplasm (m). x 12000.
Fig. b. Contracted VRM (ventral retractor muscle). The sarcoplasm (s) with mitochondria and glycogen particles, is juxta-
 261

been described in Sagitta (Duvert, 1971). The The two cells are closely joined by two types of
insertions on the supple integument (anterior junctions: mechanical ones, which are desmo-
integument surrounding the rotatory apparatus, somes and hemidesmosomes (Plate IV - a) and
or constituting the pedal articulations) are of the gap-junctions which are electrical couplings of the
same type but with less spectacular desmosomes. two muscular cells (Plate IV - a). In other cases,
the retractor is inserted on other muscles such as
the ventral retractor muscle (VRM) on the
Insertions on the ciliary of the rotatory apparatus peripheral muscles of the mastax.

Muscle insertions on the ciliary cells of the In conclusion, each skeletal muscle insertion
cingulum and of the pseudotrochus are unique. plays a precise functional role. Some mechanical
Ciliary roots or a fibrillar material which is in insertions are involved in animal deformations
close contact with them ends in the epithelial part like coronal withdrawal behavior, movements of
of the musculo-epithelial desmosomes (Plate I - the foot or of spines and paddles. More func-
d, c and Clement, 1987; Plate II, Clement & tionally complex are the insertions on the
FouiHet, 1971; Clement, 1977, 1985). With this cingulum and pseudotrochus ciliary roots in-
structure, the muscle can control the ciliary beat. volved in the control of ciliary beat.
The retractors establish classic single desmo-
somes with the anterior cells or desmosomes in
relation with the microfilamentous network which Cross-striated muscles implied in the withdrawal
lies at the base of the microvilli (Plate I - e) or behavior of T. rattus
desmosomes related with the uniformly dense
material which is at the base of the buccal cilia The most extensive study of skeletal muscles in
(Plate I - f). These musculo-epithelial desmo- rotifers was made on the retractor muscles in-
somes have a mechanical role in the retraction of volved in the withdrawal of the ciliary corona in
the rotatory apparatus by these retractor muscles. T. rattus (Amsellem & Clement, 1977, 1988).
Transversal serial ultrathin sections of one animal
were used to quantify cytological features of each
Musculo-muscular junctions muscle cell and longitudinal, oblique or trans-
versal sections on several other animals. The dif-
Rotifer skeletal muscles are generally unicellular ferent muscles implicated in withdrawal behavior,
but some are bicellular as the central retractors which is the most common in rotifers (Clement
(CRM) of Trichocerca, Brachionus and other et al., 1983; Clement, 1987) are the following.
species; the anterior cell is short and ramified. 1) Three pairs of retractors: central (CRM),

posed to a lamina of myoplasm (m) which reply on itself at the upper part of the picture. Note the supercontraction at the level
of Z line. int: rigid integument of the trunk. x 12000.
Fig. c. Contracted CRM (central retractor muscle). Axial sarcoplasm (S) in a cylindrical myoplasm (m). Supercontraction is
more pronounced at the upper part of the picture. x 12 000.
Thick myofilaments are shorter in band c than in a.
Fig. d. Axial section of the base of a CRM of Brachionus urceolaris sericus male; the myoplasm (m) is cylindrical around the
sarcoplasm (s) and attached to the integument by two desmosomes (arrows). x 20000.
Fig. e. Transverse section of a CRM of T. rattus. Thick and thin myofilaments. Note the hollow center of the thick ones.
x 100000.
Fig. fAxial section of a tonic smooth muscle of Philodina roseola. Thick myofilaments are very long. Some dense bodies can
be juxtaposed (arrow) but they are generally scattered in the myoplasm. x 19500.

C: ciliated cell; E: external medium; P: pseudocoel; ne: neurites (ventral nerve); int: integument; m: myoplasm; S: sarcoplasm;
Z: dense bodies.
262

Plate IV. Phasic cross-striated retractor muscles.

Fig. a, b, c. Trichocerca rattus. a) Junctional complex between the two muscle cells of CRM, with gap-junctions (gj), desmo-
somes (D) and hemidesmosomes (HD). x 29000. b) axial section of the supercontracted CRM. x 21 000. c) axial section of the
relaxed CRM. x 26000. Sarcoplasm is axial in a and b.
Fig. d, e,f Brachionus calyciflorus. d-e): CRM (central retractor muscle): axial sections. 0: DRM (dorsal retractor muscle): it
is more obliquely striated than the cross-striated CRM of d and e. x 8500.
Fig. g. Asplanchna brightwelli. Note the shortness of the A bands (A) and the abundance of sarcoplasmic reticulum (arrows).
This muscle is inserted to a cingulum cell by a desmosome (D) and controls swimming. x 12400.
 263

ventral (VRM) and lateral (LRM). The CRM are sarcoplasmic reticulum and the density of mito-
bicellular; VRM and LRM are monocellular but chondria. The different concepts about the struc-
the VRM are partly inserted on small peripheral ture-function relation in cross-striated muscles
muscles of the mastax or on anterior buccal has been reviewed by Josephson (1975).
muscles. The speed of shortening of a muscle cell is
2) Two anterior dorsal and ventral circular inversely proportional to the length of its A-band,
muscles constitute a spincter involved in the pro- directly proportional to the number of sarcomeres
tection of the retracted rotatory apparatus in series and to the density of dyads (sites for
(Fig. 1). excitation-contraction couplings). The density of
These four paired muscles are cross-striated sarcoplasmic reticulum surrounding the myo-
but reveal great differences in their cytological filaments is also related to the speed of con-
organization (Fig. 2). traction and relaxation. Regarding these parame-
It is generally believed that the activity of a ters, the CRM and VRM are considered fast
muscle cell is determined by the organization of its muscle cells. The circular and LRM would be of
contractile apparatus, the development of its a slower type (Table 1).

Fig. 1. Diagram of the muscles involved in withdrawal of the corona in Trichocerca rattus. Dorsal views, the brain is stippled.
a: contracted animal; b: relaxed animal. Location of the central retractors and some other muscles is indicated. C: central
retractors (bicellular) S: sphincter circular muscles; L: lateral retractors; cin: cingulum; ps: pseudotrochus; N: swimming
muscles.
264

:~~'-grid 9
,....s~ I
19",m Junctional complex between Cl and C2
I l ...
sarcolemmal invagination .···· . . .•
bended po. et'"ior
• .. end of
Ie! CI cell
he .. •

Area juxtaposi ion

•.......... •
wi h brain
20 e :t:ar-coplasaic proce •• e
to the right .--- junction between the two C2 muscles
15 C2 cell
10

15 20 25 30 35
a)cen ral muscle (Cl cell)

IIIlll ucleus
o Sarcoplasm
D
Mitochondria
Myoplasm E'ZJ
lateral extension
~~~:z::: g [" id 8
I
27",m •. . ... myoplasmic fold
... .•.... .
of the sarcoplasm

I and internal
' - grid 25 15 SR lamina
I ~~~ I
i 3",m 10

i I
~E!~~'- grif 27
, . lO",m 15 20 25 30 33
! ! I b) ventral muscle
~-g["id ))

:- grid 9
I 10
, 2S",m
i . I
~- .'Ii 25
5

10 15 20 25 30

~ s",m
c) lateral muscle
i : I
~- .d."

Fig. 2. Diagrams of the progressive characteristic levels of the central muscle (a), ventral muscle (b) and lateral muscle (c) of
T. raltus representatives of the morphological heterogeneity of muscle fibers. Quantitative cytology for the myoplasm, mitochon-
dria and sarcoplasm surface areas from serial transverse sections in the left CRM Cl cell, VRM cell and LRM celL Beyond the
curves, details about the morphology of the fiber are indicated between the precise levels where they appear and terminate.
 265

Table 1. Morphometric study. Characteristic cytological parameters of central (CRM) ventral (VRM), lateral (LRM) retractors
and anterior circular (Cir) muscles of T. rattus.

CRM VRM LRM Cir.

Speed of shortening +++ ++ + +

Maximum force + + + +++
Endurance + + ++ +

Volume of mitochondria (%) 6 8.5 10.7 3.1

volume of myoplasm
Number of sacromers 30 12-14 6-7 4
Length of the fiber (J.Lm) 81 44 37.5 20
Mean surface ofmyoplasm (J.Lm 2 ) 9 6.5 3.3 30
(transverse section)
A-band length (thick myofilaments) (J.Lm) 1.5 1.7 3 3.3
Thick myofilament diameter (nm) 15-20 15-20 15-20 15-20
Thin filaments diameter (nm) 7 7 7 7
Ratio f/F = thin (t), thick (F) 4: 1 4: 1 4: 1 3: 1
Z dense body length (nm) 90 90 140-200 90-120
Number of dyads/J.Lm2 2.7 1.5 1.7

The maximum force of a muscle cell varies with retractor fibers (80 p.m for the left). Less maxi-
the number of bridges made between the two mum shortening was found in VRM and LRM
types of myofIlaments during sliding. Force would (Table 1). In these three cases, the maximum
be proportional to the surface of the myoplasm in shortening is around half of the length of the rest-
a cross section fiber, the density of thick myo- ing fiber, due to the capacity of supercontraction
fIlaments in this section, the ratio of thin/thick where thick fIlaments cross over the Z bodies
myofIlaments and the length of these latter lines and go into the two adjacent sarcomeres
myofIlaments. Regarding these parameters, the (Plate IV - b). The network appearance (like a
three retractor muscles do not differ significantly. perforated disc) of Z material (Fig. 29, Amsellem
But the circular muscle is clearly stronger than the & Clement, 1988) in rotifers is similar to that
others which is due to the large volume of its found in supercontracting muscles of insects
myoplasm (Table 1). The function of each muscle (Hardie & Hawes, 1982).
is consistent with these characteristics. The maximum shortening is less important in
Endurance is measured by the volume of mito- the two circular anterior muscles which are
chondria/volume of myoplasm. It is very low in shorter and have a small number of sarcomeres.
the for types of muscles (3 to 11 %), compared to They are thick laminar muscles and the number of
the mastax muscles (where this relationship is sarcomeres varies from 2 to 3 depending of the
around 50%); these retractors obviously do not position of the myofibrils in the lamina.
have great endurance. This result is consistent In conclusion - muscles implicated in the with-
with the fact that the withdrawal response is gen- drawal behavior can be classified in two cate-
erally short. When contraction persists, the LRM gories: (l) the phasic muscles: CRM and VRM,
seem to be primarily involved (Amsellem & with a high speed of shortening and relaxation, the
Clement, 1977). CRM being the fastest with the maximum capa-
The maximum shortening of a muscle cell is the city of shortening, and (2) the tonic muscles:
difference in length between its resting and maxi- LRM and anterior circular muscles, with slower
mum contraction states. This value is maximum and more sustained contractions, the circular
for the Cl cell of the CRM, which has the longest muscles being the strongest and the LRM proba-
266

Plate V. Tonic skeletal muscles (a,c,d) and pedal muscles (b, e)

Fig. a. Axial section ofthe circular anterior dorsal muscle in T. rattus. Myoplasm is very abundant and organized in juxtaposed
myofibrils where the sarcomeres are well defined; thick myofilaments (A-band) are 3).1m long (arrow) and myofibrils are
 267

bly having greater endurance. Among the parame-

ters which differentiate phasic and tonic muscles,
the most useful are the thick myofilament length
and the Z dense bodies length. The presence of
invaginated sarcolemmal tubules making contacts
with the sarcoplasmic reticulum is related to a
large volume ofmyoplasm: such invaginations of 1 1
the sarcolemma are numerous in some parts of the
circular muscle (Plate V - a) and absent in LRM L •2 II
2 L
(Plate III - a).

=
Innervation of the muscles involved in the with- C C
drawal behavior

The innervation of the muscles involved in this

behavior was studied in T. rattus (Clement &
Amsellem, 1987; Clement, 1987). Results are
summarized in Fig. 3 and 4.
The two CRM are innervated into the brain by
way of the sarcoplasmic arms sent in the third
part of their longest CRM cells (Plate II - a, b).
The two cells of each CRM are coupled
(Plate IV - a) and the two CRMs are coupled by
v V
a large gap junction (Amsellem & Clement, 1988).
So, any stimulation, lateralized or not, provokes
the simultaneous contraction of this motor-unit Fig. 3. Diagram of the gap junctions present between the
(Fig. 3). muscles. A double line between 2 cells indicates that the
junctions have been observed; a single line indicates that
The other muscles (VRM, LRM, anterior
such junctions are very likely to be present (the membranes
sphincter) seem not to be directly innervated (we are in close contact) but have not been actually observed
did not find obvious synapses). They are never- because the angle of sectioning was unfavorable. Sd:
theless coupled by gap junctions and constitute sphincter circular dorsal; Sv: sphincter circular ventral; L:
one motor unit. This motor unit is innervated by lateral retractor LRM; C: central retractor muscles CRM;
V: ventral retractor muscles VRM; ml: secondary lateral
the anterior ganglia by way of a pair oflittle lateral
muscles; ii, i2: stimuli.

separated by sarcoplasm which contains mitochondria (mi), sarcoplasmic reticulum (s.r.); dyads (D) are visible between the SR
and tubular invaginations (T) of the sarcolemma int: integument; P: pseudocoeie; Z: alignment of dense Z bodies. x 21 000.
Fig. b. Transversal section of a pedal muscle of T. rattus. Note the density of longitudinal tubules of sarcoplasmic reticulum
(s.r.) and the peripheral dyads (D) x 20000.
Fig. c. Transversal section of a circular muscle in T. rattus. Note the regular arrangement of myofilaments and the hollow
structure of the thick myofilaments. x 172 000.
Fig. d. Axial section ofa tonic circular muscle of B. calyciflorus. Sarcomeres are well defined with a A-band of3.5 p,m. x 7300.
Fig. e. Axial section in a pedal muscle of T. rattus with its insertion on the integument (little arrow) and its innervation (large
arrow); the pedal ganglia innervates the other pedal muscles and the muscular lamina surrounding the pedal gland (p.g.). The
muscle is striated and the A-band length is around 3 p,m. The densities of SR (s.r.) and mitochondria (mi) are higher than in
tonic skeletal muscles of the same speci,es. x 8900.
268

weak stimuli

~
strong sti muli ___---+. i I

~
12

,
Motor unit Motor unit ..
p
small and short
retraction of lhe
2 = VRM 1= eRM rotetory apperatus
LRM
CRM
Clr. ....... strong and sustained
retraction of the
rotatory apparatus

Fig. 4. Schematic summary of the presumed involvement of successive motorunits depending of the intensity of the stimuli.

muscles, each one being innervated and coupled The cross-striated muscles which control the
to VRM and LRM. ciliary beat for swimming
In conclusion, the probable mechanism of the
withdrawal behavior can now be more precisely Swimming is a constant behavior of pelagic
explained (Fig. 4). Following stimulation, the rotifers and a frequent behavior of other rotifers.
innervation il of the CRM is activated; the con- Since the work of Viaud and others (review in
tractile response is brief and fast, followed by fast Clement et ai., 1983), it is known that different
relaxation. If the stimulus is stronger, innervation stimuli (light, vibrations, chemical substances)
i2 is also activated: coronal retraction thus influence swimming characteristics like linear
depends on the characteristics of all the muscles speed, angular speed and direction. These precise
involved in this response. The phasic CRM retract motor responses to stimuli are supported by sen-
the corona in the deepest part of the trunk; the sory organs and the nervous system (Clement
phasic VRM retract the mastax and the buccal et ai., 1983; Clement, 1987), but the ciliary cells
field, but not so deep; the tonic LRM and the which are responsible for swimming (the cingulum
anterior circular muscles then maintain this con- cells) are not innervated. Muscle cells are inserted
traction until their own relaxation. The corona on the cingulum ciliary roots (Clement, 1987 - 'N'
can then emerge because the CRM and VRM are on Fig. 1) (see above and plates I - c, d;
already relaxed, there is no antagonist muscle VI - a, c). Each of these muscle cells is mono-
working for this emergence, which seems to be a cellular and constitutes an independent motor-
passive consequence of relaxation of all retractor unit which is multi-innervated. Two independent
muscles and of internal pressure. motor-units, one at right and one at left, control
 269

swimming behavior so that the right part of the is 0.7 J-lm. This muscle is inserted on the fibrillar
cingulum can be slowed down apart from the left material of the anterior ciliary cells. We do not
part. actually know if this fibrillar material is connected
Cytologically, these muscles (Plate VI) are simi- with the ciliary roots of the cingulum. If con-
lar to small phasic muscles. In T. rattus, a nected, this muscle would be an another type of
cingulum muscle cell has the structural charac- cingulum muscle, closely related to those illus-
teristics of a CRM cell with more sarcoplasmic trated Plate VI. If not, this muscle would be a
reticulum and shorter A-bands (1.5 J-lm) retractor, faster than those found in other rotifers.
(Plate VI- d). In female Brachionus calycijlorus or plicatilis,
The cingulum muscles are more specialized in some retractors are cross-striated, but more
some rotifers. Male Brachionus have shorter irregularly than in T. rattus (Plate IV - d, e) and
A-bands (1.3 J-lm) and the sarcoplasmic reticulum the Z dense-bodies are shorter. Plate IV - d illus-
is well developed (Plate VI - c). In Asplanchna trates axial sections of B. calycijlorus CRM where
brightwelli, the swimming path is not helicoidal different states of contraction are seen in the same
and Clement (1987) described the double orienta- fiber. The supercontracted part occurred near the
tion of the cingulum ciliary roots which looks like middle of the sarcoplasmic arm of the fiber which
an inverted V at the base of each cilia. Each goes into the brain to provide innervation. In
extremity of the V is anchored in a fibrillar mate- supercontracted fibers, the superposition of the A
rial on which are inserted two opposite cingulum bands of two contiguous sarcomeres is the origin
muscle cells. The structure of these cells is excep- of a pseudostriation which is easily observed
tional in that the length of the A-band is the (Plate IV - d, upper part).
shortest known in animals (0.5 J-lm; I J-lm for the Intermediate situations exist between trans-
sarcomere length in resting state) (Plate VI - a, b). versal and oblique striation in retractor muscles.
In invertebrates, the shortest A-band lengths The dorsal retractor of the female of Brachionus
( < 2 J-lm) always correspond to fast fibers (Hoyle, (Plate IV - f) is representative of such a situation.
1983). Values like those of the cingulum A band Some small dense body alignments also exist in the
of Asplanchna occasionally have been observed. more primitive fibers of Philodina roseola where all
The thick myofilament length of the subumbrellar skeletal muscles are smooth or obliquely-striated
muscles of Scyphomedusa (Cnidaria) is 0.58 J-lm (Plate VII).
long (Chapman et al., 1962; Mackie & Singla, We need observations on the most developed
1975); that of the fast cross-striated muscles in cross-striated skeletal muscles in jumping rotifers
the tail of cercaria (Platyhelminths) is about (Polyarthra, Hexarthra, Triarthra, Filinia). This
0.5 J-lm (Rees, 1975) and in the chromatophore type of locomotion is very different than ciliary
muscle cells of Loligo (Cephalopod), this value is locomotion. Gilbert (1987) studied the escape
0.5 to 1 J-lm (Weber, 1970). Generally, in the fast response of Polyarthra with high speed cinemato-
fibers of invertebrates, A-band length averages graphy. Using this precise analysis, it would be
2.1 to 4.8 J-lm, but 1.3 J-lm was found in the mantle interesting to know the characteristics of the
of Sepia. In vertebrates, this parameter varies muscles involved in these movements.
from 1.7 to 1.9 J-lm for the same muscle type The only muscles supporting articulate move-
(Hoyle, 1983). ments in rotifers that we observed in electron
microscopy are those controling movement of the
foot. In Philodina roseola they are smooth fibers
The other cross-striated skeletal muscles of (Clement, 1987) like other smooth skeletal
rotifers muscles described below. In T. rattus, the tail is
very developed and its movements are controled
An axial section of a cross-striated muscle of by specialized original muscles (Plate V - b, e).
Asplanchna is seen on Plate IV - g; A-band length Their sarcomeres are well defined, delimited
270

Plate VI: Muscles of cingulum

Fig. a, b. Asplanchna brightwelli. a) Axial section of a muscle ending, inserted (arrow) on a cingulum cell (cc) ciliary roots
associated with mitochondria. C = cilia of cingulum. The muscle is cross-striated with a very short A band (A): 0.5 J.l.m x 14000.
 271

either by cross or oblique striation (Plate V - e).

Their myoplasm is laminar with numerous large
longitudinal tubules of sarcoplasmic reticulum
inside the myoplasm. Sarcoplasm mainly extends
from the anterior ending of the muscle cell where
1
10
II· -
1
10
1\: -
11
r
1111 . -
it contains the nucleus, glycogen particles and \II·I\! \11.:111
some mitochondria (Plate V - e). Innervation
occurs by the pedal ganglion which also inner- 1\.:\\1:
vates other muscles involved in the movement of 1\\1111111
the foot and the muscular lamina surrounding the
pedal gland (Plate V - e). Thus, all these pedal ;i
1\\\1\ \1 \\11. 1\11: 1\11
1
muscles constitute an unique motor-unit. I! .11\1 .\\\1
\\1\1. jill \1:\\1..\1 1
Obliquely striated fibers :1\·\\ \\\! \: 11\1\1
1\·\\1
Obliquely striated muscle cells are largely found in \\1·:11
Annelida, Nematoda (Rosenbluth, 1972), Cepha- III : \\1\1
lopoda (Amsellem & Nicaise, 1980), and have .:\\1
also been observed in the slow fibers of cercaria \\
(Plathyhelminths: review in Hoyle, 1983). All of III
these fibers contain paramyosin in their thick
myofilaments. In rotifers, we never observed the Fig. 5. The two types of shortening occurring in oblique
structural characteristics of the paramyosin in the striated muscle cells (sliding: -a (contracted) and -b (relaxed)
without altered degree of obliquity and shearing: -c (con-
obliquely striated fibers. In these muscle cells, the
tracted) with altered degree of obliquity; Rosenbluth, 1972)
dense bodies are obliquely lined up in relation to which permits the muscle to adjust to a wide range oflengths
the direction of the myofilaments (fiber axis). passively (from Knapp & Mill, 1971).
When a cross-striated fiber contracts, the maxi-
mum shortening is limited by the sarcomere traction; but the recent studies of Lanzavecchia
length, even if the thick myofilaments get through (1977), Lanzavecchia & Arcidiacono (1981) and
the Z material (supercontraction: Plate IV - b, d). Lanzavecchia et al. (1985) rather confirm the
The obliquely striated fibers have a greater capa- change of partner hypothesis between thick and
city of length changes than cross striated ones; in thin myofilaments (during elongation) introduced
addition to the classical sliding mechanism, by Miller (1975) to explain the length variation
Rosenbluth (1967, 1972) and Knapp & Mill range in these fibers. It is interesting to note that
(1971) (Fig. 5) suggested that the sliding would be such a length variation range is mainly found in
increased by a 'shearing' mechanism (sliding of invertebrates with soft bodies. In a previous paper
thick filaments on themselves) during con- about obliquely striated muscle in molluscs

b) Detail of the same muscle: sarcoplasmic reticulum (arrows). x 42000.

Fig. c. Male of Brachionus urceolaris sericus. The muscle is inserted (horizontal arrows) at the base of the striated ciliary roots
(R) which are associated to mitochondria (mi). The ciliary cell of the cingulum (cc) bears the cilia (c) responsible for the swimming
and is juxtaposed to the syncitial integument (int). The muscle is cross-striated, contracted, with short A-bands (1,3 Ilm),
intramyoplasmic sarcoplasmic reticulum (oblique arrows) and peripheral dyads (d). x26000.
Fig. d. Axial section of the relaxed three anterior sarcomeres of the cingulum muscle in T. rattus. Insertion on the basis of
the ciliary roots (small arrows). A-bands are 1.7 Ilm and the sarcoplasmic reticulum is abundant, particularly in the I band (large
arrows). x 16800.
272

Plate VII: Skeletal muscles of a bdelloid Philodina roseola

Fig. a, b, c. The central retractor muscle (CRM). a) Axial section showing myofilaments, dense bodies (d.b.), sarcoplasmic
reticulum (arrows), mitochondria (mi) and peripheral dyads (d). x 23000. b) Transversal section; the organisation of the
 273

(Amsellem & Nicaise, 1980), we noted that 'it is

puzzling that the most highly evolved molluscs'
Sepia and Octopus, (cephalopods) 'but not the
primitive ones, developed to that extent the use of
muscle cells which are mostly found in worms'.
Hence, this perhaps would not be a problem of
phylogeny and Lanzavecchia & Arcidiacono
(1981) considered this striation as an evolved
response to the problem of locomotion with
hydraulic systems.
An oblique striation occurs in the longitudinal
retractors of Phi/adina, where the movements of
shortening and elongation of the body seem to
overlap a variation range greater than that only
performed by the sliding mechanism in muscles.
The central retractor of Philodina inserts near the
cingulum (Fig. 6, Plate VII - b). Its contraction
provokes the withdrawal ofthe rotatory apparatus
deeply into the trunk of the animal. The dense
bodies (average length: 120 nm) are either
obliquely or transversely lined up (Plate VII -
a, c) and can be the sign of diversely contracted
myofibrils. A transverse section (Plate VII - b)
shows the typical arrangement of the myoplasm in
irregular obliquely-striated fibers. The sarcoplasm
lying between the arrays of myofilaments contains
glycogen like particles, mitochondria and sarco-
plasmic reticulum; peripheral dyads are abun-
dant. On some favorable longitudinal sections,
the thick myofilament length is around 1.3 jlm.
Fig. 6. Rotaria crurata. Skeletal muscles (modified from
Thus, this muscle seems to have the structural
Brakenhoff, 1937). CRM central retractor muscles; I to 7: characteristics of a fast muscle.
circular muscles; c: pedal muscle. Oblique striation occurs also in some retractors

contractile apparatus is visible. m: myofibrils; arrow: sarcoplasmic reticulum. x 21600. c) Axial section of the anterior part of
this CRM, showing its insertion (arrows) on the thin apical integument juxtaposed to the cingulum cells (cc). The myoplasm
is cylindrical around the axial sarcoplasm which contains the mitochondria (mi) xSOOO ..
Fig. d. Transversal section of a skeletal longitudinal muscle, located in the pseudocoel between the integument (int) and the
vitellarium (V); the myoplasmic lamina is organized in parallel myofibrils (m): this disposition is of intermediate organization
between the obliquely striated CRM (Fig. b) and the smooth circular muscles (Fig. e). Note the localization of the sarcoplasm
(s) along the myoplasmic lamina (m) and in the sarcoplasmic extensions which contain mitochondria (mi) and nucleus (N).
x 12.000.
Fig. e. Transversal section of a circular muscle (C) and axial section of a longitudinal muscle (L). The circular muscle is a
lamina, just under the integument (int) -skeletal intracytoplasmic lamina (sk) and a thin extracellular cuticle (cu). The myoplasm
of the circular lamina is not arranged in myofibrils. The cross-section aspect of thin and thick myofilaments is similar to that
found in monogononts. The dyads (D) are peripheral. The longitudinal muscle organization is similar to that seen in Fig. d). Dense
bodies (d.b.) are scattered in the myoplasm.
274

CD
CD

Plate VIII. Smooth and obliquely striated skeletal muscles of Monogononta

Fig. a. Axial section in one of the two ventro-buccal circular muscle in Asplanchna brightwelli. These two muscles are smooth:
note the irregular distribution of the dense Z bodies (vertical arrows) and SR (horizontal arrows) in this contracted muscle. The
dyads (d) are peripheral. x 15000.
 275

of monogononts, for example in male and female Philodina roseola, the circular muscles possess a
Brachionus, Rhinoglena (Plate VIII - g) and laminar myoplasm without intramyoplasmic sar-
Asplanchna (Plate VIII - 0. In Notommata copeus, coplasmic reticulum. This localization is periph-
large retractors look like smooth fibers in longi- eral with some dyadic contacts with the sar-
tudinal sections but aspects of the contractile colemma (Plate VII - e). The sarcoplasm is
apparatus in transverse section (Plate VIII - e) scarce and limited in the perinuclear zone with the
suggest that these fibers could be irregular obli- mitochondria (Plate VII - d). The dense bodies
quely-striated. are often ribbon-like, attached to the sarcolemma
and penetrating deeply in the fiber (Plate I - a).
The longitudinal retractors of Philodina roseola are
Smooth muscles more or less cylindrical fibers with a more structu-
rated contractile apparatus. Small rare mito-
In rotifers, we called 'smooth muscles' the muscle chondria are distributed along the myofilaments
cells with irregularly scattered dense bodies either and, in some fibers, dense bodies are relatively of
in transversal or longitudinal section (Plate VIII - small size (70 to 150 nm length) (Plate VII). They
-a, b). These cells possess two types of myo- can however reach more than 200 nm in others.
filaments with actin-like filaments attaching to the In this case, the thick myofilaments are more than
dense bodies. Sarcoplasmic reticulum is present, 3 J-lm long, but the micrographs did not allow
mainly peripheral, forming dyads with the sarco- more precise measurements (Plate II - 0. These
lemma. Intermediate organizations exist between differences are probably expressions offunctional
the typical obliquely striated type and the typical specializations, but we need more data about
smooth type depending of the state of con- behavior and structure.
traction. For instance, there is often a beginning Smooth fibers are observed in the long and big
of alignment of the dense bodies in super- central retractors of N. copeus (Plate VIII - e) as
contracted smooth fibers where the position of well as the short symmetrical anterior ventral cir-
thick myofilaments makes dark bands at the Z cular muscle in Asplanchna (Plate VIII - a). In
level, producing a pseudo striation (Plate VIII - c; some loricate monogononts as T. rattus, the
IV - d). smooth skeletal fibers are lacking: the only
Rotifer smooth fibers have features nearer to smooth muscles are visceral (ex: the cloacal
those of molluscan smooth fibers than those of sphincter).
coelenterates and are of different categories. In

Fig. b. Transversal section of a skeletal smooth muscle of Brachionus calyciflorus. Dense bodies (vertical arrows) and sections
of sarcoplasmic reticulum (horizontal arrows) are scattered in the myoplasm, where the myofilaments are not arranged in
myofibrils. x 28000.
Fig. c. Axial section of a contracted longitudinal muscle of the male of B. urceolaris sericus. Note the alignment of the dense
bodies and a zone of characteristic supercontraction where thick myofilaments go deeply into the neighboring sarcomeres
(arrows). x 30000.
Fig. d. Axial section of a relaxed longitudinal muscle of the male of B. urceolaris sericus. Note the insertion of the thin
myofilaments on dense bodies and the transversal (small arrows) or oblique (large arrows) orientation of dense Z bodies.
x 37000.
Fig. e. Transversal section of a CRM (central retractor muscle) of Notommata copeus. Note the cylindrical disposition of the
myoplasm around the central sarcoplasm; the myoplasm is nearly fragmented in radial myofibrils. This CRM is large because
N. copeus is a big rotifer (length = 1 mm). x 4500.
Fig. f Transversal section of a retractor muscle of Asplanchna brightwelli. The myoplasm is fragmented in radial distinct
myofibrils separated by sarcoplasm with sarcoplasmic reticulum (s.r.): this is a typical arrangement of oblique striated muscle.
mi: mitochondria; d.b.: dense Z bodies. x 43000.
Fig. g. Transversal section of a retractor muscle in the rostrum of Rhinoglena frontalis. The myoplasm displays also the typical
arrangement of an oblique striated muscle. x 14000.
276

Conclusions

Skeletal muscles in rotifers are very diverse in

form, type and function. They can be classified by
different cytological criteria as: smooth, obliquely
or cross-striated; phasic and tonic; strong or not;
endurant or not.
The precise localization of their insertions are
important data to understand their functions
which can be grouped in three behaviors: with-
drawal of the corona, control of ciliary beat in the
corona; articulated movements (foot, tail,
paddles).
Their coupling by gap-junctions and their inner-
vations allow definition of motor-units responsi-
ble of these behaviors. Each cingulum muscle is
a motor-unit which controls precisely swimming
behavior. The different kinds of phasic and tonic
retractors and the anterior circular muscles con-
stitute two motor-units. One is involved in the
quick withdrawal behavior and the other with the
complete and prolonged retraction of the corona.
All of the pedal muscles in T. rattus are a single
motor-unit which controls the movement of the •• mrrip
foot and secretions of the pedal gland.
Each muscle is either monocellular or bicellular
and functionally adapted to a precise partici-
pation in the behavior. Some specializations are
extreme as in the cingUlum muscle controling
swimming in Asplanchna (0.5 J.lm A bands).
The bdelloid Philodina roseola does not possess
typical cross-striated skeletal muscles. All are
smooth or obliquely striated muscles without
obvious phasic or tonic characteristics. Smooth
and obliquely striated skeletal muscles are also
present in monogononts, principally in those
where the skeletal integument is soft (the 'illori- Fig. 7. Brachionus. Skeletal muscles (modified from
cates'). They are present in N. copeus, but absent Stossberg, 1932) dvl to dv5 = circular muscles; mrls:
cingulum muscles; mrd: dorsal retractor muscles; mrc:
in T. rattus and more abundant in Rhinoglena than central retractor muscles; mrdp: pedal retractor muscle.
in Brachionus.
The most primitive muscles seem to be those
with a homogeneous laminar myoplasm. This ments, structurally similar to the myosin and actin
lamina could evolve by folding on itself to become of the fast fibers of arthropods and vertebrates.
a cylinder and by fragmentation of adjacent myo- Thick myoftlaments never show the structural
fibrils. aspect of p aramyo sin. Dense Z bodies are present
Rotifer skeletal muscles originate from ecto- even in smooth muscles. Some smooth muscles of
derm. They all possess thick and thin myoftla- Cnidaria (Perkins et al., 1971; Wood, 1961) and
 277

of Ctenaria (Hernandez-Nicaise & Amsellem, Chapman, R. A., C. F. A. Pantin & E. A. Robson, 1962.
1980) have thick and thin myofilaments, but no Muscle in Coelenterates. Rev. Canad. BioI., 21: 267-278.
Chien, P. & H. Koopowitz, 1972. The ultrastructure of
dense body.
neuromuscular systems in Notoplana acticola, a free-living
Platyhelminth muscles (Rees, 1975; Hoyle, polyclad flatworm. Z. Zellforsh., 133: 277-288.
1983) show some features and a diversity which Clement, P., 1969. Premieres observations sur l'ultrastruc-
can be compared with those of rotifers. Never- ture comparee des teguments de rotiferes. Vie et Milieu A,
theless, they contain paramyosin as in nematodes, 20: 461-482.
Clement, P. & X. Fouillet, 1970. Les jonctions cellulaires
annelids and other groups of worms.
d'un organisme (rotifere). 7eme Congo Intern. Micr. Electr.,
These observations are consistent with other Grenoble, 1: 7-8.
ultrastructural data, from which Clement (1981) Clement, P., 1977. Ultrastructural research on rotifers. Arch.
suggested that rotifers share a recent common Hydrobiol. Beih. Ergebn. Limnol., 8: 270.
ancestor with platyhelminths. The overall features Clement, P., 1980. Phylogenetic relationships ofrotifers and
derived from photoreceptor morphology and other ultra-
of rotifer musculature illustrate that they con- structural analysis. Hydrobiologia, 73: 93-117.
stitute a relative homogeneous group. With regard Clement, P., 1985. The relationships of rotifers, as deduced
to muscle characteristics, bdelloids seem to be from their ultrastructure and behavior. In: The origin and
more primitive than monogononts. Yet even in relationships oflower Metazoa. (eds Conway Morris et al. )
bdelloids, the observed diversity of speciali- 224-247. Clarendon Press, Oxford.
Clement, P., 1987. Movements in rotifers: correlations of
zations is related to precise functions and ultrastructure and behavior. Hydrobiologia, 147: 339-359.
behaviors. Clement, P. & J. Amsellem, 1986. Ultrastructures et com-
portements: neuro-ethologie des rotiferes. In: Neuro-
ethologie (ed. R. Campan). Comportements. 39-57.
Acknowledgements CNRS. Paris.
Clement, P., E. Wurdak & J. Amsellem, 1983. Behavior and
ultrastructure of sensory organs in rotifers. Hydrobiologia,
We acknowledge the researchers of the 104: 89-130.
Neuroethology team who worked in electron Duvert, M., 1971. Ultrastructure de lajonction myo-epider-
microscopy: E. Wurdak on Asplanchna bright- mique dans les muscles du tronc de Sagitta cetosa
welli; A. Cornillac on Brachionus calyciflorus; (Chaetognates). c.R. Acad. Sci., Paris, Ser. D, 272:
2575-2577.
Anne Luciani on Brachionus plicatilis. All the elec- Hardie, J. & c. Hawes, 1982. The three dimensional struc-
tron microscopy was done at the C.M.E.A.B.G., ture of the Z-disc in Insect supercontracting muscle.
Universite C. Bernard, Lyon I. We also ack- Tissue and Cell, 14(2): 219-231.
nowledge Terry Snell who read this manuscript to Hernandez-Nicaise, M. L. & J. Amsellem, 1980. Ultra-
correct its English. structure of the giant smooth muscle fiber of the Cteno-
phore Beroe ovata. J. Ultrastruct. Res., 72: 151-168.
Hyman, L. H., 1951. Class Rotifera. In: The Invertebrates.
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C. Ricci, T. W. Snell and C. E. King (eds), Rotiler Symposium V. 279
© 1989 Kluwer Academic Publishers.

Classical taxonomy and modern methodology

W. Koste l & RJ. ShieF

I Ludwig-Brill-Strasse 5, Quakenbriick, D-4570, F.R.G.; 2 Murray-Darling Freshwater Research Centre,
P.O. Box 921 Albury, N.S. W. 2640, Australia

Key words,' Rotifera, species definition, taxonomy, biogeography, nomenclature, ecotypes, polymorphism

Abstract

Classical rotifer taxonomy and recent approaches are reviewed. (X-taxonomy (morphology) remains the
most widely-used technique, and most prone to subjective errors. Computer technology, and genetic or
biochemical methods are not widely available in areas of most need - those with developing limnology
programs. Strict adherence to the International Code of Zoological Nomenclature, uniform treatment of
ecotypic variation and polymorphism, communication with and between systematists, and establishment
of a centralized data base are among the points discussed. A standardized nomenclature is proposed.

Introduction and B. lyratus, both Australian endemics, in the

absence of evidence to the contrary, had been
The credibility of ecological, genetic, biochemical synonymised with B.falcatus and B. angularls
and other studies relies heavily on the correct respectively (Ahlstrom, 1940; Voigt, 1957; Koste,
identification of the organisms involved. The need 1978). To resolve such synonymy on a global
for a review of the taxonomic bases ofthe Rotifera basis, in the manner of Harring's (1913)
has become increasingly urgent as a new genera- 'Synopsis' will be a daunting task.
tion of rotifer workers, particularly in areas with In the second instance, a distinct taxon may be
developing limnology programs, fmd that their masked by the range of morphological variation
taxa are not always in accord with the available (seasonal di- or polymorphism) in a co-occurring
(largely European or North American) keys. congener. Several morphs of the polytypic
Established workers, on the other hand, are fmd- Formenkreis Keratella cochlearls discussed by
ing distinct, separable morphotypes within popu- Ruttner-Kolisko (1974) are now considered valid
lations previously considered to be single, albeit species, e.g. K. tecta (cf. Ridder, 1973; Hofmann,
cyclomorphic, species. 1983; Koste & Poltz, 1984). A similar situation is
In the first instance, an indeterminate number evident in the Canadian Keratella cochlearls popu-
of good species undoubtedly has been synony- lations described by Chengalath & Koste (1987),
mised with the recognized taxon which they most and Polyarthra populations from Mali (Koste &
closely resemble, either to conform to an authora- Tobias, 1989).
titive taxonomic reference work, or by a reviewer Many species have one or more subspecies,
faced with inadequate descriptions and figures, or varieties, forms, or morphotypes. In only a few
no type material. Several of these were encoun- cases is this morphological variation attributable
tered in our review of the Australian Brachionidae to environmental variables, e.g. the responses of
(Koste & Shiel, 1987), e.g. Brachionus dichotomus brachionids and Asplanchna to predatory
280

Asplanchna (cf. Gilbert & Stemberger, 1984). evolving separately from others and with its
Comparable responses to predators or other own unitary evolutionary role and tendencies'.
environmental variables are poorly known for
most genera. In this context, the criterion of the species is the
Resolution of erroneously synonymised or genetic isolation of populations.
'hidden' taxa as noted above will add appreciably This definition encompasses the great diversity
to the rotifer record, and undoubtedly add demonstrated within apparently contiguous pop-
strength to the increasing evidence for non- ulations (e.g. King & Zhao, 1987) as well as
cosmopolitanism (cf. Dumont, 1983). Conversely, phenotypic plasticity evident in some taxa (cf.
a simultaneous intensive effort is required to deter- King, 1977; Pejler, 1980). It does, however, raise
mine the identity of species described from single the problem of the status of geographically iso-
individuals, aberrant forms, ecotypes, and inter lated populations, such as the Brachionus plicatilis
alia, preservation artefacts (see Shiel & Koste, of Soda Lake, Nevada described by King &
1985), which have been named without the Zhao, and those of western Victoria described by
slightest information on individual species' varia- Walker (1981). These populations clearly have
bility. N arning of the slightest variation from the evolved in isolation from each other, to the extent
'norm' (indeterminate!) has only promoted con- where a recognizable morph, presently identified
fusion. as the ssp. colongulaciensis Koste & Shiel occurs
In the following discussion we reiterate points in Victoria. If the evolutionary species concept is
made 25 years ago, when Ruttner-Kolisko (1963) followed to the letter, these two populations
summarised the inter-relationships of the should be accorded specific status. Comparative
Rotifera, and suggested a set of taxonomic guide- genome analysis (e.g. King, this volume) is neces-
lines. Had these been consistently followed much sary to determine the extent of interpopulation
of the present systematic uncertainty within the genetic variation, then (given that there is already
group would have been obviated, as would the lax considerable intrapopulation or clonal variation),
approach to rotifer systematics in some investi- the acceptable degree of genetic dissimilarity must
gations, noted by Dumont (1980). Attention also be defined before specific status is accorded. The
is drawn to points raised in Ruttner-Kolisko intraspecific morphological variability in B. plica-
(1974), Pejler (1977) and taxonomic/biogeogra- tilis discussed by Sudzuki (1987) may reflect such
phic papers in previous Symposium proceedings dissimilarity. Resolution, on a global scale,
(King, 1977; Dumont & Green, 1980; Pejler et af., appears to be a long-term prospect!
1983 & May et al., 1987). Because neither the expertise nor the facilities
for such genetic analyses, or those involving
biochemical specificity (e.g. Snell, this volume)
The species concept are readily available to the 'average' rotifer tax-
onomist, there is a continuing dependence on
The predominance of parthenogenetic repro- comparative morphology. Before considering
duction in rotifers renders the biological concept some of the bases of a-taxonomy, it is appropriate
of species unacceptable (i.e. groups ofinterbreeding to mention current nomenclatural considerations.
natural populations reproductively isolated from
other such groups) (see Ruttner-Kolisko, 1974, and
for bdelloid rotifers, Ricci, 1987). The evolu- The Code
tionary species of Simpson (1961) remains valid
for unisexual (bdelloid) and bisexual (Mono- The International Code of Zoological Nomencla-
gonont) rotifers: ture was established 'to promote stability and
universality in the scientific names of animals and
'An evolutionary species is a lineage (an to ensure that the name of each taxon is unique
ancestral-descendant sequence of populations) and distinct' (Ride et al., 1985: 3). To ensure uni-
 281

formity, the conventiom of the Code ~hould be (Ehrenberg), dorcas (Go~~e), spinosa (Beau-
observed, including conformity to Latin form, and champ), monstruosa (Ridder), inter alia, with inter-
the rules of Latin grammar (J effrey, 1977). Rotifer grades between some of these. Two or more of
taxonomy has not conformed consistently to the these morphs often may occur together, i.e., spine
requirements of the Code, particularly in ade- development shows intrapopulation variation,
quacy of description and/or illustration, nomi- and ssp. status clearly is inappropriate. Sub-
nation and location of type material, and con- species, as defined by Mayr (1973), are geographi-
current description of nearest related taxa with cally dermed aggregates of local populations
designation of features differentiating a new which differ taxonomically from other such sub-
taxon. Even basic errors of gender have passed divisions of the species.
referees and editors through several generations Such morphs were termed the 'calyciflorus
of major revisions, (e.g. Brachionus bidentatus is group' by Ruttner-Kolisko (1972, 1974), while
intermittently cited as B. bidentata, depending on Koste (1978), in the context of an 'Artgruppe',
the revision available). used subspecies, varietas and forma, which en-
It is beyond the scope of this paper to detail compassed morphotypic variations in sympatric
correct nomenclatural orthography, however populations. For such variable, closely-related
'rotiferologists' should be aware that incorrect ter- sympatric (sometimes allopatric) monophyletic
minations, generic and/or specific names, figures, groups, the term superspecies was applied by
authors or citations are to be found in the rotifer Mayr (1931). This is more precise than the cate-
literature. If these are not to be perpetuated, some gory 'Artgruppe' (species group) in Koste (1978).
degree of careful research is necessary when Other superspecies exhibiting wide variation
transcribing from original descriptions or previ- within populations include Anuraeopsis jissa,
ous works. Correct procedures are detailed in Asplanchna sieboldi, Brachionus angularis, B. bi-
Mayr (1973) and Jeffrey (1977), and if in doubt, dentatus, B. quadridentatus s.l., Keratella cochlea-
the Code should be consulted (Ride et aI., 1985). ris, K. slacki, Lepadella acuminata, Notholca
squamula, to name a few!
To communicate information on morphotypes
Nomenclature of a superspecies which may occur in the same
population, a more detailed nomenclature, such
The proliferation of rotifer names includes a large as that used by algologists, is desirable, e.g.:
number oftrinomina, and occasionally, especially
in the bdelloids (cf. Ricci 1987), quadrinomina. B. calyciflorus Pallas, 1766 (f. anuraeiformis
Although quadrinomina are admissable under the after Brehm, 1909)
Botanical Code, they are not under the Zoological
Code, and only one category below species (sub- uses parentheses to designate an infrasubspecific
species) is recommended under the present Code taxon, which has no systematic status per se, but
(Article 45e). In the past, trinomens have been designates which morphotype is involved. 'Form'
subspecies or infrasubspecific descriptors, i.e. in this context is ecotypic, seasonal or sexual
forma or varietas (var.). The Code recommends variation, and conforms to the recommendations
that they should take sub specific rank (with of the Code. In other than superspecies, more
exceptions, which are detailed in the Code) (Ride stringent applications of the rules of binomial
et al. loc. cit.). nomenclature are required than has been the case
This is the nomenclature used by Kutikova in the past. New names should be avoided until
(1970), however difficulties immediately become the most complete possible information on intra-
evident when considering ecotypic variation such and interpopulation variation for rotifer taxa is
as shown by Brachionus calyciflorus Pallas. This available. We believe a more detailed nomencla-
polymorphic species has variants which have ture is necessary for both ecological studies and
been named: anuraeiformis (Brehm), amphiceros better communication between rotifer workers.
282

(X-taxonomy

The description of new species of Rotifera, and

their organization into genera and higher cate-
gories often has been based on morphological
features which are at best variable, and at worst,
useless as taxonomic identifiers. In loricate taxa,
e.g. Keratella and Brachionus, lorica facettation
appears to be species-specific in some taxa, e.g.
K. cochlearis but variable in others, e.g.
B. angularis. Number and placement of anterior
lorica spines appears to be characteristic for each
species, but there is considerable variation in their
size, also of the ventrallorica margin. Similarly,
caudal spines may be highly variable or absent (cf.
Fig. 1, K. slacki from Tasmania), and of no tax-
onomic value.
Illoricate taxa present even more difficulties.
Size and shape of most species is variable, with
overlap between congeners. Comparative mor-
phology of appendages such as bristles (Filinia)
and fins or paddles (Polyarthra), and location of
lateral antennae (Testudinella), have been used
diagnostically with some success, although there
may be inter- and intrapopulation variation. Mor-
phologically similar congeneric species may
cooccur and not be identified. A more detailed
account of the difficulties in each of the mono-
gonont genera is included in Ruttner-Kolisko
(1974) and Koste (1978), and for the bdelloids, in
Donner (1965).
Trophi structure has been used successfully to Fig. 1. Ecotypic variations in lorica morphometry, Keratella
separate congeners; Guiset (1977) separated slacki (Berzins) from Tasmania, spring 1987 [see Shiel et al.
several Polyarthra species on the basis of fulcri this volume]. Scale bar 100 jlm.
length. Synchaeta species were differentiated by
Sternberger (1979). Filinia hofmanni and difficulty under oil immersion. Scanning electron
F. australiensis were distinguished from congeners microscopy (SEM) provides the necessary resolu-
by Koste (1980), and more recently, trophi struc- tion, however until recently had been used suc-
ture separated species of Polyarthra in west Afri- cessfully only with the larger trophi of Asplanchna
can waters (Koste & Tobias, 1989). Trophi struc- (e.g. Salt et al., 1978). Asplanchna trophi may
ture has been inadequately studied for many taxa, reach 340 Jlm (Koste & Shiel, 1980), are flat,
yet appears to be species-specific, and therefore a simple in structure, and can be treated for SEM
valuable taxonomic discriminator. examination reasonably easily. To treat the
The small size of most trophi has been a deter- smaller trophi of other genera requires extreme
rent to intensive study. The trophi in most taxa are care in handling (see Markevich & Koreneva,
< 100 Jlm long, in some species, e.g. Encentrum, 1981). Ideally, detailed trophi structure determi-
< 20 Jlm. These tiny structures are at the limit of nation for each species is necessary to clarify the
light microscopy, and can be resolved only with present systematic confusion. The recent work of
 283

Markevich (1985 and this volume) provides the pat ability problems between various systems have
resolution of the problem of 'lumping' morpho- yet to be resolved.
logically similar but nonetheless distinct species Computer analytical techniques have not yet
(Dumont, 1983: 21), and enables more precise become widely available in rotifer taxonomy,
diagnosis of higher categories. although their validity has been established for
field population studies. Available analytical pro-
grams permit rapid handling of amounts of data
Modern methodology which previously have been prohibitive; a number
of programs are applicable to species morphome-
Technological advances in optics, computing, tric analysis. An example of these methods
analytical techniques, inter alia, have been in evi- applied to rotifer community data is found in
dence in each successive symposium volume. We Nogrady (1988). Details of methodology are given
see these as valuable adjuncts to morphological in Romesburg (1984).
taxonomy which will continue to enhance rather Examples of biochemical and molecular tech-
than replace traditional methodology. Improved niques which represent future directions from
methods have played a significant role in the which taxonomy must benefit are given by Snell
renaissance of systematics in this generation (cf. (this volume) and King (this volume).
Mayr, 1973), permitting the finer resolution or It is clear from the range of papers presented at
information processing capacity previously lack- this symposium that the expertise exists to resolve
ing. SEM methods, for example, will undoubtedly the taxonomic uncertainties of the Rotifera, but it
clarify the systematic problems; the recent work is widely dispersed. It cannot be expected that
of Kutikova & Markevitch (this volume) repre- those with the expertise and facilities to run gels,
sents a major impetus to a revision of current extract DNA, operate SEM, TEM and computer
rotifer taxonomy. SEM is not widely available analytical systems can do so on a global basis.
however, and its use is in practice restricted to Comparative morphology must, therefore, remain
research institutions. Similarly, image-analysis the principal tool of taxonomic workers while the
systems enable rapid morphometric compu- specialized techniques become more widely dis-
tations, with image and data storage on disk facili- seminated. We are hopeful that the global coope-
tating exchange of information between workers, ration which results from the rotifer meetings, will
but again are not widely available. include more communication between widely-dis-
More accessible for image/data storage and persed taxonomists. To this end we would like to
exchange are increasingly inexpensive video sys- see a centralised data base established which ulti-
tems. The resolution obtainable by video record- mately could include morphological and ecologi-
ing is approaching that of a microscope-mounted cal information for all rotifer taxa. Such a data
35 mm camera; we have used a high resolution base would extend beyond the lifetime of individ-
National F-lO system in ecological studies on uals contributing to it; housing in a research insti-
rotiferJbacteria and rotiferJalgae interactions (e.g. tution would be desirable, with provision for con-
Tetrasiphon hydrocora grazing on Staurastrum) in tinued contributions. We consider the benefits of
Murray-Darling waters, but see a wider use as a centralized records to far outweigh the costs of
potential means of communication between setting up a workable system.
rotifer workers. For a taxonomist located far from
source material (which generally arrives preserved
and less than life-like), a recording of the organ-
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ficulties encountered with preservation artefacts,
Ahlstrom, E. H., 1940. A revision of the Rotatorian genera
provide behavioural, ecological information and Brachionus and Platyias with descriptions of one new
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Although video is more widely available, com- 148-184.
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Chengalath, R. & W. Koste, 1987. Rotifera from North- May, L., R. Wallace & A. Herzig, 1987. Rotifer Symposium
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Donner, J., 1965. Ordnung Bdelloidea. Bestimmungsb. posium. Hydrobiologia 147: 1-381.
Bodenf. Eur. 6: 1-297. Mayr, E., 1931. Birds collected during the Whitney South
Dumont, H. J., 1980. Workshop on taxonomy and biogeo- Sea expedition. XII. Notes on Halcyon chloris and some
graphy. Hydrobiologia 73: 205-206. of its subspecies. Am. Mus. Nov. 469: 1-10.
Dumont, H. J., 1983. Biogeography ofrotifers. Hydrobiologia Mayr, E., 1967. Artbegriff und Evolution. Paul Parey,
104: 19-30. Hamburg.: 1-617.
Dumont, H. J. & J. Green, 1980. Rotatoria. Proceedings of Mayr, E., 1973. Priciples of Systematic Zoology.
the 2nd International Rotifer Symposium. Hydrobiologia McGraw-Hill, New Delhi.: 1-428.
73: 1-263. Nogrady, T., 1988. The littoral rotifer plankton of the Bay of
Gilbert, J. J. & R. S. Sternberger, 1984. Asplanchna-induced Quinte (Lake Ontario) and its horizontal distribution as
polymorphism in the rotifer Keratella slacki. Limnol. indicators of trophy. Arch. Hydrobiol. Suppl. 79: 145-165.
Oceanogr. 29: 1309-1316. Pejler, B., 1977. On the global distribution of the family
Guiset, A., 1977. Some data on variation in three planktonic Brachionidae (Rotatoria). Arch. Hydrobiol. Suppl. 53:
genera. Arch. Hydrobiol. Beih. 8: 237-239. 255-306.
Harring, H. K., 1913. Synopsis of the Rotatoria. Bull. am. Pejler, B., P. Starkweather & T. Nogrady, 1983. Biology of
Nat. Hist. Mus. 81: 1-226. Rotifers. Proceedings of the 3rd International Rotifer sym-
Hofmann, W., 1983. On temporal variation in the rotifer posium. Hydrobiologia 104: 1-396.
Keratella cochlearis (Gosse): the question of ' Lauterborn- Ricci, c., 1987. Ecology of bdelloids: how to be successful.
cycles'. Hydrobiologia 101: 247-254. Hydrobiologia 147: 117-127.
Jeffrey, c., 1977. Biological Nomenclature. 2nd edit. E. Ridder, M. De, 1973. Atlas provisoire de Rotifert':s de
Arnold, London: 1-70. Belgique. Fac. Sci. Agron., Gembloux.
King, C. E., 1977. Proceedings of the First International Ride, W. D., C. W. Sabrosky, G. Bernardi & R. V. Melville
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King, C. E., 1989. Molecular genetics of rotifers: preliminary ture. # 3rd edit. Univ. of Calif. Press, Berkeley.: 1-321.
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King, C. E. & Y. Zhao, 1987. Coexistence of rotifer Ruttner-Kolisko, A., 1963. The inter-relationships of the
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Hydrobiologia 147: 57-64. Calif. Press.: 263-272.
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Osnabrucker Naturwiss. Mitt. 11: 91-125. morphology relative to food habits in six species of rotifers
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characteristics of the rotifer fauna of Australia (Notogaea). Shiel, R. J. & W. Koste, 1985. New species and new records
Hydrobiologia 73: 221-227. of Rotifera (Aschelminthes) from Australia. Trans. r. Soc.
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Rotatoria). Senckenbergiana biologic a 69 (4/6) (in press). logia, this volume.
Kutikova, L. A., 1970. Rotifer fauna of the U.S.S.R. Subclass Sternberger, R. S., 1979. A guide to the rotifers of the
Eurotatoria. Fauna CCCP 104: 1-744. Laurentian Great Lakes. E.P.A., Ohio.: 1-186.
Kutikova, L. A. & G. I. Markevich. 1989. The basic prin- Sudzuki, M., 1987. Intraspecific variability of Brachionus
ciples of constructing a system of rotifers. Hydrobiologia, plicatilis. Hydrobiologia 147: 45-47.
this volume. Voigt, M., 1957. Rotatoria. Die Radertiere Mitteleuropas.
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evolution of rotifers: Jaws. IN Proceedings of the 2nd Walker, K. F., 1981. A synopsis of ecological information on
All-Union symposium on rotifers, Leningrad.: 17-37. the saline lake rotifer Brachionus plicatilis Muller, 1786.
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Hydrobiologia 186/187: 285-289, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Roti/er Symposium V. 285
© 1989 Kluwer Academic Publishers.

Mastax morphology under SEM and its usefulness in reconstructing

rotifer phylogeny & systematics

G.!. Markevich 1 & L.A. Kutikova

1 The Institute for the Biology Inland Waters, USSR Academy of Sciences, Borok, 152742, USSR; The
Zoological Institute, USSR Academy of Sciences, Leningrad, 199034, USSR

Abstract

Scanning electron micrographs have been prepared of mastaxes of more than 120 species of rotifers. The
basic directions of the evolutionary transformation of mastax sclerites and their significance for recon-
structing rotifer phylogeny are illustrated.

Introduction What are the characters that are best for clas-
sifying rotifers and revealing their phylogeny?
Dissatisfaction with the current taxonomy of Kutikova (1970, 1983) suggested that these
rotifers is growing since it is constructed as should be characteristics of the structure and
phenetic classification based solely upon the typo- functioning of the corona and mastax (locomo-
logical species concept (Dumont, 1980; Kutikova, tory-trophic approach). Other rotiferologists,
1983). So far, it has been difficult to apply the however, do not expect to find answers about
techniques of phylogenetic systematics to rotifers. rotifer evolution by using conventional morpho-
This is despite the advantages of this approach for logical analysis of females. Rather, they pin their
describing the evolutionary history of this group. hopes on male morphology (Sudzuki, 1964, 1977)
Analysis of the prevailing classification systems or on tissue ultrastructural organization (Clement,
for rotifers suggests that, in spite of some dif- 1980). Previous efforts to reconstruct rotifer
ferences, they have much in common. For exam- phylogeny with morphological methods have not
ple, the separation of rotifer classes by gonad succeeded because of the insufficient resolving
number is used by both de Beauchamp (1907) and power of optical microscopy. In principle, mor-
Remane (1929-1933). The main groups ofrotifers phological analysis is sufficient to describe phylo-
on the order level are actually identical in both genetic relationships within this phylum. Our
classification systems. The main disagreements work uses the greater resolution of scanning elec-
among these taxonomies result not from the clas- tron microscopy to examine the mastaxes of
sification of rotifers, but from different interpreta- several rotifer species and employs this informa-
tions of their evolution due to different assump- tion to describe a phylogeny.
tions about ancestral groups. Previous attempts
to reconstruct the phylogeny of rotifers on the
basis of comparative, anatomical and morpho- Results
logical data have lead to contradictory con-
clusions even by such eminent zoologists as de Methods for preparing inner sclerite parts of the
Beauchamp and Remane. mastax for scanning electron microscopy (SEM)
286

Fig. 1. Sclerite systems of mastaxes of some rotifers and details of their structure. a - Rotaria tardigrada; b,h,i - Conochiloides
natans; c - Brachionus calyciflorus; d - Notommata allantois; e - Trichocerca sulcata; f - Asplanchna herricki; g - Floscularia decora;
j - Epiphanes brachionus; d - Euchlanis dilatata; 1 - Dicranophorus Jorcipatus. -a-d,f,l- sclerite systems; e,g-i - rami; j - unci; k
 287

- manubrium. - al- alula; apc - apophysis cardalis; bf - basifenestra; ca - camera anterior; cap - camera apicalis; cb - camera
basalis; cdm - cauda manubrialis; cm - camera medialis; cn - constrictor; cp - camera posterior; cr - crista; csb - camera
subbasalis; fn - fenestrula; fu - fulcrum; iu - impressio uncinalis; Ih -lamina hilaris; lu -lamina uncinalis; m - manubrium;
mu - metamerae unci; pu - praeuncus; r - ramus; rs - rostellum; sa - sutura anterior; sbf - subbasifenestra; se - sutura externa;
sl- satelliti; sp - scleropili; sju - sutura jugalis; tr - trabecula. Scale bars, marked by one quadrant - 1 mkm; by two - 10 mkm;
by three - 100 mkm.
288

B c
Fig. 2. The main directions of evolutionary transformation of sc1erites ofthe mastax. A) Uncus: 1 - a polymeric row of separated
metameres; 2 - a consolidated row of metameres with different derivatives of the uncus row (the latter is shaded); 3 - modes
offormation ofuniapical uncus - contraction, confluence, abduction; 4 - mono-elementary uncus. B) Manubrium: 1 - navicular
form ofthe lamina; 2 - growth oflamina and formation of chambers; 3 - formation ofthe cauda by the medial chamber. C) Rami:
1 - are separated lamellar; 2 - growth of ramus trabecules and formation of sc1eropilar fulcrum.

are available (Markevich & Koreneva, 1981 ; trabecula, fenestrula, basifenestra, subbasifenes-
Markevich, 1984). These allowed Markevich tra, constrictor; camera: apicalis, posterior,
(1987a, b) to successfully apply morpho-func- medialis, anterior; and lamina: hilaris, uncinalis
tional analysis to the evolution of the mastax. A (Markevich, 1989, Fig. 7).
rotifer phylogeny based on these data favors a Comparative analysis of mastax fine structure
close phylogenetic relationship between bdelloid suggests that the reduction in mastax elements
rotifers and cercomer worms (Monogenea). The common in Eurotatoria are derived from an
SEM method was used to study the sclerite com- ancestral polymeric sclerite system, present only
plexes of the mastax in 120 species of freshwater in the Bdelloida (Markevich, 1983, 1985a&b,
rotifers representing practically all morpho- 1987b). These transformations of the mastax can
functional types (Markevich, 1983, 1985a & b, be accomplished by changes in single elements
1987b). The detailed study of sclerites of the rather than a fundamental change in mastax
mastax has revealed the following structures organization.
which were previously unknown: sutura: exterior, The following basic trends of morphological
jugalis, anterior; rostellum, crista, scleropilli, transformations of jaws of the mastax have been
 289

revealed: oligomerization of rows of uncus References

metamers, 'ramatization' of the sclerite system,
i.e. a progressive transfer of functional load from Beauchamp, P. de, 1907. Morphologie et variations de
unci to rami, intersclerite integration of elements l'appareil rotateur dans la serie des Rotiferes. Arch. Zool.
exptl. et gen. 4,6: 1-29.
in the mastax, and development of close spatial Beauchamp, P. de, 1965. Classe des Rotiferes. Traite de
and functional relations between the mastax and Zoologie, Anatomie, Systematique, Biologie IV, 3:
corona (intra- and intermodule integration) 1225-1379.
(Fig. 2). The latter is accompanied by a shift ofthe Clement, P., 1980. Phylogenetic relationships of rotifers, as
mastax closer to the mouth, turning on its axis in derived from photoreceptor morphology and other ultra-
structural analyses. Hydrobiologia 73: 93-117.
the pharynx and other complex allometric trans- Kutikova, L. A., 1970. Rotifers of the USSR. Leningrad.
formations. Combinations of in tram as tax charac- 744 pp. {In Russian).
ters provides significant information about trends Kutikova, L. A., 1983. Pecularities of diagnostics of taxa in
in sclerite system transformation and is useful in rotifers. In: The Rotifers. Proceedings of the Second All-
rotifer systematics in general. Union Symposium on Rotifers. Leningrad, pp. 4-17. (In
Russian).
Markevich, G.I., 1984. Ultrastructure of chitionoid struc-
tures of the coracidia Triaenophorus nodulogus (Pallas,
Conclusions 1781). DAN 278, 4: 1022-1024. (In Russian).
Markevich, G.I., 1985a. The main trends of idioadaptive
On the basis of our analysis, the phylogeny of evolution ofrotifers. Jaws. In: Proceedings of the Second
All-Union Symposium on Rotifers. Leningrad 17-37. (In
rotifers has been revised. We suggest that the Russian).
Bdelloidea be placed in a new subclass called the Markevich, G.I., 1985b. Ultrathin morphology of mast ax of
Archeorotatoria. We also suggest that the Ploima rotifers.1. Bdelloida. Bulletin of the Institute of the Biology
be divided into four new orders: Trans- Inland Waters 68: 31-35. {In Russian).
versiramida, Saltiramida, Saeptiramida, Antror- Markevich, G. I., 1987a. The main trends of idioadaptive
evolution in rotifers, Locomotion and locomotory
siramida. behavior. In: Fauna and biology of freshwater organisms.
Therefore, the fIrst results of a comparative Leningrad. Proceedings of the Institute of the Biology
SEM morphological study of the mastax support Inland Waters: 54(57): 175-190. (In Russian).
the usefulness of morphological analysis and Markevich, G.I., 1987b. Functional morphology of the jaw
Remane's criteria (1956). The morphological apparatus in rotifers. Synopsis of a thesis for the degree of
candidate of biological sciences. Leningrad. 23 pp. (In
approach to rotifer phylogeny based on SEM of Russian).
mastax fIne structure of the complex sclerite Markevich, G.I., 1989. (in press). Morphology and the prin-
muscular apparatus appears to be sufficient for ciple organization of sc1erite system of the mastax in
reconstruction of a rotifer phylogeny. A successful rotifers. In: Biologia and functional morphology of fresh-
water animal. Leningrad. Proceedings of the Institute of
application of SEM methods to morphological
the Biology Inland Waters (In Russian).
analysis in a study of rotifer polygeny suggests Markevich, G.1. & E. A. Koreneva, 1981. On the technique
that further analysis of their locomotory and tro- of preparing of the mastax of rotifers for electron micro-
phic systems (corona, mastax) is likely to yield scopy. Zool. zhurn. 60(10): 1562-1564. (In Russian).
interesting insights into rotifer evolution. Remane, A., 1929-33. Rotatorien. Dr. H. G. Bronn's Klassen
and Ordnungen d.Tierreichs, Leipzig IV,II,I,I-4: 576 S.
Remane, A., 1956. Die Grundlagen des naturlichen Systems
der vergleichenden Anatomie une der Phylogenetik.
Leipzig. 364 S.
Sudzuki, M., 1964. New Systematical Approach to the
Japanese Planktonic Rotatoria. Hydrobiol. Acta Hydro-
bioI., H ydrogr. et Protist. 23: 1-124.
Sudzuki, M., 1977. Classification based on the male. Arch.
Hydrobiol. Beih. Ergebn. Limnol. 8: 221.
Hydrobiologia 186/187: 291-298, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 291
© 1989 Kluwer Academic Publishers.

Problems in taxonomy of rotifers, exemplified by the Filinia longiseta -

terminalis complex

Agnes Ruttner-Kolisko
Biological Station of the Austrian Academy of Science

Key words: rotifers, taxonomy, variation, genetics, parthenogenesis, resting eggs, ecology

Abstract

The validity of taxonomical categories in parthenogenic groups is discussed. Special problems of rotifer
taxonomy are caused by: facultative or obligatory parthenogenic reproduction, high morphological and
genetic variability and paucity of morphological characteristics.
Examples of these problems are provided by the Filinia terminalis-longiseta group. The different
ecological properties of various populations belonging to this group are emphasized. Suggestions concern-
ing the creation of new taxa are made; in particular, the importance of using ecological data is stressed.

Taxonomy is an artificial grid, imposed by chy of the animal and plant kingdoms, we would
scientists on the continuous flow of adaptations, be hopelessly lost in a bewildering diversity whose
variations and mutations, which is characteristic direction of flow we understand poorly or not at
of nature and results in evolution. There is only all. But it must be kept strictly in mind that our
one point where this artificial grid has a biological taxonomical grid - whether based on morphology,
meaning; that is the concept of species, which is physiology, genetics or ecology, and whether
defined biologically as 'a group of organisms, coarse or fine - is not a property of nature as such.
which breed to produce viable offspring' (Mayr, It is, in fact, only our necessary means to converse
1942). All other categories, above or below the with each other about the facts, that we have
species level, are arbitrary conventions. From an found in nature.
evolutionary perspective (Simpson, 1951), a I don't want to dwell any longer on these ideas;
species is 'a lineage, an ancestral-descendent they are not new or original. Instead I shall try to
sequence of populations, evolving separately from apply these ideas to the special conditions of
others'. This latter definition applies better to rotifers and use, as an example, the terminalis-
rotifers or other facultative or obligatory asexual longiseta complex of the genus Filinia.
organisms, where breeding, in the sense of ex- The problems of rotifer taxonomy have various
change of genes, occurs rarely or not at all. inter-related sources, namely:
So, if taxonomy is artificial and 'does not corre- 1. The facultative (heterogonic) or obligatory
spond with anything in nature' (Simpson, 1967, parthenogenetic reproduction;
3.ed. pg. 17), why do we accept it as part - and 2. The short generation time and high mutation
even an important part - of biological sciences? rate;
The answer is obvious: without taxonomical 3. The paucity of morphological characteristics;
names and categories and the man-made hierar- 4. The high variability.
292

1. Parthenogenetic reproduction implies trans- nalis was scarce in the whole water column, but,
fer of the parent's genotype to offspring without when thermal stratification and oxygen depletion
change. New mutations, if not lethal, may be started to build up, the population became denser
immediately multiplied within the population. in the bottom layer. On May 15th we found an
Both facts are partly responsible for the high enormous increase just above the lake bottom
degree of variability observed in rotifers. Obliga- (14 m) that could not be explained by repro-
tory parthenogenetic groups are potentially even duction at the low temperature of 9 0 C (Fig. 1).
more variable than heterogonic ones, since they Vertical migration was out of the question
lack the sexual recombination of genes during because there was no epilimnic population. The
mixis, which unifies the genetic pool of a hetero- only explanation was an explosive hatching of
gonic population. Such groups are: the Bdel- resting eggs, but what triggered the hatching is not
loidea, some planktonic populations which have known.
never been found to undergo a mictic period, and Such an explosive hatching implies a massive
populations which produce pseudosexual resting production of resting eggs during the life cycle of
eggs, a phenomenon that probably occurs more the population. There are, in fact, reports of very
often than we know. high resting egg production; among others,
Very likely, Filinia terminalis produces Hofmann (1979) mentions 36-45 %, Loose (1987)
pseudo sexual resting eggs in Esthwaite Water, a reports 50-100 % females carrying resting eggs
eutrophic lake of 15 m depth in the English Lake (referred to as 'mictic rate') in the PluBsee. Pejler
District. A peculiar situation concerning the verti- (1961) indicates several mictic events on his graph
cal distribution of this population was found in of F. terminalis in Osbysj6n, but in the text he
1979. At spring homothermy, in April, F. termi- speaks of resting eggs only. Neither mictic females

Fig. 1. Vertical distribution of Filinia terminalis in Esthwaite Water, English Lake District, on 15.5.1979. N = number of
individuals; T = Temperature; O2 = Oxygen content.
 293

carrying the small eggs from which males hatch, posture of the caudal seta in preserved condition.
nor males as such are mentioned by these authors. The tooth formula is similar to F. terminalis
Moreover, a resting egg ratio of over 50% makes (15/16) and the teeth are not easy to count; seta
it very unlikely, and of 100% virtually excludes a measurements vary widely. The posture of the
'normal' mictic phase. Both the unusual high per- caudal seta, whether it is straight or abduced,
centage of resting eggs and the lack of males depends on the insertion of the retractor muscles
corroborate my suspicion that the resting eggs of on the body wall, behind or before the seta
F. terminalis are not fertilized, but pseudo sexual. (Fig. 2). It is also influenced by the intensity of
If this hypothesis is correct, the expression 'mictic contraction during preservation; if a specimen is
females' and 'mictic rate' are inadequate. More- narcotized, the caudal seta is not abduced. There-
over, the likelihood of extensive variation - geneti- fore the new taxon is difficult to distinguish. Even
cal as well as morphological - would, theoreti-
cally, increase.

2. The short generation time of rotifers, which

allows for approximately 100 generation in one
year under favorable conditions (for instance in
the tropics), multiplies the spreading of any
genetic change in the population. King (1980)
made a rough estimate of mutations per locus/ r m.
year; he gives a figure of approximately 4000 for
F. terminalis, which, in my opinion, is too high
since F. terminalis, living at 4-6 0 C has a genera-
tion time about four to five times longer than
assumed in that paper.

3. The paucity of useful morphological charac-

teristics poses - together with variability - the
most serious difficulties for rotifer taxonomy.
Rotifers are small and their morphological
features, below the generic level, are hard to
detect, at least with the means that ecologists are
forced to apply in their work. Encouraged by
modern microscopical techniques, particularly
scanning electron microscopy, taxonomists have
recently tended to use more and more minute
morphological features to create new taxa in order b
to cope with an increasing diversity of biological
data. In practice, such inconspicuous features are
ascertained on very few specimens (often only
one) and the results are applied to the whole
population. a
An example is again available from the F. term i-
nalis-longiseta complex. Koste (1980) described a
Fig. 2. Insertion of the retractor muscles on the body wall.
new taxon, F. hofmanni, from the PluBsee, N. a = Filinia terminalis (Plufisee, 11.5.1970); b = Filinia termi-
Germany, on the grounds of a symmetrical tooth naZis = hoJmanni (Lunzer Obersee, 20.8.1979). r.m. =
formula (15/15), setae measurements, and the retractor muscle.
294

Loose (1987), working also on PluBsee material, even found that the setae of Filinia increased in
could not distinguish F. hofmanni from F. termi- length by about 50 %within a few days in cultures
nalis neither 'living nor dead' (pg.21). In fact, containing the well-known 'Asplanchna sub-
most investigators rely on the dubious, but easily stance'. Slonimski (1962) has shown experi-
visible, criterion of seta posture. mentally that the insertion of the caudal seta is
shifted ventrally with increasing temperature.
4. The taxonomical difficulties arising from vari- To add one more set of data to the host already
ability are convincingly exemplified in the genus existing, Fig. 3 shows the variation of F. termi-
Filinia. To differentiate F. longiseta and F. term i- nalis x in 4 lakes in the English Lake District. The
nalis, absolute measurements or ratios of body result of these measurements indicates no clear
and appendices have until recently been used, trend, except for a decrease of size from spring to
although the variability of these features has been summer and from eutrophic to oligotrophic lakes.
documented for at least 30 years. Ratios oflateral In summary, the efforts of many scientists over a
to caudal setae and the insertion of the latter have long time have shown that measures of body or
been computed in graphs and tabulations by appendices are poor means to distinguish taxa or
Pejler (1957a), Parise (1961), Larrson (1971), to describe new ones within the genus Filinia. A
Hofmann (1974), Sternberger (1977), and number of measurements sufficient for statistical
Schaber & Schrimpf (1984) among many others. analysis is, nonetheless, helpful; mean setae ratios
Taken as a whole these data show the wide range may differ, and regression lines on plots show
of variation and overlapping of criteria used for different slopes for various popUlations or taxa
identification. Moreover, the data vary independ- (Schaber & Schrimpf, 1984).
ently and in opposed directions. Pourriot (1964) In contrast to the picture that emerges from

A Filinia terminalis

BI.1969 0 BW. B. l. C. s. l. s. C/l V:fJ3.10 5

x ,
: min. max. ,
1. 3.-15.4.
+ ~:' I ~ ,,, 7·9

....... ,-----+>-+
15.4.- 1.6. f--t :
, I
~ , 5·8
1.6.- 1.8. 1- , , , ~:, 2 8
, , , ,
,I , , , :, , , ,
I
, , , , , , ,
~ 0 10 20 3~ I ill lW 1111 1~0 180 310 360
, I '
I,[)() 340 380 410 460 500 0.60 O<lO 1'~

B
0 BW B. l. c.s l. s. CIL V:fJ3.10 5
March 1979 ,, ,
+
I
BI. ..f >--t + I
+- 10·6 I

+
I I
~ ~ >--f-----< >-f ~ , 118
Est. I
I
I
I
WN.
+ ,, ,
~ + I
I
+ +I 7-4

, , , +
WS.
, + r-r----r-. : , ,
I
, , , , ,
I
I
,
, I
>---t---
, , , , , , , , , ,
~:I 6·3
, ,
3~0
t I
0 10 10 30 70 90 110 130 110 100 340 380 , : 460 500 0.60 l'od
f 160 160 300 380 ~10 G80

Fig. 3. Measurements of Filinia terminalis in 4 lakes of the English Lake District. A. Variation in time in one lake; B. Variation
in different lakes at the same time (March 1979). Bl = Blelham Tarn; Est. = Esthwaite Water; W.N. = Windermere North Basin;
W.S. = Windermere South Basin. D = Distance of caudal seta to end of body; B.W. = Body width; B.L. = Body length;
C.S. = Caudal seta; L.S. = Mean oflateral setae; CjL = Ratio of caudal to lateral setae; V = Volume calculated from mean body
length; Each graph represents at least 10 measurements.

x Determination according to Pourriot (1965) and Pejler (1957a) and confirmed by Hofmann (letter of 11.7.79).
 295

morphological variation, from the ecological the two eutrophic lakes the population maximum
standpoint, at least in palaearctic lakes, two c1ear- occurs in the nearly deoxygenated layer just above
cut groups occur within the genus Filinia. One lake bottom. In the two much deeper meso- to
group lives in summer at temperatures above oligotrophic lakes, the maximal abundance occurs
approximately 18 °C in the epilimnion of lakes, in the metalimnion at oxygen saturation and is
ponds and slowly flowing rivers. The other group about 2-3 orders of magnitude lower. Only the
lives at temperatures below approximately 12 °C temperature required is identical in all 4 lakes.
in the hypolimnion of eutrophic or meromictic Obviously, food availability regulates the location
lakes. The latter group can tolerate, but is not of the maximum of these bacteria-feeding rotifers.
restricted to very low or nearly zero oxygen levels. This assumption corresponds fairly well with the
The epilimnic, thermophilous group, with all its quantitative distribution of bacteria known from
variations, is comprised under the name the respective lakes (Jones, 1972).
F. longiseta (sensu lato). The predominantly hypo- In meromictic lakes the Filinia populations
limnic, cold stenothermous group with all its show a different distribution, as depicted in Fig. 4;
variations goes under the name of F. terminalis similar situations are described by Pejler (1957a)
(sensu lato). and Schaber & Schimpf (1984). In these cases the
Within the cold stenothermous group the situa- population is perennial and stays in a water layer
tion is rather complicated. Various populations of approximately 4 ° C and between 1 mg I and
occupy definite ecological niches according to the zero 02-content. This layer is just above the plate
lake type, as we have shown in the 4 lakes of the of chemotrophical bacteria. In the Lunzer
English Lake District (Ruttner-Kolisko, 1980). Obersee (Fig.4A) this layer moves up to 3-4 m
The ecological behaviour of these populations at depth under ice and down to 11-12 m in the short
their maximal density is computed in Table 1. In summer period. In Blankvatn (Fig.4B) low

Table 1. Ecological data of Filinia terminalis populations at maximal density.

Blelham Esthwaite Windermire Windermire

Tarn Water South-Basin North-Basin
(14 m) (15 m) (40 m) (60 m)
(eutrophic) (eutrophic) (mesotrophic) (oligotrophic)

Abundance at max. density

(Indiv./l) 1800 1500 100 3

Time of max. density May-June May-June June June-Juley

Depth of max. density 9-12 12-15 10-15 10-15

(m) (hypolimn. ) (hypolimn. ) (metalimn.) (metalimn. )

Temperature at max. <10 <10 10 10-12

density (0 C)

Oxygen content 10-40 10-40 >90 >90

( %-saturation)
Mean indiv. body size 10.6 11.8 6.3 7.4
(11 3 • WS = V)

Occurrence of the popu- spring/ spring/ perennial perennial

lation during the year summer summer
 296

A LUNZER OBERSEE
ice
o (
\ '. .' I /
/
\ ". "
m ....... - .... / "" \
\
,
5

1 mg 02/1
0.2mg 02/1
10

14
J
1972 1973 1974

I= 50 0/0

RUTTNER - KOLISKO 1974

FILINIA TERMINALIS = F. HOFMANNI

B BLANKVATN
1965 - 1966

N o J N
O+-__- L_ _ _ _L -_ _- L_ _ _ _L -_ _~--~k---~--~----~--~----~--~----
4
8
12
16
20

25
30
m
35

~ illill
26-50
~
51 -100
i nd. pro 31 : 6-25 LARSSON 1971
Fig. 4. The vertical distribution of Filinia terminalis = hofmanni in two meromictic lakes. A. Lunzer Obersee, Austria (after
Ruttner-Kolisko 1974); black bars indicate the location of50% of the population. B. Blankvatn, Norway (after Larsson 1971);
the black line indicates zero oxygen, the dotted line 4.6 temperature, the striped areas show the number of individuals per
0

3/1.
 297

02-content and low temperature are found con- grid and the question of who imposes the grid.
tinuously between 18-20 m depth. Under such Geneticists use the finest grid; their mesh size is
ecological conditions Filinia populations x can defined as a succession of genetical units which
occur all year, while they are eliminated because can change in time and some consider a popula-
of too high temperature and zero 02-level in tion, in the sense of ecologists, not as a unit, but
Esthwaite Water (Ruttner-Kolisko, 1980) and as an assemblage of units with no more affinity to
other eutrophic lakes, where the anoxic layer each other than 'griffins, unicorns and mermaids'
reaches temperatures above 12-16 °C in summer. (King, 1980). The grid of ecologists, like me, is
Hofmann investigated the PluBsee in N. much coarser; they call the mesh size a population
Germany over many years (l972-1978). This - at least in the case of predominantly
eutrophic lake completely overturns only at irregu- parthenogenetic groups. A population consists of
lar intervals and stays meromictic in between. a considerable number of similar individuals of
Hofmann found both types of Filinia distribution the same, clearly discernible unit, living together
in this lake and also a part of the population in the same ecological niche. The niche must be
showing a deduced caudal seta. xx Reluctantly defined by as many environmental parameters as
Hofmann called it F.c.f. longiseta, but fortunately are available. In my opinion, this discernible unit
we have now the name F. hofmanni for these popu- is, in the case of rotifers, what is called 'genus'.
lations, thanks to Koste (1980). Although the The mesh size of taxonomists is arbitrary and
taxon is not discernible on morphological criteria depends entirely on the investigator. Because of
only, the name is now, rightly, used for any popu- this arbitrariness I have purposely used only the
lation of meromictic distribution. general term 'taxon' in this paper, to avoid any
The taxonomical difficulties and errors in the position in the taxonomical hierarchy.
F. terminalis/longiseta group have been described With my criticisms of creating taxa, I can rely
by Pejler (1957b) from their beginning until the on a very renowned authority in hydrobiology,
time of his own investigations. At present, the Wesenberg-Lund, who wrote as early as the year
situation is such that it is sometimes impossible to 1900 about non-scientific species making, in this
discern what someone has seen when he uses a own German words 'diese recht unwissenschaft-
particular name. The more papers that are written, liche Artmacherei'. Although I do not want to go
the more confused the situation becomes. There that far, I would like to make a few suggestions,
already exist approximately 50 publications on which - I hope - may be considered when new
the F. terminalis/longiseta problem; they all go rotifer taxa are created.
around in more or less the same circles. This is an
incredible waste of time and effort, not resulting Suggestions:
in any new knowledge of natural facts. The state
of affairs is not much better in the genus Hexarthra 1. Preserved material from an ecologically
and will be the same - I am quite certain - in the unknown biotop should not be considered suf-
genus Kellikottia as soon as rotiferologists start to ficient to describe a new taxon.
look closer into the ecological data already availa- 2. The description of a new taxon should be
ble on what we call today Kellikottia longispina. based on easily discernible features. Minute
Coming back to my view of taxonomy as an characteristics, which cannot be determined
artificial grid imposed on the continuous flow of during ecological work, should be avoided or
nature, we must still discuss the mesh size of the used only as supplemental evidence.

x Named F. terminalis by both authors according to the nomenclature prevailing at that time.
xx The same phenomenon is described by Schaber (1980), who found up to 25% of the so called F.c.f.longiseta in some of his
Tyrolian lakes.
298

3. In order to ascertain possible variation of the Loose, C, 1987. Populationsakologische Untersuchungen an

hypolimnischen Riidertieren des PluBsee. Diplomarbeit,
criteria used to characterize the new taxon, a
Kiel.
fair number of individuals (50-100) should be Mayr, E., 1942. Systematics and the origin of species.
available. Columbia Univ. Press, N. York.
4. The description of a new taxon using only Parise, A., 1961. Sur les genres Keratella, Synchaeta, Poly-
variable morphological features should not be arthra et Filinia d'un lac italien. Hydrobiologia 18:
121-135.
considered; if the variation overlaps with
Pejler, B., 1957a. On variation and evolution in planktonic
already existing taxa, the criterion is useless. rotatoria. Zool. Bidr. Uppsala Univ. 32: 1-66.
5. Ecological data are to be incorporated in the Pejler, B., 1957b. Taxonomical and ecological studies on
description of the new taxon and should be planktonic rotatoria from northern Swedish Lapland.
given a special emphasis. Kung!. Svenska Vetens. Akad. Hand!. 6: 1-67.
Pejler, B., 1961. The zooplankton of Osbysjan I. Seasonal
6. A new taxon should only be created if discrimi-
and vertical distribution. Oikos 12: 225-248.
nation from already known taxa is needed for Pourriot, R., 1964. Etude experimentale des variations mor-
some scientific reason - never just for the sake phologiques chez certaines especes de rotiferes. Bul!. Soc.
of a new name. Zool. France 89: 555-561.
Pourriot, R., 1965. Notes taxonomiques sur quelques
rotiferes planktoniques. Hydrobiologia 16: 579-604.
References Ruttner-Kolisko, A., 1974. The vertical distribution of plank-
ton rotifers in a small alpine lake. Verh. int. Ver. Limno!.
Hofmann, W., 1972. Zur Produktionsbiologie des Zoo- 19: 1286-1294.
planktons im PluBsee. Verh. int. Ver. Limno!. 18: 410-418. Ruttner-Kolisko, A., 1980. The abundance and distribution
Hofmann, W., 1974. Zur Taxonomie und Verbreitung von of Filinia terminalis in various types of lakes. Hydro-
Filinia-Arten ..... Faun. ako!. Mitt. 4: 437-444. biologia 73: 169-175.
Hofmann, W., 1979. Untersuchungen zur akologischen Schaber, P., 1980. Die Riidertiergattung Filinia in Tiroler
Nische zweier nahe verwandter Rotatorien Arten des Badeseen. Jb. Abt. Limno!. Innsbruck 6: 159-171.
PluBsees. Habilitationsschrift, Kie!. Schaber, P. & Schrimpf, A., 1984. On morphology and
Hofmann, W., 1982. On the coexistence of two pelagic Filinia ecology of the Filinia-terminalis-Iongiseta group in
species in lake PluBsee. Arch. Hydrobio!. 95: 125-137. Bavarian and Tyrolian lakes. Arch. Hydrobiol. 101:
Hofmann, W., 1987. Population dynamics of hypolimnetic 147-257.
rotifers in the PluBsee. N. Germany. Hydrobiologia 147: Simpson, G. G., 1951. The species concept. Evolution 5:
197-201. 285-298.
Jones, J. G., 1972. Studies of Freshwater bacteria ..... 1. Eco!. Simpson, G. G., 1967. Principles of Animal Taxonomy. 3. ed.
60: 59-75. Columbia Univ. Press, N. York.
King, C E., 1980. The genetic structure of zooplankton popu- Sionimski, P., 1926. Sur la variation saisoniere chez Filinia
lations. Evolution and Ecology of Zooplankton Com- longiseta. C R. Soc. BioI. 94: 543-545.
munities. Univ. Press of New England. Sternberger, R. S., 1976. Systematics of the Rotatorian gen-
Koste, W., 1980. Dber zwei Plankton-Riidertiertaxa, Filinia era Synchaeta and Filinia. Proposal to National Science
australiensis, n.sp. und Filinia hofmanni, n.sp. Arch. Foundation. Manuscript.
Hydrobiol. 90: 230-256. Wesenberg-Lund, C, 1900. Von dem Abhangigkeitsverhaltnis
Larsson, P., 1971. Vertical distribution of planktonic rotifers zwischen dem Bau der Planktonorganismen und dem
in a meromictic lake. Norw. J. Zoo!. 19: 47-75. spez. Gewicht des Wassers. BioI. Zentr. Blatt 20: 606.
Hydrobiologia 186/187: 299-310, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 299
© 1989 Kluwer Academic Publishers.

Systematics, reproductive isolation and species boundaries in

monogonont rotifers

Terry W. Snell
Division of Science, University of Tampa, Tampa, FL 33606, USA

Key words,' mating, reproductive isolation, Rotifera, sexuality, systematics

Abstract

The typological concept of rotifer species and the morphological basis of rotifer systematics is reviewed
and alternatives proposed. Occasional sexuality in the cyclical parthenogenetic life cycle of monogononts
permits application of the biological species concept to this group. Data from cross-mating experiments
with Asplanchna, Brachionus and Epiphanes illustrate the usefulness of reproductive isolation as a criterion
for species boundaries. Populations from different geographic regions are often interfertile indicating that
rotifer species are genetically integrated over wide areas. The main types of isolating mechanisms
operating in monogononts are reviewed. The role of behavioral reproductive isolation in maintaining
species boundaries is examined. The use of a mate recognition bioassay which estimates the probability
of copulation and quantifies the degree of isolation is described. Recent work of the mechanism of mate
recognition is reviewed. It is concluded that the biological species concept is applicable to rotifers and
that a more experimental approach to determining species boundaries is both feasible and desirable.

Introduction native views. He concludes that the biological

species concept provides an objective means for
Although the concept of species is essential in separating species and represents the causal fac-
systematics and evolution, it lacks a universal tor which produces and maintains discrete evolu-
definition. The most widely accepted species con- tionary units. Hauser refutes the major criticisms
cept was developed by Dobzhansky and Mayr of the biological species concept which are based
beginning in the 1930's (see Dobzhansky, 1970 on its lack of universality, impracticality and
and Mayr, 1970 for reviews). The hallmark of the inapplicability to species separated in time. It is
biological species concept is that species are dis- concluded that the biological species concept
tinguished by their reproductive isolation from does not need to be changed or rejected on the
other species. Because of its prominent role, basis of the prevailing criticisms.
reproductive isolation has been the subject of Perhaps the strongest stimulus for re-examining
intense study (Littlejohn, 1981). Despite its wide- the species concept has come from Paterson
spread acceptance for several years, the biological (1982, 1985). He argues that the prevailing bio-
species concept recently has been vigorously logical species concept is actually an 'isolation
attacked by a number of authors. Hauser (1987) concept'. Paterson reasons cogently that specia-
reviews the objections to the biological species tion occurs as a side-effect of adaptations that
concept and examines 15 recently published alter- promote fertilization and is therefore an evolu-
300

tionary effect rather than a cause. Selection acts approach. A first attempt to apply cladistic
to enhance fertilization systems in populations, a methodology to rotifers appears in this volume
consequence of which is speciation. Defined in (Wallace & Colburn, 1989).
this way, species are the most inclusive popula- For the most part, rotifer genera are morpho-
tion of biparental organisms which share a com- logically well defined as taxa, but establishing
mon fertilization system. Paterson's 'recognition species boundaries has been a very difficult prob-
concept' of species more clearly separates evolu- lem for rotifer systematists (Ruttner-Kolisko,
tionary causes from effects and does not rely on 1963). The use of morphology to define rotifer
relational characteristics to define species. The species is confounded by extensive morphological
recognition concept of species is gaining wide variation, like cyclomorphosis, that is typical of
attention, but it is not without critics (Templeton, the group (Hutchinson, 1967). In spite of these
1987). difficulties with morphology, rotifer systematics
Against this background of controversy, the has not employed approaches that have been
classification of rotifers proceeds, as it does in all helpful in establishing species boundaries in other
organisms, without a unanimous notion of animals. Reproductive isolation, perhaps the
species. Yet the question of how to best organize most natural marker of a species boundary, has
rotifer diversity into meaningful taxonomic and seldom been utilized in rotifers. While theoreti-
evolutionary units persists. There is a basic cally attractive, there is some question whether
impression that some populations are similar due the criterion of reproductive isolation is applica-
to genetic integration, probably resulting from ble to rotifers. The problem is rooted in the
gene flow, stabilizing selection, developmental question asked by Pejler (1977a): is partheno-
homeostasis or some combination of these genesis essentially obligatory in monogonont
factors. Other populations are decidedly different, rotifers? Obligatory parthenogenesis is well
probably due to evolutionary divergence in iso- known in the class Bdelloidea where males have
lation. But how can these ideas be forged into a never been observed and the cytological evidence
systematics of rotifers consistent with modern supports the idea that bdelloids are agamospecies.
evolutionary theory? If most monogononts are also obligatorily par-
As presently conceived, rotifer species are thenogenetic, reproductively definable
groupings based on morphological similarity of boundaries between species do not exist and
easily measured characters. These are for the rotifer systematics can only be based on subjec-
most part visually distinguished at the light or tive decisions. Whether sexual reproduction
electron microscope level. Data are available for occurs occasionally, rarely, or never in mono-
only a few species on biochemical traits or on gononts therefore will determine if reproductive
reproductive isolation from cross-mating experi- isolation is useful as a criterion for rotifer species
ments. Discontinuities among rotifer species are boundaries. Holman (1987) has compared the
identified by the judgement of systematists, so recognizability of species in bdelloids and mono-
rotifer taxonomy is based on the classical typo- gononts.
logical concept of species. Species boundaries My objective in this paper is to explore the
occur where phenotypic variation between groups feasibility of establishing a more experimental
is too large to fit comfortably into one group. In definition of rotifer species. The use of cross-
theory, rotifer taxonomy is based on evolutionary mating experiments and mate recognition bio-
systematics, rather than numerical phenetics or assays as reproductive isolation tests is demon-
cladistics (Mayr, 1981). But in practice it is based strated. It is argued that these tools make it possi-
almost entirely on morphology. Rotifer sys- ble to derme rotifer species boundaries empiri-
tematics is therefore more phenetic than evolu- cally. The small data base from cross-mating
tionary or cladistic, but has not utilized the experiments between species and among strains
modern numerical taxonomic methods of this within species is reviewed. The most common
 301

types of reproductive isolating mechanisms detected to verify the completion of sexual repro-
operating in rotifers is characterized and the role duction. Cyst bearing female density exceeded 1
of behavioral isolation in defining and per liter for 8 days in April and 5 days in June.
maintaining species boundaries discussed. The These observations clearly illustrate the ephem-
use of the probability of copulation for quantifying eral nature of sexual reproduction in natural
the degree of isolation is described as is recent rotifer populations and the rigorous sampling pro-
work on mechanisms of mate recognition. gram required for detection. It is therefore not
surprising that males and cyst-bearing females are
not often observed in natural rotifer populations.
Do monogononts reproduce sexually? Although sexual reproduction has not been
observed in many natural rotifer populations, sex
Males have not been observed in many mono- is common in lab populations, especially when
gonont species, but are they really absent or are rotifers are first isolated from the field. Males and
sampling programs so inadequate as to make cyst production have been observed for many
male detection improbable? Long term studies by species of Asplanchna and Brachionus. More
Carlin (1943) provide some insight into the inter- species will have to be cultured, however, before
mittent nature of male production by Polyarthra it is known whether these genera typify mono-
and Notholca in the Motala River in Sweden. gononts. At present, it seems safe to conclude that
Male production did not occur in every popula- at least many Asplanchna and Brachionus species
tion in every year. In P. vulgaris, for example, male are not obligate parthenogens and that sexual
production did not occur in 1935 and occurred reproduction is an important feature of their life
during only one week in 1936, despite the near cycle.
continuous presence of this species. Similar pat-
terns were observed for P. major and N. caudata.
Of the 23 monogonont species closely monitored Defining species boundaries with mating experi-
by Carlin, 52 % produced males at some time ments
during the five years of sampling. Although many
species eventually produced males, these were The classical approach to defming species
only detectable with a long-term sampling pro- boundaries as prescribed by the biological species
gram with frequent observations. concept is the mating experiment. Several mating
The difficulties of confirming sexual repro- experiments using monogonont rotifers have been
duction in natural rotifer populations were further completed, verifying the feasibility of this
elucidated by King & Snell (1980) who quantita- approach with species amenable to laboratory
tively sampled Asplanchna girodi in Golf Course culture.
Pond (Florida) daily or bidaily from April through Early attempts to cross different strains of a
July of 1977. In that study, hundreds ofliters were single rotifer species suggested that strains from
sampled each time, providing one of the most distant geographical locations could be mated.
finely detailed views of population dynamics for Shull (1911) crossed an Epiphanes senta strain
any natural rotifer population. Males were present from New York with one from Maryland and
(> 0.001 per liter) for 15 days in April, 35 days in later (Shull, 1915) crossed a strain from England
May-June and 12 days in July. Male density with one from Lincoln, Nebraska. Also working
exceeded 1 per liter for only 13 days in April and in the same area of Nebraska, Hertel (1942)
5 days in June. If the sampling program had not successfully crossed four E. senta strains col-
been so intensive, it is unlikely that males would lected from surrounding ponds.
have been detected in this population. The pres- Gilbert (1963) attempted to mate three
ence of males only suggests sexual reproduction. brachionid species, all collected from a small,
Resting egg (cyst) bearing females also were slightly alkaline pond near New Haven, Con-
302

necticut. Brachionus calyciflorus and B. angularis utilized precluded quantification of post-mating

were planktonic whereas B. quadridentatus was barriers. Differences in the outcome of reciprocal
sometimes planktonic, but usually attached to crosses illustrates the asymmetry common in
submerged objects. All three species commonly post-mating reproductive isolation (Giddings &
co-occur in alkaline ponds in temperate regions Templeton, 1983). The species boundary between
throughout the world. Gilbert found that the B. uroeolaris and B. quadridentatus therefore is
sexual periods of B. calyciflorus and B. angularis not clearly defined. Even though these species
overlapped, but all attempts to mate these species appear to be morphologically distinct, further
failed. Interspecific matings with B. quadriden- analysis of their reproductive isolation is
tatus also failed indicating that the reproductive warranted.
isolation of these species is complete. Similar experiments have been conducted with
Mating experiments between geographically A. brightwelli, A. intermedia and A. sieboldi (Gil-
separated Asplanchna brightwelli populations were bert et al., 1979). Crosses between A. brightwelli
carried out by Birky (1967). Populations collected females and A. intermedia or A. sieboldi males
from Bloomington, Indiana were mated with yielded a 39% fertilization rate in 74 attempts
populations from Paris, Tennessee and produced indicating little if any pre-mating isolation among
viable F l , F2 and backcross progeny. These these species. However, since none of the cysts
populations clearly share a common gene pool, hatched, zygote inviability was the likely cause of
but intra- and interpopulational crosses were not the complete post-mating isolation. The recipro-
equally fertile. Intrapopulational crosses (self cal cross with A. intermedia or A. sieboldi females
fertilization) were 77% successful, whereas and A. brightwelli males yielded a fertilization rate
crosses between Indiana and Tennessee popu- of 47% in 47 crosses, but again none of the cysts
lations had only a 48 % success rate, suggesting hatched. Crosses between A. intermedia and
partial reproductive isolation. Birky also A. sieboldi indicated no pre-mating isolation, but
attempted to cross A. brightwelli from Tennessee too few crosses were completed to conclude
with A. girodi from the same pond. No inter- anything about post-mating barriers. The general
specific matings were ever observed although conclusions from this study were that A. bright-
both species self fertilized with > 60 % success. welli is reproductively isolated from A. intermedia
Asplanchna brightwelli andA. girodiwere therefore and A. sieboldi by real, quantifiable post-mating
completely reproductively isolated despite the fact barriers. Additional crosses are necessary to
that both species occupied the same region of the determine if A. intermedia and A. sieboldi are like-
pond and both reproduced sexually at the same wise completely reproductively isolated. Only a
time. No hybrids were observed verifying the few strains were used in this study so more popu-
effectiveness of the isolating mechanisms. lations need to be examined to assess the range of
Mating barriers between rotifer species are not intraspecific variation for reproductive isolation.
always complete. Ruttner-Kolisko (1969) suc- Results of interspecific crosses among rotifer
cessfully crossed B. uroeolaris with B. quadriden- species are summarized in Table l.
tatus. Males of both species copulated with con- Two unpublished works provide additional
specific as well as heterospecific females, information on reproductive isolation among
although the probability of copulation was not geographically separated Asplanchna strains.
quantified so male preferences may have gone Birky (unpublished, reported in Birky & Gilbert,
undetected. However, while pre-mating barriers 1971) attempted to cross his Tennessee and
were absent, post-mating barriers were observed. Indiana strains of A. brightwelli with a German
Crosses between B. quadridentatus females and strain which was morphologically identical and
B. uroeolaris males produced cysts but none physiologically similar. No cysts were produced
hatched. The reciprocal cross, in contrast, was from any of these crosses. Snell (unpublished,
fertile but the small number of mictic females reported in King, 1977) attempted to cross
 303

Table 1. A summary of interspecific crosses attempted with monogonont rotifers. Brachionus species: caly - calyciflorus. ang
- angularis. quad - quadridentatus. plic - plicatilis, rubens - rubens, urcea - uroeolaris. Asplanchna species: bright - brightwelli, girodi
- girodi, inter - intermedia, sieboldi - sieboldi.

Cross Pre-mating barrier Post-mating barrier Reference

0' V Brachionus
caly X ang no mating Gilbert 1963
caiy X quad no mating Gilbert 1963
caiy X Synchaeta no mating Gil bert 1963
caly X Euchlanis no mating Gilbert 1963
ang X caly no mating Gilbert 1963
plic X Synchaeta no mating Snell & Hawkinson 1983
plic X rubens no mating Snell unpublished
urceo X quad none* no cyst hatching Ruttner-Kolisko 1969
quad X urceo none* none? Ruttner-Kolisko 1969
Asplanchna
bright X girodi ? no cysts produced Birky 1967
bright X inter none* no cysts hatching Gilbert et al. 1979
bright X sieboldi none* no cysts hatching Gilbert et al. 1979
sieboldi X inter none* ? Gilbert et al. 1979

* Probability of copulation not quantified, subtle differences in male mating preferences could exist.

A. brightwelli from Florida with Birky's strains incompatibilities can be due to either males or
from Tennessee and Indiana and the German females. Results of intraspecific crosses among
strain. None of these crosses produced cysts, yet strains of a single rotifer species are summarized
all strains were successfully self fertilized. in Table 2.
Reproductive isolation among geographically These studies permit several conclusions about
separated Brachionus plicatilis strains was investi- reproductive isolation in rotifers. (1) It is possible
gated by Ruttner-Kolisko (1983, 1985). A strain to identify absolute species boundaries, con-
cultured in Ruttner-Kolisko's lab in Austria for 20 sistent with the biological species concept, among
years was crossed with strains from Scotland and closely related rotifer species. Cross-mating
Colorado. Austria, Scotland and Colorado males experiments have accomplished this for B. calyci-
did not attempt to mate with Austria females. jlorus, B. angularis and B. quadridentatus,
Austria females apparently have lost the ability to A. brightwelli, A. sieboldi and A. intermedia. These
elicit male mating responses after many years of taxa are good biological species which are not
parthenogenetic reproduction in the laboratory. exchanging genes and are pursuing independent
Austria males, in contrast, were able to fertilize evolutionary courses. (2) Absence of hybrids sug-
Scotland and Colorado females and produce via- gests that reproductive isolation is usually com-
ble cysts. Crosses between Scotland males and plete even in species with overlapping sexual
Colorado females produced viable cysts, but the periods (e.g. B. calyciflorus and B. angularis,
reciprocal cross with Scotland females and A. brightwelli and A. girodi). (3) Populations
Colorado males was infertile. Self fertilization in within a species often are genetically integrated
Scotland and Colorado strains was highly over a wide range and retain the ability to
successful. Gene flow therefore is possible among exchange genes. This is demonstrated with
all three strains, but all are at least partially repro- A. brightwelli from Indiana and Tennessee and
ductively isolated from one another. These experi- B. plicatilis from Austria, Scotland and Colorado.
ments illustrate that reproductive isolation is often (4) Postmating barriers effectively separate close-
asymmetrical between strains and reproductive ly related species as in A. brightwelli, A. sieboldi
304

Table 2. A summary of intraspecific crosses attampted with monogonont rotifers. Asplanchna abbreviations: Ind - Indiana, Tenn
- Tennessee, Fla - Florida, 4 temporal pops - all from Lake Thonotosassa, Florida. Brachionus abbreviations: 3 temporal pops
- all from McKay Bay, Florida; geographic pops - from different countries worldwide.

Cross Pre-mating barrier Post-mating barrier Reference

Epiphanes senta
New York X Maryland none* none Shull 1911
England X Nebraska none* none Shull 1911
4 Nebraska pops none* none Hertl 1942
Asplanchna brightwelli
Ind X Tenn none* partial Birky 1967
Ind X Germany complete isolation, barrier unknown Birky unpublished
Tenn X Germany complete isolation, barrier unknown Birky unpublished
Fla X Ind complete isolation, barrier unknown Snell unpublished
Fla X Tenn complete isolation, barrier unknown Snell unpublished
Fla X Germany complete isolation, barrier unknown Snell unpublished
4 temporal pops none* partial King 1977
Brachionus plicatilis
Austria X Scotland none* partial Ruttner-Kolisko 1983
Austria X Colorado none* partial Ruttner-Kolisko 1983
Scotland X Colorado none* partial Ruttner-Kolisko 1983
3 temporal pops none Snell & Hawkinson
1983
8 geographic pops partial Snell & Hawkinson
1983

* probability of copulation not quantified, subtle differences in male mating preferences could exist.
and A. intermedia. (5) Sibling species may be sterility. Examples of seasonal reproductive iso-
common in monogont rotifers as suggested by the lation in rotifers are found in Carlin (1943) who
reproductive isolation of Tennessee and Indiana quantitatively sampled the Motala River for five
A. brightwelli from morphologically identical consecutive years. Polyarthra vulgaris and
strains from Florida and Germany. P. remata produced males from September
through November in each of the five years
sampled. Polyarthra major consistently produced
Reproductive isolating mechanisms males July through October and P. dolichoptera
produced males April through June. Even though
Mating experiments illustrate that many mono- females of these species sometimes co-occurred in
gonont rotifer species can be defined as reproduc- the plankton, the sexual periods of P. major and
tively isolated evolutionary units. An important P. dolichoptera did not overlap one another or
consideration is how reproductive isolation those of P. vulgaris and P. remata. Non-over-
among species arises and how it is maintained. lapping sexual periods enforce reproductive iso-
Several examples of reproductive isolating lation by preventing males and mictic females
mechanisms in rotifers will provide insight into from meeting.
this question. In some types of environments seasonal repro-
Reproductive isolating mechanisms are ductive isolation can breakdown. Pejler (1956)
broadly classified as pre-mating or post-mating investigated the reproductive isolation between
(Mayr, 1963). Pre-mating barriers include seaso- P. vulgaris and P. dolichoptera in Lake
nal, habitat, behavioral and mechanical mecha- Luossajarvi in Sweden. Polyarthra vulgaris was
nisms; post-mating barriers include gametic eurythermal, preferred oxygen rich surface waters
incompatability, hybrid inviability and hybrid and had a sexual period stretching from summer
 305

to early winter. Polyarthra dolichoptera, in con- reported for rotifers, but there is some doubt
trast, preferred deep, cold water that was oxygen about their significance. Ruttner-Kolisko (1983)
poor and reproduced sexually in early spring. This described a 'series of mating experiments with
seasonal and ecological isolation was very effec- B. plicatilis where some interstrain crosses failed.
tive in maintaining species boundaries, as no She attributed this failure to differences in lorica
hybrids were ever detected in the lake. These thickness of newborn females and the males
species also co-occurred in tarns, small ponds 1-3 ability to penetrate the lorica with hyperdermic
meters deep that are abundant in the region. Here, insertion of the penis. If this accurately represents
P. vulgaris dominated, but intermediate forms copulation, it would be an example of mechanical
were common. Pejler argued that the environment reproductive isolation based on lorica thickness.
in the shallow tarns broke down the reproductive There is some doubt, however, whether males
isolation between these two species leading to penetrate the lorica since most copulations occur
hybridization. This example illustrates the impor- in the coronal region which is not covered by the
tance of the environment in maintaining species lorica (Gilbert, 1963; Snell & Hoff, 1987).
separation. Examples of post-mating reproductive iso-
Examples of habitat isolation can be found in lation in rotifers are not as abundant as pre-
various Brachionus species. Ruttner-Kolisko mating barriers. There is some evidence of hybrid
(1974) described the ecological characteristics of inviability in crosses among A. brightwelli,
several brachionid species in the uroeolaris group. A. sieboldi andA. intermedia (Gilbert et aI., 1979).
Brachionus uroeolaris is found primarily in fresh- King (1977) reported partial post-mating isolation
water where it is mainly benthic, often attaching among A. brightwelli populations collected at dif-
to submerged vegetation; B. plicatilis is found in ferent times from Lake Thonotosassa, Florida.
brackish waters and soda lakes where it is usually Four discrete populations were collected in 1974:
planktonic; and B. rubens is also found in fresh- April, June, September and November. The
water, often epizooic on Daphnia. The habitat hatchability of cysts produced from intrapopula-
differences of these three species promotes their tional crosses of was much higher than that of
reproductive isolation. Additional examples of interpopulational crosses.
habitat isolation among six species of Brachionus
are provided by Miracle et al. (1987). They
analyzed the distribution of these species using Mate recognition and species boundaries
discriminant analysis of 17 physical and chemical
parameters. Separation was achieved on the basis Mate recognition is a crucial behavior with wide
of sulphate, temperature, chloride, alkalinity, con- ranging evolutionary consequences. Mate recog-
ductivity and O 2 concentration, suggesting con- nition systems restrict gene exchange between
siderable habitat specialization by these brachio- species and promote divergence among popu-
nids. Studies like this and that of Bogdan & lations. Divergence in mate recognition systems
Gilbert (1987) suggest that rotifers finely partition can be an important component of speciation
the environment providing ample opportunity for (Paterson, 1982; Thornhill & Alcock, 1983;
habitat isolation. Giddings & Templeton, 1983; Nevo & Capranica,
Behavioral reproductive isolating mechanisms 1985; Ryan & Wilczynski, 1988). Incorrect recog-
are among the most important in animals nition leads to gamete wastage which is a serious
(Dobzhansky, 1970). Mate recognition in mono- error considering the small number of gametes
gononts and its role in the development of repro- produced by male rotifers (Snell & Hoff, 1987).
ductive isolation is an area of active research. Inviable or reduced fertility hybrids lower repro-
This work will be described in detail in the next ductive output and overall fitness. As a result,
section. strong selection for precise mate recognition sys-
Examples of mechanical isolation have been tems is expected.
306

Mate recognition in rotifers is based on coronal quantitatively comparing homogamic and hetero-
contact chemoreception by the male of a chemical gamic pairings (Snell & Hawkinson, 1983). The
on the body surface of females (Gilbert, 1963). absence of mating behavior is therefore sufficient
Mating begins when a male makes head-on con- evidence for reproductive isolation, but if mating
tact with a con specific female. Chemosensory occurs additional experiments are necessary to
receptors in the male's coronal region (Clement establish species boundaries.
et aI., 1983) sense a glycoprotein present on This approach to determining species
females (Snell et aI., 1988). If a male senses the boundaries is appealing because it relies on rotifer
correct glycoprotein, he begins copulatory behav- males rather than human systematists to discrimi-
ior by circling the female, skimming over her body nate species. Mate recognition is a natural bio-
while maintaining coronal contact. After several assay for species boundaries and can be carried
seconds of circling, the male moves toward the out quickly with high reliability. The protocol is
female's corona where he then inserts his penis simple and requires minimal equipment. The
into her pseudocoelom. The entire copulatory application of this bioassay to laboratory popu-
sequence takes an average of 1.2 minutes (Snell & lations should not be difficult because male pro-
Hoff, 1987). Males are smaller, faster swimming duction is common in many cultured species. The
than females and actively discriminate con specific mating bioassay will be most useful in sorting out
females. Females swim about randomly, taking sibling species where morphological criteria are
no apparent role in mate recognition or copula- not discriminating. Sibling species are a common
tion. problem in cladocera (Hebert, 1987) which are
Mate recognition behavior of male rotifers can sexually similar to rotifers. Another application
be a useful tool for establishing species will be in cases where cyclomorphosis confuses
boundaries. Males discriminate con specifics phenotypic variation. Although, the mate recogni-
using a precise biochemical mechanism that has tion bioassay is not a panacea, it is a useful tool
evolved over thousands of generations. This that, when properly applied, can uncover biologi-
recognition system can be used as a simple bio- cally meaningful discontinuities in rotifer diver-
assay to probe the dimensions of a species' gene sity.
pool and its boundaries with adjacent species. U sing mate recognition to discriminate rotifer
Mate recognition occurs at the instant of male- species is appealing for other reasons. Mate recog-
female contact. A con specific contact usually nition is a central element of the recognition con-
results in circling of the female by the male. cept of species (Paterson, 1985) and is gaining
Circling behavior is easily detected as it differs considerable attention. Specific mate-recognition
markedly from normal male swimming behavior. systems (SMRS) (Paterson 1978, 1980) are
A heterospecific contact results in males changing involved in signaling between mating partners and
direction and swimming away. Observation of are therefore a subset of the fertilization system.
50-100 encounters provides enough data to calcu- In Paterson's view, species boundaries are set by
late a probability of copulation {PC>, the liklihood the limits of shared fertilization systems, a primary
that males will initiate mating behavior upon con- component of which is the SMRS. Species are
tacting a female. If no mating attempts are genetically cohesive units that are resistent to
observed after a suitable number of encounters change (homeostatic) because of stabilizing selec-
(Pc = 0), reproductive isolation is assumed to be tion of the SMRS. Speciation occurs as an
complete between the populations tested. If Pc is incidental effect when the SMRS breaks down in
greater than 0, the species are potentially able to allopatry. In the case of rotifers, the limits of the
exchange genes, but further tests are necessary to SMRS can be defined experimentally by the mate
determine if mating is consumated or if post- recognition bioassay. The recognition concept of
mating barriers exist. Calculation of Pc allows species makes several predictions (Lambert &
detection of partial reproductive isolation by Paterson, 1983), many of which are testable with
a rotifer model.
 307

Mate recognition bioassays have been used to species be discriminated. Asplanchna males also
examine species boundaries in several brachionid often attempt to copulate with con specific males,
rotifers. Gilbert (1963) mated Brachionus calyci- a phenomenon that is not observed in brachio-
jlorus and B. angularis males with B. calyciflorus, nids. This further suggests that the mating
B. angularis, B. quadridentatus, Synchaeta sp. and reaction in Asplanchna may not be as specific as
Euchlanis sp. females. Males of these two species in brachionids.
attempted to mate only with con specific females
demonstrating the strict species specificity of the
mating reaction. Brachionus plicatilis males are Molecular aspects of mate recognition
also strictly species-specific in their mating
activity when exposed to Synchaeta bicomis The female's signal and the male's reception are
females (Snell & Hawkinson, 1983) or B. rubens the basis of mate recognition on rotifers. More
females (Snell, unpublished). Snell and Hawkin- knowledge of the chemical nature of the signal
son also examined mating reactions among tem- and how it varies among rotifer species will pro-
porally isolated B. plicatilis populations from the vide insight into how reproductive isolation
same bay as well as spatially and geographically develops.
separated populations. No differences in Pc were Gilbert (1963) showed that mate recognition in
detected among the temporally separated popu- rotifers is based on contact chemoreception and
lations. However, spatially and geographically provided initial biochemical observations of the
separated populations showed mating prefer- mate recognition factor on female B. calyciflorus.
ences. Then strains from around the world were U sing biochemical techniques considered crude
characterized for probabilities of copulation. Pc by today's standards, Gilbert concluded that the
values ranged from 78 % to 6.2 % in all possible mate recognition factor is a small, amphoteric
pairings. In no case was Pc = 0, indicating that all molecule less than 4000 daltons with aromatic
strains have retained their mating compatability characteristics. More recently, Snell et al. (1988)
despite wide geographical separation. This sug- examined the biochemical basis of mate recog-
gests that a globally distributed species like nition in B. plicatilis. They showed that females
B. plicatilis (Pejler, 1977b) can maintain an inte- heated to 100 0 C for 5 minutes lost their ability to
grated gene pool. Post-mating barriers need to be elicit a male mating reaction. In contrast, freeze-
investigated, but an intriguing question is raised. killed females retained their ability to elicit male
How is mate recognition maintained in geographi- mating activity at a level indistinguishable from
cally isolated populations separated for live females. Snell et al. (1988) also showed that
thousands of generations? Theoretical models of the mate recognition factor was destroyed by the
the evolution of reproductive isolation suggest proteases proteinase K, pronase E and chymo-
that this is sufficient time for the establishment of trypsin and the glycohydrolase p-amylase. These
reproductive barriers (Nei et al., 1983). Several observations clearly demonstrated that the mate
possible causes for the maintenance of species recognition factor is a heat labile glycoprotein and
integrity in the absence of gene flow have been that the carbohydrate portion of the molecule is
postulated (e.g. Hutchinson, 1968 ; Van Valen, necessary for mate recognition.
1982). The mate recognition glycoprotein (MRG) of
Strict species specificity of the mating reaction B. plicatilis females has been further characterized
may not be characteristic of all rotifers. In by Snell & Nacionales (1989a). They showed that
Asplanchna, Gilbert et al. (1979) report that male male mating activity is blocked by exposing
A. brightwelli, A. intermedia and A. sieboldi females to the lectin concanavalin A (con A) and
attempt to mate with females of all three species. a lectin isolated from the lentil Lens culinaris.
Only when post-mating barriers were examined, These lectins selectively bind to glycoproteins
along with six morphological traits, could these containing terminal-D-mannosyl, -D-glucosyl or
308

-N-acetylglucosamine residues. Their binding to Future research

the MRG suggests the presence of these residues
and their blocking of mating activity reinforces the In my view, there are several areas that would
conclusion that the carbohydrate portion of the benefit from a focused research effort. These
MRG is important in mate recognition. When include the following:
males are exposed to these lectins, mate recog- 1) There is more information to be extracted
nition also is blocked, suggesting that lectins bind from cross-mating experiments. Although
to the male receptor of the MRG, causing dis- these are tedious, they yield a large amount
ruption of mate recognition. of information. A better understanding of
Using fluoresence microscopy, localization of intraspecific variation in reproductive
the MRG on females and its receptor on males barriers is needed. Do intraspecific barriers
has been accomplished (Snell & Nacionales, to gene flow exist and how do they compare
1989b). Con A conjugated to the fluorochrome to interspecific barriers? Enough strains
FITC (fluoroisothiocyanate) produced fluo- within a single species should be tested to
resence at sites where Con A bound to the MRG. characterize these barriers. We need to
On females the MRG was highly localized in the clearly establish species boundaries for at
buccal field and corona, dorsal and lateral anten- least one model rotifer species using the cri-
nae, pedal sphincter and tip of the foot. The MRG terion of reproductive isolation. Brachionids
receptor on males is also highly localized in the are good candidates for this.
coronal region, dorsal and lateral antennae, tip of 2) The mate recognition bioassay should be
the foot, tip of the penis and the accessory gland. used as a rapid screening technique to
The sites of highest MRG concentration on examine species boundaries in as many
females correspond to the sites where the most rotifer species as possible. Closely related
copulations occur (Snell & Hoff 1987). It appears species in the families Brachionidae (genera
that the MRG guides males to a site on the like Epiphanes, Brachionus, Euchlanis) or
female's body surface that is favorable for pene- Synchaetidae (Synchaeta, Polyarthra) are
tration. good candidates because several species
The goal of research on the biochemical can be cultured, males are common and
features of mate recognition in rotifers is to better many species have been described. Several
understand the molecular basis of reproductive predictions of the recognition concept of
isolation. Since mate recognition is the corner- species could be tested.
stone of reproductive isolation, it is important to 3) More work is needed on mate recognition in
know how barriers to cross-mating develop. rotifers. Are glycoproteins the basis of
What kind of molecular changes cause reproduc- recognition phenomena in all species? What
tive isolation? Are the mutations responsible molecular changes in the carbohydrate
simple or complex? This could indicate how dif- moieties lead to reproductive isolation? Is it
ficult it is for behavioral reproductive isolation to possible to construct a phylogeny of mono-
evolve. Isolation and characterization of the gononts using molecular differences in mate
MRG will provide opportunities to develop recognition glycoproteins?
biochemically based, rapid screening assays of
reproductive isolation. Information like this will Acknowledgments
make it easier to sort out discontinuities in rotifer
diversity and provide better tools for reconstruct- I thank Mike Childress and Lisa Nacionales for
ing rotifer evolution. expert technical assistance. Bob Wallace made
several suggestions that improved the manuscript.
This material is based upon work supported by
the National Science Foundation under Grant
 309

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Holman, E. W., 1987. Recognizability of sexual and asexual Beitrag zur Fortpflanzungbiologie der heterogonen Rota-
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King, C. E. & T. W. Snell, 1980. Density-dependent sexual experiments in three distinct strains of Brachionus plicatilis
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adaptation in rotifers. Arch. Hydrobio!. Beih. 8: 187-201. frogs. Science 240: 1786-1788.
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Shull, A. F., 1915. Inheritance in Hydatina senta. IV. Charac- Snell, T. W. & M. A. Nacionales, 1989b. Localization of the
ters offemales and their parthenogenetic eggs. J. expo Zoo!. mate recognition glycoprotein on the rotifer Brachianus
18: 145-186. plicatilis. Submitted.
Snell, T. W. & c. A. Hawkinson, 1983. Behavioral reproduc- Templeton, A. R, 1987. Species and speciation. Evolution
tive isolation among populations of the rotifer Brachianus 41: 233-235.
plicatilis. Evolution 37: 1294-1305. Thornhill, R & 1. Alcock, 1983. The Evolution of Insect
Snell, T. W. & F. H. Hoff, 1987. Fertilization and male Mating Systems. Harvard University Press, Cambridge,
fertility in the rotifer Brachianus plicatilis. Hydrobiologia MA.
147: 329-334. Van Valen, L., 1982. Integration of species: Stasis and
Snell, T. W., M. J. Childress & B. C. Winkler, 1988. Charac- biogeography. Evo!. Theory 6: 99-112.
teristics of the mate recognition factor in the rotifer Wallace, R L. & R A. Colburn, 1989. Phylogenetic relation-
Brachianusplicatilis. Compo Biochem. Phys. 89A: 481-485. ships within the phylum Rotifera: orders and genus
Snell, T. W. & M. A. Nacionales, 1989a. Sex pheromone Nathalca. Hydrobiologia 186/187: 311-318.
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C. Ricci. T. W. Snell and C. E. King (eds). Rotifer Symposium V. 311
© 1989 Kluwer Academic Publishers.

Phylogenetic relationships within phylum Rotifera: orders and genus

Notholca

Robert Lee Wallace & Rebecca Arlene Colburn

Biology Department, Ripon College, Ripon, WI, 54971-0248, U.S.A.

Key words,' cladistics, computers, evolution, evolutionary trees, Notholca, orders, phylogeny, Rotifera,
synapomorphies

Abstract

We investigated evolutionary relationships among orders in phylum Rotifera and among species in genus
Notholca (Rotifera) by computing parsimonious cladograms. All of the most-parsimonious cladograms
generated for the ordinal level confrrm the view that class Monogononta, superclass Eurotatoria, and
phylum Rotifera are monophyletic. Species within the genus Notholca were separated into six groups
(clades), but some species have been defmed based on highly variable characters not reliably studied using
cladistics. Therefore, phenetic studies are warranted, especially for species possessing caudal processes.

Introduction Variation in body form and size has led to

confusion in the defmition of species in some
Cladograms (phylogenetic trees or networks) gen- genera. For example, the taxonomy of Notholca
erated with the aid of computer programs are has been described as hopelessly confused and
powerful tools for investigation of plausible evolu- chaotic, and the morphological variation as extra-
tionary relationships among organisms. However, ordinarily great (e.g., Bj6rklund, 1972; Koste &
to our knowledge these techniques have never Shiel, 1987; Pejler, 1977; Ruttner-Kolisko,
been applied to taxa within phylum Rotifera 1974:77). To help rectify this problem we initiated
below the level of class although they could help a cladistic analysis of the genus Notholca by devel-
elucidate difficult taxonomies: especially because oping parsimonious cladograms with the aid of
cladistical analyses focus on the evolution of several computer programs. Notholca was selected
shared derived characters (synapomorphies) among other genera that suffer from difficult tax-
(Ridley, 1986). onomies for two reasons. (1) Notholca was the
In general, distinctions among taxa within the subject of a detailed study in which 53 morpho-
Rotifera are well-defmed, leaving little ambiguity. logical characters plus seven ecological and
However, as Ruttner-Kolisko (1963) indicated, biogeographical features were analyzed (Kuti-
variation below the level of genus can be so great kova, 1980). (2) About 40 putative species and
that clear designation of some species has become subspecies have been described for this genus,
extremely difficult. This problem is due, in part, to thereby providing sufficient information for a
phenotypic variation (e.g., dietary- and predatory- cladistical study without the necessity of manipu-
induced polymorphisms, differences in resting egg lating an intractable data set. We began our study
hatchlings from subsequent generations, and by analyzing the phylum Rotifera at the ordinal
dwarfing). level as a simple test of our procedures.
312

Methods Results and discussion

At the ordinal level information about eight char- Phylogeny of rotiferan orders
acters (ovaries, vitellarium, males, resting eggs,
desiccation ability, type of trophi, spermato- Our phylogenetic analysis of rotifer orders pro-
phores, and prostate glands) was obtained from duced several equally parsimonious cladograms.
four sources (Edmondson, 1959; Gilbert, 1988; We selected one to display here to illustrate three
Koste, 1978; Ruttner-Kolisko, 1974). For 37 points (Fig. 1). (1) Three characters created some
Notholca species (including subspecies and varia- difficulty in our study (type oftrophi, presence or
tions) data on nine characters (four types of absence of vitellaria, and presence or absence of
spines, spurs, keels, ornamentation of the surface, prostate glands) because we were unsure how to
type of caudal process, and habitat) were obtained define their ancestral character states and/or
from 18 reports (Amren, 1964; Bjorklund, 1972; transformation types. Therefore, we decided to
Carlin, 1943; Chengalath, 1978; Chengalath & use an unordered transformation type for the tro-
Koste, 1987; Daday, 1913; Dartnall & Hollow- phi and report alternative positions on the clado-
day, 1985; Focke, 1961; Koste, 1974; Koste, gram where evolutionary events may have
1978; Koste& Shiel, 1987; Kutikova, 1980, 1986; occurred for the other two characters; neither
Lair & Koste, 1984; de Paggi, 1982; Pejler, 1977; compromise alters our basic conclusions.
Ruttner-Kolisko, 1972; Sternberger, 1976). (2) Treelength was independent of the transfor-
U sing these data, cladograms were calculated mation type used for all of the most parsimonious
by employing computer programs available in cladograms: 11 steps were required using either
PHYLIp© (Phylogeny Inference Package, v3.0) Wagner parsimony for all characters (except
which use numerical procedures that are applied trophi, unordered) or a combination of transfor-
to morphological data (character states), ordering mation types (Mix-I) based on assumptions of
them into the most parsimonious cladogram(s). how each character might evolve. (See the legend
Cladograms were then examined more closely of Fig. 1 for a list of these assumptions.) (3) No
(corroboration) for treelength and homoplasy characters demonstrated homoplasy using either
(Wiley, 1981 :12) using MacClade© (v2.1). For a Wagner or Mix-l parsimony.
review of PHYLIp© and MacClade© and infor- All available information was not used in con-
mation concerning their availability consult Fink structing this cladogram (e.g., certain ultrastruc-
(1986). During corroboration we amended our tural information, physical development of males
assumptions concerning possible evolution within class Monogononta, and presence or
among character states by using a combination of absence of sessile forms). However, inclusion of
the following transformation types: unordered such data does not significantly improve discrimi-
(u), any change in character state is possible nation among the five orders. Nevertheless, dif-
(Fitch parsimony); reversible (r), forward and ferences in trophi, coronae, and feeding types in
reverse changes may occur in binary characters orders Collothecaceae and Flosculariaceae sug-
(Wagner parsimony); Dollo (d), increases in state gests that the sessile condition has evolved twice,
may occur only once; irreversible (i), no losses are at least once in each order. In some forms, sub-
possible (Camin-Sokal parsimony) (Maddison & sequent evolution has led to planktonic existence
Maddison, 1987). All characters were weighted once again (e.g., Collotheca mutabilis, Ptygura
equally throughout both analyses. Copies of the libera).
data sets used in both analyses are available from Our analysis of the phylogeny of rotifer orders
the senior author. is very similar to one proposed by Epp & Lewis
(1979), except that their phylogeny emphasizes
different taxonomic levels within the Mono-
gononta. In contrast, Kutikova (1983) proposed
 313

Rotifera

Eurotatoria

Monogononta

CIS CIS
(I)
(I)
"0' "0
'2 CIS
0 :2 E
,!!2 Q)
'15
(I) "0
CJ) a:I a:::

6a -r-6e
(-8 )*

(-2)*-~ ~<--- 2

---T-~ Ancestral rotifer

pseudocoelomate; corona;
mastax with trophi; males
present; paired gonads;
vitellarium present (2*);
prostate glands present (8*)
Fig. 1. Cladogram of phylum Rotifera at the ordinal level. Numbers refer to changes in character state as gains or losses ( - ).
Numbers with asterisks (*) indicate alternative positioning of evolutionary events when the status of the ancestral character state
has not been ascertained. Characters and transformation types used (Mix-I) are as follows: I = paired gonads (i); 2 = vitellarium
(r); 3 = male (i); 4 = resting egg innovation (d); 5 = desiccation (r); 6 = trophi (u): a - aberrant, b - ramate, c - others, d -
malleoramate, e - uncinate; 7 = spermatophores (d); 8 = prostate glands (r). See text for an explanation of the transformation
types. (NB: Four steps are required for the evolution of the five types of trophi described here, assuming one is ancestral.)

a very different phylogeny based on use of the (Seisonidea, (Bdelloidea, (Ploima, (Floscu-
corona in locomotion and feeding, and direction lariaceae, Collothecaceae»); (eq. 2).
of ciliary metachronal movement. Kutikova's We suggest that our cladogram (Fig. 1, eq. 2) is
phylogeny may be represented using the standard more plausible for several reasons. It is based on
convention for tree representation as follows: a larger number of unequivocal anatomical
(Seisonidea, (Ploima, «Bdelloidea, Floscu- features and is more parsimonious: 11 (eq. 2)
lariaceae), Collothecaceae»); (eq. 1). verses 14 steps (eq. 1), using these eight charac-
Our view of phylogeny at this taxonomic level ters and the transformation type Mix-I. Even
(Fig. 1) differs significantly from Kutikova's view. using Wagner parsimony (except trophi, un-
Whereas Kutikova's tree (eq. 1) suggests that the ordered), eq. 1 requires 13 steps. Further, eq. 1
Bdelloidea and Flosculariaceae are sister groups, entails homoplasous evolution of both ovarian
ours places bdelloids more distant, thus: number and resting egg innovation and predicts
 w
.j:>.
-
.j:>.
~
....... / cinetura
~
1::0
;:::
\0
psammarina
~
haueri ~ / lamellifera
-:J
...til 0- n
b.. Pi"
::T foliacea ~ p. ~ ~ lapponica
.'"
o· ~.
gaigalasi ~ .;:.. I\>
::I
~ niL
s:.
- n labis
~ corn uta "'1::0 caudata 1::0
~
<= n ;:::
<:l Vl
;;.
~
intermedia Pi"
p.
~ .;:..
lyrata
A;1koZhoVi
~acundnata
;s
;;.
.;:.. 1::0
Q
" I\> ~~
~
I -:J
n
---
'-'
J '\.. I\>
manna

caudata clade ./ beta

./ ~ I~
'" latistyla
~ ~ acuminata - labis clade
0
en LJ ~ orbiculata
co I
triarthroides clade .;:.. I~
PI' ..., '-'
(12
jugosa 0.1::) I ~ verae
;:::
e:
!!. squamula
1::0
;s
~
;::: . / ..j"' walterkostei
:0
00 Q "-
-.I

0 n
- Pi"
p.
.::: ~
&; / )l j. japonica
'-' .;:..
p.
laurentiae ---l
'" ....
~.
salina ..., balkalensis
-
....
....
~.
- :....:-
-<::)
st. striata ~ jasnitzkii
~
n triarthroides "'...,
liepetterseni Pi"
p.
~
angulata
st. bipalium I~
olchonensis
 315

polyphyly for class Monogononta. Based on the cladogram over other equally parsimonious ones
number of evolutionary steps required and the because it separated species into clades of 4-13
lack of homoplasy, our analysis provides strong taxa, a size convient for further comparisons.
evidence for monogonont monophyly. Suprisingly, we found that the first four clades can
The cladogram shown in figure 1 is only one of be combined into a single larger one or separated
several most parsimonious cladograms possible: further without increasing treelength or homo-
minor variation in placement of monogonont plasy. These clades are defined based on a few
orders within that clade is possible without important characters, including presence or
increasing the number of evolutionary steps. Fur- absence and type of lateral spines (movable or
ther differentiation among cladograms will not be rigid) and presence or absence and type of caudal
possible without additional information, espe- processes. Other characters that were useful in
cially of the type provided by ultrastructural separating these clades included number of ante-
research (e.g., Clement, 1980, 1985). Presence of rior spines and habitat (fresh- or salt water).
a unique syncytial epidermis comprised of a intra- Characters concerning size and shape of anterior
cytoplasmic lamina (Clement, 1985; Lorenzen, spines were much too variable to be of value in
1985) suggests a relationship between the Rotifera this study and therefore were not included.
and the Acanthocephala, perhaps with the Three synapomorphies were congruent and
acanthocephalan line diverging before the easily traceable: movable and rigid, lateral spines
ancestral rotifer. However, we disagree with (striata and triarthroides clades, respectively), and
Lorenzen's view that the lemnisci and proboscis reduced number of anterior spines (i.e.,japonica).
of acanthocephalans are synapomorphic with However, homoplasy was found in the charac-
seemingly similar structures in bdelloids, and that teristic of inhabiting salt water, indicating multiple
acanthocephalans and bdelloids are sister groups. invasions of the marine habitat (Fig. 2).
Two characters clearly require greater study
before they may be resolved. Both type of caudal
Phylogeny of Notholca process and ornamentation on the surface of the
lorica routinely are provided in descriptions of
U sing a user-defined transformational type species, but neither have been defined precisely.
(Mix-2), our analysis produced several equally Once these characters have been examined more
parsimonious cladograms for the 37 putative critically, cladistic studies may eventually prove
species of Notholca examined here (Fig. 2). This useful in separating species possessing them.
cladogram separates the taxa into six groups of However, in those species possessing a caudal
species or clades, named for prominent mem- process misintrepretation of exuberent morpho-
ber(s) contained in each: four clades possessing types for solid species, especially in the acuminata
caudal processes (Joliacea, caudata, acuminata - -labis clade, should not be overlooked as a possi-
labis, triarthroides), and two lacking caudal ble explanation for some forms.
processes (squamula, striata). We selected this Differences in the phenetic characters of size

Fig. 2. Cladogram of the genus Notholca. Numbers refer to changes in character state as gains or losses ( - ). Numbers with
asterics (*) indicate alternative positioning of evolutionary events when the status of the ancestral character state has not been
ascertained. Characters and transformation types used (Mix-2) are as follows: 1 = moveable lateral spines (d); 2 = rigid lateral
spines (d); 3 = central spur (d); 4 = caudal processes (u): a - variable spatulate, b - thin, c - broad, d - forked, e - crescent,
f - moveable,? - unknown; 5 = six anterior spines (i); 6 = keeled (d); 7 = lorica surface (u): a - stripes, b - dotty, c - granular,
d - ridges, e - cup-like, f - smooth; 8 = ventral plate appendage (d); 9 = marine (r). See text for an explanation of the
transformation types. Polymorphic forms for character states within characters 4 and 7 are present in some taxa (e.g., kostei
possesses both characters 7a and b). Taxa above open rectangles are separated based on characters best studied using phenetic
analysis.
316

and shape of the body or spines account for the senario is correct, then invasion of the marine
separation of 19 taxa, especially species within the environment took place in these forms before the
acuminata - labis, squamula, and striata clades evolution of moveable, lateral spines by members
(Fig. 2). Because these characteristics do not lend of the striata clade. Placement of cornuta in the
themselves to cladistic analysis, a detailed triarthroides clade and angulata in the acuminata -
phenetic analysis is required before these species labis clade (Kutikova, 1980) requires two or more
may be assigned a permanent position. However, evolutionary steps and a concomitant increase in
size and shape may vary according to nutritional homoplasy.
status and season (Bjorklund, 1972), therefore, We do not suggest that all species shown in
the examinations of a few samples from natural Figure 2 are 'good' species, rather our analysis
populations may lead to improper separation of suggests several places where additional study
species. Any phenetic study must examine speci- might lead to combining several taxa or separating
mens collected throughout the year and should them. For example, acuminata and marina differ
include populations from laboratory cultures only in habitat and development of the caudal
grown under a wide variety of conditions. process (see Bjorklund, 1972). Thus, the funda-
Several authors have separated Notholca into mental question remains; to what extent do
groups comprising various species (Bjorklund, species within the various subgroups represent
1972; Koste, 1978; Kutikova, 1980; Ruttner- true species, incipient speciation within a broadly
Kolisko, 1974), but each has emphasized slightly distributed population, or simply phenotypic
different criteria, the most frequently used being responses to different environments (i.e., eco-
type of caudal process and lateral spines, and size types)? Further differentiation of the Notholca will
and shape ofthe lorica. Our analysis is not directly not be possible without additional information,
comparable to any of these investigations, as none especially concerning the caudal processes, sur-
of them attempted a phylogenetic study, but a face ornamentation, and size and shape of the
general comparison shows that we differ signifi- lorica. Data concerning interspecific mating will
cantly from these other works only in the place- be exceedingly important, if sexuality can be
ment of six species. Koste (1978) and Ruttner- induced in laboratory cultures [e.g., Snell, in this
Kolisko (1974) place psammarina in the striata volume]. Finally, little information is available on
clade and Kutikova (1980) places verae in the the trophi of Notholca species. This inadequacy
squamula clade. Both relationships are possible also must be rectified before significant progress
without increasing the number of evolutionary may be made in clearing up confusion within this
steps required or homoplasy. However, we genus.
arbitrarily elected to place psammarina in the
caudata clade to emphasize the presence of a
caudal process in psammarina, and we placed Conclusions
verae in the acuminata - labis clade to stress the
putative relationship between verae and walter- The computer programs used in this study can
kostei (Dartnall & Hollowday, 1985). Koste greatly expedite phylogenetic analyses, so that
(1978), Kutikova (1980), and Ruttner-Kolisko relationships among rotifers may be examined
(1974) place lapponica in the squamula clade, but using cladistic analysis even with a recondite data
this requires an extra evolutionary step. The salt set containing more than 100 elements. However,
water form salina is routinely considered to be species with great morphological variability are
related to squamula (e.g., Bjorklund, 1972), but not well resolved with cladistics. Further, such
surprisingly an extra evolutionary step is required efforts are only as good as the data available.
for it to be placed there. Placement of salina as a Therefore, we recommend that taxonomists pay
sister species within the striata clade, therefore, is close attention to the suggestions of Ruttner-
more parsimonious and less homoplasous. If this Kolisko (1963) concerning rotifer taxonomy and
 317

we restate them here in a slightly revised form. References

(1) Formal descriptions of new species should not Amren, H., 1964. Ecological and taxonomical studies on
be based on fragmentary information of' ... a few zooplankton from Spitsbergen. Zool. Bidrag. Uppsala Bd
individuals ... ' or the' ... naming of morphological 36: 209-276.
Bjorklund, B. G., 1972. Taxonomic and ecological studies of
pecularities', or even morphological analysis of species of Notholca (Rotatoria) found in sea- and brackish
specimens from a single habitat. Such practices water, with description of a new species. Sarsia 51: 25-66.
only serve to obfuscate, not elucidate taxonomic Carlin, B., 1943. Die Planktonrotatorien des Motalastrom.
relationships. (2) Detailed statistical analyses of Zur Taxonomie und Okologie der Planktonrotatorien.
morphological characteristics ought to be pro- Medd. Lunds Univ. Limnol. Instn. 5: 1-256.
Chengalath, R., 1978. A new species of the genus Notholca
vided along with taxonomic descriptions when- Gosse, 1886, (Brachionidae: Rotifera), from Great Slave
ever appropriate (i.e., phenetic analyses [e.g., Lake, N.W.T. Can. J. Zool. 56: 363-364.
Bjorklund, 1972; Sternberger, 1976]). (3) If at all Chengalath, R. & W. Koste, 1987. Rotifera from North-
possible, several typical specimens from each western Canada. Hydrobiologia 147: 49-56.
season should be cultured under a wide variety of Clement, P., 1980. Phylogenetic relationships of rotifers, as
derived from photoreceptor morphology and other ultra-
conditions to define the developmental plasticity structural analyses. Hydrobiologia 73: 93-117.
of the species in question. (4) Where possible Clement, P., 1985. The relationships ofrotifers. In S. Conway
physiological, ecological, and genetic par<l:illeters Morris,J. D. George & H. M. Platt (eds.), The Origins and
must be investigated along with morphological Relationships of Lower Invertebrates. System. Assoc. Vol-
ume 28. Clarendon Press, Oxford, U.K.: 224-247.
ones. Finally, although we may be accused of
Daday, E., 1913. Beitrage zur Kenntnis der Mikrofauna des
adopting a reductionist position, we suggest that Kossogol-Beckens in der Nordwestlichen Mongolei.
studies of form and function (i.e., functional mor- Mathmatische und naturwissenschaftliche Berichte aus
phology) be undertaken with the goal of better Ungarn 26: 274-360.
understanding the structural significance of the Dartnall, H. J. G. & E. D. Hollowday, 1985. Antarctic
characters used in species designation. rotifers. Brit. Antarctic Sur. Sci. Reports 100: 1-46.
Edmondson, W. T., 1959. Rotifera. In W. T. Edmondson
(ed.), Fresh-Water Biology, 2nd ed., John Wiley & Sons,
Inc., N.Y.: 420-494.
Acknowledgements Epp, R. W. & W. M. Lewis, Jr., 1979. Sexual dimorphism in
Brachionus plicatilis (Rotifera): evolutionary and adaptive
significance. Evolution 33: 919-928.
We wish to thank Dr. Joe Felsenstein (Depart-
Fink, W. L., 1986. Microcomputers and phylogenetic
ment of Genetics, University of Washington) who analysis. Science 234: 1135-1139.
supplied us with a copy of PHYLIp© and Drs. Focke, E., 1961. Die Rotatoriengattung Notholca und ihr
Wayne and David Maddison (Museum of Com- Verhalten im Salzwasser. Kieler Meeresforschungen 17:
parative Zoology, Harvard University) who 109-205.
Gilbert, J. J., 1988. Rotifera. In K. G. & R. G. Adiyodi (eds.),
supplied us with a copy of MacClade©, version
Reproductive Biology ofInvertebrates, Volume III. Acces-
2.1. We also thank Drs. Terry Snell, George sory Sex Glands. Oxford & IBH Publ., Co., New Delhi.:
Wittler, and William Brooks who read and 73-80.
improved this manuscript. Help from Oussama Koste, W., 1974. Das Radertier-Portrat. Die Radertier-
El-Hilali, Larry Fahnoe, Mike Post, Tom Oyster gattung Notholca. Mikrokosmos 63: 48-52.
Koste, W., 1978. Rotatoria. Die Radertier mitteleuropas. 2
(Computer Services, Ripon College), especially
vols, Gebriider Borntraeger, Berlin-Stuttgart, 673 pp, 234
their loan of a Macintosh Plus™, was greatfully plates.
appreciated. This work was supported, in part, by Koste, W. & R. J. Shiel, 1987. Rotifera from Australian
a grant from Ripon College funds for Faculty Inland Waters. II. Epiphanidae and Brachionidae
Development. (Rotifera: Monogononta). Invertebr. Taxon. 7: 949-1021.
Kutikova, L. A., 1980. On the evolutionary pathways of
speciation in the genus Notholca. Hydrobiologia 73:
215-220.
Kutikova, L. A., 1983. Parallelism in the evolution ofrotifers.
Hydrobiologia 104: 3-7.
318

Kutikova, L. A., 1986. Taxonomic review ofthe rotifer fauna Pejler, B., 1977. On the global distribution of the family
of the Lake Baikal. USSR Acad. Sci. Proc. Zool. Instit. Brachionidae (Rotatoria). Archiv fUr Hydrobiologie
152: 89-105. (in Russian, with English summary) (Suppl.) 53: 255-306.
Lair, N. & W. Koste, 1984. The rotifer fauna and population Ridley, M., 1986. Evolution and Classification. Longman,
dynamics of Lake Studer 2 (Kergule1en Archipelago) with New York, 201 pp.
description of Filinia terminalis kergueleniensis n.ssp. and a Ruttner-Kolisko, A., 1963. The interrelationships of the
new record of Keratella san eta Russel 1944. Hydrobiologia Rotatoria. E. C. Dougherty (ed.), The Lower Metazoa,
108: 57-64. Comparative Biology and Phylogeny. Univ., California
Lorenzen, S., 1985. Phylogenetic aspects of pseudo coelom ate Press, Berkeley, CA.: 263-272.
evolution. In S. Conway Morris, J. D. George & H. M. Ruttner-Kolisko, A., 1974. Planktonic rotifers biology and
Platt (eds.), The Origins and Relationships of Lower taxonomy. Binnengewasser (Suppl.) 26: 1-146.
Invertebrates. System. Assoc. Volume 28. Clarendon Snell, T. W., 1989. Systematics, reproductive isolation and
Press, Oxford, u.K.: 210-223. species boundaries in monogonont rotifers. Hydrobiologia
Maddison, W. P. & D. R. Maddison, 1987. MacClade, (in this volume).
version 2.1. An interactive, graphical program for analyz- Sternberger, R. S., 1976. Notholea laurentiae and N. miehi-
ing phylogenies and studying character evolution. Harvard ganensis, new rotifers from the Laurentian Great Lakes
Univ., Cambridge, MA. (distributed by the authors). region. 1. Fish. Res. Bd Can. 33: 2814-2818.
de Paggi, S. B. J., 1982. Notholea walterkostei sp. nov. y otros Wiley, E.O., Phylogenetics: the Theory and Practice of
Rotiferos dulceacuicolas de la Peninsula Potter, Isla 25 de Phylogenetic Systematics. John Wiley and Sons, NY, 439
Mayo (Shetland del sur, Antartida). Rev. Asoc. Cienc. pp.
Nat. Litoral 13: 81-95.
Hydrobiologia 186/187: 319-324, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 319
© 1989 Kluwer Academic Publishers.

A re-appraisal of two members of the genus Notholca from the Andes,

with a note on the fine structure of the lorica of N. foliacea (Ehrenberg)

Eric D. Hollowday 1 & Charles G. Hussey2

145, Manor Road, Aylesbury, Buckinghamshire HP20 1JB, England; 2 British Museum (Natural History),
Cromwell Rd., London SW7 5BD, England

Key words: Rotifera, Notholca, fme structure, Andes, South America, taxonomy

Abstract

Rotifers described from the Andes by Murray (1913) and De Beauchamp (1939) as Notholcafoliacea
(Ehrenberg) are reviewed and re-assessed as Notholca walterkostei De Paggi, 1982. They are compared
with N. foliacea and details of the lorica of this species seen with the scanning electron microscope are
presented.

Introduction Materials and methods

A rotifer collected in 1912 from Lake Titicaca, In addition to the published literature, the follow-
Peru/Bolivia, was described by Murray (1913) as ing material was examined:
Notholca foliacea (Ehrenberg, 1838). In 1937 the (1) A permanent slide preparation labelled
Percy Sladen Trust Expedition to Lake Titicaca 'Notholca foliacea (Ehr.) sensu Murray. Exped.
brought back a Notholca from Lagunilla Titicaca 225/4. Lag. Saracocha 4.1X.37.' in the
Saracocha, a lake 80 km east of Lake Titicaca at collections of the British Museum (Natural His-
an altitude of 4130 m. This single specimen, virtu- tory), London. Registration No. 1940.4.1.6.
ally an empty lorica, was briefly described, (2) Living and preserved specimens of
without a figure, by De Beauchamp (1939), also N. foliacea from Aylesbury, Buckinghamshire.
as N. foliacea. He noted its similarity to Murray's The preserved specimens were stored in 2 %
material but expressed uncertainty regarding its formalin and mounted unstained. Light micro-
true identity. scopy included use of differential interference
De Beauchamp's specimen has been located by contrast (Nomarski). Specimens of N.foliacea
the senior author in the collections of the British examined by scanning electron microscopy were
Museum (Natural History). With access to this dehydrated in acetone, critical point dried, coated
material and the more recent literature on with gold/palladium and viewed in a Hitachi S800
Notholca, we are in a position to redetermine these Field Emission Scanning Microscope operated at
speCImens. an accelerating voltage of 8kv.
320

Systematic section dorsal midline slope down to the lateral margins,

but in N. foliacea this is accentuated by the occur-
Both Murray and De Beauchamp appear to have rence of a three-ridged median dorsal crest, the
identified their specimens with N. foliacea outer pair of which arise on the dorsal surface of
because of the presence of numerous small circu- the antero-dorsal median spines, converging
lar depressions or fossettes on the dorsal plate. In slightly below these spines and then proceeding
considering the taxonomic status of the Titicaca along a fairly parallel course until a little over half
and Saracocha specimens, it is necessary to con- way along the dorsal plate, almost level with the
sider this, and other, characteristics of N. foliacea. apertures of the lateral antennae. Depending on
the degree of compression or crumpling of the
lorica, these outer ridges may then unite to form
Taxonomic position of N. foliacea
a single ridge extending as far as the base of the
postero-dorsal spine, or they may remain separate
We wish to endorse the current view that this
as far as the base of this spine (Fig. la). From the
species belongs in the genus Notholca. Gillard
margin of the antero-dorsal median sulcus, two
(1948) erected a new genus, Argonotholca, for this
more ridges arise (Fig. lc), uniting about 10 flm
species, chiefly because of the presence of a ven-
below the sulcal margin, and running between the
tral keel and a median dorsal ridge. The former
two outer ridges as far as the point at which these
structure is not however, a permanent feature. It
widen, and then unite just below the median
appears when the flexible posterior portion of the
transverse line.
ventral plate expands to allow for the displace-
Examination with the scanning electron micro-
ment of the internal organs when the head is
scope shows that the three ridges forming the
retracted within the lorica. When swimming and
median crest consist of closely-packed smoothly-
feeding, the ventral plate appears little different, if
rounded tubercles, varying in size from approxi-
at all, to that of other species we have observed
mately 0.4-0.8 flm (Fig. lc). Extensive areas on
alive.
each side of the median dorsal crest are free of
In species such as N. squamula (Milller, 1786),
longitudinal ridges, but have long been known to
N. labis Gosse, 1887, N. bipalium (Milller, 1786)
be decorated with randomly scattered pimples.
and, to a slightly lesser extent, N. acuminata
SEM examination shows these also to be
(Ehrenberg, 1832) such displacement is accom-
smoothly-rounded tubercles of similar size to
modated by the bilateral expansion of the lorica.
those forming the median dorsal crest. They also
It would seem that N. foliacea, together with other
occur, more sparingly, on the ventral plate.
species which exhibit a postero-ventral keel, (i.e.
Examination of these areas between the median
N. walterkostei De Paggi, 1982; N. verae Kuti-
and lateral ridges of the dorsal plate with the light
kova, 1958), lack this bilateral flexibility. Unless
microscope at high magnification, using
preserved specimens have died with the head fully
brightfield illumination, reveals what appears to
extended (as does happen on very rare occasions),
be a finely reticulate pattern. SEM examination
such material will invariably exhibit this so-called
shows this pattern to consist of very minute
'keel'. We have not seen living material of
dimples or fossettes varying in diameter from
N. walterkostei or of N. verae but believe our
approximately 0.8-1.75 flm (Fig. 1d). In addition,
remarks on this phenomenon also apply to these
the whole surface of the lorica, both dorsal and
speCIes.
ventral, is densely covered with extremely small
tubercles no larger than 0.1 flm (Fig. Id).
Aspects of the structure of N. foliacea Towards the lateral margins on each side of the
median dorsal crest are at least four dorso-lateral
Most members of the genus are strongly convex longitudinal ridges, two and sometimes three of
in cross-section. The areas on each side of the which can usually be seen from the dorsal aspect
 321

Ia

(b)

Fig. 1. Notholca foliacea (Ehrenberg). (a) Dorsal aspect (drawn from Aylesbury specimen); (b) Entire animal, dorsal aspect,
S.E.M. x 500; (c) Antero-dorsal area of lorica S.E.M. x 2000; (d) Small fossettes and tubercles. S.E.M. x 12000.
322

ID

[ IOpm

Fig. 2. Notholca walterkostei De Paggi, from L. Saracocha. (a) Dorsal aspect; (b) Ventral aspect; (c) Left ventro-Iateral aspect.
(d) Postero-dorsal extension, detail of base, dorsal aspect; (e) Postero-dorsal extension, ventral aspect. (f) Shallow fossettes on
anterior of ventral plate. (g) Notholca walterkostei from Lake Titicaca. (after Murray 1913); (h) N. walterkostei from South
Shetland. (Unpublished drawing by Walter Koste). Note: Figs. 2a-f drawn from British Museum (Nat. Rist.). slide Figs. 2e and
f share common scale.
 323

(Fig. la), and these also consist of unbroken rows also be assigned to N. walterkostei. There are,
of rounded tubercles, like those forming the however, some peculiarities that may make it a
median dorsal crest. The ventral plate has at least distinct form or even subspecies, although this
eight longitudinal ridges, although these might cannot be determined on the basis of a single
more accurately be described as pleats, the edges specimen. The arrangement of depressions, or
of which are furnished with tubercles, more fossettes, is unique. They extend backwards along
widely spaced than those forming the dorsal and the dorsal surface, in decreasing size, from the
dorso-Iateral ridges. It lacks the extensive dim- anterior dorsal margin for nearly a quarter of the
pling so characteristic of the dorsal plate. length of the dorsal plate and re-occur on the
Although it has not been possible to make a posterior tenth of the dorsal plate, continuing, in
comparative SEM study of the Andean speci- reduced size, on all surfaces of the posterior exten-
mens, it would appear, on the basis oflight micro- sion and some way along the ventral surface of the
scopy alone, that neither can be considered to dorsal plate. They also occur on the antero-dorsal
belong to N. foliacea. median spines. These spines are long (34 ,um) and
are separated by a deep sulcus. The intermedian
spines are obliterate and the dorso-Iateral spines
Titicaca specimens are approximately 17 ,um long. There is no median
dorsal crest but at least sixteen longitudinal
From Murray's figure (Fig. 2g), one can see that ridges, similar to N walterkostei. The posterior
the antero-dorsal intermediate spines are almost extension is a bluntly terminated narrow tube,
obliterate and although he shows the two central 23 ,um long. Total length is 200 ,um and maximum
longitudinal ridges uniting in the posterior half of width is 75 ,um.
the dorsal plate, there is no central ridge com-
parable with the crest of N foliacea. Instead, the
whole of the dorsal plate is ridged, with no rela- Discussion
tively smooth areas between the median longitudi-
nal line and the lateral margins. The posterior Koste & De Paggi (1982), in their checklist of
prolongation of the dorsal plate is widened and Monogononta from Neotropis, list De Beau-
flattened at its extremity and is spatula-like rather champ'S record from L. Saracocha as Nfoliacea
than a spindle-shaped spine as in N foliacea. without comment and do not include any records
Most strikingly, the grooves between the longi- from Murray (1913). N walterkostei was de-
tudinal ridges are pitted with rows oflarge circular scribed by De Paggi (1982) from the South
fossettes extending the whole length of the dorsal Shetland Islands. It is also known from the South
plate and along its posterior extension. Orkney Islands (Dartnall, 1983 - as Notholca sp.),
All these features agree with N. walterkostei De South Georgia (Dartnall & Hollowday, 1985) and
Paggi, 1982. It is unfortunate that Murray pro- as far south as Alexander Island off the
vides no indication of scale. There are slight dif- Grahamland Peninsular (Hollowday, unpub-
ferences in the shape of the postero-dorsal exten- lished). A subspecies, N. walterkostei reducta
sion and in the length of the antero-dorsal median Dartnall & Hollowday, 1985, occupies arestricted
spines, but we have no hesitation in assigning distribution on the South Orkney Islands.
Murray's material to this species. Our redetermination of the Andean specimens
therefore considerably extends the known range
of N walterkostei. Assuming a Laurasian origin
Saracocha specimen for the genus (Dumont, 1983), and a gradual
southerly migration into South America after that
This specimen differs from N foliacea in much the continent had attained its present position, it is
same details as the Titicaca material and should not difficult to envisage a further southward
324

movement of N. walterkostei. Although the References

Andean specimens do exhibit some variation with
regards to N. walterkostei sensu stricto, it is not Dartnall, H. J. G., 1983. Rotifers of the Antarctic and sub-
antarctic. Hydrobiologia 104: 57-60.
unreasonable to suppose that they may have had
Dartnall, H. J. G. & E. D. Hollowday, 1985. Antarctic
a common origin. Even so, parallelism cannot be rotifers. Scient. Rep. Br. Antarct. Surv. 100: 1-46.
entirely ruled out (Kutikova, 1983). There is a De Beauchamp, P., 1939. The Percy Sladen Trust Expedition
striking resemblance between the postero-dorsal to Lake Titicaca in 1937. V. Rotiferes et turbellaries. Trans.
extension of Notholca lamellifera jashnovi Kuti- Linn. Soc. Lond. 3rd ser., 1(1): 51-79.
De Paggi, S. 1., 1982. Notholca walterkostei sp. nov. y otros
kova, 1986 from Lake Baikal and that of
rotiferos dulceacuicolas de la Pensinsula Potter, isla 25 de
N. walterkostei. Mayo (Shetland des Sur, Antartida). Rev. Asoc. Cienc.
Another Antarctic species, N. verae Kutikova, nat. litoral 13: 81-95.
1958, with its apparently bilaterally rigid dorsal De Paggi, S. J. & W. Koste, 1984. Checklist of the rotifers
plate and postero-ventral flexibility seems to recorded from Antarctic and Subantarctic areas. Sencken-
have affinities with the group, especially the sub- bergiana bioI. 65: 169-178.
Dumont, H. J., 1983. Biogeography ofrotifers. Hydrobiologia
species reducta, wherein the postero-dorsal exten- 104: 19-30.
sion is almost obliterate and the antero-dorsal Gillard, A. A. M., 1948. De Brachionidae (Rotatoria) van
spines exceedingly short. Kutikova (1980) tends Belgie met Beschouwingen over de Taxonomie van de
to associate N. verae with the N. squamula species Familie. Natuurwet. Tijdschr. 30: 159-218.
group but a recent analysis by Wallace & Colburn Koste, W. & S. 1. De Paggi, 1982. Rotifera of the Super Order
Monogononta recorded from Neotropis. Gewass. Abwass.
(this volume) suggests a relationship between
68/69: 71-102.
N. verae and N. walterkostei. Furthermore, their Kutikova, L. A., 1980. On the evolutionary pathways of
work separates N. foliacea and N. walterkostei as speciation in the genus Notholca. Hydrobiologia 73:
occurring in different clades. 215-220.
Kutikova, L. A., 1983. Parallelism in the evolution ofrotifers.
Hydrobiologia 104: 3-7.
Murray, J., 1913. Notes on the natural history of Bolivia and
Acknowledgements Peru. Scottish Oceanographical Laboratory, Edinburgh,
45 pp.
Wallace, R. L. & R. A. Colburn, 1989. Phylogenetic relation-
We wish to thank Dr. Walter Koste for per- ships within the phylum Rotifera: orders and genus
mission to use his unpublished drawing of Notholca. Hydrobiologia 186/187: 311-318.
Notholca walterkostei De Paggi (Fig. 2h).
Hydrobiologia 186/187: 325-330, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 325
© 1989 Kluwer Academic Publishers.

Protein patterns in rotifers: the timing of aging

Maria Jose Carmona, Manuel Serra & Maria Rosa Miracle

Departamento de Ecologia, Facultad de Biologia, Universidad de Valencia, 46100-Burjasot, Valencia,
Spain

Key words: aging, rotifers, protein electroforetic patterns, individual growth

Abstract

Single rotifer individuals have been characterized biochemically to obtain a fmgerprint of their physio-
logical state using a modified ultrasensitive silver-stain procedure to detect total proteins in poly-
acrylamide gels. Patterns are completely uniform for young isogenic individuals raised in the same culture,
but they start to change when these individuals reach a certain age. This age is close to the mean lifespan
and to both the cessation of body growth and reproduction. Variability is greatest among individuals of
the same chronological age, thus the rate of aging is different even among individuals having identical
genotypes and experiencing the same environment.

Introduction cally the different ages, except for a few data on

the age-variation of certain enzyme activities
Aging is the age-dependent acquisition of altera- (Fanestil & Barrows, 1965; Meadow & Barrows,
tions in both morphology and physiology. It is a 1971). The aim of this paper is to obtain a charac-
complex problem which can be more easily inves- terization of aging through the study of protein
tigated by studying the lower metazoa. Rotifers patterns.
are ideal for this kind of study because they are
simple eutelic organisms with a few cells, a short
lifespan, and parthenogenetic reproduction. High Methods
numbers of isogenic individuals can be obtained
for experimental purposes. For many years A clone CD of Brachionus plicatilis (Serra &
rotiferologists have observed aging effects, like the Miracle, 1983 and 1985) was cultivated since
maternal age effect on offpring senescence 1981 in our laboratory at 25°C, 12 ppt salinity,
reported by Jennings & Lynch (1928) and Lansing under continuous illumination (PAR: approx.
(1947, 1948). These studies have stimulated a 35 JlE/m2 s), and fed with Tetraselmis sp. Experi-
large number of investigations, not only on rotifers ments were initiated by collecting 500 ovigerous
(reviewed by King, 1969, 1980 and 1983), but also parthenogenetic females from log phase cultures.
on other organisms (reviewed by Lints, 1978). These females produced 430 newborn females
Most rotifer aging studies have been based on which were isolated in individual glass wells and
laboratory popUlation dynamics and almost no transfered daily into fresh medium with 5.3 x 105
attempt has been made to characterize biochemi- Tetraselmis cells ml- 1. Twenty F 1 females, all
 326

parthenogenetic, were collected at random 2, 4, 6, protein dissolving solution containing SDS and
8, and 9 days after birth. A total of 100 individuals mercaptoethanol (Semancik, 1976). The samples,
were collected, and the proteins of each were once checked for complete dissolution, were col-
separated electrophoretic ally. This procedure was lected and boiled for 5 minutes. Molecular weight
repeated twice over one year. protein markers were treated in the same way.
For protein extraction, single individuals of Proteins were separated by vertical SDS-discon-
B. plicatilis were rinsed with diluted sea water tinuous polyacrylamide gel electrophoresis on a
(12 ppt salinity), separated from their eggs, and 1 mm gel. Gels made according to established
isolated in 5 ,ul of this water on a hydrophobic procedures (Conejero & Semancik, 1977) had the
surface. Rotifers were then treated with 20 ,ul of a following acrylamide concentrations: 4 % (pH:

2 DAYS 4 DAYS

66

45
36
29
24
20.1
I I

14
j I

8 DAYS 6 DAYS

66

45
36
29

24
20.1 -
'I I I
I
I
14 I. I II
Fig. 1. Protein patterns of Brachionus plicatilis (clone CU at 25 C at 12 % salinity). Molecular weight protein markers are shown
0

at the left.
 327

y 0 8), 6.5% (pH: 8.8) and 14% (pH: 8.8) in the

n Y 0
1
stacking, spacer and resolution gels, respectively.
2 - 1 0 Since an ultrasensitive method is required to
3 -
4 -
4
4 detect the proteins of a single rotifer, an improved
5 - 5 1 silver stain technique was applied, after pre-
6 -
7 -
4
3
staining with Coomassie Brilliant Blue. We have
8 - 4 1 modified the ultrasensitive silver stain procedure
9 - 3 1 described by Oakley et al. (1980) to obtain both
66 --- 10
11
-
-
4 1
1 0 lower levels of background and maximum
12 - 1 0
staining sensitivity. Proteins were fixed overnight
with a solution of trichloroacetic acid (12.5%,
w/v) and isopropyl alcohol (25%, v/v). It was
===~
=== ~~ pre-stained in 5 %Coomassie Brilliant Blue over-
night and then washed with a solution of 10 %
===~ acetic acid and 10% isopropyl alcohol until the
19 - 3 1
45 20
21
background was colourless. The gel was then
stained using the modified silver stain procedure.
22 - 3 0
---23-63
The gel was soaked in a solution of 40 %methanol
36
- - - 24
---25-3 and 10% acetic acid for 3 hours and then washed
---26-2
twice for 15 minutes in a solution of 10%
- - - 27
methanol and 5 % acetic acid. Once fixed with
- - - 28
29 29 -
- - - 30
5 0 10% glutaraldehyde for 30 minutes (washed after-
---31-13 wards with water overnight and 3 additional times
---32-31
---33-15 of 1 hour each), the gel was stained with a freshly
34 - 2 0
---35-15
prepared ammoniacal silver solution (0.4%, w/v
- - - 36 AgN0 3 ; 0.35%, v/v NH 4 0H; 0.072%, w/v
---37-14 N aOH) for one hour. Afterwards, three washes in
water for 2 minutes were performed. Then, a citric
24 36 - 1 0
acid-formaldehyde developer (Oakley et aI., 1980)
39 - 2 0
was applied at 40 0 C until the bands appeared.
- - - 40
(The agitation of the gel is mandatory especially
in staining and developing.) Development was
20.1 41 - 1 0
- - - 42 stopped by discarding developer and adding
acetic acid (l %, w/v), for 5 minutes. The gel was
43 - 1 0
---44

45 - 1 0 then washed with water and hot-vacuum dryed.

- - - 46 The basic modifications to obtain both lower
---47
---48-14
levels of background and maximum staining sen-
49 - 1 5 sitivity are: (1) a longer application of the
ammoniacal silver-stain solution (1 hour, instead
of 15 minutes) but at half the AgN0 3 concen-
14
tration, (2) more washes after application of the
Fig. 2. Outline of the electrophoretograms corresponding to
young (Y) and old (0) females. Position and molecular
weight (Kd) of protein markers is shown at the left. The registered for young and old patterns. Intensity is a subjective
pattern of a young individual consists of 49 bands. The bands number, ranging from I to 6 for increasing intensity, esti-
are numbered according to their mobility, at the right hand mated for each band independently (this estimation is
side. For each of the bands that showed differences, a number derived from many electrophoretograms from this and other
indicating their absence (0) or intensity, when present, is works on protein variability different clones and females).
328

above mentioned solution, and (3) a much higher geneity among young individuals (Fig. 1). How-
developing temperature (40 0 C, instead of room ever as age increases, the patterns change and
temperature). show great variability among individuals of the
Moreover, in order to evaluate the individual same chronological age. Individuals of 2 and
somatic growth, 100 ovigerous parthenogenetic 4 days of age characteristically have the same
females were collected and their offspring were juvenile pattern. To check this homogeneity we
cultured individually. They were transferred daily conducted an experiment to compare the protein
into 0.5 ml offresh culture medium with 5.3 x 105 patterns of juvenile individuals (aged 2 days)
Tetraselmis sp. cells ml- 1. At 4, 6, 8, 12 hours of coming from eggs layed by young and old mothers
life and daily from then on, 10 females, randomly (aged 2 and 6 days). No maternal age effect was
collected, were fixed with lugol's and measured detected in these juvenile individuals, as all of
with a Zeiss Invertoscop at 40 x magnification. them showed identical protein patterns. But at
The number of measured females was slightly less age 6 days, protein patterns become variable and
for the last three days of the experiment due to the the variability increases with age. The major
survival structure of the population. trends of the age-dependent changes can be seen
in Fig. 2 where the two banding patterns are pres-
ented for comparison. The juvenile pattern con-
Results sists of 49 bands (the same pattern has been con-
firmed by samples containing 5 and 10 individ-
The protein patterns of single individuals of clone uals). The senescent pattern shows a reduction in
CU in the two replicated experiments show homo- the number of bands; 11 bands are lost, dis-

pm

300
____ t ------+------t ------f ---- _J ------,,--_ .. ~- --....r---_
. r". +. . . .,
250
/V . r .... r.......fL . r . . .··t···t .... r . ··+·f
200 , f···· i r
J f +"
~f
150 • LENGTH
• WIDTH

100

50

1 2 3 4 5 6 7 8 9 10 11 12
AGE (DAYS>
Fig. 3. Growth oflength and width during the life of Brachionus plicatilis at 2S C. The vertical bands represent 9S %confidence
0

limits calculated from Student's t distribution.

 329

tributed among the whole range of relative by Luciani et al. (1983) and the fertility decrease
molecular mass (Mr). These bands represent with the senescence is a well-known (e.g. King,
more than the 22 % of the total number of poly- 1967; Aronovich & Spektorova, 1974; Snell &
peptide subunits. Moreover, 13 bands show a King, 1977; Ricci, 1983). The reduction of high
remarkable decrease in intensity, corresponding Mr proteins in senescent individuals may be
to polypeptides of high Mr, usually bands over Mr related to the reduction of gonads in senescence
66 KD and a few others over Mr 24 KD. On the and degenerative processes of other kinds. On the
other hand, 6 bands oflow Mr, almost impercepti- other hand, the intensification of some low Mr
ble in the juvenile pattern, are intense in senescent bands could be associated with post-translational
individuals. These are four 24-29 KD bands and changes in proteins, such as spontaneous racemi-
the two fastest proteins. The most distinctive zation or deamination in certain amino acid
features of the senescence patterns are the two residues, leading to increased proteolytic degrada-
intense protein bands at 26 and 27 KD, respec- tion, (McKerrow, 1979; Finch, 1987).
tively, and the reduced intensity of the highest Mr Aging patterns were observed from age 6 days
protein components. in our experiment, the same age when growth
Additionally, some direct observations were ceases. This agrees with Lansing's conclusion
carried out on the old females during the experi- (Lansing, 1948) that aging is a consequence of
ments. Aged females had degenerate digestive growth cessation in rotifers, which is also true for
organs and gonads and very low or nonexistent other organisms (Lints, 1978). The effects of
egg production. Also evident was decreased senescence, leading to death, became increasingly
movement of cilia and increased pigmentation. apparent after growth cessation. Thus, the mean
Study of the individual somatic growth (Fig. 3) lifespan is almost coincident with the average age
showed that growth, with increasing age, follows of last reproduction. Nevertheless, there is great
approximately the Bertalanffy function (Ber- variability in individual lifespans and the maxi-
talanffy, 1948), as found in other rotifer species mum lifespan is almost double the mean. The
(Lebedeva & Gerasimova, 1985). The individuals cessation of reproduction is much less variable. In
of clone CU have at birth already 58 % of their accordance with this, our data illustrate the lack
maximum length and 54 % of their maximum of relationship of physiological age and chrono-
width. They reach adult size - 90 % of maximum logical age, even among isogenic individuals under
length, 85 %of maximum width - between the first the same environmental conditions. Individuals
and second day of life, which is when they begin of the same chronological age show different pro-
reproduction. Maximum size is more or less tein patterns. In spite of this individual variability,
reached by the sixth day of life, and the maximum life-history traits respond to environmental fac-
reproductive age is 8 days, while maximum tors in predictable ways. Thus, if these eutelic
longevity is 12 days. organisms are grown under lower temperature,
Results from population dynamic experiments the lifespan is longer (Miracle & Serra, in this
(Serra, 1987) showed that clone CU has a mean volume) and body size is larger (King & Miracle,
lifespan of 6.86 days and a mean age oflast repro- 1980; Levedeva & Gerasimova, 1985; Serra &
duction of 6.5 days old. Miracle, 1987), but the relation between mean and
maximum lifespan, reproductive and growth
periods, is maintained.
Discussion From these results, it may be inferred that
rotifer aging is a consequence of development,
The directly observable features associated with growth and/or differentiation, which implies that
senescence that we have found have been the number of genes potentially capable of
described by others authors (Lansing, 1947). expressing themselves becomes less and less. This
Slower swimming with aging has been quantified could be associated with the reduction in the num-
330

ber of bands. However, considerable variability in King, C. E., 1969. Experimental studies of ageing in rotifers.
the timing of developmental aging events can Exp. Gerontol. 4: 63-79.
King, C. E., 1983. A re-examination of the Lansing Effect.
occur, which is due not only to genetic or general
Hydrobiologia 104: 141-146.
environmental conditions, but also to other King, C. E. & M. R. Miracle, 1980. A perspective on aging in
causes. These causes could be: (1) random fac- rotifers. Hydrobiologia 73: 13-19.
tors involved in the reduction of gene expression Lansing, AI., 1947. A transmissible, cumulative and reversi-
in non-dividing cells, (2) other factors such as ble factor in aging. 1. Gerontol. 2: 228-239.
Lansing, A.I., 1948. Evidence for aging as a consequence of
maternal age or infective agents. Both causes
growth cessation. Proc. nat. Acad. Sci. U.SA 34:
could be due to small differences between the 304-310.
individuals' environment and its effects on gene Lebedeva, L.1. & T. N. Gerasimova, 1985. Peculiarities of
expression or to random allocation of non- Philodina roseola (Ehrbg.) (Rotatoria Bdelloida). Growth
nuclear genetic material among isogenic individ- and reproduction under various temperature conditions.
uals. Aging could amplify these differences Int. Revue. ges. Hydrobiol. 70: 509-525.
Lints, F. A., 1978. Genetics and aging, Interdisciplinary
dramatically. Topics in Gerontology-Karger, S., Basel 129 pp.
Luciani, A, J. Chasse & P. Clement, 1983. Aging in
Brachionus plicatilis: The evolution of swimming as a
Acknowledgements function of age a two different calcium concentrations.
Hydrobiologia 104: 141-146.
McKerrow, J. H., 1979. Non-enzymatic, post-translational
We are most grafetul to Dr. C. Dawson who anmino acid modifications in ageing. A brief review. Mech.
corrected the English text. M. J. Carmona was Ageing Dev. 10: 371-377.
supported by a grant from 'Ministerio de Edu- Meadow, N. D. & c. H. Barrows, 1971. Studies on aging in
caci6n y Ciencia'. a bdelloid rotifer I. The effect of various culture systems on
longevity and fecundity. J. expo Zool. 176: 303-313.
Oakley, B. R., D. R. Kirsch & N. R. Morris, 1980. A simpli-
fied ultrasensitive silver stain for detecting proteins in
References polyacrylamide gels. Anal. Biochem. 105: 361-363.
Ricci, c., 1983. Life histories of some species of Rotifera
Aronovich, T. M. & L. V. Spektorova, 1974. Survival and Bdelloidea. Hydrobiologia 104: 175-180.
fecundity of Brachionus calycijlorus in water of different Semancik, J. S., 1976. Structure and replication of plants
salinities. Hydrobiol. J. 10: 71-74. viroids. In: Animal Virology, ICN-UCLA, Symposia on
Bertalanffy, L., 1948. Das organische Wachstum und seine Molecular and cellular Biology. Baltimore, D., A S.
Gesetzmssigkeiten. Experientia 4: 255-269. Huang & c. F. Fox (eds.), pp. 529-545 Acad. Press, New
Conejero, V. & J. S. Semancik, 1977. Analysis of the proteins York: 523-545.
in crude extracts by polyacrylamide slab gel electro- Serra, M., 1987. Variaci6n morfometrica, isoenzimatica y
phoresis. Phytopathology 66: 1424-1426. demografica en poblaciones de Brachionus plicatilis:
Fanestil, D. D. & c. H. Barrows, 1965. Aging in the rotifer. diferenciaci6n genetic a y plasticidad fenotipica. Ph. D.
J. Gerontol. 20: 462-469. Thesis, Universidad de Valencia.
Finch, C. E., 1987.The ordely decay of order in the regulation Serra, M. & M. R. Miracle, 1983. Biometric analysis of
of aging processes. In F. E. Yates (ed.), Self-organizing Brachionus plicatilis ecotypes from Spanish lagoons. Hydro-
systems. The emergence of order. Plenum Press, New biologia 104: 279-291.
York: 213-236. Serra, M. & M. R. Miracle, 1985. Enzyme polymorphism in
Jennings, H. S. & R. S. Lynch, 1928. Age, mortality, fertility Brachionus plicatilis populations from several Spanish
and individual diversities in the rotifer Proales sordida lagoons. Verh. int. Ver. Limnol. 22: 2991-2996.
Gosse. I. Effect of the age of the parent on characteristics Serra, M. & M. R. Miracle, 1987. Biometric variation in three
of the offspring. 1. expo Zool. 50: 345-407. strains of Brachionus plicatilis. Hydrobiologia 147: 83-89.
King, C. E., 1967. Food, age, and the dynamics of a labora- Snell, T. W. & c. E. King, 1977. Lifespan and fecundity pat-
tory population of rotifers. Ecology 48: 111-128. terns in rotifers: The cost of reproduction. Evolution 31:
882-890.
Hydrobiologia 186/187: 331-337, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 331
© 1989 Kluwer Academic Publishers.

Brachionus plicatilis tolerance to low oxygen concentrations

Angeles Esparcia, Maria R. Miracle & Manuel Serra

Dep. de Ecologia, Fac. C. Biol6gicas, Universidad de Valencia, 46100-Burjasot (Valencia), Spain

Key words,' rotifers, oxygen, extreme conditions, population dynamics

Abstract

Tolerance to low oxygen concentrations is expected in Brachionus plicatilis, a rotifer adapted to live in
saline warm waters. The population dynamics of a clone of this species, isolated from an endorreic saline
lake, was studied under controlled laboratory conditions. Although their growth and metabolism is
extremely reduced, B. plicatilis populations are able to maintain relatively high-density populations (a
mean of 35 ind ml- 1) in oxygen concentrations below 1 mg 1- 1, for more than one month. Major features
of population growth related to oxygen are discussed.

Introduction aim of the present work is to evaluate the tolerance

of Brachionus plicatilis to limited oxygen levels and
Oxygen concentration may play an important role to characterize the main features of such toler-
in distribution of the rotifers both in different ance.
lakes and in the vertical profile of stratified lakes,
where oxygen is frequently correlated with tem-
perature. Rotifer communities adapted to both Material and methods
low oxygen and low temperature (Pejler, 1957;
Edmondson, 1965; Larsson, 1971 ; Miracle, Clone CU of Brachionus plicatilis (Serra &
1976; Hoffman, 1977) are found in deep waters. Miracle, 1983) was cultivated in diluted sea water
Oxygen could also be a limiting factor in brackish with a salinity of 12 g 1- 1, at 25 0 C, and under
waters of warm latitudes, as well as in deep waters light. The rotifers were fed with Tetraselmis sp.
of meromictic lakes near the oxycline (Pourriot, thermally treated at 65-70 0 C, for 90 min, in
1965; Ruttner-Kolisko, 1980; Miracle & Vicente, order to inactivate them. The efficiency of both
1983). The studied species, Brachionus plicatilis, is thermic treatment and feeding with the obtained
adapted to high salinity and temperature, condi- food has been studied by Esparcia & Serra
tions that determine low oxygen solubility (1988). All cultures were grown in flasks with
coefficients. Moreover it is cultivated in mass cul- three mouths. Two of these inputs were for N 2 gas
tures for applied purposes, where its extremely inflow into the culture and into the gas chamber
high densities may cause oxygen depletion. over the culture; the third opening was for access
There are very few experimental studies on to the culture (see Fig. 1). A magnetic stirrer was
rotifer tolerance to low oxygen concentrations used to stir the flasks.
(e.g., Ruttner-Kolisko, 1980; Schluter, 1980). The Experiment 1 was initiated by placing 500
332

9<l'ml 80

6 -it
.!.
.. ffi
20
">
)(

eo
DAY
991ml 80

c
60

40
e
6 -CII
.!.
z
20 .. w

2 0
>"
)(

O~=---~T-------~-----'-------T------~Q-----~
o 20 40 60
DAY
 333

99/ml 80 1
60

-
40

6 :::
CII
S
20 4ffi
20
">
)(

O~~--~------.-~~-'L-----.------r------~
20 40 60
DAY
991ml 80

c
60

40

6 -
eo
CII
!.
z
20 4 w

2
"
> )(
0

0
60
DAY
Fig. 1. Daily variation in population density of Brachionus plicatilis (open symbols) and oxygen concentration (closed symbols).
A, Band C are populations grown in hermetically closed flasks; the control is a population not grown in a hermetically closed
flask. The four populations were followed until their extinction except for population A. The control population died at the 81st
day. A drawing of the culture flask is shown in the upper right corner.
334

W'm180

60

40
8

6
-!-
20 z
4 w

2
"
>
><
0

0
60
DAY

991m180

B
60

40
8

6
-!.
::
01

20 z
4 !AI

2
">-S·
0
60
DAY
Fig. 2. Daily variations in population density of Brachionus plicatilis (open symbols) and oxygen concentration (closed symbols).
The oxygen concentrations were daily adjusted to 1-2 mg 1-1 (A) and to 0-1 mg 1-1 (B). The arrows indicate the times offood
supply; dilutions ofrotifer density were 1: 1.03 (10th day), 1: 1.02 (18th day), 1: 1.03 (29th day), 1: 1.13 (39th day) and 1: 1.12
(51st day) respectively.
 335

females, from a population previously adapted to the culture medium. The cultures were fed periodi-
the above indicated conditions, into each of four cally (arrows in Fig. 2), by adding medium with
flasks. These flasks contained 400 ml of culture 0.8 x 106 cells ml- 1 of Tetraselmis sp. The experi-
medium with 0.8 x 106 cells ml- 1 treated algae, ment ended on the 53rd day.
(food concentration was determined with a ZM Instantaneous growth rates (r) were estimated
Coulter Counter). Three flasks were hermetically using the regression equation lnNt = (lnNo ) + rt,
closed and the oxygen was reduced by the respira- where No and Nt are the number of individuals at
tion of the rotifers themselves, the fourth flask was the initiation and the end, respectively, of the
maintained as a control and stoppered with a exponential growth phase at the beginning of the
cotton plug. experiment. The integration of the number of
Both population density and oxygen concen- rotifers with respect to time, was calculated as
tration were monitored daily. Rotifers from one or I (N; + N; + 1 )/2, where N; and N; + 1 are the daily
two samples of 1 ml were counted using a dissect- i
measurements of population density.
ing microscope. Oxygen was measured with an
electrode (Orion 97-08-99), placed directly into
the medium. During culture handling, oxygena- Results
tion was prevented with N 2 flow into the air
chamber, except in the control. Food was never Experiment 1 showed that Brachionus plicatilis is
added and the experiment ended when the popu- able to tolerate oxygen concentrations lower than
lation died, (except for population A which could 1 mg 1- 1, at least for 13 days (Fig. 1). The three
not be followed to completion). replicates behaved differently. The B population
In experiment 2, two cultures were initiated reached a high density with a maximum similar to
each with 6000 females (15 females ml- 1) and that of the control, but after this maximum, the
food in excess. The oxygen level was adjusted to oxygen level dropped quickly due to rotifer respi-
approximately 1-2 mg 1-1 in one culture and to ration and the popUlation died when the oxygen
0-1 mg 1- 1 in the other by bubbling N 2 through level reached zero. However, the A and C popu-

Table 1. Some population parameters and oxygen concentrations for low aeration cultures. The values in parentheses indicate
the oxygen concentrations (mg 1- I) at the time interval when the corresponding parameter was evaluated. r is calculated for
the exponential growth phase at the begining of the experiment. N x time is the integrated number of rotifers through time.

r, days - I Maximum Average N x time till

density, N x time population
Ind 'ml- I per day, extinction,
Ind'ml- I Ind' d· ml- I

Experiment 1
(low initial
density)
CONTROP 0.773 (4.77) 78 (8.90) 22.7 (8.70) 1836.7
A 0.701 (0.95) 29 (0.30) 12.3 (0.91)
B 0.528 (0.90) 71 (2.30) 26.3 (1.20) 787.9
C 0.564 (0.93) 27 (2.70) 13.4 (2.18) 698.8

Experiment 2
(high initial
density)
A 0.147 (1.20) 65 (3.57) 38.4 (1.95)
B 0.336 (0.78) 55 (1.00) 37.1 (0.79)

(a). Without N2 flow.

336

lations did not reach high densities, while the In contrast, high salinities could decrease (Epp &
oxygen concentration was maintained at a lower Winston 1978) Brachionus plicatilis oxygen con-
level it was not exhausted and the populations sumption as has been observed in other osmo-
survived longer. conformers. However no difference was observed
Oxygen concentration appeared to have an by Ruttner-Kolisko (1972) between 17 and 36 g
effect on both instantaneous rates of population 1- 1 salinity.
increase and maximum densities (Table 1); both Our results give experimental proof of
decrease when oxygen levels go down. However, Brachionus plicatilis tolerance to low oxygen con-
the r values were measured in the initial culture centrations for long time periods, independent of
phase and a time lag from the oxygenated cultures other enviromental factors. However, the cultures
could be present. For a closed culture, without living at low oxygen levels grow at very low rates.
addition of food, the integration of rotifers with A decrease in metabolic rate seems to be the main
respect to time, till the extinction of the population feature induced by low oxygen availability, as has
(Table 1), is an estimation of the efficiency of been observed in other invertebrates (e.g., Zwaan
exploitation of the medium's resources (this is & Wijsman, 1976; Gade, 1984; Hochachka,
similar to the carrying capacity concept). This 1986). Because oxygen depletion slows down
parameter has been calculated and it shows a growth and thus increases the survival time of the
clear decrease with the oxygen concentration population, it could function as a regulating
decrease (Table 1). mechanism. In one of the studied cases (Experi-
Experiment 2, with addition offood, confirmed ment 1, population B) in which the density of
that rotifer populations are able to live at oxygen Brachionus plicatilis reached high values, the
concentrations lower than 1 mg 1- 1 for long population died off immediately after the period
periods of time (Fig. 2). For the oxygen concen- of uncontrolled growth.
trations assayed, the calculated population Finally, a poorer use of resources has been
parameters also showed a clear inverse relation- found at lower oxygen concentration, as could be
ship with oxygen concentration. derived from the integrater number of rotifer
through time, supported at the different condi-
tions.
Discussion In conclusion, Brachionus plicatilis could be
maintained at reduced metabolic rates in micro-
Brachionus plicatilis inhabits waters where low aerophilic conditions. However individual re-
oxygen concentrations might be expected because placement occurred at low rates due to poor
of its strong tolerance of high salinities and tem- reproduction and extremely long generation
peratures (Ito, 1960; Ruttner-Kolisko, 1974; times. This species also seems to be capable of
Walker, 1981). In fact, abundant populations of activating fermentative metabolic pathways
this species have been found at oxygen concentra- (Esparcia, unpublished data), thus reducing the
tions lower than 0.37 mll- 1 in saline and brackish oxygen requirements but also reducing the energe-
waters (Serra, 1987), and in saline meromictic tic efficiency.
lakes near the oxycline (Swift & Hammer, 1979;
Miracle & Vicente, 1983).
Salinity and temperature have different effects Acknowledgments
on oxygen requirements. High temperature in-
creases the metabolic rate and the oxygen con- The authors wish to thank Dr. C. A. Dawson for
sumption in rotifers (e.g., Pourriot & Deluzarches, language revision. A. Esparcia is the recipient of
1970; Galkovskaya, 1987). Thus, as temperature a grant from the Ministerio de Education y
increases, an opposition between oxygen availa- Ciencia.
bility and oxygen requirements might be expected.
 337

References Pejler, B., 1975. Taxonomical and ecological studies on

planktonic Rotatoria from Northern Swedish Lappland.
Kung!. Svensk. Vetensk. Hand!. 6(5): 1-68.
Edmondson, W. T., 1965. Reproductive rate of planktonic
Pourriot, R, 1965. Recherches sur l'ecologie des rotiferes.
rotifers as related to food and temperature in nature. Eco!.
Supplement 21 a 'Vie et milieu'. Masson, Paris. 224 pp.
Monogr. 35: 61-111.
Pourriot, R & M. Deluzarches, 1970. Sur la consommation
Epp, R W. & P. W. Winston, 1978. The effect of salinity and d'oxigene par les rotiferes. Ann. Limno!. 6: 229-248.
pH on the activity and oxygen consumption of Brachionus
Ruttner-Kolisko, A., 1972. Der Einfluss von Temperatur und
plicatilis. Compo Biochem. Physio!. 59A: 9-12. Salzgehalt des Mediums auf Stoffweschsel-und Ver-
Esparcia, A & M. Serra, 1988. Efecto del alimento tratado
mehrungs-intensitat von Brachionus plicatilis (Rotatoria).
termicamente en el crecimiento poblacional de Brachionus
Dt. Zoo!. Ges. 65: 89-95.
plicatilis Miiller 1786 (Rotifera: Brachonidae). Inv. Pesq. Ruttner-Kolisko, A, 1974. Plankton rotifers. Biology and
52: 345-353.
taxonomy. Biennengewasser 26, 146 pp.
Gade, G., 1984. Effects of oxygen deprivation during anoxia
Ruttner-Kolisko, A., 1980. The abundance and distribution
and muscular work on the energy metabolism of the cray-
of Filinia terminalis in various types of lakes as related to
fish Orconetes limosus. Compo Biochem. Physio!. 77 A: temperature, oxygen, and food. Hydrobiologia 73:
495-502. 169-175.
Galkovskaya, G. A, 1987. Plancktonic rotifers and tem- Schliiter, M., 1980. Mass culture experiments with
perature. Hydrobiologia 147: 307-317. Brachionus rubens. Hydrobiologia 73: 45-50.
Hochachka, P. W., 1986. Defense strategies against hypoxia Serra, M. & M. R Miracle, 1983. Biometric analysis of
and hypothermia. Science 231: 234-241. Brachionus plicatilis ecotypes from spanish lagoons. Hydro-
Hofmann, W., 1977. The influence of abiotic environmental biologia 104: 279-291.
factors on population dynamics in planktonic rotifers.
Serra, M., 1987. Variabilidad morfol6gica, isoenzimatica y
Arch. Hydrobiol. Beih. 8: 77-83.
demognl.fica en poblaciones de Brachionus plicatilis.
Ito, T., 1960. On the culture ofmixohaline rotifer Brachionus Diferenciaci6n genetica y plasticidad fenotipica. Tesis
plicatilis O. F. Miiller in the sea water. Rep. Fac. Fish. doctoral. Universidad de Valencia.
prefect. Univ. Mie. 3: 708-740. Swift, M. C. & U. T. Hammer, 1979. Zooplankton popula-
Larsson, P., 1971. Vertical distribution of planktonic rotifers tion dynamics and Diaptomus production in Waldsea lake,
in a meromictic lake, Blankvatn near Oslo, Norway. a saline 'meromictic' lake in Saskatchewan. J. Fish. Res.
Norwerg. J. Zoo!. 19: 47-75. Bd Can. 36: 1431-1438.
Miracle, M. R, 1976. Distribuci6n en el espacio y en el Walker, K. F., 1981. A synopsis of ecological information on
tiempo de las especies del zooplancton del lago de the saline lake rotifer Brachionus plicatilis O. F. Muller
Banyoles. Instituto Nacional para la Conservaci6n de la 1786. Hydrobiologia 81: 159-167.
Naturaleza. (Ministerio de Agricultura). Monografias 5, Zwaan,A. De & T. C. M. Wijsman, 1976. Anaerobicmetabo-
Madrid 270 pp. lism in bivalvia (Mollusca). Characteristics of anaerobic
Miracle, R & E. Vicente, 1983. Vertical distribution and metabolism. Compo Biochem. Physio!. 54B: 313-324.
rotifer concentrations in the chemocline of meromictic
lakes. Hydrobiologia 104: 259-267.
Hydrobiologia 186/187: 339-346, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 339
© 1989 Kluwer Academic Publishers.

Analysis of protein, carbohydrate and lipid in rotifers

Castor Guisande & Laura Serrano

Departamento de Ecologia, Facultad de Bilogia, Universidad de Sevilla, 41080 Sevilla, Spain

Key words: rotifers, protein, carbohydrate, lipid

Abstract

Protein, carbohydrate and lipid amounts were determined for several rotifer species collected directly
from the field. Brachionus calyciflorus was the most abundant species; therefore making possible more
data for it. An increase in protein content of this species occurred when its concentration in food (Jlg
protein/ml) also increased. Keratella tropica showed a similar pattern, but Asplanchna brightwelli did not.
Carbohydrate proved to be the main form of storage in rotifers. In Brachionus calyciflorus females
bearing no egg, 8 %of the total biomass was carbohydrate; in females bearing one egg, 15 %carbohydrate
was found. Lipid does not appear to be used for storage since no increase in the amount of lipid was
detected in females bearing eggs or embryos. This suggests that lipid has a structural function. Finally,
a relationship between rotifer body volume and protein content at a given food concentration was
obtained. The cladoceran Daphnia magna follows the same pattern.

Introduction recently differences between food threshold levels

have been examined (Sternberger & Gilbert,
Rotifers can be viewed as the most significant 1985). These factors may help to explain rotifers
community within the freshwater zooplankton in species distribution and community structure in
regard to the shifts that take place in species nature.
dominance. Temperature plays an important role The aim of this research was to determine the
in the separation of species into ecological niches relationship between existing food concentration
and the time of year may determine their presence and nutritive state of rotifer populations. A total
or absence. Feeding plays a significant part in this protein analysis was performed on both rotifers
separation as species differ in their ability to ingest and their natural medium. Carbohydrate and lipid
different food sizes (Bogdan & Gilbert, 1987). contents were also quantified in order to estimate
Energy requirements also vary among species rotifer investment in stored reserves and the rela-
(Guisande & Toja, 1988). The importance of tive importance of each component.
rotifer feeding has produced a wealth of published
material, such as clearance rate experiments
(Starkweather & Gilbert; 1977; Starkweather, Material and methods
1980; Bogdan & Gilbert, 1982; Walz, 1983) and
feeding selectivity studies (Dumont, 1977 ; Animals were collected with a 35 Jlm closing net
Pourriot, 1977; Gilbert & Bogdan, 1984). More from several ponds throughout the year. Samples
340

Table 1. Minimum and maximum values of protein content (protein range) and each mean range value for different rotifer
species, measured at several food concentrations.

Species Food concentration Protein Mean range N

range value
Body volume (J1m 3 ) (J1g protein ml- I)
Body lenght (J1m) < 10 J1m <0.2 J1m (J1g ind- I ) (J1g ind- I )

Asplanchna brightwelli 27.25 26.94 0.40-0.82 0.61 10

22.45 19.94 0.28-1.40 0.84 12
V = 1.725* W 15.95 0.26-2.55 1.40 10
12.86 9.66 0.85-1.25 1.05 8
I = 491.41 9.09 0.45-1.47 0.96 13
8.78 8.06 0.11-0.36 0.24 21
Hexarthra fennica 7.25 0.09-0.24 0.16 14

V= 7.63*W

L = 156
Brachionus urceolaris 25.57 0.16-0.51 0.33 25

V= 6.99*W

I=171.1
Brachionus calycijlorus
f anuraeiformis 27.25 26.94 0.38-0.45 0.41 7
22.23 11.5 0.21-0.60 0.40 17
V= 4.68*W 18.06 0.33-0.55 0.44 18
15.95 0.13-0.54 0.34 26
L = 150.35 12.90 0.04-0.63 0.33 32
12.86 9.66 0.17-0.58 0.37 20
8.78 8.06 0.06 0.06 4
7.28 7.85 0.04-0.18 0.10 15
Keratella tropica 27.25 26.94 0.26-0.44 0.35 10
22.45 19.94 0.29-0.49 0.39 14
V= 2.57*W 22.23 11.5 0.43-0.46 0.44 8
18.06 5.3 0.20 0.20 3
L = 103.31 15.95 0.10-0.27 0.18 18
14.56 10.94 0.12-0.41 0.26 14
Filinia terminalis 22.23 11.5 0.08-0.17 0.12 15
18.06 5.3 0.02-0.18 0.10 18
V= 1.72*W 15.95 0.05-0.21 0.13 8

I = 102.43
Trichocerca sp. 27.25 26.94 0.17-0.18 0.17 9
22.45 19.94 0.15-0.35 0.25 8
V=9.47*W

I = 105.86
 341

were also collected from Dofiana National Park uals for each sample, making the use oflaboratory
(D.N.P.). After examination under a binocular cultures unnecessary; the samples were taken
microscope, the animals were transferred to dis- directly from the field.
tilled water so that errors from protein, carbo- Experimental animals were grouped into dif-
hydrate and lipid in the natural water could be ferent categories depending on their reproductive
reduced. Under a Nikon inverted microscope, the state: CO (females bearing no egg), C1 (females
animals were taken from the distilled water with bearing one egg), C2 (females bearing two eggs),
a micropipette in order to get the same volume of etc. Neither males nor females with mictic eggs
10 Jll for each individual in protein and carbo- were collected. Asplanchna brightwelli fell into only
hydrate analysis and 5 Jll in lipid analysis. three categories: CO (females with no embryo);
Measurements of rotifer body lengths were taken C1 (females with embryo) and C2 (females with
from living, field-collected individuals. The sensi- resting egg).
tivity ofthe analysis allowed the use offew individ- In the protein analysis 2-5 individuals were

o
o Om

"0
C 0
.-
'-c:
'G)
-0
I...
0...
O'l
::t
.-

Kt
Ft

.-
0

Fig. 1. The relationship of volume (V) and the mean value of the protein range (P) for various zooplankton species at a given
food concentration. Vertical lines designate the protein range between the minimum and maximum protein content values for
each species. Solid line designates the predicted log-log least squares linear regression, which fits the equation: P = 3.6 * 10 - 4
*VO.5036. Ab = Asplanchna brightwelli; Bc = Brachionus calyciflorus; Dm = Daphnia magna; Ft = Filinia terminalis; Kt = Keratella

tropica.
 342

used; 2-10 were used for carbohydrate and undegraded protein was disregarded determining
15-20 for lipid analysis. Lowry's et al. method total protein content.
(l951), modified by Markwell et al. (l976) was Protein and carbohydrate analysis of the
used for the protein analysis, reaching a final medium were also performed in order to estimate
volume of 1.075 ml. BSA up to 18 J,lgwas used for the amount of food available for rotifer popu-
the linear standard. For the carbohydrate experi- lations. The water was passed through a 10 J,lm
ments Dubois et al. method (1956) was followed, net and a 0.2 J,lm filter in some cases to determine
reaching a final volume of 2.410 ml; glucose was the portion of dissolved protein and carbohydrate
used for the linear standard. Boeringher's stand- in the medium.
ard was used for lipid quantifications, using a final It was found to be more convenient to calculate
volume of2.5 ml and BOeringher's solution for the the rotifer body volume using Ruttner-Kolisko's
linear standard. method (l977), since body volume as measure-
In the case of protein analysis, SDS was ment of biomass is more reliable than body size.
employed to degrade the rotifers. This detergent Temperature was also measured in every pond.
did not affect the mastax and this portion of Values varied between 15 and 23 0 C.

CXJ
o

"0 <0
c 0
"-c
......
Q)

0
'-
0...
01
~ V
0

N
o

f
o
o 5 10 15 20 25 30

Food concentration (fJ,9 protein/m l)

Fig. 2. The relationship offood concentration (C) and the mean value of the protein range (P) for Brachionus calyciflorus. Vertical
lines designate the protein content range between the minimum and maximum protein content values at each food concentration.
Curve fits the equation: P = - 0.446 + 0.282*Ln (C).
 343

Results Daphnia magna (Guisande, unpublished) matched

the equation estimated for rotifers: P = 3.6 * 10 - 4
Table 1 lists the protein values obtained for each *VO.5036 (r2 = 0.99; p < 0.001). When body size,

species. Some other species were collected in the excluding spines, was considered, instead of body
samples, such as B. angularis, B. budapestinensis, volume, the equation was: P = 4.33* 10- 5 *L1.758
Polyarthra major and Synchaeta pectinata, but the (r2 = 0.99; p < 0.001), which is very similar to the
populations of these species were not sufficiently one given by Sternberger & Gilbert (1987)
large for the experiment. between body mass (M) and body length (L) for
At a food concentration of 15.95 jlg protein several rotifer species at optimum food level:
ml- 1 reached in one of the ponds (Table 1), four M= 3.1*10- 5 *L1.574.
species appeared in sufficient numbers for the B. calyciflorus was the most widely-represented
analysis. A relationship between body volume (V) species in the ponds tested. Figure 2 shows the
and mean protein value (P) was found (Fig. 1). variation of protein content in B. calyciflorus for
But even more significant was that the protein several food concentrations. Individual mean
amount predicted at this food concentration for protein content (P) as a function of protein con-

to
o

"0 co
c 0
"-c
.......
0
Q)

a.
0'1
::t .q-
0

I
N
o

o
o 5 10 15 20 25 30
Food concentration (J.i.g protein /ml)

Fig. 3. The relationship offood concentration (C) and the mean value of the protein range (P) for Keratella tropica. Vertical lines
designate the protein content range between the minimum and maximum protein content values at each food concentration.
Curve fits the equation: P = - 0.225 + O.l87*Ln (C).
344

Table 2. Relation between protein and carbohydrate (ch) content at several food concentrations for different rotifers. This food
concentration mentioned here, is the one measured in the water filtered through 10 I'm. co: rotifers with no egg; Cl: rotifers
with one egg; C2: rotifers with two eggs; C3: rotifers with three eggs. N = number of individuals tested.

Species Food concentration Mean N Mean N CH

protein CH
(Jtg protein ml- I ) (Jtg ch ml- I ) content content (%)

(Jtg ind- I ) (Jtg ind- I )

Asplanchna 15.95 13.00 CO: 1.173 8 0.059 5 4.7

brightwelli CI: 0.768 2 0.116 2 13.1
12.86 7.33 CO: 0.850 4 0.056 5 6.2
Cl: 1.190 4 0.167 9 12.3
Hexarthra 7.25 8.46 CO: 0.166 10 0.029 22 15.1
fennica
Brachionus 26.38 23.28 CO: - 0.027 17
calycijlorus CI: - 0.052 16
f anuraeiformis C2: - 0.123 3
22.23 10.01 CO: 0.260 8 0.018 7 6.5
C3: 0.600 4 0.062 8 9.4
15.95 13.00 CO: 0.275 12 0.028 16 9.2
Cl: 0.380 14 0.066 24 14.8
12.90 11.46 CO: 0.108 20 0.039 30 26.5
CI:0.191 11 0.045 24 19.1
C2: 0.631 1 0.113 2 15.1
12.86 7.33 CO: 0.230 8 0.022 13 8.7
Cl: 0.405 8 0.047 6 10.4
Filinia 22.23 10.01 CO: 0.086 9 0.002 8 2.3
terminalis Cl: 0.170 8 0.020 6 10.5

centration existing in the medium (C) fits the bility that these components could be used as
equation: P = - 0.446 + 0.282 * LN C stored reserves. In this work we suggest that
(r2 = 0.76, p < 0.001). A similar relationship was rotifers do not accumulate lipid as a stored
predicted for Keratella tropica (Fig. 3) which reserve. The quantity oflipid in B. calyciflorus was
matched the equation: P = - 0.225 + 0.187* LN
C (r2 = 0.66, p < 0.01). Table 3. Lipid mean range value for two rotifer species.
A. brightwelli, however, did not show the same N = number of individuals tested.
type of relationship. This could be due to their
feeding habits or to their greater body-size varia- Species Mean range N
----------value
bility.
In regard to stored reserves, carbohydrate
revealed itself to be the most significant storage
component. Table 2 shows protein and carbo- Asplanchna brightwelli CO: 0.025 15
hydrate contents at different food concentrations. CI: 0.019 15
CO: 0.023 15
Only copepodes and cladocerans, among 15
C2: 0.010
zooplankton species, are known to accumulate
lipid (Goulden & Henry, 1984). Wurdak et al. Brachionus calycijlorus CO: 0.009 25
f anuraeiformis CI: 0.003 17
(1978) found lipid droplets in B. calyciflorus and C2: 0.008 15
Asplanchna sieboldi eggs and suggested the possi-
 345

the same regardless of the presence or absence of hydrate content is similar in both species
eggs; this was also the case for A. brightwelli (Table 2). There is not enough information to
(Table 3). Lipid represents 1.06 % of the total assess the relative expenditure of stored reserves
biomass of B. calyciflorus and approximately for either species.
1.5% of A. brightwelli. Carbohydrate analysis of B. calyciflorus was
only performed at food concentrations above
12 J.lg protein ml- 1. At low food levels, individ-
Discussion uals were scarce and few animals had eggs. No
data are available below this food concentration,
The results of protein analysis for B. calyciflorus thus whether or not a further decrease in stored
(Fig. 2) suggest that upon increasing food concen- reserves exists at lower food levels has not been
tration, the individual protein content increases as determined. It would be extremely interesting to
well, though this increase becomes less pro- know if there is a decrease in the stored reserves
nounced when the food level is increased in large invested in reproduction at food limiting condi-
amounts. Keratella tropica shows a similar pattern tions or if, on the contrary, the amount of carbo-
(Fig. 3). Although there is not enough available hydrate allocated stays constant and a reduction
data for the rest of species, a similar relationship in the number of eggs then occurs.
could be expected. In regard to the relationship obtained between
These results indicate that the food concen- body volume and the amount of protein existing
tration required to reach the optimum protein in the environment (Fig. 1), it would be interesting
content is lower for smaller species. This is seen to determine if other filtering species, of different
by observing the behaviour of Filinia terminalis sizes, also fit this equation. Larger species show
(Table 1). Its protein range does change when a greater change under varying food concentra-
food concentration varied from 15.95 to 22.23 J.lg tions than smaller species. The existence of this
ml- 1. However, K. tropica varied from 0.293 to kind of relationship in filtering species could indi-
0.355 and B. calyciflorus from 0.335 to 0.428 J.lg cate that feeding strategy variability for different
protein ind - 1 at those food concentrations. species does not reflect protein content variation.
Therefore, larger species required more food to A variability of the amount stored reserves
reach their optimum nutritive state and their pro- invested in reproduction and of the consumption
tein content changed over a greater food range of these compounds by embryos may also occur
than smaller species. This was also found by (Goulden & Henry, 1987).
Stemberger & Gilbert (1985; 1987), who show To summarize, our results suggest that protein
that the food level required to increase population content increases with increasing food level. On
was lower for smaller species. reaching the optimum food level, further protein
Carbohydrate is the main storing substance in increase is less pronounced. When food concen-
rotifers. As can be seen in Table 2, the average tration increases greatly, a decrease in the protein
B. calyciflorus carbohydrate content for females content can be expected, since above a certain
bearing no egg is 0.027 J.lg ind - 1 (SE = 0.003); food concentration the animals spend more
for females bearing one egg is 0.052 J.lg ind - 1 energy and time rejecting excess food than they
(SE = 0.0004) and 0.118 J.lg ind - 1 (SE = 0.025) obtain through ingestion. Carbohydrate is the
for females bearing two eggs. The average main stored reserve in the rotifers tested, as no
Asplanchna brightwelli carbohydrate content is significant lipid accumulation was detected.
0.057 J.lg ind -1 (SE = 0.001) for females with no In future research, it would be interesting to
embryo and 0.141 J.lg ind -1 (SE = 0.025) for estimate the minimum food concentration
females with embryo. Also A. brightwelli showed required for each species, as well as determining
a greater amount of carbohydrate than whether stored reserves decrease or disappear at
B. calyciflorus, the relationship of protein/carbo- low food levels. Studies on the determination of
346

the amount of protein and carbohydrate appear to postembryonic yolk, and survival ability. Oecologia, 72:
be a valuable approach to the study of rotifer 28-31.
Guisande, C. & J. Toja, 1988. The dynamics of various
feeding processes. species of the genus Brachionus (Rotatoria) in the
Guadalquivir River. Arch. Hydrobiol. 112: 579-595.
Lowry, O. H., N. J. Rosenbraugh, A. L. Farr & R. 1. Randall,
Acknowledgement 1951. Protein measurements with the Folin phenol
reagent. 1. BioI. Chern. 193: 256-275.
Markwell, M. A. K., S. M. Haas, L. L. Bieber & N. E.
This research was supported by CAICYT Project Tolbert, 1978. A modification of the Lowry procedure to
no. 0940/87. simplifY protein determination in membrane and lipo-
protein samples. Analyt. Biochem. 87: 206-210.
Pourriot, R. 1977. Food and feeding habits of Rotifera. Arch.
References Hydrobiol. Beih. Ergeb. limnol. 8: 98-122.
Ruttner-Kolisko, A., 1977. Suggestions for biomass calcula-
tion of plankton rotifers. Arch. Hydrobiol. Beih. 8: 71-76.
Bogdan, K. G. & J.1. Gilbert, 1982. Seasonal patterns of
Starkweather, P. L. & 1. J. Gilbert, 1977. Feeding in the
feeding by natural populations of Keratella, Poly~rth~a, and
rotifer Brachionus calyciflorus II. Effect of food density on
Bosmina: Clearance rates selectivities and contnbutlOns to
feeding rates using Euglena gracilis and Rhodotorula glutinis.
community grazing. Limnol. Oceanogr. 27: 918-934.
Oecologia (Berlin) 28: 133-139.
Bogdan, K. G. & 1. 1. Gilbert, 1987. Quantitative comparis~n
Starkweather, P. L., 1980. Aspects of the feeding behaviour
offood niches in some freshwater zooplankton. OecologIa.
and trophic ecology of suspension feeding rotifers. Hydro-
(Berlin), 72: 331-340.
biologia 73: 63-72.
Dubois, M., K. A. Gilles, 1. K. Hamilton & F. Smith, 1965.
Sternberger, R. S. & 1. 1. Gilbert, 1985. Body size, food con-
Colorimetric method for determination of sugars and
centration, and population growth in planktonic rotifers.
related substances. Anal. Chern. 28: 350-356.
Ecology 66(4): 1151-1159.
Dumont, H.1., 1977. Biotic factors in the population
Sternberger, R. S. & 1. 1. Gilbert, 1987. Rotifer threshold
dynamics of rotifers. Arch. Hydrobiol. Beih. Ergeb.
food concentration and the size-efficiency hypothesis.
Limnol. 8: 98-122.
Ecology 68(1): 181-187.
Gilbert, J. J. & K. G. Bogdan, 1984. Rotifer grazing: in situ
Walz, N., 1983. Continuous culture of the pelagic rotifers
studies in selectivity and rates. In D. G. Meyers & 1. R.
Keratella cochlearis and Brachionus angularis. Arch. Hydro-
Strickler, editors. Trophic interactions within aquatic eco-
bioI. 98: 70-92.
systems. AAAS Selected Symposium series, 85: 97-133.
Wurdak, E. S., J. 1. Gilbert & R. Jagels, 1978. Fine structure
Goulden, C. E. & L. Henry, 1984. In D. G. Meyers & 1. R.
of resting eggs of the rotifers Brachionus calyciflorus and
Strickler, editors. Lipid energy reserves and their role in
Asplanchna sieboldi. Trans. Amer. Micros. Soc. 97(1):
cladocera. AAAS Selected symposia series 85: 167-185.
49-72.
Goulden, C. E., L. Henry & D. Berrigan, 1987. Egg size,
Hydrobiologia 186/187: 347-354, 1989.
C. Ricci. T. W. Snell and C. E. King (eds), Rotifer Symposium V. 347
© 1989 Kluwer Academic Publishers.

A laboratory study of phosphorus and nitrogen excretion of Euchlanis

dilatata lucksiana

R. D. Gulati\ J. Ejsmont-Karabin2 , J. Rooth! & K. Siewertsen!

! Limnological Institute, Vijverhof Laboratory, Rijksstraatweg 6, 3631 AC Nieuwersluis, The Netherlands;

2 Polish Academy of Sciences, Institute of Ecology, Hydrobiological Station, ul. Lesna 13, 11-730
Mikolajki, Poland

Key words: rotifers, Euchlanis, phosphorus, nitrogen, excretion, body size

Abstract

Phosphorus (P0 4-P) and nitrogen (NH4-N) excretion rates of Euchlanis dilatata lucksiana, a rotifer,
isolated from Lake Loosdrecht (The Netherlands) and cultured in the lake water at 18-19 DC, were
measured in the laboratory.
In a series of experiments, the effects of experiment duration on the P and N excretion rates were
examined. The rates measured in the first half-hour were about 2 times higher for P and 2-4 times for
N than the rates in the subsequent three successive hours which were quite comparable.
Eight experiments were carried out in triplicate, 4 each for P and N excretion measurements, using animals
of two size ranges: 60-125 p,m and > 125 p,m. The specific excretion rates varied from 0.06 to 0.18 p,g
P.mg- 1 DW.h - 1 and 0.21 to 0.76 p,g N.mg- 1 DW.h -1. Generally an inverse relationship was observed
between the specific excretion rates and the mean individual weight. The excretion rates of Euchlanis
measured by us are lower than those reported for several other rotifer species, most of which are much
smaller than Euchlanis.
Extrapolating the excretion rates of Euchlanis to the other rotifer species in Lake Loosdrecht, and
accounting for their density, size and temperature, rotifer excretion appears to be a significant, potential
nutrient (N,P) source for phytoplankton growth in the lake. The excretion rates for the rotifers appear
to be about two thirds ofthe total zooplankton excretion, even though the computed rotifer mean biomass
is about one-third of the total zooplankton biomass.

Introduction zooplankton, mainly the crustaceans. Studies on

the composition, densities and biomass of
In a recent study on phosphorus and nitrogen zooplankton of the Loosdrecht lakes (Gulati, in
excretion rates of the zooplankton community press) and calculations of P and N regeneration
from the Loosdrecht lakes, it was demonstrated rates of zooplankton based on Ejsmont-Karabin
that the P excreted by zooplankton is substantial (1983) show that the rotifers in summer, when
in comparison with P-demand of phytoplankton they are abundant, may contribute from 45 to
(den Oude & Gulati, 1988). This study was, how- 95 % of the P regeneration by total zooplankton.
ever, restricted to the large-sized (> 150 p,m) These release rates, if included in the P-flow
348

scheme of Lake Loosdrecht (van Liere et al., in exclusive food source in the cultures. The animals
press), will largely meet the primary production were removed from the cultures using 60 Jim and
requirements of phytoplankton. 125 Jlm sieves and categorized as small and large,
The mean annual densities of rotifers in the respectively. On termination of each experiment,
Loosdrecht lakes are between 3 and 8 times the lengths of the animals used in the replicates
higher than the crustacean densities. Because the were measured and the mean dry weights com-
mean individual weight of the rotifers (0.05 Jlg C puted from the length-weight regression formula
ind - 1) in the lakes is an order of magnitude lower given by Dumont et al. (1975). For deriving the
than that of the crustaceans (0.44 Jlg C ind - 1 ) carbon biomass, the computed dry weight was
(Gulati, in press), the mean specific excretion multiplied by a factor of 0.427 (Gulati et al.,
rates of rotifers will be about 2.5 times higher than 1987).
the rates of crustaceans, based on the studies of
Ejsmont-Karabin (1984). Excretion experiments
Considering the aforementioned arguments,
the role of P and N regeneration by rotifers in the The excretion rates of Euchlanis were measured in
Loosdrecht lakes needs to be investigated, and its the dark at 18-19 DC, the culture temperature.
quantitative importance in the lake ecosystem About 200 animals were transferred to a Winkler
assessed. This paper deals with Nand P excretion bottle containing 50 ml fresh, filtered (G FF
rates of Euchlanis dilatata lucksiana and is the first Whatman filter) lake water. The experiments were
attempt to examine rotifer excretion in these carried out in triplicate; the controls contained
lakes. The choice of Euchlanis sp. for measuring only the filtered lake water.
excretion rates, like in the feeding studies of this In the 'time-course experiments' the effect of
euchlanid (Gulati et al., 1987), is based on the incubation time on the N and P excretion rates
ease of culturing and handling this species, mainly was studied in separate experiments using two
because of its relatively large size. The animals size classes of Euchlanis. The excretion rates were
used for the excretion studies were obtained from measured after 30 min of incubation in the filtered
the same cultures as for the studies on feeding and lake water and in the subsequent 1st, 2nd and
assimilation rate (Gulati et al., 1987). Besides the 3rd h, i.e. after each hour removing the animals
excretion rates experiments, the effect of experi- and re-transferring them into the freshly filtered
mental duration on the excretion rates was lake water in the Winkler bottles.
measured. The results obtained are discussed in In the second set of experiments the excretion
relation to: 1) the Nand P eontents of the body rates were measured. The animals were incubated
and food of Euchlanis; and 2) their significance in for 30 min in the glass-fibre filtered lake water to
the nutrient regeneration by the zooplankton allow them to clear their guts of the unassimilated
community as a whole. food, and thereafter transferred to fresh filtered
lake water again and excretion rates measured
after 2 h of incubation. Four such experiments
Material and methods were carried out each for N as well as P excretion
measurements separately, using two size cate-
Isolation, culture and biomass of Euchlanis gories. At the end of each experiment the animals
were separated by filtering, killed and fixed in 4 %
Euchlanis was isolated from Lake Loosdrecht and formalin for counting and length measurements.
identified as E. dilatata lucksiana (Gulati et aI.,
1987). It was cultured in the laboratory at Nand P analyses
18-19 DC in lake water filtered with a 33 Jlm mesh
sieve; Therefore, the in situ food ( < 33 Jlm), domi- The water from which the animals had been
nated by filamentous cyanobacteria, was the removed was used for N and P analyses.
 349

Ammonium nitrogen (NH4-N) was determined Table 1. A comparison of the specific NH4-N and P0 4-P
by the bis-pyrazolone method (Prochazkova, excretion rates (J.lg N or P' mg - I DW· h - I) of Euchlanis
1964) in which the coloured complex is extracted dilatata lucksiana in the first 30 min of incubation (A) with the
means of the three successive one-hour incubations (B);
in chloroform and its extinction measured at coefficients of variation (%) are given in the parentheses.
450 nm. Phosphorus (P0 4-P: molybdenum-blue
reactive phosphate) was measured according to Animal size A B AlB
Stephens (1963), using isobutanol as the extract- (J.LgDW·ind- l )
ing solvent for the coloured complex, and the N excretion rates
1.07 0.61 0.26 2.35
extinction values at 690 nm as measure of the P (12.1 ) (14.2) (9.7)
concentration. Both these methods have been 0.51 3.31 0.77 4.30
used successfully in recent other excretion (33.7) (24.7) (5.3)
studies of zooplankton (e.g. Blazka et al., 1982;
Ejsmont-Karabin, 1984). P excretion rates
1.01 0.17 0.078 2.05
For calculating the nutrient recycling rates, (17.7) (34.3) (22.3 )
body nitrogen and phosphorus contents of both 0.79 0.72 0.18 4.00
Euchlanis and its food in the cultures were (25.7) (20.8) (16.4 )
measured. Micro-Dumas method was used for
the analysis of N using an automated C,H,N
analyzer (Perkin Elmer, No. 240); for P, a known assimilation efficiencies of Euchlanis are rather
amount of the dry matter was combusted to ash, high, invariably 100% (Gulati et a!., 1987). Small
hydrolyzed and the concentration measured animals excreted N and P 5 times and 4 times
according to Murphy & Riley (1962). faster, respectively, than the large animals. Since
there was no trend in the fluctuations in the 3
successive one-hour measurements, it was
Results implied that the excretion rates stabilized after
transfer to the fIltered lake water within the fIrst
Time course experiments 30 min.

The observed excretion rates of both N and P, for

large as well as small Euchlanis, are markedly Excretion rates
higher in the fIrst 30 min than in the subsequent
3 h (Fig. 1; Table 1). The excretion rates in the The mean rates of N excretion in the first three of
fIrst half-hour can be apparently higher if the the four experiments were quite comparable, with
egestion products contribute signifIcantly to the daily recycling via excretion of 5-6 % body N
excretion losses. The egestion, however, appears (Table 2). The specifIc excretion rates for the
to contribute negligibly to the excretion because smallest animals (0.514 jlg DW.ind - 1: Exp.4)

Table 2. Specific NH4-N excretion rates (J.Lg N· mg- I DW· h - I) of Euchlanis dilatata lucksiana of different sizes; coefficients
of variations (%) of the replicates are given in parentheses; also daily recycling of the body N is given.

Exp. No. Size of the animal Excretion rate Recycling (d - I)

(J.Lg DW 'ind- I )

1 1.09 ( 8.6) 0.24 ( 5.1) 0.06

2 1.06 (l2.1) 0.25 ( 0.6) 0.06
3 0.62 (26.7) 0.21 (15.7) 0.05
4 0.51 (33.7) 0.76 ( 5.3) 0.18
350

were, however, about 3 times the rates of largest The P recycling rates of about 14 %d - 1 for the
animals. Interestingly, these N excretion rates are animals weighing about 1 /-lg DW.ind - 1 were on
almost identical to the rates measured in the time- the average 2.4 times faster than of N recycling
course experiments excluding the first half-hour (Tables 2 and 3: Exp. 1 and 2). Such a com-
(Fig. 1 and Table 1). parison of the Nand P recycling is, however, not

3
I
.c.
j
0
.--
I
C'I
E 2
z
C'I
:1

o~--~~--~~--------~~------~-U~--

1.00

.--
~ 015
:3
o
.--
I

~ 0.50
a..
C'I
:1

0.25

0.0 0 '--_......L.-I~_..L.....L..a...-_-:--_--L-~_ _ _--J-L.L.'--_ _

0-0.5 0.5-1.5 1.5 - 2.5 2.5-3.5
Hour of in(u bation
Fig. I. Specific NH4-N and P04-P excretion rates of Euchlanis dilatata lucksiana in the course of 3.5 h; incubation time of the
first experiment was 30 min and of the subsequent three experiments 1 h each. The two bars for each experiment represent: blank
bar, large animals; and hatched bars, small animals; the asterisk represents the + S.D. For more details see Table 1.
 351

Table 3. Specific P04 -P excretion rates (J.tg P' mg- 1 DW· h -1) of Euchlanis dilatata lucksiana of different sizes; coefficients of
variation (%) of the replicates are given in parentheses; also daily recycling of the body P is given.

Exp. No. Size of the animal Excretion rate Recycling (d - 1 )

(J..Lg DW'ind- l )

1 1.01 (17.7) 0.068 ( 6.8) 0.14

2 0.95 ( 7.3) 0.063 (24.8) 0.14
3 0.81 (25.7) 0.182 (13.8) 0.39
4 0.65 (30.4) 0.170 (17.3) 0.36

possible for the smaller animals: first, because of temperature; b) animal size; c) feeding history;
the greater weight differences between the animals d) concentration or extent of crowding; and
used for N and P in the Exp. 3 and 4 (Tables 2 e) length of the incubation period.
and 3); and secondly, also due to the high rates of The temperature which has generally a signifi-
P excretion for the smaller animals, both in Exp. 3 cant and direct effect on rotifer excretion, with a
and 4 (Table 3) but of N excretion only in Exp. 4 QlO of about 2.6 (Ejsmont-Karabin, 1984), was
(Table 2). kept at 18.5 ± 0.5 DC both in the culture medium
The coefficients of variation in the replicates of and during the incubations for excretion measure-
Nand P excretion rate measurements, with one ments. The excretion rates tended to increase with
exception (Exp. 2, Table 3), were less than the the decreasing weight of Euchlanis. The signifi-
variations in the size replicates of Euchlanis. The cance of the differences, however, cannot be
replicates of N excretion rates varied less than the tested because of the limited number of size
replicates of P excretion rates in 3 out of the 4 classes studied. This narrow size range within a
experiments. clone of a rotifer species in culture may be because
of the short life span and constancy of cell number
in rotifers after the embryonic development is
Discussion completed (Ruttner-Kolisko, 1972). Moreover,
our computed dry weights of Euchlanis based on
the L-W regressions of Dumont (1975) are about
This study restricts the excretion measurements
twice the weights based on the volume-to-weight
to P04-P and NH4-N. The measured rates of P
conversion (Ruttner-Kolisko, 1977), assuming
excretion will include, besides the orthophos-
dry weight to be 10% of the wet weight (see e.g.
phates, the phosphates formed by hydrolysis of
Bottrell et al., 1976). Consequently, our specific
dissolved organic phosphorus compounds. These
excretion rates calculated using the regression-
latter comprise about 20 %of the total P excretion
based dry weights are half as high as the rates
(Peters & Lean, 1973; Ejsmont-Karabin, 1984).
based on the volume. Such discrepancies will
For N, ammonia is the predominant break-down
affect the comparability ofliterature data adverse-
product of aquatic invertebrates including the
ly, like the errors relating to methodological and
zooplankton, which are ammoniotelic.
behavioural differences.
As regards feeding history and starving of the
animals during the experiments, the stability of
Factors affecting the excretion rates excretion rates after the first 30 min shows that
starving most likely did not alter the physiology
Several factors may affect the rate of zooplankton such that the rates were reduced (see also Ikeda,
excretion as well as comparability of the different 1971; Mayzaud, 1976). Also, release of N and P
studies. Important among these are: a) the water through bacterial activity and increased minerali-
352

zation, which may mask the decrease in excretion incubation were not only comparable with one
rates (Le Borgne, 1979), does not appear to have another (Fig. 1), but also with those measured
played a role because we used fIltered lake water subsequently on the basis of 2-h incubations
and corrected our results for changes in the con- (Tables 2 and 3).
trols. Ejsmont-Karabin (1984) has clearly demon-
In our experiments we used concentrations (ca. strated that the mean specific excretion rates of
4000 ind.l- I) of Euchlanis that are quite similar to rotifers increase markedly with decreasing weight,
the mean density (3770 ind.l- I) of total rotifers e.g. P excretion rates increase from 0.19 to
encountered in the Loosdrecht lakes (Gulati, in 35.81lg P.mg- I DW.h - 1 corresponding to a
press). Moreover, they are much lower than the decrease in individual weights from 0.776 to
densities (40000 ind.l- I) in our Euchlanis stock 0.013 Ilg DW. The range of variations is even
cultures (Ejsmont-Karabin et ai., 1989). There- much larger if all the extreme values are included.
fore, crowding-related factors do not appear to be P excretion rates of Euchlanis measured by us
relevant in our study. (Table 3) are very comparable with the minimal
The exposure-time related changes in the speci- rates (range: 0.06-0.42 Ilg P.mg- 1 DW.h - I; and
fic excretion rates of Euchlanis differed in their mean: 0.19 mg P.mg- I DW.h -I) reported for a
magnitude for the small and large animals. Both, rotifer, namely Asplanchna priodonta (Ejsmont-
the N and P excretion rates are higher during the Karabin 1984: Table 1). Asplanchna has a range
first 30 min than in the subsequent 3 h, roughly 4 of weights, 0.5-1.07Ilg DW.ind - I (Ejsmont-
times for the small animals and 2 times for the Karabin, 1984), that are quite similar to the
large animals (Table 1). In the first 30 min the weights of our Euchlanis (0.5-1.0 Ilg DW.ind - I).
small animals excrete both N and P 4-5 times Interestingly, these Euchlanis weights converted
more rapidly than the large animals. The size- to carbon do not differ much from the mean
dependent difference in the animal behaviour is weight (0.44 Ilg C.ind - I) of crustacean com-
too great to be explained on the basis of metabo- munity of the Loosdrecht lakes, in which small
lism alone. cladoceran species of Bosmina and Chydorus pre-
For the stabilization of the excretion rates after dominate (Gulati, in press). Also P excretion rates
the first 30 min there are two possible explana- of crustaceans in these lakes of 0.17 p.g P.mg- I
tions. First, the stress effects (Le Borgne, 1979) DW.h - 1 (Den Oude & Gulati, in press: Table 1)
on transferring the animals from the cultures to are almost identical with the rates of Euchlanis
fresh, fIltered lake water without food may stimu- measured by us (Exp. 3 and 4: Table 3).
late the animal to fIlter maximally in a medium
devoid of food. However, this increased activity Elemental ratios
is short-lasting, tending to stabilize even before
the 3 successive hourly transfers into the fresh The N: P ratios of 3.4-3.6 for the large animals
food-less medium each time. Second, the animals and 4.2-4.5 for the small animals, based on
apparently get quickly adapted to the short-term ammonia and phosphate excretion, were constant
starvation, with no more decrease in metabolic and comparable with the ratios derived from the
activity than that observed in the first hour after generalized regression formulae for the rotifers of
the first 30 min in the time course experiments. In Ejsmont-Karabin (1984), namely 3.6 for the large
short, like in several other studies on zooplankton and 4.0 for the small animals. The higher N: P
excretion, but especially those on the rotifers (e.g., ratios for the smaller animals may be because of
Ejsmont-Karabin, 1983, 1984; Ejsmont-Karabin the relatively higher protein catabolism of these
et ai., 1983), no symptoms of starvation, or other animals; moreover, the smaller animals (0.51 p.g
adverse effects, were observed, which may lead to DW.ind - 1) in the N-experiment were much
decreased excretion in the initial few hours of smaller than the animals (0.79 p.g DW.ind - 1) in
incubation. Lastly, the rates in the 0.5-3.5 h of the P experiments (Table 1).
 353

On comparing the elemental ratios, namely the (ca. 65%) to the mean zooplankton excretion
C : N : P ratio of the Euchlanis food, Euchlanis rates, with a range of values between 45 and 90%
body and the excretion products summarized in summer (Gulati, in press; Van Liere et al., in
below, some interesting discrepancies emerge: press).
The excretion rates of Euchlanis found by us,
Ratio C :N :P though in the range of literature values that vary
Food 100: 15.4 : 0.9 widely, are low. This, like for the crustacean excre-
Euchlanis 100: 23.7: 2.6 tion rates (Den Oude & Gulati, 1988), may be
Excretion products 100: 16.0: 4.0 attributed to high C : P ratios (± 125) of sestonic
lake food of Euchlanis, because of P-limitation of
Euchlanis contains about 3 times more P and cyanobacteria that dominate the seston. Evidence
1.5 times more N than its food. Also, per unit of the direct effects of P-content of zooplankton
carbon, the excretion products contain about food on excretory turnover rate of P has been
50% less N, but 50% more P than N and P in the presented in some recent studies (e.g. Lehman &
animal body. Evidently, the animal is able to Naumoski, 1985).
maintain a lower C : N ratio in the body than its Finally, our study on Euchlanis and some com-
food; however, P losses as indicated by the both parisons of the literature data on rotifer excretion
lower C : P and N : P ratios of the excretory pro- have demonstrated: 1) the specific excretion rates
ducts - than of the animal and its food - are of rotifers - even within a species - vary
difficult to explain. enormously because of the inverse relationship
between the rates and individual weights; 2) the
range of excretion rates is very wide if one com-
Role of rotifer excretion pares the different rotifer species, which differ in
weight about 60 fold; and 3) in shallow and eutro-
The Euchlanis excretion rates are not quite repre- phic lakes like Loosdrecht Lake, both the much
sentative of the rotifer community of the smaller size and relatively higher densities of
Loosdrecht lakes. The rotifers in these lakes are rotifers than of the crustaceans may contribute
dominated by Keratella cochlearis and Anuraeop- significantly to the importance of rotifers in
sis jissa, with individual weights (0.01-0.02 J.lg nutrient regeneration, especially in summer.
DW.ind -I) that are an order of magnitude lower
than that of Euchlanis. Their excretion rates
(Ejsmont-Karabin, 1984: Table 1) are, therefore, Acknowledgements
probably two orders of magnitude greater than the
rates of Euchlanis measured under similar tem- We thank Drs. B. Z. Salome, Dr. J. Vijverberg &
peratures (± 20 0 C), using the same methods. Miss A. L. Wilms for critical reading and Miss
Our computed individual mean weight of rotifers Cecilia Kroon for typing of the manuscript. The
in the Loosdrecht lakes of ca. 0.05 J.lg DW.ind - 1 second author (J. Ejsmont-Karabin) is grateful to
(Gulati, in press) is about one-tenth the individual the Limnological Institute (Nieuwersluis) for
mean weight of crustaceans. One would, there- funding her travel to and short stay in The N ether-
fore, expect the rotifer specific excretion rates to lands in 1984 when the Euchlanis investigation
be much higher and their contribution to the P was carried out.
and N excretion of total zooplankton in the lakes
much greater than based on the crustacean:
References
rotifer mean biomass ratio of ca. 2: 1 calculated
from the lake data of 1982-'87. Thus, the rotifers Blaika, P., Z. Brandl & L. Prochazkova, 1982. Oxygen con-
that contribute about 35% to the total zooplank- sumption and phosphate excretion in pond zooplankton.
ton biomass will contribute roughly twice as much Limnol. Oceanogr. 27: 294-303.
354

Bottrell, H. H., A. Duncan, Z. M. Gliwicz, E. Grygierek, A. ratory study of feeding and assimilation in Euchlanis
Herzig, A. Hillbricht Ilkowska, H. Kurasawa, P. Larrson dilatata lucksiana. Hydrobiologia 147: 289-296.
& T. Weglenska, 1976. A review of some problems in Ikeda, T., 1971. Changes in respiration rate and in com-
zooplankton production studies. Norw. J. Zool. 24: position of organic matter in Calanus cristatus (Crustacea,
419-456. Copepoda) under starvation. Bull. Fac. Fish. Hokkaido
Den Oude, P. & R. D. Gulati, 1988. Phosphorus and nitrogen Univ. 21: 280-298.
excretion rates of zooplankton from the eutrophic Lehman, J. T. & T. Naumoski, 1985. Content and turnover
Loosdrecht lakes, with notes on other P sources for phyto- of phosphorus in Daphnia pUlex: Effect of food quality.
plankton requirements. Hydrobiologia 169: 379-390. Hydrobiologia 128: 119-125.
Dumont, H. J., I. van de Velde & S. Dumont, 1975. The dry Le Borgne, R. P., 1979. Influence of duration of incubation on
weight estimate of biomass in a selection of Cladocera, zooplankton respiration and excretion results. J. expo mar.
Copepoda and Rotifera from the plankton, perifyton and BioI. Ecol. 37: 127-137.
benthos of continental waters. Oecologia 19: 75-97. Mayzaud, P., 1976. Respiration and nitrogen excretion of
Ejsmont-Karabin, J., 1983. Ammonia nitrogen and inorganic zooplankton. IV. The influence of starvation on the
phosphorus excretion by the planktonic rotifers. Hydro- metabolism and biochemical composition of some species.
biologia 104: 231-236. Mar. BioI. 37: 47-58.
Ejsmont-Karabin, J., 1984. Phosphorus and nitrogen excre- Murphy, J. & J. P. Riley, 1962. A modified single solution
tion by zooplankton (rotifers and crustaceans) in relation method for the determination of phosphate in natural
to individual bodyweights of the animals, ambient tem- waters. Anal. Chim. Acta 27: 31-36.
perature and presence or absence of food. Ekol. pol. 32: Peters, R. & D. Lean, 1973. The characterization of soluble
3-42. phosphorus released by limnetic zooplankton. Limnol.
Ejsmont-Karabin, J., R. D. Gulati & J. Rooth, 1989. Is food Oceanogr. 18: 270-279.
availability the main factor controlling Euchlanis dilatata Prochilzkovil, L., 1964. Spectrophotometric determinations
lucksiana Hauer abundance in a shallow, hypertrophic of ammonia as rub azoic acid with bispyrazolone reagent.
lake? Hydrobiologia, 186/187: 29-34. Anal. Chem. 365: 865-871.
Ejsmont-Karabin, J., L. Bownik-Dylinska & W. A. Ruttner-Kolisko, A., 1972. Rotatoria. In: Das Zooplankton
Godlewska-Lipowa, 1983. Biotic structure and processes der Binnengewasser Teill. Schweizerbart, Stuttgart,
in the lake system of R. Jorka Watershed (Masurian 1972. (Series) Die Binnengewasser 26 Teill. P 99-234.
Lakeland, Poland). VII. Phosphorus and nitrogen regen- Ruttner-Kolisko, A., 1977. Suggestions for biomass calcula-
eration by zooplankton as the mechanism of the nutrient tion of plankton rotifers. Arch. Hydrobiol. Beih. 8: 71-76.
supplying for bacterio- and phytoplankton. Ekol. pol. 31: Stephens, K., 1963. Determinations of low phosphate con-
719-746. centrations in lake and marine waters. Limnol. Oceanogr.
Gulati, R. D., in press. Zooplankton structure in Loosdrecht 8: 361-362.
lakes in relation to the trophic status and the recent resto- Van Liere, L., R. D. Gulati, F. G. Wortelboer & E. H. R. R.
ration measures. Hydrobiologia. Lammens, in press. Phosphorus dynamics following resto-
Gulati, R. D., J. Rooth & J. Ejsmont-Karabin, 1987. A labo- ration measures in Loosdrecht lakes. Hydrobiologia.
Hydrobiologia 186/187: 355-361, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 355
© 1989 Kluwer Academic Publishers.

A new method to estimate individual dry weights of rotifers *

Hans-Rainer Pauli
Limnologisches Institut, Universitiit Konstanz, FRG

Key words: rotifers, dry weight, biomass, productivity

Abstract

Body and egg volumes of 12 rotifer species of Lake Constance were calculated from more than 18000
individually measured animals using geometric approximations. Body (VA) and egg (VE) volumes of
various rotifer species have an allometric relationship described by: VE = 1272· VAO.3379, while the
relation VElVA for each species was constant ranging from 0.016 to 0.87. Dry weight of adult rotifers
was assumed to be the sum of the egg dry weight plus a corrected weight increment from egg to adult.
It was estimated with the formula WA = WE '(1 + a'FA/FE ) where (a) was varied between 0 and 0.1.
Egg dry weight content was assumed to be 35% of fresh weight. The estimates of dry weight to wet weight
ratios ranged from 0.57% (Asplanchnapriodonta) to 29.4% (Kellicottia longispina). Dry weight estimates
obtained with this method are comparable to those obtained with direct measurements of dry weight or
carbon content.

Introduction When estimating biomass or production of

natural rotifer populations from field data the
A reliable estimate of productivity of natural most disputable step is the conversion of body
rotifer communities is still difficult. While final volume to dry weight or other biomass units. Only
calculation of production is relatively simple to very few direct measurements of dry weight or
perform (Downing & Rigler, 1984), the collection carbon content exist. The common practice to
of the appropriate data needed for these calcu- derive individual body mass from values pub-
lations poses several problems. The estimate of lished in the literature is unsatisfactory, because
rotifer production requires the following data: size variation within a given rotifer species can be
(1) popUlation census in short temporal and significant (Dumontet al., 1975; Latja& Salonen,
dense spatial intervals (including numbers of indi- 1978; Salonen & Latja, 1988). Similarly, transfor-
viduals as well as number of eggs, sorted by mation of body volumes, which are commonly
species categories), (2) conversion of numbers of used as an estimate of fresh weight, with a general
individuals and eggs to appropriate biomass conversion factor of 10 percent dry weight of body
units, (3) determination of rates for somatic volume, does not reflect the differences among
growth and growth of the popUlation. rotifer species. Large bodied genera like

*This investigation was supported by the Deutsche Forschungsgemeinschaft within the Special Collaboration Program (SFB)
'Cycling of Matter in Lake Constance'.
356

Asplanchna contain more than 90 percent water calculated from more than 18000 measured ani-
(Salonen & Latja, 1988) while small genera like mals:
Keratella contain less (Walz, 1987).
This study proposes a method for conversion of (1)
volumes to biomass incorporating some simple
but probably not unrealistic assumptions. The equivalent graphic regression line is shown in
Fig. 1. The portion of explained variance is 73 %,
standard error of the mean is 0.7% for the inter-
Material and methods
cept and 1.3 % for the slope, respectively. The
square symbols represent mean values for dif-
The material for this study was collected during
ferent rotifer species from Lake Constance. Addi-
two years (1984, 1985) from Lake Constance-
tional values for other species are included in this
Uberlingersee (West Germany). The sampling
figure, recalculated from literature data on length
intervals ranged from two days during spring to
and width of animals and of eggs (e.g.
14 days during winter. The upper 20 meters of
Ruttner-Kolisko, 1972; Koste, 1978). Even these
water were sampled completely with 10 samples
very rough estimates fit well into the confidence
using a 2 meter long sampling device. Three addi-
limits of the calculated regression line. Only one
tional samples were taken at 30, 40, and 50 meters
value, reported for Keratella quadrata from Walz
depth. Samples were filtered through steel gauze
(1987), falls outside the confidence limits for
of 50 f.lm mesh size and preserved in 4 % sugar
predicted values. Equation (1) was used to esti-
formalin (Haney & Hall, 1973). Counting ofindi-
mate the fresh weight of eggs of those species
viduals and eggs was carried out following the
where no eggs could be found or assigned to
Utermohl method (1958). Measurements ofmor-
without doubt.
phometric parameters (body length, body width,
In Fig. 2, the relationship between body and
le~gth of spines and appendices, egg length, egg
egg volumes for species of different size is com-
WIdth) were performed using an inverted micro-
pared with four relationships calculated for four
scope combined with a semi-automatic image
groups of species of similar dimensions. The
analysis system. Body and egg volumes were cal-
culated from these individual size measurements
using geometric approximations as proposed by 6r-------------------------~
Ruttner-Kolisko (1978). These volumes were then v =1272*V. .3379
(I)
used as estimates of fresh weight, assuming a E egg bOd; 0 .•..••.•..•.• •·•·· 0 ••..••••.
density of 1. o
::l
,"0 * ......................
The following abbreviations and definitions are > ""l~f
I
used in this paper: V (f.lm 3 ): volume, F: fresh g:S """*
___>Ie 0 ..•....,
w ..••.. ~ ..-*.,
weight, W (f.lg): dry weight, a: increment factor b ....·"""" ............-;....
- '
E: ~gg, A: adult, I: increment, p: proportion dry o
weIght of fresh weight (W/F), F = V, W = F·p.
O'l ••••••••••••••
.2 ...
~or the conversion of carbon content to dry
4~ ____~______~____~~____~
weIght, the carbon content of organic material
4 5 6 7 8
was assumed to be 50 %of dry weight, according
log10 of Body-Volume
to Latja & Salonen (1978).
Fig. 1. Volumes of rotifer eggs versus body volumes: Regres-
sion line of the base-l0 logarithms of egg and body volumes
Results
(,um 3 ) of 12 rotifer species from Lake Constance-Uber-
linger see (squares). Stars represent values recalculated from
An allometric relationship (y = a' x b ) between literature data. The dashed lines represent 95% confidence
egg and body volume of 12 rotifer species was limits for the predicted single values.
 357

6r-----------------------------~~ weight increment (WI)' a correction factor (c) was

introduced to obtain:

(3)

Replacing (WI) with (WA - WE) and FI with

(FA - FE) we get:
o
O'l
o (4)
4~--~--._----~_.------_.------~
4 5 6 7 8 Under the simplified assumption that the amount
log 1 0 of Body-Volume of biomass accumulated from egg to adult is
related to the final body size, FI in equation (3)
Fig. 2. Volumes ofrotifer eggs versus body volumes: Regres-
can be replaced by FA' with a modified correction
sion line of the base-IO logarithms of egg and body volume
(J.lm 3 ) of 12 rotifer species from Lake Constance-
factor (a):
Dberlingersee and separately calculated regression lines for
four groups of rotifer species with similar body size. For (5)
composition of the groups see text.
The final dry weight of the adult animal (WA ) can
be calculated from:
groups are composed of the species: (1) Keratella
cochlearis, Kellicottia longispina, Pompholyx sul- (6)
cata, (2) Polyarthra vulgaris, P. dolychoptera,
P. major, Brachionus angularis, Filinia terminalis, and the relative proportion of dry weight of the
Keratella hiemalis, K. quadrata, (3) Synchaeta adult (p A) as
pectinata, (4) Asplanchna priodonta. Body size rela-
tionships for each group were calculated sepa- (7)
rately and their slopes were tested for significant
differences. The slopes of the regression lines for
the four groups ranged from 1.065 to 0.905. None 4.0.---------------,
was significantly different from any other at the
95 % significance level. These slopes with numeri- ~ 3.5
3:
cal values close to one indicate a linear proportio- ~3.0
nality of egg to body volumes among rotifers of -J
similar size or within species. (f)
Q)
2.5
-+J

The following formulas and assumptions are o

E 2.0
proposed for the transformation offresh weight to :;:;
(f)
w 1.5
dry weight. Dry weight of an adult rotifer (WA )
'0
was assumed to be the sum of egg dry weight ( WE) 0 1.0
:;:;
and a weight increment (WI) from egg to adult: 0 0.0 0.5 1.0 1.5 2.0
0::

(2)

Fig. 3. Ratios WA(a)/WA(o) of dry weight (WA ) of adult

Under the assumption that dry weight to wet
rotifers estimated with WA = WE' (1 + -FA/FE) among
weight ratios of adult animals are smaller than species with different VAlVE ratios. WA(a) was calculated for
those of their eggs (WA/FA < WE/FE)' and there- different values of (a) (0.02, 0.05, 0.07, 0.1), WA(o) was calcu-
fore also the dry weight to wet weight ratio of the lated for a = O.
358

Table 1. Mean body volumes, mean egg volumes and mean than 1 - FE/FA' A value for (a) equal to
egg to body volume ratios for 12 rotifer species from Lake 1 - FE/FA would result in an identical value for
Constance-Uberiingersee (W-Germany).
the proportion of dry weight in both eggs and
Species Body Egg VElVA
adults. To obtain a reasonable value for (a), it was
volume volume varied arbitrarily between 0 and 0.1, which means
a volume dependent weight increase between 0
J-Lm 3 J-Lm 3 and 10 percent from egg to adult. The values
Asplanchna priodonta 27270000 424600 0.016
852000 192500 0.23
obtained with equation (4) differ from those esti-
Brachionus angularis
Filinia terminalis 443000 100700 0.23 mated with equation (6) by 0 to 10 % of the egg
Kellicottia longispina 99700 86600 0.87 dry weight. Figure 3 shows the effect of different
Keratella cochlearis 87500 61000 0.70 values of (a) on dry weight estimates (WA ) calcu-
Keratella hiemalis 350200 92600 0.27 lated with equation (6). The curves represent the
Keratella quadrata 733700 183500 0.25
Polyarthra dolychoptera 407800 103500 0.25
ratio of WA for a = 0.02, a = 0.05, a = 0.07,
Polyarthra major 826000 140300 0.17 a = 0.1 to WA for a = 0 among species with dif-
Polyarthra vulgaris 443200 105800 0.24 ferent VAlVE ratios. The effect on the estimates
Pompholyx sulcata 85600 60200 0.70 when varying (a) is greater in species with large
Synchaeta pectinata 2338000 188000 0.08 values for the ratio VAlVE' Table 1 summarizes
egg and body volumes and ratios VElVA' which
cover the range fram 0.016 (Asplanchna priodonta)
Because the dry weight to fresh weight ratio to 0.87 (Kellicottia longispina). Table 2 contains
(WA/FA) of an adult rotifer is expected to be dry weight estimates obtained under the assump-
smaller than that of its eggs (WE/FE)' the value of tion that egg dry weight content of ratifers is com-
the weight increment factor (a) has to be smaller parable to that of Daphnia in which it is 35 %

Table 2. Measurements and estimates ofrotifer dry weights. All values are transformed to dry weight. Original units were dry
weight (DW), body volume (V), and carbon content (C).

Species or Genus % Dry weight Dry weight Original Reference

in fresh weight J-Lg/lnd unit

Asplanchna brightwelli 3.90 0.28 -1.50 DW

Asplanchna herricki 0.34- 0.96 2.10 -3.20 C 2
Asplanchna priodonta 10.00 V 3
Asplanchna priodonta 1.70 V 4
Asplanchna priodonta 0.60 DW 5
Asplanchna priodonta 0.48 -0.52 DW 1
Asplanchna priodonta 0.44 DW 6
Asplanchna priodonta 0.12 -0.50 C 7
Asplanchna priodonta 0.42- 1.9 0.30 -1.30 C 2
Asplanchna priodonta 0.57- 4.1 0.04 -3.80 V 11
Brachionus angularis 0.40 -0.54 DW 1
Brachionus angularis 0.05 DW 6
Brachionus angularis 15.1 0.11 C,V 8
Brachionus angularis 8.3 -11.8 0.08 -0.12 V 11
Brachionus calyciflorus 0.11 -0.47 DW 1
Brachionus calyciflorus 0.20 DW 6
Brachionus calyciflorus 2.2 - 5.7 0.06 -0.37 V 11
Conochilus unicornis 0.084 C 7
Conochilus unicornis 10.4 -13.9 0.03 -0.06 V 11
Kellicottia longispina 0.03 -0.06 C 7
Kellicottia /ongispina 43.2 0.04 C, V 8
 359

Table 2. Continued.

Species or Genus % Dry weight nry weight Original Reference

in fresh weight ,ug/Ind unit

Kellicottia longispina 25.9 -29.4 0.02 -0.04 V 11

Keratella cochlearis 0.038 DW 9
Keratella cochlearis 0.11 DW 1
Keratella cochlearis 0.07 DW 6
Keratella cochlearis 0.04 -0.05 C 7
Keratella cochlearis 43.2 0.04 -0.05 C,V 8
Keratella cochlearis 24.2 -27.7 0.02 -0.04 V 11
Keratella quadrata 11.9 0.075 DW,V 10
Keratella quadrata 0.32 -0.35 DW 1
Keratella quadrata 0.07 DW 6
Keratella quadrata 7.7 -11.2 0.03 -0.11 V 11
Polyarthra vulgaris 0.043 DW 6
Polyarthra vulgaris 0.066 DW 7
Polyarthra vulgaris 8.7 -12.2 0.02 -0.06 V 11
Polyarthra spp. 0.74 DW 1
Polyarthra dolychoptera 9.4 -12.9 0.03 -0.06 V 11
Pompholyx sulcata 22.8 -26.4 0.02 -0.03 V 11
Synchaeta spp. 0.26 -0.27 DW 1
Synchaeta spp. 9.8 -13.3 0.03 -0.06 DW 11
Synchaeta oblonga 0.02 DW 6
Synchaeta stylata 0.20 DW 6
Synchaeta stylata 2.7 - 6.2 0.06 -0.15 V 11
Synchaeta pectinata 3.2 - 6.7 0.03 -0.23 V 11
Ascomorpha ecaudis 12.4 -15.9 0.03 -0.04 V 11
Ascomorpha ovalis 16.7 -20.2 0.02 -0.03 V 11
Filinia terminalis 9.2 -12.7 0.03 -0.06 V 11
Gastropus stylifer 9.0 -12.5 0.02 -0.04 V 11
Keratella hiemalis 9.5 -13.0 0.02 -0.06 V 11
Notholca caudata 4.1 - 7.6 0.05 -0.08 V 11
Polyarthra major 5.7 - 9.2 0.04 -0.11 V 11
Trichocerca birostris 12.2 -15.7 0.03 -0.04 V 11
Trichocerca capucina 5.2 - 8.7 0.05 -0.08 V 11
Trichocerca porcellus 10.2 -13.7 0.03 -0.05 V 11
Trichocerca pusilla 24.2 -27.7 0.02 V 11

Reference: (1) Dumont et al. (1975), (2) Salonen & Latja (1988), (3) Naulaplili (1966), (4) Aasa(1970), (5) Narita & Mori (1975),
(6) Bottrell et al. (1976), (7) Latja & Salonen (1978), (8) Walz (1987), (9) Schindler & Noven (1971), (10) Doohan & Rainbow
(1971), (11) present study

(Bikar, 1986). Included in this table (2) are litera- Discussion

ture values of direct measurements. The lower
values of the given ranges are obtained by setting The common use of allometric equations in bio-
(a) to 0, the upper bound reflects a value for (a) logy in general has good theoretical grounds
of 0.1. The resulting estimates of adult dry weight (Schmidt-Nielsen, 1984; Peters, 1983). Even if
content of fresh weight ranges from 0.57% not completely understood in a particular case,
(Asplanchna priodonta) to 29.4 % (Kellicottia empirical use of such equations can be very
longispina) among the species included in this helpful.
study. In rotifers, egg volume obviously represents a
360

constant fraction of the body volume in a given absolute dry weight or carbon content of single
species, where larger animals carry larger eggs. specimens are of little use without additional
For rotifer species of increasing size, the relation information on size or fresh weight of the analyzed
of egg volume to body volume decreases. The individuals. Body size of defined species can vary
exponent of 0.34 in equation (1) is relatively small in a range of one order of magnitude. Therefore,
when compared to those for other groups of dry weight or carbon content should be expressed
invertebrates. Peters (1983) suggested that the in relation to body size.
reproductive investment of invertebrates rises The assumption of 10% dry weight of body
with body mass to the power of 0.75. The con- volume for all rotifer species can no longer be
siderable difference between these two values is accepted. Large, soft-bodied and transparent
possibly due to the fact that rotifer species differ genera like Asplanehna and Synehaeta have dry
more in their dry biomass content than species of weight content lower than 10%, while the values
other taxa. are higher in smaller genera with thick loricas or
Constant egg to body volume ratios of particu- long spines like Keratella or Kellieottia. For
lar rotifer species have been reported by Walz several species of Asplanehna dry weight to fresh
(1987). These values found by Walz are com- weight ratios between 3.9% (Dumont et al., 1975)
parable to those of the present study although his and 0.34% (Salonen & Latja, 1988) have been
values are smaller for Kellieottia longispina (0.75) measured, while the proposed method for the esti-
and Keratella eoehlearis (0.56) and higher for mation of dry weight results in values between
Keratella quadrata (0.50) and Polyarthra (0.27) 0.57 % and 4.1 %, depending on the value for the
species. Because these values are based on a rela- factor (a). For Keratella eoehlearis a carbon con-
tively small number of observations, they are tent of21.4% was measured by Walz (1987). This
probably biased by seasonal and individual varia- value suggests a dry weight content of 40% under
tions. They fall, however, into the limits of the the general assumption that 50% of dry weight
observed range of egg to body volume relation- consists of carbon. The values of 24.2-27.7%
ships of these species. estimated in the present study agree better to that
A comparison of dry weight estimates obtained value than to the common 10% concept. Latja &
by the method proposed here with the few existing Salonen (1978) reported for singular individuals
direct measurements of dry weight or carbon con- of Keratella eochlearis carbon weights of 0.024 Jig
tent is difficult for several reasons. The necessity and for singular individuals of Kellieottia
of expressing the results in equal units is not yet longispina carbon weights between 0.015 and
realized by most authors. Some results are 0.03 Jig. When recalculating these values, assum-
expressed as dry weight or carbon weight for ing a mean body size similar to those found by
single individuals, others as dry weight recalcu- other authors, carbon content amounts to almost
lated from body volumes with the general assump- 50% for Keratella cochlearis and it ranges from
tion of 10% dry weight content, and in few cases 30% to 60% for Kellicottia longispina. Although
as percentage of body volume. Body volume, even these values seem to be rather high, they support
when calculated from a large number of measure- the assumption of a higher dry weight content
ments of linear size with suitable geometric than 10 % for these species. The estimated
models, is only a rough estimate of fresh weight. weights for particular individuals in this study,
Nevertheless, it is the best available one. Body ranging from 0.02 to 0.04 Jig for both Keratella
size measurements are easy to perform and pro- cochlearis and Kellicottia longispina, agree well.
vide additional information on seasonal changes The value of 11.9% dry weight content of
and individual variation among species. Data on Keratella quadrata reported by Doohan &
dry weight or carbon content of individuals Rainbow (1971) was calculated from the given
should be applicable for the conversion of abun- mean length of 142 Jim using Ruttner-Kolisko's
dance data to biomass. Therefore, data on (1977) formula and the given individual dry
 361

weight of 0.075 J.lg. The corresponding values of Downing, J. A. & F. H. Rigler, 1984. A manual on Methods
this study cover the range between 7.7 % and for the Assessment of Secondary Productivity in Fresh
Waters, 2., Blackwell Scientific Publications, Oxford,
11.2 %, the dry weights of single individuals calcu- 501 pp.
lated from this value are similar to the 0.075 J.lg Dumont, H. J., 1. Van de Ve1de & S. Dumont, 1975. The dry
from Doohan & Rainbow (1971) and to the weight estimate of biomass in a selection of Cladocera,
0.07 J.lg given in Bottrell et al. (1976). The only Copepoda, and Rotifera from the plankton, periphyton,
values which do not match well are those reported and benthos of continental waters. Oecologia (Berl.) 19:
75-97.
by Dumont et al. (1975), which are considerably Haney, J. F. & D. J. Hall, 1973. Sugar-coated Daphnia: A
higher than most other values in the literature. For preservation technique for Cladocera. Limno!. Oceanogr.
example, their value for Keratella quadrata 18: 322-333.
(0.35 J.lg) exceeds by seven times those of other Koste, W., 1978. Rotatoria. Die Radertiere Mitteleuropas. 1.
authors (0.07 J.lg). For Brachionus angularis, their Textband. Gebriider Borntrager, Berlin, Stuttgart, 673 pp.
Koste, W., 1978. Rotatoria. Die Radertiere Mitteleuropas. 2.
values (0.4-0.54 J.lg) are about 10 times higher Tafelband. Gebriider Borntrager, Berlin, Stuttgart, 234
than the value of 0.05 J.lg cited in Bottrell et al. Tafeln.
(1976). Latja, R & K. Salonen, 1978. Carbon analysis for the
The method proposed in this study does not, of determination of individual biomasses of planktonic ani-
course, estimate 'true' values of dry weights in mals. Verh. int. Ver. Limno!. 20: 2556-2560.
Naulapaa, A., 1966. Eraiden Suomessa esiintyvien plank-
rotifers, but the estimates are quite comparable to tereiden tilavuuksien keskiarvoja [Mean volumes for some
those obtained by direct measurements and can plankters found in Finland]. Vesiensuoje1utoimiston
be calculated for any species, with a defined, con- tiedonantoj a 21: 1-26.
sistent method. Peters, R H., 1983. The ecological implications of body size.
Cambridge University Press, Cambridge, England.
Ruttner-Kolisko, A., 1972. Rotatoria. In: Elster, H.-J. & W.
References Ohle (eds.), Die Binnengewasser. Bd. 26. Das Zooplankton
der Binnengewasser. 1. Tei!. Schweizer-bart'sche Verlags-
Aasa, R, 1979. Plankton i Lilla Ullevifjarden. Nat. Swedish buchhandlung, Stuttgart: 99-234.
Envir. Prot. Bd, Limno!. Surv. Uppsala, Rep. 33: 1-62. Ruttner-Kolisko, A., 1977. Suggestions for biomass calcula-
Bikar, K., 1986. Dauer der Eientwicklung von Plankton- tion of planktonic rotifers. Arch. Hydrobio!. Beih./Ergebn.
crustaceen des Vberlingersees (Bodensee) in Abhangigkeit Limno!. 8: 71-76.
von der Saisonalen Variation der Umgebungstemperatur, Salonen, K. & R. Latja, 1988. Variation in the carbon content
der AdultgroBe und des Eigewichtes. Diplomarbeit, of two Asplanchna species. Hydrobiologia 162: 79-87.
Universitat Freiburg (W-Germany). Schmidt-Nielsen, K., 1984. Scaling: Why is animal size
Bottrell, H. H., A. Duncan, Z. M. Gliwicz, E. Grygierek, important? Cambridge University Press.
A. Herzig, A. Hillbricht-Ilkowska, H. Kurasawa, P. Lars- Utermohl, H., 1958. Zur Vervollkommnung der quantita-
son & T. Weglenska, 1976. A review of some problems in tiven Phytoplankton-Methodik. Mitt. int. Ver. Limnol. 9:
zooplankton production studies. Norw. J. Zoo!. 24: 1-38.
419-456. Walz, N., 1987. StotTumsatz und Kinetik von Regulations-
Doohan, M. & V. Rainbow, 1971. Determination of dry prozessen bei Zooplankton-Populationen. Habilitations-
weights of small Aschelminthes «.1 J-Lg). Oecologia schrift, Universitat Miinchen, W-Germany, 225 pp.
(Ber!.) 6: 380-383.
Hydrobiologia 186/187: 363-369, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 363
© 1989 Kluwer Academic Publishers.

Temperature aspects of ecological bioenergetics in Brachionus angularis

(Rotatoria)

Norbert Walz, Tanja Gschloessl, Ulli Hartmann

Zoologisches Institut, Seidlstr. 25, D-8000 Miinchen 2, FRG

Key words: rotifers, temperature, food concentration, bioenergetic parameters

Abstract

The influence of temperature and food quality was studied on the following energy balance parameters
of B. angularis: ingestion, production, growth and mortality. The ingestion rate rises to an optimum at
15 and 20°C and decreases at 25 DC. The other rates increase continuously over the 5-25 °C range.
The QlO-values of production rate are higher than those of ingestion rate. Temperature also modifies the
relationship between food concentration and bioenergetic rates. They react according to a Monod
function (production at all temperatures, growth at 10 ° C) or decrease at high concentrations (growth
at 15° and 20°C).

Introduction modifies the dependence of some bioenergetic

parameters on food density.
Bioenergetic metabolism is ecologically important
for all species in their environment. Two factors
have especially important influences on energy Methods
balance: food conditions and temperature. From
studies on energy balance we can draw con- Brachionus angularis was cultured with the unicel-
clusions on the food and temperature demands a lular green alga Stichococcus bacillaris. The culture
species makes to its environment. conditions were further described by Walz (1983)
The energy balance can be formulated as and Walz (1987). Rotifers were acclimated to ex-
follows (Walz, 1987b): perimental temperatures for at least three weeks
before the tests. The ingestion experiments were
ingestion - feces = production + respiration
conducted by 14-C-Iabelled Stichococcus ac-
Production = growth + mortality
cording to Walz & Gschloessl (1988). The rotifers
(parameters in italics were measured). in the growth-experiments were transferred daily
Studies of the food demands of Brachionus into 20 ml glass vessels with fresh suspensions of
angularis, a rotifer common in eutrophic lakes algae of defined concentration. To avoid sedimen-
were made by Walz (1987b) and Walz & tation the vessels were fixed on a rotation wheel
Gschloessl (1988). Temperature has such a great (1 rpm) and exposed to the desired temperature
influence on this eurythermic species, that it even ('rotating cultures'). The number of rotifers and
364

their eggs were controlled daily. The specific pro- food concentrations between 2 and 5 Jlg C ml- 1
duction rate was measured as birth rate according (Walz & Gschloessl, 1988). Therefore, the 2.5 and
to Paloheimo (1974) and Romanovsky & 5 Jlg C ml- 1 values are not significantly different.
Polishchuck (1982). For more details see If temperature is increased to 25 ° C, the 5 Jlg C
Hartmann (1987). ml- 1 ingestion rate remains unchanged. The de-
crease of the 2.5 Jlg C ml- 1 ingestion rate at
25 ° C is not significant, but the 25 ° C ingestion
Results rates at both food concentrations differ signifi-
cantly (P = 0.05). At temperatures < 20 ° C no
Bioenergetic rates plateau was found.
In contrast to these curves of food ingestion,
At the experimental food concentrations, the in- the output bioenergetic rates all increase with tem-
gestion rate of a B. angularis population increases perature (Fig. 1, b-d). Specific production
with temperature (Fig. la). The 5 Jlg C ml- 1 in- achieves a maximum value of about 0.7 d - 1,
gestion rate reaches a maximum at 15 °C, double growth rate about 0.5 d -1 and mortality rate
the value at 2.5 Jlg C ml- 1, and decreases further about 0.3 d - 1. Food concentration also has a
on. At 2.5 Jlg C ml- 1, the maximum is achieved distinct influence on these rates. In general, rates
at 20 ° C. The 20 ° C rates are on a plateau at all rose with increasing food concentration (see

--I
-g a) spec . ingestion rate b) spec . production rate
CII
~ 1.01--1-- poe l,ugC/ml l

..
'-
u ....12
,.. ...19
CII '·· 5
0\ ·· ···· 0.5
'-
CII ""' 0.25
c:
CII
0.5
o
.c
CII
>
..IV O~----~----+-----~---+-r+---~-----T-----r----T1
c) spec . growth rate d) spec. mortality rate
1.0 ~------+-----~----~~----4-+-r-----~-----T------r------r1

PO C [,u 9 (1m II
...... .s
··· ··· 12
0.5t-+- ------+- .. ' ···19
poe I I-I g (lmlJ
····· 0.5 .....19
·····-0.25 ',.• 05
_ __ _~~'":::::;-""
..' .. 12
0.25
·····5

o 5 2S S 20 5
temperature tOC)

Fig. 1. Temperature relationship of individual bioenergetic parameters at different food concentrations (POe = particulate
organic carbon = J1.g e ml - I). ± 95 % confidence intervals. Solid squares: 0.25 J1.g e ml- I, solid circles: 0.5 J1.g e ml- I, open
triangles: 2.5 J1.g e ml- I, open circles: 5.0 J1.g e ml- I, solid triangles: 12.0 J1.g e ml- I, open squares: 19.0 J1.g e ml - I.
 365

Table 1. The food utilization efficiencies of B. angularis. different rate with increasing temperature
(PE = production/ingestion, Y = growth/ingestion. (Table 2). Production rate shows the greatest in-
Food concentration Temperature (0C)
crease with a mean QIO of 4.0. The temperature
dependencies of some coupled life cycle parame-
10 15 20 ters, when treated as reciprocals to time, are
similar. Life span QIO is 3.3, embryonic develop-
2.5 Jlg C ml- 1 PE 0.49 0.42* ment QIO is 4.0, juvenile development QIO is 4.9,
Y 0.48 0.48*
and development of egg laying interval Q10 is 3.1
5.0 Jlg C ml- 1 PE 0.22 0.49 0.28* (according to Walz, 1987a).
Y OJO 0.44 0.18*
Population growth rate and mortality rate have
* From chemostat experiments (Walz 1987b). lower QIOS of 2.7 and 3.0 respectively. Respir-
ation rate QlOS, calculated from Galkovskaya
(1987) are also 2.7. The ingestion rate QIO is still
section: kinetics), but this effect was not found for lower = 2.3. Although it is possible that the as-
mortality rate. similation quotient increases with temperature,
the energy available for utilization is lower at
higher temperatures. The QIO also is dependent
Food utilization efficiency on the food supply and, in general, it is higher at
lower food densities.
The efficiencies (production efficiency, PE;
yield = growth/ingestion, Y) at 5 jlg e ml- 1 in-
crease with temperature, climbing from 10 to Temperature modification offood - bioenergetic rate
15°C where a turnover of little less than 0.5 is kinetics
observed (Table 1). With a temperature increase
to 20 °e the efficiencies decrease again. Food With rising food concentration ( = poe = parti-
utilization in the lower food concentration (2.5 jlg culate organic carbon) the specic production rate
e ml- 1) is higher than at 5 jlg e ml- 1 (see also (b) increases at all studied temperatures according
Walz, 1987b) and is a little less than 0.5 at 10 and to a Monod saturation function
20°C. At 15 °e, no experiments were made at (b = bmax ' POC/Ks + POC) (Fig. 2a). In the low-
this food concentration. er concentration range the rate accelerates steeply
and reaches a plateau where additional food does
not produce further rate increases. Plateau values
increase with temperature and the increase is
higher between 10 and 15°C than between 15
The QIO values in the temperature range of 10 to and 20 ° e, corresponding to the slightly flattened
20 ° e were different for each individual bioener- curve in Fig. 1c.
getic parameters. Each parameter increased at a Monod function, regression and correlation

Table 2. The QIO-values of bioenergetic parameters between 100 and 20 0c. SE = Standard error.

Parameter Food concentration (Jlg C ml- 1) mean QIO

± SE
0.5 2.5 5.0 8.5 12.0 15.5 19.0

Ingestion rate 2.6 2.0 2J ± OJ

Production rate 4.7 4.4 4.2 3.8 3.5 4.1 3.4 4.0 ± 0.2
Growth rate 3.1 4.1 2.5 2.7 2.4 1.9 2.0 2.7 ± OJ
Mortality rate 3.6 4.8 2.6 2.2 3.0 ± 0.6
366

-0·20 5 10 15 20
food concentr.tion, poe l~gCI lJ

Fig. 2. Modification of the kinetics of individual bioenergetic rates in dependence on temperature and food concentration. The
dashed lines are fitted by eye. ± 95 % confidence intervals.

coefficients are shown in Table 3. They were fitted growth rate accelerates at the lower concentra-
with a non-linear regression according to Bliss & tions up to a maximum and decreases linearly
James (1966). With rising temperature the bmax thereafter. The linear regression coefficients
values (plateau levels) increase and the Ks values (Table 3) at 15 0 and 20 0 C are significantly differ-
decrease. ent from zero, but they do not differ from each
The specie growth rates (Fig. 2b) take con- other (P = 0.05).
trasting courses. Only the 10 0 C curve is in accor- The mortality rate decreases with increasing
dance with a Monod model and shows a satu- food concentration in the lower concentration
ration function. At higher temperatures the range and increases further at 15 0 and 20 0 C.
 367

Table 3. Coefficients of the Kinetics of production and growth rate. * coefficients of Monod function, ** coefficients of linear
regression. poe, food concentration (Jlg e ml- 1), bioenergetic rate (d - 1), R = correlation coefficient.

Temperature Spec. production rate Spec. growth rate

'rotating cultures'
b* = 0.196· poe 0.239· poe
lOoe r*
0.313 + poe 1.344 + poe
R= 0.94, n = 29 R= 0.89, n = 29
15°C poe> 2.5 Jlg C/ml:
b* = 0.529· poe
r** = 0.504 - 0.008 . poe
0.097 + poe

R= 0.99, n = 17 R= -0.99, n = 8
20°C poe> 2.5 Jlg C/ml:
b* = 0.712' poe r** = 0.676 - 0.Dl5 . poe
0.073 + poe

R= 0.73, n = 49 R= -0.73,n=37

ehemostat culture
b* = 0.548· poe r*= 0.579·POe
20°C
(WALZ 1987b) 0.775 + poe 4.045 + poe
R= 0.84, n = 22 R= 0.94, n = 25

This increase is in contrast to chemostat experi- species, which is the basis for calculation of pro-
ments (Walz, 1987b), where the mortality rates duction rate (Paloheimo, 1974), has a maximum
only decline asymptotically with food concen- at intermediate temperatures and decreases
tration from high values at low concentrations. thereafter. This is also the case for B. calyciflorus
(Halbach, 1970). The decreasing egg-ratio of
B. angularis and K. cochlearis at higher tempera-
Discussion tures is probably more than compensated by the
increased rate of embryonic development. In con-
The individual bioenergetic rates of B. angularis trast to the above species, the egg-ratio of
show characteristic differences in their tempera- B. rubens declines with climbing temperature
ture course. They either increase continuously (Vuckovic, 1981).
with temperature or reach a maximum and then According to the portion of production going
decrease at higher temperatures. For B. angularis into mortality or growth (see equation in the Intro-
ingestion rates, a maximum was reached at 15 0 or duction), growth rate increases continuously, if
20 0 C. In contrast, all other rates increase con- mortality rate rises constantly (Fig. 1) (B. calyd-
tinuously, but this is not always the case for other j1orus, Halbach, 1970; B. dimidiatus, Pourriot &
species. In B. calyciflorus, ingestion rate increases Rougier, 1975). Alternatively, in cases mortality
continuously with temperature (Galkovskaya, rate increases progressively, growth rate declines
1987). Respiration rate seems to be the only rate after reaching a maximum (K. cochlearis, Walz,
that consistently increases continuously with tem- 1983; B. plicatilis, Hirayama & Kusano, 1972).
perature in all species studies (Galkovskaya, B. rubens follows a more complicated pattern. At
1987; Lampert, 1984). As with B. angularis, high food levels growth rate decreases with tem-
Keratella cochlearis production rate also con- perature and increases at low food conditions
tinuously rises with temperature (Edmondson, (Vuckovic, 1981).
1965; Walz, 1983). In contrast, egg-ratio in both In B. angularis we showed that the energy
368

balance grows more unfavorable at higher tem- adults was not affected. However, the O 2 content
peratures (higher QIO values for production than in 'rotating cultures' was not measured. Though
ingestion). At 25°C, our population approached we tried to prevent O 2 depletion by leaving air
its thermal tolerance limits. At this point, it was bubbles in the vessels, this may not have been
difficult to keep the culture going and no growth sufficient at high temperatures and high algae den-
experiments were carried out. B. calyciflorus, an sities. Although in chemostats good aeration is
even more pronounced warm-eurythermal rotifer, provided.
was possible to be cultured up to 40 ° C Alternatively, the algae in 'rotating cultures'
(Galkovskaya, 1987). In contrast, the net repro- could have clumped together in higher concentra-
ductive rate (Ro) of the cold-stenothermal tions and become unavailable as food for the
Notholca caudata falls dramatically at tempera- young rotifers. Currently, no decision can be
tures above 10 0 C (Laxhuber & Hartmann, made between these hypotheses. Differences
1988). Also N. squamula could be maintained in between chemostat and 'rotating culture' produc-
culture only under 10 °C (May, 1987). tion coefficients at 20°C (Table 3) was discussed
Temperature effects on bioenergetic rates are by Walz (1987b).
modified by food concentration. In rotifers, this
relationship is often represented as a saturation
function according to the Monod model (e.g. the
References
production rates in B. angulans at all tempera-
tures and growth rate at 10 0 C). The egg-ratio of Bliss, C. I. & A. T. James, 1966. Fitting the rectangular
B. rubens increases either according to a satu- hyperbola. Biometrics 22: 573-602.
ration function (Vuckovic, 1981) or continuously Edmondson, W. T., 1965. Reproductive rates of planktonic
(Rothhaupt, 1985). In contrast, production rate is rotifers as related to food and temperature in nature. Ecol.
Monogr. 35: 61-112.
often found to decrease at higher food concentra-
Galkovskaya, G. A., 1987. Planktonic rotifers and tempera-
tions (e.g. B. rubens, Pilarska, 1977; and B. plica- ture. Hydrobiologia 147: 307-317.
tilis, Hirayama et al., 1974). The same is also true Gschloessl, T., 1985. Die Nahrungsaufnahme von Brachionus
for growth rates (Euchlanis dilatata, King, 1967 angularis (Rotatoria) in Abhiingigkeit von der Futterart,
and B. calyciflorus, Halbach & Halbach-Keup, der Futterkonzentration, der Temperatur und der Fiitte-
1974). rungsdauer. Diplomarbeit Faculty of Biology, Univ.
Munich, 127 pp.
Usually this decrease is explained by algae Halbach, U., 1970. EinfluB der Temperatur auf die Popu-
toxins, but this cannot explain our observations in lationsdynamik des planktischen Riidertierchens Bra-
B. angulans. First, a toxic effect also would have chionus calyciflorus Pallas. Oecologia 4: 176-207.
affected production rate. Second, growth rate at Halbach, U. & G. Halbach-Keup, 1974. Quantitative Be-
ziehungen zwischen Phytoplankton und der Populations-
10 0 C follows a Monod function. Only at 15 ° and
dynamik des Rotators Brachionus calyciflorus Pallas. Be-
20 ° C did a decline of growth rate at higher food funde aus Laboratoriumsexperimenten und Freilandun-
concentrations occur. tersuchungen. Arch. Hydrobiol. 73: 273-309.
This decrease of growth rate was not found in Hartmann, u., 1987. Die Populationsdynamik der pela-
chemostat experiments at 20 ° C with the same gischen Rotatorien Brachionus angularis und Notholca
caudata in Abhiingigkeit von der Futterkonzentration und
food algae (Walz, 1987b) (Table 3). Only in long-
der Temperatur. Diplomarbeit Faculty of Biology, Univ.
term experiments with residence times longer than Munich, 91 pp.
the daily transfer-times of the 'rotating cultures' Hirayama, K & T. Kusano, 1972. Fundamental studies on
could such a toxic effect be detected. physiology of rotifer for its mass culture. II. Influence of
As rotifer growth rates decreased at higher tem- water temperature on population growth of rotifers. Bull.
Jap. Soc. Sci. Fish. 38: 1357-1363.
peratures, increasing respiration of algae (dark
Hirayama, K, K Watanabe & T. Kusano, 1973. Fundamen-
experiments) could have led to a critical O 2 supply tal studies on physiology of rotifer for its mass culture. III.
for the rotifers. Young rotifers seemed to have Influence of phytoplankton density on population growth.
suffered more from this, as the egg laying of the Bull. Jap. Soc. Sci. Fish. 39: 1123-1127.
 369

King, C. E., 1967. Food, age, and the dynamics ofa laborato- approach to calculation of secondary production at the
ry population of rotifers. Ecology 48: 111-128. population level. Int. Revue. ges. Hydrobiol. 67: 341-359.
Lampert, W., 1984. The measurement of respiration. In: Rothhaupt, K. 0., 1985. A model approach to the population
Downing,J. A. & F. H. Rigler (eds),Amanuai on methods dynamics of the rotifer Brachionus rubens in two-stage
for the assessment of secondary production in fresh chemostat culture. Oecologia 65: 252-259.
waters. 2nd ed. Blackwell Scientif. Publ., Oxford. IBP- Vuckovic, M., 1981. EinfluB der Temperatur und der
Handbook 17: 413-468. N ahrungsquantitiit auf die Populationsdynamik des
Laxhuber, R. & u. Hartmann, 1988. The influence of planktischen Riidertiers Brachionus rubens Ehrenberg.
temperature on the life-cycle of the cold-stenothermal Diplomarbeit Dep. Biology, Univ. Frankfurt/M., 75 pp.
rotifer Notholca caudata. Verh. int. Ver. Limnol. 23: Walz, N., 1983. Individual culture and experimental popula-
2016-2018. tion dynamics of Keratella cochlearis (Rotatoria). Hy-
May, L., 1987. Culturing freshwater planktonic rotifers on drobiologia 107: 35-43.
Rhodomonas minuta var. nannoplanktica Skuja and Sticho- Walz, N., 1987a. Comparative population dynamics of
coccus bacillaris Niigeli. J. Plankton Res. 9: 1217-1223. Brachionus angularis and Keratella cochlearis. Hydrobio-
Paloheimo, 1. E., 1974. Calculation of instantenous birth logia 147: 209-213.
rates. Limnol. Oceanogr. 19: 692-694. Walz, N., 1987b. Stoffumsatz und Kinetik von Regulations-
Pilarska, 1., 1977. Ecophysiological studies on Brachionus prozessen bei Zooplankton-Populationen. Analysen und
rubens Ehrbg. (Rotatoria). II. Production and respiration.- Modelle in Rotatorien-Chemostaten und im Plankon eines
Pol. Arch. Hydrobiol. 24: 329-341. Sees. Habilitationsschrift Faculty of Biology, Univ.
Pourriot, R. & c. Rougier, 1975. Dynamique d'une popula- Munich, 225 pp.
tion experimentale de Brachionus dimidatus (Bryce) Walz, N. & T. Gschloessl, 1988. Functional response of
(Rotifere) en fonction de la nourriture et de la temperature. ingestion and filtration rate of the rotifer Brachionus angu-
Ann. Limnol. 11: 125-143. laris to the food concentration. Verh. int. Ver. Limnol. 23:
Romanovsky, Yu. E. & L. Polishchuck, 1982. A theoretical 1993-2000.
Hydrobiologia 186/187: 371-374, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 371
© 1989 Kluwer Academic Publishers.

Biomass and elemental composition (C.H.N.) of the rotifer Brachionus

plicatilis cultured as larval food

M. YUfera & E. Pascual

Instituto de Ciencias Marinas de Andalucia (C.S.l.c.), Apdo. oficial, 11510 Puerto Real. Cadiz. Spain

Abstract

Changes in dry weight, carbon, nitrogen, hydrogen and ash content of the rotifer Brachionus plicatilis were
investigated during population growth in culture. Also caloric equivalent was estimated from carbon
content. Body dry weight and weight of C, N, and H increased with increasing the egg/female ratio of
the population, while ash content remained constant. As percentage of dry weight, C content increased
with fecundity, H and N remained constant and ash content decreased. The caloric equivalent ranged
from 1278 ,ucal to 4406 ,ucal for rotifers from 314,ug to 846,ug of dry weight.

Introduction out to determine the variations in body mass and

composition. Cultures A, B and C were fed on the
Rotifer Brachionus plicatilis is widely used as first Eustigmatophyceae Nannochloropsis gaditana,
food for marine fish larvae in aquaculture. The and the culture D was maintained with bakers
strains commonly cultured range from yeast Saccharomyces cerevisiae.
90-140,um to 200-300,um in length and differ Dry weight (DW) was determined by drying
considerably in female body mass. However, samples of about 400-800 individuals (females
variations of biomass may also be found in a with eggs) at 85 ° C to constant weight. Elemental
single strain as a consequence of the culture sys- analyses (C.N.H.) were carried out with an Ele-
tem and the demographic characteristics of the mental Analyzer Model 1106 (Carlo Erba
population. Such variations may affect the nu- Science) using cyclohexane 2.4-dinitrophenyl-
tritional quality of rotifers as prey due to its caloric hydrazone as standard; samples weighed about
content. Knowledge of this energy value is needed 1 mg. Ash content was determined by incinerating
in feeding and metabolic studies oflarval fish and dried samples at 550°C during 24 h. Three repli-
crustacea. This paper studies changes in biomass cates per sample were used to determine ash and
and C, N, H content through population growth C.N.H. content. Energetic content was estimated
of B. plicatilis in culture. from carbon content using the factor of 10.9 cal
per mg C (Salonen et al., 1976).

Materials and methods

Results
Brachionus plicatilis strain S-1 (YUfera, 1982) was
cultured at 25°C. Two batch cultures (A and B) Batch cultures A and B had a characteristic peak
and two semicontinuous cultures (C and D) with of fecundity on day 2 and 3 due to synchronous
a partial harvest of 10-20% daily, were carried deposition of the first egg (Fig. 1). The semicon-
372

1
~

A B c ",
I'
o
I

200 tII
I

~
I
•It
,/I , ,
.
,,
fI I ' 0,6
,\ t~ ' ', I
,
I \
'\
I

,'' ',,,
I

..,,
rl
I
\ ~
,I
~ 150
I \
, ,, !-
, ,
"
I \

,, / '.
I
\
a:
I..L.J ,
I
\
I '. 0. 4
I..L.J
~
u..
, I ,I :IE:
I..L.J
5100
a: , I
I
I u..
I
I I

,,
I
I ,, I
, ~ "' '''' '''' ., .. . t!l
: ' ...... ~ . ' 0, 2 ~
50
I
\
I
\
\
, ,.
I
,, ,,
, I

,,
I
I \ I ' , I
I \ ~/
,
•
•""
o 5 0 5 o 5 10 0 5 10
D AY S
Fig. 1. Population growth curves. Cultures A, B, and C with Nannochloropsis gaditana and culture D with Saccharomyces
cerevisiae as food. Arrows indicate the exhaustion of food.

tinuous cultures (C and D) had more erratic yeast have been combined because no significant
variation in fecundity. Changes in individual dry differences were found between them.
weight were related to fecundity. Body dry weight The H and N contents were practically con-
were positively correlated to eggs/female (E/F) stant through population growth (Fig. 3). H
ratio (Fig. 2). In contrast, ash content varied little, accounted for about 6.3 ± 0.3 % and N
ranging from 60-120 ng (average 86 ng ± SD 19) 9.2 ± 0.65 % of the body dry weight, indicating a
and was not correlated with E/F ratio (r = 0.189). mean protein level (N x 6.25) of about 57.5%.
Data of rotifers fed micro algae and those fed on The C percentage increased with the ElF ratio

60~~---,--~---.---r---r---r--'
Body ma ss: Y : 31 4 .5 + 664.4 X c ( . 1: Y ~ 46.80 xO.0 55 r - 0 . 754
r • 0.901 •
• • •
-;, 800
c • •
• •
~ 600 • • Ash ( _ I: Y 13.08 x-o. l72 r -0.176
N ( A): Y = 9.59 xO.0
24 r 0.421 =

:t: :-_
UJ
:.: • >- _ H ( ) Y 6.61 xO.0 26 r = 0.417
•
~ ~ *"
>-
:5 400 20 - = _
ASh : Y - 81.7 + 15.3 X
r - 0.189
o~""_ ____~~~_'-~T-____~____ I
-•
'tt • ••

200
••
• •
0.2 0.4 0.6 0.2 0.4 0.6
EGG / FEMAlE EGG / FEMAlE

Fig. 2. Body dry weight (circles) and ash weight (squares) in Fig. 3. Regressions of percent C, N, H and ash on egg/female
relation to egg/female ratio. Open symbols = rotifer fed on ratio. Open symbols = rotifer fed on algae; dark sym-
algae; dark symbols = rotifer fed on yeast. bols = rotifer fed on yeast.
 373

Table 1. Caloric data of rotifer biomass as a function of egg/female ratio. Values estimated from regression equations.

E/F ratio 0.0 0.1 0.2 0.4 0.6 0.8

cal/rotifer 1278 1669 2060 2842 3624 4406
cal/mg DW 4.06 4.39 4.61 4.89 5.08 5.21
caljmg AFDW 5.49 5.61 5.69 5.77 5.83 5.87

according to a power curve. This increase occurs Discussion

until ElF reaches 0.2 egg per female. Percentage
of ash increases markedly when the fecundity is Rotifer dry weight obtained in this study is higher
very low and it may reach up to 32 % of body than that obtained for the same strain fed on the
weight. Chlorophyceae Nannochloris oculata (YUfera
The C, N and H contents in weight per et al., 1983). This agrees with the bigger size of
individual follow a similar pattern that the body females and eggs observed when fed on N. gadi-
dry weight and show high correlations with ElF tana at the same culture conditions (YUfera,
ratio (Fig. 4). The value of these parameters are 1987).
three or four times higher in the samples showing Relationships between biomass and ElF ratio
highest fecundity than in the populations without has been reported by Scott & Baynes (1978) and
eggs. YUfera et al. (1983). The present work reveals that
The calorimetric data estimated from carbon C content, and therefore its caloric equivalent, are
content has values ranging from 1278 to also related to ElF. This supports the usefulness
4406 Ilcal per individual and between 4.06 and of ElF as a good index to assess the nutritional
5.21 cal per mg of dry matter (Table 1). quality and growth potential of population
(YUfera & Pascual, 1984; Snell et aI., 1987).
Theilacker & McMaster (1971) determined a
caloric content of 5.34 cal' mg - 1 ash free DW for
B. plicatilis (females without eggs). Doohan
(1973) measured 790-1290 Ilcal per rotifer
(DW = 158-250 ng' ind - \ assuming 50% of C
C ( .): Y ~ 117 .28 + 358.65 X content). From C data reported by Schlosser &
N (Al: Y ~ 28 .45 + 64 .48 X
1100 H ( el: Y ~ 18 .12 + 49 .70 X Anger (1982), a caloric content of 1264 Ilcal per
rotifer (DW = 280 ng' ind -I) and 4.5 cal' mg- 1
'"c
DW can be estimated. More recently Theilacker
• • & Kimball (1984) determined 440-2070 Ilcal
• (4.4 cal, mg- 1 DW and 4.8 cal' mg- 1 ash free
200
• DW, assuming 8.7% of ash content) for non-ovi-
gerous females ranging from 100 to 470 ng' ind - 1
100
DW. In general, values reported in this study
(4.06-5.21 cal' mg- I DW) are similar to those
• • presented for these authors and agree with
0.2 0.£1 0.6
5.4-5.7 cal' mg- 1 AFDW estimated for aquatic
EGG / FEAALE invertebrates (Wacasey & Atkinson, 1987). The
differences among caloric values presented by
Fig. 4. Regressions of C, H, and N content per rotifer (dry
weight· individual- 1) on egg/female ratio. Open sym- different authors are attributable to variations in
bols = rotifer fed on algae; dark symbols = rotifer fed on ash content.
yeast. It is interesting to note that caloric equivalent
374

of the organic matter (cal· mg- 1 AFDW) in- the rotifer Brachionus plicatilis. Helgoliinder wiss.
Meeresunters. 35: 215-225.
creases with increasing fecundity, which is asso-
Scott, A. P. & S. M. Baynes, 1978. Effect of algal diet and
ciated with the yolk content in the population. In temperature on the biochemical composition ofthe rotifer,
routine semicontinuous mass culture the ElF Brachionus plicatilis. Aquaculture 14: 247-260.
ratio usually ranges between 0.2-0.4. Caloric Snell, T. W., M. 1. Childress, E. M. Boyer & F. H. Hoff, 1987.
content per rotifer in these conditions ranged Assessing the status of rotifer mass cultures. 1. World
from 1871 to 2574/lcal (4.61-4.89 cal· mg- 1 Aqua. Soc. 18: 270-277.
Theilacker, G. H. & A. S. Kimball, 1984. Comparative
DW). When food was exhausted fecundity fell quality of rotifers and copepods as food for larval fishes.
quickly and the caloric content decreased CalCOFI Rep. 25: 80-86.
1275 /lcal per individual and 4.06 cal' mg- 1 DW. Theilacker, G. H. & M. F. McMaster, 1971. Mass culture of
the rotifer Brachionus plicatilis and its evaluation as a food
for larval anchovies. Mar. Bio!. 10: 183-188.
Wacasey, J. W. & E. G. Atkinson, 1987. Energy values of
Acknowledgements marine benthic invertebrates from the Canadian Artic.
Mar. Eco!. Prog. Ser. 379: 243-250.
We thank Olimpio Montero and Jose Ma Espi- YUfera, M., 1982. Morphometric characterization ofa small-
gares for technical assistance and Manuel Arjon- sized strain of Brachionus plicatilis in culture. Aquaculture
27: 56-61.
illa for its collaboration in the CRN analyses.
YUfera, M., 1987. Effect of algal diet and temperature on the
embryonic development time of the rotifer Brachionus pli-
catilis in culture. Hydrobiologia 147: 319-322.
References YUfera, M. & E. Pascual, 1984. Influencia de la dieta sobre
la puesta del rotifero Brachionus plicatilis en cultivo. Inv.
Salonen, K., J. Sarvala, I. Hakala & M. L. Viljanen, 1976. Pesq. 48: 549-556.
The relation of energy and organic carbon in aquatic inver- YUfera, M. & L. M. Lubitm & E. Pascual, 1983. Efecto de
tebrates. Limno!. Oceanogr. 21: 424-730. cuatro algas marinas sobre el crecimiento poblacional de
Schlosser, H. J. & K. Anger, 1982. The significance of some dos cepas de Brachionus plicatilis (Rotifera: Brachionidae)
methodological effects on filtration and ingestion rates of en cultivo. Inv. Pesq. 47: 325-337.
Hydrobiologia 186/187: 375-380, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotiler Symposium V. 375
© 1989 Kluwer Academic Publishers.

Molecular genetics of rotifers: preliminary restriction mapping of the

mitochondrial genome of Brachionus plicatilis

Charles E. King
Dept. of Zoology, Oregon State University, Corvallis, OR 97331, U.S.A.

Key words: mitochondrial DNA, genetic variation, restriction mapping, rotifers, Brachionus plicatilis

Abstract

Methods are presented to extract and purify mitochondrial DNA from the rotifer Braehionus plicatilis.
The mtDNA obtained is of sufficient purity for digestion with restriction endonucleases. EeoR I restric-
tion patterns are presented for 4 geographically separated clones. A restriction map based on digestion
with 5 different restriction enzymes is included for one of these clones. Finally, use of mtDNA analysis
for studies on the population structure and biogeography of rotifers is discussed.

Introduction ducts are restricted to 2 rRNAs (12S and 16S), 22

tRNAs and 13 polypeptides that serve as subunits
Over the past decade a change of revolutionary for the enzymes of the inner mitochondrial
proportions has taken place in evolutionary membrane (Brown, 1985; Garesse, 1988). Rates
biology. The methodology of molecular genetics of mtDNA evolution in vertebrates appear to be
has profoundly impacted the study of such diverse substantially higher that those of single-copy nu-
areas as biogeography, interspecific gene flow, clear DNA, but this relationship may not hold for
systematics, phylogenetic relationships and the some invertebrates (Caccone et ai., 1988).
genetic structure of populations. Problems in The present paper is the first to present infor-
these areas that have long been sequestered in the mation on the mtDNA of rotifers. In fact the
realm of speculation are now yielding to objective entire literature on the molecular biology of roti-
analysis (Avise eta!., 1987; Hillis, 1987; Moritz fers consists of only two papers; Kumazaki et al.
et al., 1987). Much of this progress has been made (1982) sequenced the 5S rRNA of Brachionus pli-
possible by molecular analysis of mitochondrial catilis and Ishikaea (1977) studied the thermal
DNA (mtDNA). stability of the rRNA of the same species. An
The mtDNA of metazoans is composed of important qualification is required for the present
double-stranded, circular molecules that vary paper. Successful extraction of mtDNA and its
from about 15.5 to 20 kbp (kilobase pairs) in size. digestion with restriction enzymes has only re-
Mitochondria are maternally inherited and their cently occurred and additional replication and
DNA lacks introns and has few or no spacers confirmatory tests are necessary before the de-
between genes. The mtDNA base sequences of tailed quantitative results of this research can be
humans, cattle, mice and 2 species of Drosophila accepted with complete confidence. Never-the-
are now known and in each case the gene pro- less, the power of this approach and its potential
376

applicability to many of the problems being dis- combined with the first and the pellet resuspended
cussed at this symposium make publication of the in 10 ml H medium and centrifuged a third time.
techniques and the preliminary results appro- The combined supernatants from the three centri-
priate. fugations were then centrifuged at 3000 rpm for
4 min and the supernatant was transferred to a
clean Oak Ridge tube. The pellet was discarded.
Preparation of mtDNA This procedure was repeated 3 to 5 times until no
visible pellet was formed.
Mitochondrial DNA was extracted and purified The clean supernatant was then centrifuged at
using a modification of the methods employed by 7500 rpm (6780 x g) for 15 min to pellet the mito-
MacNeil & Strobeck (1987). Because the success chondria. The mitochondrial pellet should be a
and yield of the extraction is dependent upon uniform pale yellow color - a brown ring in the
closely following the protocol, procedures will be pellet indicates that additional slow spins should
presented in detail. Except as noted, all steps were be made to further clean the preparation before
carried out at 4 ° C using autoclaved labware. proceeding. The supernatant from this centrifu-
Brachionus plicatilis clones were cultured for gation was discarded and the mitochondrial pellet
mtDNA analysis in 20 I glass carboys in a 25°C was resuspended in 10 ml H medium using a pas-
constant temperature room. Saccharomyces cere- teur pipette. The mitochondrial suspension was
visiae was used as food and autoclaved 2/3 Pacific then centrifuged for 10 min at 13 000 rpm
Ocean sea water as the medium. When ready for (20200 x g) and the supernatant was again dis-
DNA extraction, individuals were removed from carded.
their medium by filtration through a 60 f.1m nitex The pellet from this second high speed spin was
screen and resuspended overnight in 2 I sea water resuspended in 1.4 ml H medium and divided
without food to permit the rotifers to clear pre- between two 1.5 ml microfuge tubes. To digest
viously ingested food. The following morning the any contaminating nuclear DNA in the mitochon-
rotifers were again filtered from the medium, a wet drial suspension, MgCl 2 (1 M stock) and DNase
weight was taken, and the animals were washed 1(10 mg ml- 1 stock) were added to final concen-
off the filter and into a 15 ml Dounce tissue grin- trations of 0.01 M and 40 f.1g ml- 1 and incubated
der with approximately 10 ml of ice cold pH 8.5 on ice for 30-60 min. At this point the mitochon-
H (homogenization) medium (0.07 M sucrose, dria should still be intact thereby protecting the
0.21 M manitol, 1.9 mM HEPES, 0.5% bovine mtDNA from degradation by the DNase. Follow-
serum albumen). Approximately 2-3 g wet weight ing the incubation the tubes were microfuged at
of rotifers were used for each extraction. 4 ° C for 10 min and the pellets were resuspended
Animals were homogenized on ice with five up in 1 ml H medium and again pelleted as above.
and down strokes of the grinder. A small aliquot The washed pellets were gently resuspended
of the homogenate was then checked under a with a pasteur pipette in 950 f.11 of pH 7.5 L (lysis)
dissecting scope to ensure adequacy ofhomogeni- buffer (0.1 M EDTA, 0.1 M Trizma-HCI, 0.2 M
zation. The homogenate was transferred to a NaCl) and placed on ice for 10 min. Proteinase K
15 ml corex tube and centrifuged in a Sorvall from a 10 mg ml- 1 stock was then added to a
SS-34 rotor at 3000 rpm (1085 x g) for 4 min to concentration of 50 f.1g ml- 1 and incubated at
pellet debris and nuclei. Mitochondria remain 37°C for 1 hr. SDS from a 30% stock was added
suspended in the H medium during this centrifu- to a final concentration of 1% and the incubation
gation because of their density. The supernatant continued for another 30 min. The microfuge
was transferred to a 50 ml Oak Ridge tube and tubes were then placed on ice for 15 min and
held on ice while the pellet was resuspended in microfuged for 5 min. The resulting pellet was
10 ml H medium and recentrifuged as above. The discarded. Contaminating proteins and SDS
supernatant from this second centrifugation was were removed from the supernatants by three ex-
 377

tractions with 0.1 M, pH 8.0 Trizma-HCI bufTer- the recommended temperature for at least 3 hr
saturated phenol. An additional extraction was (and usually overnight). Three JLI of stop mixture
performed with a 1: 1 mixture of phenol: chloro- were then added and the electrophoresis was
form (containing 4% isoamyl alcohol here and carried out at 30-40 V with a 0.7-0.9% agarose
subsequently) and another extraction was per- gel.
formed with the chloroform. Traces of phenol This protocol differs from most DNA isolation
were then removed with two ether extractions. procedures not employing density-gradient centri-
After residual ether was removed using a gentle fugation in two regards. The first of these is the
stream of air, the DNA was precipitated over- DNase treatment. In the absence of this step it is
night with 1/10 volume of 3 M NaOAc at pH 8.0 not unusual to see substantial quantities of nu-
and two volumes of 100 % ethyl alcohol at clear DNA contaminating the preparation. This
- 20 °C. The DNA was then pelleted by micro- contamination is either from nuclei that ruptured
fuging for 15 min and the pellets were washed early in the procedure or from nuclei that were not
twice in 70% EtOH and desiccated under va- removed by the initial 3000 rpm centrifugations.
cuum. 100 JLI ofTE buffer (10 mM trizma, 1 mM While this nuclear DNA may not interfere with
EDTA, pH 8.0) were added to each of the dried the analysis, its removal by DNase treatment was
pellets and the tubes were placed at 4 ° C until the desirable since specific mitochondrial probes
DNA dissolved. have not been used in this study.
RNase T1 (from a stock of 1000 units ml- 1) The second and more critical difference is the
and RNase A (from a stock of 10 mg ml- 1) were use of spermidine prior to addition of the restric-
added to the samples at concentrations of 40 units tion enzymes. A potent inhibitor of restriction
and 100 JLg per ml and incubated for 1 hr at enzyme activity is liberated from the rotifers dur-
50 ° C. Samples were then extracted once each ing the isolation procedure. This inhibitor co-
with phenol, phenol-chloroform, chloroform and precipitates with the mtDNA and produces a
then twice with ether. A second overnight complete or partial inhibition of the digestion. An
mtDNA precipitation was carried out with 1/2 alternative method to remove the inhibitor - sper-
volume of 7.5 M NH 4 0Ac and 2 volumes of mine precipitation - was also successful, but was
100% EtOH at - 20 0c. After pelleting, washing judged to be more time consuming than the sper-
twice with 70% EtOH, and desiccation, the midine treatment.
mtDNA in each microfuge tube was redissolved
in 30 III of water.
To check for purity and yield of mtD N A, a 2 JLI
Results and discussion
aliquot was electrophoresed on a 0.75% agarose
minigel. Hind III digested bacteriophage lambda
DNA was used for a size marker. Ideally, nuclear Preliminary analyses have been carried out on
DNA should be absent and the rotifer mtDNA four clones of Brachionus plicatilis. Clone Q 16 was
should be seen as two bands, i.e., one for the collected in October, 1984 from Dong Feng Sal-
relaxed and one for the supercoiled mtDNA. Re- tern on the Shandong Peninsula near Qingdao,
striction digests were performed according to the People's Republic of China (PRC). Additional
recommendations of the enzyme supplier with the information on this site is available in King et al.
following important exception. Each digest was (1988a, b). Clone G8 was collected at Shenzhen
carried out by diluting the desired volume of Saltern near Guangzhou (Canton) in the
mtDNA solution to 20 JLI with water, adding Southern PRC. Clone AS is from Scotland (but
2.5 JLI of 10 x buffer, and then adding 1 JLI of was not mentioned by May, 1983) and was given
0.1 M spermidine. This solution was placed on ice by 1.M. Scott to A. Ruttner-Kolisko who
for 10 min and then 1 JLI of restriction enzyme was supplied it to me. This clone was designated S in
added. Incubation for the digestion took place at Ruttner-Kolisko (1983). Clone BS3 was collected
378

by the author in December, 1987 from Big Soda While the RFLP patterns in Fig. 1 are obvious-
Lake in Nevada, U.S.A. (King & Zhao, 1987). ly quite different, for such comparisons to be use-
Figure 1 presents restriction patterns ofthe four ful intrapopulation variability must also be
clones obtained after digestion with the enzyme measured. This step will require identification of
Eeo R I. Variation among clones is of two types. more map sites, i.e., genetic markers, so that the
First, there are obvious restriction fragment clonal structure of a given population can be
length polymorphisms (RFLPs) within clones specified with greater confidence. Two methods
that can be used to distinguish between clones. are available for increasing the number of map
Second, the estimated sizes of the four mitochon- sites per clone. Eeo R I and the other restriction
drial genomes vary from 17.5 kb (QI6) to 19.9 kb enzymes used in this study have hexanucleotide
(AS). As discussed by Brown (1985), most size targets - that is, a specific sequence of six base
variation in mtDNA is attributable to changes in pairs is required for cleavage. Under the assump-
the displacement loop (D-Ioop) region that is pre- tion of random ordering of four bases, hexanu-
sumed to act as the origin of H strand replication cleotide targets are expected to occur at intervals
in vertebrates. The same region also appears to of 46 = 4096 bases. A mitochondrial genome of
have more base sequence variation. Thus the 18.6 kb is therefore expected to have 4.5 target
variability among clones seen in Fig. 1 may be sites. An obvious way to increase the number of
nonrandomly distributed over the entire mito- genetic markers is to use restriction enzymes that
chondrial genome. In particular each of the four recognize smaller targets. Enzymes that recognize
clones has a fragment of approximately 5.4 kb in pentanucleotide targets are expected to produce
size (the apparent variation is less than 300 bp). 18.2 cleavages in the same molecule, those that
Mitochondrial probes could be used to determine recognize tetranucleotide targets are expected to
whether or not these bands are the same. Unfor- produce 72.7 cleavages.
tunately, however, a mitochondrial genome libra- An alternative method to obtain more genetic
ry has not been constructed for B. plieatilis or any markers per clone is to increase the number of
other rotifer. restriction enzymes employed. This latter tech-
nique also has the advantage of making it possible
G8 AS Q16 BSa to physically map the mitochondrial genome.
14_
Of the four clones discussed in this paper, only
12 -
G8 has been studied with more than a single
10- mtDNA preparation. To date, the mitochondrial
genome of G8 has been treated with five restric-
8-
tion enzymes in both single and double digestions.
The resultant data have been used to construct
6-
the preliminary restriction map presented in
kb Fig. 2. Twenty-two sites have been identified for
the five enzymes: four for BamH I, six for BglIl,
4-
three for EeoR I, four for Hind III, and five for
Xba I.
The map in Fig. 2 has sufficient detail to permit
reasonable determinations of clonal identity. The
2-
strategy to be employed in future work is to deter-
SUM=18.6 19.9 17.5 17.9 mine the amount of map variability among clones
from the same lake. Findings based on allozyme
Fig. 1. EcoR I restriction patterns for the mtDNA of four
strains of Brachionus plicatilis. The vertical scale is fragment differences suggest relatively little variation within
size in kilobases (kb) and the total mitochondrial genome populations of Asplanehna girodi (King, 1977,
sizes are given at the bottom of the figure. 1980), but substantially more variation within the
 379

BamH I compare the mtDNA patterns of nearby and dis-

tant (intercontinental) populations to determine
patterns of colonization. That is, homogeneity of
popUlations within an area and heterogeneity
between areas would support a vicariance model
based on little effective gene flow. The counter
BamH I
rmding - as much variation within an area as
BgllI between areas - would provide powerful evidence
for the role of gene flow in tying together disjunct
Hind III populations of a species.
Xba I
The role of dispersal in determination of
species distributions constitutes but one example
of an important problem in rotifer biology that
might yield to mtDNA analysis. Many others oc-
cur in fields as diverse as hybridization and eco-
typic vs genetic determinants of morphological
variation (Pejler, 1980; Serra-Galindo, 1987). It
Fig. 2. Mitochondrial DNA restriction map of Brachionus thus seems likely that the revolution referred to in
plicatilis clone G8 based on digestions with five restriction the first sentence of this paper will likewise
endonucleases.
profoundly impact the study of rotifer biology.

B. plicatilis of Soda Lake (King & Zhao, 1987;

References
Zhao & King in press). If the mtDNA restriction
maps of this latter species show a comparable Avise,1. c., 1. Arnold, R M. Ball, E. Bermingham, T. Lamb,
amount of variation, it is likely that the Soda Lake J. E. Neigel, C. A. Reeb & N. C. Saunders, 1987. Intra-
rotifers had multiple independent origins. On the specific phylogeography: The mitochondrial bridge
other hand, homogeneity of the mtDNA would between population genetics and systematics. Ann. Rev.
Ecol. Syst. 18: 489-522.
suggest a small number of founders. Thus the
Brown, W. M., 1985. The mitochondrial genome of animals.
analysis of mtDNA variation is potentially a In: MacIntyre, R J. (Ed.). Molecular Evolutionary
powerful tool to unravel details of population Genetics. Plenum Press, New York: 95-130.
structure and dispersal. Caccone, A., G. D. Amato & J. R. Powell, 1988. Rates and
At our 1982 U ppsala symposium, Dumont patterns of scnDNA and mtDNA divergence within the
(1983) presented an elegant treatment of rotifer Drosophila melanogaster subgroup. Genetics 118: 671-683.
Dumont, H. 1., 1983. Biogeography ofrotifers. Hydrobiologia
biogeography. The core of this paper was a con- 104: 19-30.
sideration of the relative significance of dispersal Garesse, R, 1988. Drosophila melanogaster mitochondrial
vs. vicariance in determining the distributions of DNA: Gene organization and evolutionary con-
rotifers. That rotifers have a remarkable capacity siderations. Genetics 118: 649-663.
for passive dispersal via their resting eggs is Hillis, D. M., 1987. Molecular versus morphological
approaches to systematics. Ann. Rev. Ecol. Syst. 18:
beyond question. At an earlier rotifer symposium, 23-42.
Lair (1980) reported collecting Brachionus Ishikaea, H., 1977. Comparative studies on the thermal sta-
havanaensis (a Caribbean species) and Lecane pa- bility of animal ribosomal RNA's. IV. Thermal stability
puana (restricted to tropical and subtropical areas and molecular integrity of ribosomal RNA's from several
of the eastern hemisphere) immediately down- protostomes (rotifers, round-worms, liver-flukes, and
brine shrimps). Compo Biochem. Physiol. 56: 229-234.
stream from the heat exchangers of nuclear power King, C. E., 1977. Genetics of reproduction, variation, and
plants in central France. If such dispersal is both adaptation in rotifers. Arch. Hydrobiol. Ergebn. Limnol. 8:
frequent and successful it should be possible to 187-201.
380

King, e. E., 1980. The genetic structure of zooplankton MacNeil, D. & C. Strobeck, 1987. Evolutionary relationships
populations. In: Kerfoot, W. e. (Ed.). Evolution and among colonies of Columbian ground squirrels as shown
Ecology of Zooplankton Communities. New England by mitochondrial DNA. Evolution 41: 873-881.
University Press: 315-329. May, L., 1983. Rotifer occurrence in relation to water tem-
King, e. E. & Y. Zhao, 1987. Coexistence of rotifer perature in Loch Leven, Scotland. Hydrobiologia 104:
(Brachionus plicatilis) clones in Soda Lake, Nevada. Hy- 311-315.
drobiologia 147: 57-64. Moritz, C., T. E. Dowling & W. M. Brown, 1987. Evolution
King, e. E., Y. Zhao, X. Liu & M. R. Li, 1988a. Poly- of animal mitochondrial DNA: relevance for population
acrylamide gel electrophoresis of isozyme variation in biology and systematics. Ann. Rev. Ecol. Syst. 18:
Artemia from Chinese salterns. J. Shandong College of 269-292.
Oceanography 18: 64-69. Pejler, B., 1980. Variation in the genus Keratella. Hydrobio-
King, e. E., Y. Zhao, X. Liu & M. R. Li, 1988b. Genetic logia 73: 207-213.
variation in parthenogenetic Artemia from the Shandong Ruttner-Kolisko, A., 1983. The significance of mating pro-
Peninsula, P.R.e. Oceanology and Limnology 6: 179-185. cesses for the genetics and for the formation of resting eggs
Kumazaki, T., H. Hori, S. Osawa, N. Ishii & K. Suzuki, in monogonont rotifers. Hydrobiologia 104: 181-190.
1982. The nucleotide sequences of 5S rRNA's from a Serra-Galindo, M., 1987. Variaci6n morfometrica, isoenzi-
rotifer, Brachionus plicatilis, and two nematodes, Rhabditis matica y demografica en poblaciones de Brachionus
tokai and Caenorhabditis elegans. Nucleic Acids Res. 10: plicatilis: Diferenciaci6n genetica y plasticidad fenotipica.
7001-7004. Ph.D. Thesis, Universitat de Valencia.
Lair, N., 1980. The rotifer fauna of the river Loire (France), Zhao, Y. & c. E. King. Ecological genetics of the rotifer
at the level of the nuclear power plants. Hydrobiologia 73: Brachionus plicatilis in Soda Lake, Nevada, U.S.A. Hy-
153-160. drobiologia in press.
Hydrobiologia 186/187: 381-386, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 381
© 1989 Kluwer Academic Publishers.

Size variation in Brachionus plicatilis resting eggs

Laura Serrano, Manuel Serra & Maria R. Miracle

Dep. Ecologia. Universidad de Valencia, 46100-Burjasot (Valencia), Spain

Key words: biometric analysis, resting eggs, rotifers, salinity, temperature, genetic variation

Abstract

The effect oftemperature and salinity on resting egg size of two Brachionus plicatilis (Rotifera) clones was
investigated. Clones were selected according to their different behaviour in laying resting eggs: one clone
ejects them, whereas they remain inside the females body in the other clone. The difference in resting eggs
size between the two clones is noticeable, although the difference is not as great as that between female
body size. An important temperature-salinity interaction on resting egg size has been observed. The
general inverse relationship between size and temperature is only true at lower temperatures. At high
temperatures size varies around the mean although could be greater than at intermediate temperatures.
This is more evident at the intermediate salinity tested which is considered to be the closest to the optimum
in our experiments. This pattern of variation suggests that mean size is bigger than expected, in relation
to temperature and salinity, when these factors have values close to the extremes of their range, normally
found in nature, and to which adaptative mechanisms can evolve. Size is bigger at the salinity -
temperature low -low and high - high combinations which are the most commonly found in the temperate
environments.

Introduction persal capabilities. Moreover, the evaluation of

these effects may be useful in exploring the recent
In a previous paper (Serra & Miracle, 1986) history of natural rotifer populations by monitor-
several clones of Brachionus plicatilis, a cos- ing resting eggs in the sediment.
mopolitan species from a fluctuating environ-
ment, were used to clarify the degree of morpho-
logical plasticity that this genome expresses. We Materials and methods
examined plasticity by studying the effect of tem-
perature and salinity on body size. Biometrical experiments on resting eggs were
We have extended that work by analyzing the carried out on two clones of Brachionus plicatilis,
direct response of the same clones to the same CU, collected from a continental saline lake, and
sets of experimental conditions, but this time SPO, collected from a coastal marsh, which have
studying the size ofthe resting eggs produced. Our been maintained in laboratory culture since 1981.
intention is to evaluate the extent of genetic con- The clones have been characterized by electro-
trol and of environmental variation on the size of phoretic analysis as well as morphometric ally
the sexual cyst, which has dormancy and dis- (Serra & Miracle, 1983, 1985 and 1986).
382

The two clones of B. plicatiUs were cultured in the lorica containing the eggs was also measured
diluted sea water enriched with f/2 medium of (Fig. 1). All measurements were made with a Wild
Guillard & Ryther (1962) in flasks of 300 ml, M40 microscope at 40 x . Egg volume was esti-
using Tetraselmis sp. as food at concentrations mated representing the egg as a composite figure
greater than 106 cells ml- 1. Rotifers were cultured of two spheres connected by a cylinder of the
at different salinities (9, 12 and 24%0) and tem- same diameter. This shape is more similar to that
peratures (20, 25 and 30 0 C). of a resting egg than a revolution ellipsoid. Volume
For each salinity, temperature and clone, two was computed as V = (n/12)(3 WL - W) where L
replicate flasks (blocks) were prepared; more is the length and W the width of the egg.
than three months elapsed between the replicate To determine the main effects and interactions
experiments. The cultures were kept under the of the different experimental conditions and
experimental conditions for three or more weeks clones on resting egg size, a multivariate analysis
until resting eggs appeared. Each week, half of the of variance (MANOVA) was performed and their
culture volume was renewed with new medium global significance was tested with Wilkis' cri-
containing more than 106 cells ml- 1. Then, 25 terion (Cooley & Lohnes, 1971; Kres, 1983). The
resting eggs were collected from each replicate significance of each effect on individual measure-
flask at random. ments was determined with a univariate analysis
Resting egg size was determined by measuring of variance (AN OVA) as described by Cooley &
length and width, and for clone SPO the width of Lohnes (1971).

'-

. eu SP.O
o

o
o

Fig. 1. Photomicrograph of clone CU and SPO. Bar is 40 JLm.

 383

Results ,um
SPO
LENGTH
130
Two clones were selected to study the effect of
environmental conditions on resting egg size, in 124=m,
clone CU which lays resting eggs outside the
120
female and SPO which deposits resting eggs
inside the animal where they remain until its
death, with release occuring when the lorica
decomposes (Fig. 1). Resting eggs of both clones 110

are viable and their hatching levels (40-70 %) were 20 25 30

similar to that obtained by Minkoff et al. (1983).
Figures 2, 3 and 4 show means and standard ,um WIDTH

100

,um LENGTH
cu eo
_ _ _T

--- --- --- ...

170

20 25 30

160
,um LORICA
\

--
\ 150
T

\ i.. r
\ ...... T ___ ~.J
--__ 1 __ ---
150 \
\ --~,
\
.. "',
:\-\~-- j
140
"
140 \ ~ 24%0
\
\
\;l __ _ --
T

--~ 130

130 20 25 30

20 25 30 °C Fig. 3. Length and width means and standard error of rest-

ing eggs of clone SPO and width of the lorica containing a
,um WIDTH resting egg (ordinates) as a function of temperature (20, 25
and 30°C, abscissas) and salinity (9%0' 12%0 and 24%0)' The
120
global means for each measurement are also indicated (m!,
m2 , m3 ).

110

~.L_ errors of resting egg length, width and volume at

"" . the different temperatures and salinity conditions.
100 '\----------~ . The results of the multivariate analysis of
variance (Table 1) show significant differences
20 25 30 °C
between clones and the significant effect of tem-
Fig. 2. Length and width means and standard errors ofrest- perature and blocks in all measurements. The
ing eggs of clone CU (in ordinates) at different conditions of
interactions clone-temperature and temperature-
temperature (20, 25 and 30°C, abscissas) and salinity (9%0'
12%0 and 24%0)' The global means for each measurement are salinity are also significant in terms of length. So
also indicated (m!, m2 ). the genotype largely determines the response to
 384

The effect of temperature on the width of the

.. m3 xl0
1.4
\
cu mictic females of clone SPO was different from
the effect on parthenogenetic females of this
\

1. 3
\ clone. Parthenogenetic females showed a de-
\
\ crease in width at higher temperatures at all salini-
\
\ ties and an increase at higher salinities (Serra &
1. 2
\
, Miracle, 1986). In contrast, the response to tem-
\ perature-salinity conditions of mictic females with
1.1 \
~ \ resting eggs had a complex pattern similar to that

'"
\
of resting eggs (Fig. 1). The differences in the
1.0 = m1 ~~ width of these females were much more attenu-
ated than those of amictic females.
0 .9 The general effects of temperature on resting
egg size for both clones can be summarized in the
O.B following points: (1) variation in size is mainly
due to variation in length, width being more con-
20 °C
stant, (2) at the highest salinity tested (24%0) rest-
ing egg size remains close to the mean and the
.. m3 x108
SPO variations due to temperature are small, (3) at the
0.8 lowest salinity (9%0) the expected and confirmed
..... "-
"-
asymptotic inverse relationship in active animals
0 .7 between size and temperature (Serra & Miracle,
1986) is also true for resting eggs, (4) at the
0 .8 medium salinity (12%0) temperature effect on size
is very complex, resting egg size decreases with
0 .5 temperature to a point and then increases to
20 25 30 values near the mean size, in most cases exceed-
ing it.
Fig. 4. Volume means and standard error of resting eggs of The effect of salinity is not significant in the
both clones CU and SPO (ordinates) at different tempera- statistical analysis, perhaps due to the great
tures (20, 25 and 30 C, abscissas) and salinity (9%0' 12%0 0
influence of temperature on size. At the lowest
and 24%0)' The global means are also indicated (ml, m2 ).

Table 1. MANOV A and ANOV A results of three factorial

temperature, and salinity influences this response fixed effects (clone, temperature and salinity) and a
greatly. random nested effect (blocks) on two resting eggs
measurements (N = 25,* is p < 0.05 and ** is p < 0.01).
Differences in resting eggs size between the two
clones are noticeable, the average length and Source Wilks' A df F ratio
width of the CD resting eggs being 1.18 and 1.16
times bigger respectively than the average length nl n2 Length Width
and width of the SPO resting egg. However, these
Clone (e) 0.0547** 1 18 298.3** 75.7**
differences are much smaller than the differences
Temperature (T) 0.0848** 2 18 80.0** 8.5**
between amictic female body sizes. CD females Salinity (S) 0.7900 2 18 0.9 OJ
are approximately 1.4 times bigger than SPO exT 0.4183* 2 18 10.4** 1.3
females (the relationship between the average CxS 0.8120 2 18 0.8 0.2
lengths of the two clones, at different conditions TxS 0.2862** 4 18 8.8** 1.9
exSxT 0.7243 4 18 1.4 OJ
is 1.41, and between their widths it is 1.38 (Serra 0.06830** 18 3.4** 15.6**
Blocks 864
& Miracle, 1986).
 385

Table 2. Linear correlations between egg length and width at width is greater than on length, while in resting
different temperatures and salinities.
eggs width shows little variation.
Clone Salinity Temperature

20°C 25 °C 30°C Discussion

CU The two clones differ much more in amictic female

9%0 0.361 0.856 0.699
12%0 0.181 0.676 0.796 size than in resting eggs size. However resting
24%0 0.625 0.557 0.367 eggs size in each clone is only slightly less variable
SPO than body size of parthenogenetic females. This
9%0 0.152 0.788 0.489 variation in resting eggs size has been observed in
12%0 0.171 0.529 0.460 nature and on laboratory cultures whenever it has
24%0 0.301 0.551 0.661
been measured. For instance, polymorphism has
been observed in rotifer resting eggs (Gilbert,
1974), and a significant size variation of
temperature, resting eggs are bigger at the lowest
B. plicatilis resting eggs in relation to their vertical
salinity levels and smaller at the highest in both
location in sediments has been recorded (Snell
clones. However, at the highest temperature they
et aI., 1983).
a:e smaller at the lowest salinity level and they are
The two clones selected for this study origi-
bIgger at the intermediate optimum salinity 12%0.
nated from contrasting environments: CU comes
Since the ratios length/width of the two clones
from a shallow highly fluctuating, endorreic saline,
are rather similar, we investigated the degree of
inland lagoon, which shows an euryoic behaviour.
correlation between these measurements under
Clone SPO comes from a less fluctuating karstic
different conditions (Table 2). The two measure-
coastal lagoon (Serra, 1987). Resting eggs of SPO
ments are similar, but the correlation coefficients
from a more stable environment are bigger in
are not very high, approximately 0.5. In a few
relation to the size of the maternal females than
cases correlation is very low. These cases are the
those produced by clone CU. Moreover, the
same for both clones and correspond to rotifers
variation in resting eggs size of the euryoic clone
reared at the lowest assayed temperature, where
CU with salinity and temperature variation is
length greatly exceedes the mean, but width
higher. When body size was analyzed (Serra &
closely approximates the mean.
Mir~cle, 1986), clone CU again showed an impor-
The comparison of these results with those
tant Increase after a decrease with rising tempera-
derived from the effect of temperature and salinity
ture when salinity was high. Clone SPO showed
on the body size of active parthenogenetic females
the expected inverse size-temperature relation-
(Table 3) shows that the effect of these factors on
ship at all tested salinities.
resting egg length is similar to the effect on body
The tendency to follow the inverse size-tem-
length. However, in active females the effect on
perature relationship has been confirmed in a
Table 3. Ratios of the maximum and minimum mean values great variety of organisms, including amictic eggs
at the different temperatures and salinities (L: length; W: of ~~achionus calyciflorus (Pourriot, 1973). High
width). sallmty reduces this response and the volume of
CU SPO resting eggs closely approximates the clone mean.
At intermediate salinities, high as well as low
Body Resting Body Resting Lorica temperatures determine a bigger size. Intermedi-
size egg size egg ate salinity (12%0) is common in spanish mediter-
ranean wetlands (Miracle et aI., 1986) and is the
L 1.28 1.29 1.15 1.14
W 1.36 1.16 1.22 1.11 1.11 salinity we used in maintaining all clones. Accord-
ing to the literature, half-strength sea water is
386

optimal for B. plicatilis (Ruttner-Kolisko, 1974; References

Walker, 1981), as well as for resting egg hatching
(Minkoff et aI., 1983), who recorded a optimal Cooley, V. W. & P. R Lohnes, 1971. Multivariate data
analysis. John Wiley & Sons, N.Y., 364 p.
salinity of 16%0, among the 9, 16,24,32 and 40%0
Gilbert, 1. 1., 1974. Dormancy in rotifers. Trans. Amer.
tested. On the other hand, population dynamics Micros. Soc., 93 (4): 490-S13.
ofthe same clones (Serra, 1987) showed that SPO Guillard, R L. & 1. M. Ryther, 1962. Studies of marine
and CU are better adapted to the low range of planktonic diatoms. Ca. 1. Microbiol. 8: 229-239.
salinities studied (9%0 and 12%0)' Kres, H., 1983. Statistical tables for multivariate analysis.
At the latitudes when the clones were collected, Springer-Verlag. New York. S04 p.
Minkoff, G., E. Lubzens & D. Kahan, 1983. Environmental
high temperatures correspond to summer periods factors affecting hatching of rotifer Brachionus plicatilis
of reduced food or desiccation. Furthermore, resting eggs. Hydrobiologia 104: 61-69.
hatching levels, according to Minkoff et al. Miracle, M. R, M. Serra, E. Vicente & c. Blanco, 1987.
(1983), show an inverse relationship with tem- Distribution of Brachionus species in the Spanish
perature, which has been attributed to lower dis- Mediterranean Wetlands. Hydrobiologia 147: 7S-81.
Pourriot, R, 1973. Rapports entre la temperature, la taille
solved oxygen concentration at higher tempera- des adultes, la longueur des reufs et Ie taux de developpe-
tures. For all these reasons, larger resting egg size ment embryonneire chez Brachionus calyciflorus Pallas
at high temperatures could be adaptive. High tem- (Rofitere). Ann. Hydrobiol. 4: 103-lIS.
peratures correspond, in temperate aquatic eco- Ruttner-Kolisko, A., 1974. Plankton Rotifers. Biology and
systems, to high salinities, the tolerance to extreme taxonomy. Binnengewasser. 26: 146 p.
Serra, M., 1987. Variabilidad morfol6gica, isoenzimatica y
values of the latter parameter being highly affected demografica en poblaciones de Brachionus plicatilis.
by the other. Diferenciaci6n genetica y plasticidad fenotipica. Tesis
From the global observation of the results, two doctoral. Universidad de Valencia.
conclusions seem logical with respect to resting Serra, M. & M. R Miracle, 1983. Biometric analysis of
eggs: (1) width is less variable because egg is Brachionus plicatilis ecotypes from spanish lagoons. Hydro-
biologia 104: 279-291.
oriented longitudinally at its passage through Serra, M. & M. R Miracle, 1985. Enzyme polymorphism in
oviducts which are restrictal narrow openings; Brachionus plicatilis populations from several spanish
(2) mean size is bigger in extreme environments: lagoons. Verh. int. Ver. Limnol. 22: 2991-2996.
low salinity -low temperature, high salinity - high Serra, M. & M. R Miracle, 1987. Biometric variation of
temperature. On the other hand, mean size is Brachionus plicati/is strains as a direct response to abiotic
parameters. Hydrobiologia 147: 83-89.
smaller in favorable conditions where hatching is Snell, T. W., B. E. Burke & S. D. Messur, 1983. Size and
not delayed by diapause. At intermediate salini- distribution of resting eggs in a natural population of the
ties rotifers seem to be able to regulate egg size rotifer Brachionus plicatilis. Gulf Research Reports. 7 (3):
against direct environmental effects such as those 28S-287.
Walker, K. F., 1981. A synopsis of ecological information on
due to temperature.
the saline lake rotifer Brachionus plicatilis, Muller, 1786.
Hydrobiologia 81: IS0-167.
Hydrobiologia 186/187: 387-400, 1989.
c. Ricci, T. W. Snell and C. E. King (eds), Rouler Symposium V. 387
© 1989 Kluwer Academic Publishers.

Rotifers as food in aquaculture

E. Lubzens 1, A. Tandler2 & G. Minkoff3

I National Institute of Oceanography, Israel Oceanographic & Limnological Research, Tel-Shikmona,

P.O.B. 8030, Haifa 31080, Israel; 2National Center for Mariculture, Israel Oceanographic &
Limnological Research, P.O.B. 1212, Eilat, Israel; 3 Tinamenor S.A., Pesues, Cantabria, Spain

Key words: rotifers, marine larvae, nutrition

Abstract

The rotifer Brachionus plicatilis (O.F . Muller) can be mass cultivated in large quantities and is an important
live feed in aquaculture. This rotifer is commonly offered to larvae during the fIrst 7-30 days of exogenous
feeding. Variation in prey density affects larval fIsh feeding rates, rations, activity, evacuation time, growth
rates and growth efficiencies. B. plicatilis can be supplied at the food concentrations required for meeting
larval metabolic demands and yielding high survival rates. Live food may enhance the digestive processes
of larval predators. A large range of genetically distinct B. plicatilis strains with a wide range of body size
permit larval rearing of many fIsh species. Larvae are fIrst fed on a small strain of rotifers, and as larvae
increase in size, a larger strain of rotifers is introduced. Rotifers are regarded as living food capsules for
transferring nutrients to fIsh larvae. These nutrients include highly unsaturated fatty acids (mainly 20 : 5
n-3 and 22: 6 n-3) essential for survival of marine fIsh larvae. In addition, rotifers treated with antibiotics
may promote higher survival rates. The possibility of preserving live rotifers at low temperatures or
through their resting eggs has been investigated.

Introduction were ciliates, dinoflagellates, trochophores and

wild plankton (Okamoto, 1969; Harada, 1970).
The history of larval rearing of marine fIsh can The second phase started when rotifers were
clearly be divided into two distinct phases. Lack found to be suitable as fIrst feed for marine fIsh
of adequate fIrst feeds caused limited success in larvae in Japan in the late sixties and early
larval rearing of marine fIsh up to the sixties seventies. Rotifers became ubiquitous in all mass
(Dannevig, 1897; Blaxter, 1968; Lasker et al., rearing trials after their successful use in the mass
1970). This was before rotifers, primarily the rearing of the red seabream (Pagrus major) in
species Brachionus plicatilis, were established as Japan (Fujita, 1973, 1979). The introduction of
fIrst larval feed. Most of the effort in this fIrst rotifers marked the fIrst regular successes in the
phase was limited to small scale experiments of mass larval rearing of several marine species of
rearing larvae of different species for identifI- economic value such as grey mullet (Mugi/
cation purposes or for fIsh stock assessment in cephalus) (Nash et al., 1974), sole (Solea solea)
fIsheries (Houde & Palko, 1970). In these experi- (Howell, 1973; Girin, 1974; Fuchs, 1978, 1982;
ments, small prey organisms for the early larvae Dendrinos & Thorpe, 1987), gilthead seabream
388

(Sparus aurata) (Person-Le Ruyet & Verillaud, except for a few species, their size ranges between
1980, 1981; Tandler & Helps, 1985), sea bass 2-7 mm. Rotifers offered to them must meet their
(Dicentrarchus labrax) (Barnabe, 1974; Girin, nutritional requirements for optimization of
1975), turbot (Scophthalmus maxim us) (Kuhl- growth and survival. These include: (1) the size;
mann et aI., 1981; Olsen & Minck, 1983; Witt (2) the distribution and concentration of rotifers
et al., 1984), and flounder (Paralichthys olivaceus) in the larval tanks; (3) the total amount available;
(Fukusho et al., 1985), milkfish (Chanos chanos) (4) digestability and absorption; and (5) nutri-
(Liao et aI., 1979; Juario et al., 1984) and others tional quality.
(Morales, 1983). These successes resulted from
the development of rotifer culturing methods (Ito,
1960; Theilacker & McMaster, 1971; and reviews Size of rotifers
by Hirata, 1980; Fukusho, 1983; Hirano, 1987;
Lubzens, 1987). Today, most marine fish are The size of the prey eaten by the fish larvae is a
raised on basically the same methods, whereby function of the larval mouth width. Within a fish
B. plicatilis is provided as the first food during the species, mouth width is related to length, but it
first days of exogeneous feeding. The length of the varies greatly between species (Arthur, 1976;
period depends on the fish species. Beyer, 1980; Hunter, 1980; Hunter & Kimbrell,
Marine fish larvae are usually small at hatching 1980). Although mouth width limits the maximum
(Theilacker & Dorsey, 1980; Kissil, 1984/85), and prey size, in nature the mean diameter of the prey

0.50.
u
~
o.Ao. 9
Width
'-
Q)
0.30.
~ 90. - 10.0. urn
a..
0..20. 6 .... ... .. . .
' • • • • • • Q. . • • • • • • <l 70. - 90. urn
0..10.

E 0. i::-=------,=--~.....:.;;~----'"'-+-~....:...---I-=--o.-I-=-20':"'"/-"1-"""'I
70 3..,..---1
0 4-:0---I50=-=---...,.
leo-=--- Age (hours)
" / '(Y
-0.,10. ~~ \
'l!!" '\ \
\
-0.20. '\ \ -0--- ---050-70. urn
" Q) \ b---------
-0.30.
"'6> \
-o.Ao.
«
-0.50.
\
--,--- _ -e- ' - ' - -- 30.-50. urn

Fig. 1. Changes in the e1ectivity indices (E) for larval sea bream (Sparus aurata) feeding on rotifers of various sizes as a function
of larval age (redrawn from Helps [1982)). Electivity index (E) was calculated, based on Ivlev [1961], in the following way:
E = (r - p )/(r + p)
where rand p are the relative occurrence of a given rotifer size in populations exposed and unexposed to larval predation pressure,
respectively.
 389

consumed by fish larvae was only 38 % of their strains of different sizes was examined in the labo-
mouth width (Hunter, 1980). There is a tendency ratory (Helps, 1982). In this study, larvae showed
for larvae in the sea to feed on progressively larger a clear age effect on their feeding preference of
prey as they grow. In the laboratory it was found different size rotifers (Fig. 1). Young larvae, up to
that the size of the prey fed to larvae must increase 85 h after hatching, preferred small rotifers
for larvae to grow at maximum rates (Lasker (30-70 f.1m in width) and avoided feeding on large
et aI., 1970; Hunter, 1980; Hunter & Kimbrell, rotifers (90-100 f.1ID in width), while in older lar-
1980). Beyer & Laurence (1981) concluded that vae (160 h) the pattern was reversed. The effect of
as larvae reach certain sizes, the energetic cost of rotifer size in long-term experiments was further
each attack on prey exceeds the gain from ingest- tested, and it was found that the presence of small
ing smaller food particles. This size depends upon rotifers for the first 12 days of feeding of gilthead
the larva's metabolic requirements, which are seabream larvae was associated with an improved
imposed both genetically and environmentally. It growth rate (Tandler, pers. com.). This was espe-
means that the larvae develop selection related to cially pronounced in the size structure of the
particle size both in natural populations and labo- population of 32-day-old larvae. In these larvae,
ratory-reared species (Stepiens, 1976; Hunter, the percent of the population with a wet weight
1980). greater than 12 mg was 51.5 and 36, respectively,
The effect of age of gilthead seabream larvae when small and large rotifers were offered as first
(Sparus aurata) on their preference for rotifer food. In these studies, two genetically different

o.s
\.:)

<D 0.6
~
<D
>
0..4
u
:::J

e
U

0.
Q) 0..2
a::

0.
I JA'A I TAHITI I JS I I JL I JAVA I TAHITI I JS, I JL
NRE NRE SPAIN EIL/!J JS2 NRE SPAIN E ILAT JS2
SMALL SMALL

Strains
Fig. 2. The effect of temperature on the amictic reproductive rates of eight rotifer strains or clones cultured on Nannochloris sp.
Reproductive rate was calculated from the equation:

G = I/T In Nt/No,
where Go = experimental growth coefficient,
No = initial estimated population size,
Nt = final estimated popUlation size,
T = time in days.
The mean values of five replicates and the standard deviations are presented by vertical bars.
Strains Java and Tahiti were obtained from INRA/IFREMER, France; strain Spain from Dr. E. Yufera; clones N.R.E. and Eilat
Small from Israel; and clones JS l' JS 2 and JL were hatched from resting eggs collected by Dr. M. Okauchi in Japan (from Lubzens
et al., in preparation).
390

size strains of rotifers were used. There is interest cally different size rotifers offers a better solution
in these different size strains because they allow (see Lubzens, 1987). However, this requires aqua-
different larval stages to select prey of the most culturists to maintain stocks of at least two rotifer
effective size (Fukusho, 1983; Korunuma & strains in the hatchery.
Fukusho, 1987). Obtaining optimal mass cultures of different
Two methods of obtaining different sizes of rotifer size strains may mean culturing them at
rotifers have been considered; (1) sieving rotifer different temperatures and offering them different
cultures through small size meshes which will types of food. For example, the small size strain
allow the separation of small size rotifers from the in Japan (S-type) occurs in the summer when high
larger ones; (2) culture of rotifer strains whose temperatures prevail, while the large size rotifer
size is genetically determined. The first method strain (L-type) predominates in the winter
does not guarantee full control over rotifer size. (Fukusho & Iwamoto, 1980; Fukusho, 1983).
By sieving a rotifer culture, the small neonates will Laboratory studies under controlled conditions of
be separated from the adults, but within a short food and temperature show that the reproductive
period they will eventually increase in size, while rate of rotifer strains is not always related to size
staying in the larval tanks. The culture of geneti- (Fig. 2). Some of the small size strains (Java,

Clone : N. R. E Clone: JAVA

lc.
1.0 1.0
If.
,,!!

C)
0.8 k/ Tc. C)
0.8

(l)

...
0 0.6 - -...
(l)

0 0.6

(l)
> 1tl I:
(l)
>

u 04 u
0.4 Tt
,,
:J
:J I
- U

,r/ A
U
...
0 ...
0
a.
T
a.
(l) T I (l)
0: /1
0: 02 0.2
i /1
I' / ~
T T /
I II
(f" I 1 )f
.!. IT"
0 0 I {/
1/
b'"

0.10 0.25 0.50 1.0 0 2.00 0.)0 0.25 0.50 LOO 2.00

Concentration ( :d0 6 Cells ml- I )

Fig. 3. The effect of two algae differing in size (Tetraselmis tetrathele and T. chuii) on the reproductive rates oflarge (clone N.R.E.)
and small (strain Java) rotifers cultured at 25 0c. The average dimensions (in I'm) of the algae were 15.3 x 8.5 x 10.5 and
12.0 x 6.5 x 6.0 for T. tetrathele and T. chuii, respectively. The average length and width (in I'm) of the rotifers were 196.0 x 139.2
and 173.5 x 115.6 for clone N.R.E. and the Java strain, respectively (from Lubzens et al., in preparation). The average
reproductive rates and standard deviations were calculated as described in Fig. 2.
 391

Tahiti) reproduce equally well at 20 0 , 25 0 or life stage and temperature. First feeding larvae
30 °C while others (Spain, Eilat small, lSI and have relatively slow swimming speeds (Fukuhara,
IS2) clearly prefer high temperatures. Similarly, 1983) and low capture success at onset offeeding
the large size strains (N.R.E. and lL) higher (2-10%) (Hunter, 1980), so they may require a
reproductive rates were found at 25 a C or 30 a high rotifer concentration. This concentration
than those at 20 a C. Furthermore, a positive cor- could be reduced up to the stage where the larvae
relation was found between body size and maxi- may demand higher quantities in order to meet
mum size of particles ingested (Hino & Hirano, energetic requirements. Up to a rotifer concen-
1980). It is likely that large size algae (e.g. tration of 10 ml- 1, a direct relationship between
Tetraselmis tetrathele), which are currently popu- survival, growth and food concentration was
lar in feeding rotifers, will not easily be ingested by shown in gilthead seabream (Tandler & Sherman,
small rotifers. Small rotifers cultured on such 1981; Peguin, 1984), and in bream (Archosargus
algae will probably have low reproductive rates rhomboidalis) (Dowd & Houde, 1980). Beyond
(Fig. 3). Consequently, choosing the appropriate this concentration, a sharp drop in both growth
rotifer strains to be mass cultured will depend on and survival of seabream larvae was observed
the environmental conditions prevailing at the (Tandler & Sherman, 1981; Peguin, 1984). These
hatchery and the size of larval predators. More- results, however, were not related to an increase
over, in offering different rotifer size strains, their in the metabolite concentration (NH 3 - NH4 +) in
dry weight must be taken into account in evaluat- the water body. In a study in which rotifers,
ing the number of rotifers required by the preda- labelled with 14C, were offered to seabream larvae,
tory larvae (see below). at concentrations of 10 and 40 ml- 1, it was
shown that feeding rate of these larvae was
directly correlated to rotifer concentration
The distribution and concentration of rotifers in lar-
(Tandler & Mason, 1984). Therefore, it can be
val tanks
proposed that the negative effect of elevated
The distribution of the rotifers in the water column rotifer concentrations on growth were associated
oflarval tanks depends mainly on the salinity and with reduced efficiency of the digestive process at
to a lesser extent on water quality. Exposing high rotifer concentrations. Similar observations
rotifers to sudden changes in salinity will result in were made by Boehlert & Yoklavich (1984), who
their adhesion to the bottom or sides of containers found that the assimilation efficiency of herring
(Gatesoupe & Luquet, 1981; Lubzens, 1987), larvae (Clupea harengus) was negatively and loga-
making them unavailable to the larvae. Other fac- rithmically correlated to rotifer concentrations
tors affecting the distribution of rotifers are oxy- between 1 and 10 ml- 1.
gen and ammonia levels which affect the swim- Almost no information is available on the effect
ming speed (Epp & Winston, 1978; Snell et al., of larval density in the culture tanks on rotifer
1987) and the type of food offered to rotifers. concentration. Okamoto (1969) showed that sur-
The nutrition of fish larvae depends primarily vival of larvae is clearly related to larval stocking
on the probability of encounter between the food density, but it is not known whether this is related
and the larva as well as its suitability in terms of also to the shortage in food which may occur at
size and nutrient composition. The probability of high larval densities.
encounter between the larva and the rotifer Marine fish larvae depend primarily on vision
depends on concentration. In tum, the perception to find their food, as a result of the pure cone
of food organisms affects larval swimming retina found in the majority of marine larvae
behavior (Hunter, 1980). Several studies showed (Blaxter & Staines, 1970; Blaxter, 1975). Photo-
a direct correlation between food concentration period, light intensity and color of background of
and larval survival (Theilacker & Dorsey, 1980). rearing containers are therefore of paramount
The optimal food concentration depends on the importance for their hunting success.
392

The effect of photoperiod in combination with sudden shortages in supply as a result of unfore-
rotifer concentration is intimately related to the seen collapses. Preventive measures may be
probability of encounter between the larva and its taken, to some extent, by routine daily monitoring
food (Dowd & Houde, 1980; Peguin, 1984). In of swimming speed, proportion of eggs (Snell
both studies it was found that at low rotifer con- et ai., 1987), and physical parameters (pH, oxygen
centrations, there was a direct relationship level, etc.). Preserved rotifers could be helpful in
between the duration of the photoperiod and the management of supply and demand in the
growth. At high rotifer concentrations of 25 ml- 1 hatchery. In this respect, the possible use of artifi-
in gilthead seabream (Peguin, 1984) and 100 to cially produced rotifer resting eggs has been evalu-
5001- 1 in seabream (Dowd & Houde, 1980), ated in recent years (Lubzens, 1981, 1989). It was
there was a mid-range photoperiod which sup- found to be too expensive for routine use, in the
ported best growth, beyond which a further way that Artemia cysts are used today in aqua-
increase was associated with no improvement in culture. In extreme cases, these eggs could be used
the former and with reduced growth rates in the to initiate small-scale new cultures. Preservation
latter species. A similar response of growth to a of live rotifers at low temperatures has been
mid-range photoperiod of 18 h was reported by examined recently (Berghahn et al., 1989;
Barahona-Fernandes (1979) for sea bass (Dicen- Lubzens, 1989; Lubzens et al., 1989). This
trarchus labrax). On the other hand, Fuchs (1978) method would permit the preservation of different
obtained best larval growth in sole (Solea solea) strains or clones, including those that do not pro-
under continuous illumination. Survival of both duce resting eggs. In those cases where B. plicatilis
gilthead seabream (Peguin, 1984) and seabream is abundant, it could also be used for culturing of
(Dowd & Houde, 1980) peaked at mid-range freshwater fish larvae, e.g. ayu (Plecoglossus
photoperiods of 15 and 13 h, respectively. In altivelis) or cyprinids (Cyprinus carpio) (Kanazawa
seabream, the effect of photoperiod was inde- et ai., 1982; Lubzens et al., 1987).
pendent of rotifer concentration, while in gilthead
seabream, a direct correlation between photo-
period duration and survival was observed only at Ingestion, digestion and conversion rates of
low food concentrations of 1.0 ml- 1. rotifers by fish larvae
The number and concentration of rotifers
supplied to fish cultures is expected to vary with Ingestion
temperature. Temperature will affect several
processes in the growth and survival of larvae, As already mentioned above, fish larvae tend to
among them metabolic rate and swimming activity increase both the quantity and the size range of
(Theilacker & Dorsey, 1980). This will result in particles upon which they feed throughout their
higher predation and energy (food) requirements. ontogeny. The increase in the number of food
items ingested daily by fish larvae is an exponen-
tial function of larval age (Stepiens, 1976;
Availability of rotifers Buckley & Dillmann, 1982) or length (Okauchi
et al., 1980; Barahona-Fernandes & Conan,
The large quantities of rotifers required for raising 1981). In absolute terms, larval daily rations
marine fish larvae (40000-173000 per surviving increase as a linear function of their weight
larva) (Okauchi et al., 1980 Kafuku & Ikenoue, (Laurence, 1977; Houde & Schekter, 1983;
1983) are of constant concern to the aquaculturist. Klumpp & von-Westernhagen, 1986; Minkoff,
The development of high density mass culture 1987). The numbers of rotifers consumed daily
techniques (Hirata, 1980; Lubzens, 1987; (Rn) as a function of larval length (L) has been
Yamasaki et ai., 1988) allows for constant pro- determined for porgy (Acanthopagrus schlegeli) as
~uction of rotifers, but does not insure against Rn = 2.43 L 3 .05 (Okauchi et ai., 1980) and red sea
 393

bream (Pagrus major) as Rn = 0.39 L 3 .67 (Fujita, efficiency in the early larval stages is that of food
1979). Likewise, the dry weight of rotifers con- density. As already discussed before, larvae,
sumed daily, Rw (daily ration) as a function of being 'number maximizers', tend to consume prey
larval dry weight (Wt, in mg) has been evaluated in relation to availability rather than according to
for the blenny (Blennius pavo) as satiation. Therefore, at high prey densities they
will tend to consume food at a rate which
Rw = 314.5 Wt + 26.3
decreases its residence time in the gut with a con-
(Klumpp & von-Westemhagen, 1986), herring comitant reduction in the assimilated efficiency
(Clupea harengus) as (Boehlert & Yoklavich, 1984).
Rw = 0.13 Wt + 49.67
and turbot (Scophthalmus maximus) as Nutrition
Rw = 0.43 Wt + 38.45
The current knowledge pertaining to marine fish
(Minkoff, 1987). Concomitant to the increase in larval nutrition has been derived through the
feeding rates, the growing larvae tend to require analysis of the performance of the larvae while
larger food particles. This factor has led to the use feeding on different rotifers 'enriched' with var-
of different rotifer strain sizes in rearing of fish ious nutrients and through testing of formulated
larvae (see above). inert diets. It was found that rotifers cultivated on
baker's yeast alone could not support larval
growth or survival. The failure of these rotifers
Conversion was attributed to their lack of the long chain
highly unsaturated fatty acids (HUFA) on the n-3
Rotifers were found to contain acid proteinase, series, mainly 18: 3 n-3, 20: 5 n-3 and 22: 6 n-3
alkaline proteinase and two kinds of alkaline pro- (Owen et aI., 1975; Yone & Fujii, 1975; Cowey
teases (Hara et al., 1979a, b, 1984). All the cur- et al., 1976; Fujii & Yone, 1976; Gatesoupe et aI.,
rent data show that larvae at first feeding seem to 1977; Watanabe et al., 1983; Dendrinos &
possess the necessary complement of enzymes for Thorpe, 1987). These fatty acids are found in
digesting their prey (e.g. Govoni et al., 1986). some of the algae of marine origin. This has led
The assimilation of ingested rotifers, or any to an extensive search for species of algae which
other food organism, by fish larvae has been are most suitable for enrichment of rotifers. From
shown to be very rapid. 14C originating from a practical aspect, using yeast to raise rotifers
labeled rotifers has been found to be respired by instead of algae reduces greatly the production
fish larvae 3 h following their ingestion (Govoni costs in the hatchery. Furthermore, replacing the
et al., 1982). algal enrichment step with inert rotifer diets will
The gross growth efficiency, which is the pro- assist in curtailing the production price of nutri-
portion of the ingested food invested in growth tionally adequate rotifers.
(Ki) (Brett & Groves, 1979), has been evaluated
for larvae feeding on rotifers; for herring at
20-60%, turbot 20-40% (Minkoff, 1987) and Rotifer biochemical composition in relation to
blenny 50-56% (Klumpp & von-Westemhagen, larval requirements
1986). The values obtained for the first two
species are similar to values reported for other fish The nutritional value of rotifers to larvae depends
species feeding on other types of prey (Govoni on their dry weight, caloric content and bio-
et aI., 1986; Klumpp & von-Westemhagen, chemical composition. The dry weight of a single
1986). rotifer, excluding eggs, as measured by Doohan
One factor which influences the assimilation (1973), Tandler & Mason (1984), Theilacker &
394

Kimball (1984), Yufera & Pascual (1984) and and the values reported fall within the range of the
Minkoff (1987), ranges between 120-360 ng/ind, experimental error. Both Scott & Baynes (1978)
depending on the nutritional state and body size. & Minkoff (1987) have found protein in the range
Scott & Baynes (1979) estimated that during of 50-58% of the rotifer's dry weight. Watanabe
periods of high growth rates and ample food et al. (1983) have set protein levels at 65%, while
supplies, the rotifer's weight could increase up to Dendrinos & Thorpe (1987) found these levels to
620 ng/ind. Furthermore, the latter authors have be 34-36 %. Contrary to this, Ben-Arnotz et al.
also demonstrated that at 24-26 °C, in the (1987) have reported a much wider range of crude
absence of food, the rotifer loses daily between protein from 28-51 %. Lipids usually range from
18-26% of its body weight. The influence of the 9-16% of the dry weight (Scott & Baynes, 1978;
dietary condition on the rotifer's dry weight has Minkoff, 1987), and up to 23.5% and 28%
also been noted by Minkoff (1987), who found (Watanabe et aI., 1983; Dendrinos & Thorpe,
that rotifers raised on baker's yeast alone 1987) (Table 1). One observation seems to be
increased their dry weight by 12-23 % following consistent throughout the literature: rotifers fed
prolonged (48 h) enrichments with algae. Con- only on baker's yeast are poorer in their total lipid
versely, rotifers lost 30% of their body weight content than their counterparts that had either
following a 24 h starvation period at 20 ° C. The been cultivated only on algae or had received
rapid loss of organic material from rotifers which some algae in their diet. This low level of lipids
are deprived of food is generally perceived as one can be elevated by prolonged feeding on algae
of the main factors causing poor growth and high (Minkoff, 1987) or by feeding rotifers with lipid-
mortalities in fish larval cultures. This is probably enriched yeasts (Dendrinos & Thorpe, 1987).
the reason for the use ofthe 'green water' by most Inasmuch as marine fish require a diet contain-
larval culturists, which maintains the rotifers in a ing 40-60 % protein (Castell et al., 1986) and
healthy nutritious state. 13-16% lipid, there is no doubt that rotifers fulfill
The other prominent factor influencing the these requirements. However, for the rapidly
rotifer's weight is that of the animal's size, which, developing larval stages, it is probable that the
as pointed out previously, is mainly strain- balance of dietary amino acids, fatty acids,
dependent. Theilacker & Kimball (1984) have vitamins and minerals determines the success of
described the relationship between the width (X, organogenesis rather than the gross composition
in Jim) and dry weight (Y, in ng) of B. plicatiUs to of the diet.
be Y= 1.4 x 10- 5 XZ. The amino acid profiles of rotifers enriched on
Direct caloric measurements show that the ash- a variety of marine algae or yeast have been deter-
free caloric value of a rotifer ranges from mined by a number of researchers (Watanabe
4.8 cal/mg (Theilacker & Kimball, 1984) to et al., 1983; Dendrinos & Thorpe, 1987; Minkoff,
6.46 cal/mg (Minkoff, 1987). Again, the value per 1987; Rezeq & James, 1987). These analyses
individual will depend on both the size and nutri- show no significant differences in rotifer amino
tious state of the organism. Moreover, it was acid profiles and therefore fail to explain the
reported (Kentouri & Divanach, 1982; Minkoff, advantage of certain algae used for rotifer enrich-
1987), and also found in Sparus aurata cultures ment. Furthermore, the amino acid profiles of
(Tandler, pers. com.), that rotifer eggs and the rotifers do not seem to deviate significantly from
rotifer lorica are not digested by the fish larvae. those of other zooplanktonic organisms either
Therefore, it is essential to correct for egg/female cultured or of wild origin, which are recognized as
ratios in the estimates made of the nutritional being beneficial for growth oflarvae (Castell et aI.,
requirements of larvae (Minkoff, 1987). 1986).
Most researchers have noted that the species of Undoubtedly, the most significant dietary fac-
algae or yeast had only minor effects on the tors to influence the growth and survival of marine
biochemical composition of rotifers fed on them, fish larvae are the highly unsaturated fatty acids
 395

Table 1. Proximate dry weight values of B. plicatilis cultured on different diets

Diet Protein Lipid Carbo- Ash Authors

% % hydrate %
%
Isochrysis galbana 36.3 21.1 Dendrinos & Thorpe, 1987
Nannochloris oculata 34.5 21.4 Dendrinos & Thorpe, 1987
Saccharomysis cerevisiae 41.6 14 Dendrinos & Thorpe, 1987
Saccharomysis cerevisiae L4 33.17 24 Dendrinos & Thorpe, 1987
Saccharomysis cerevisiae L5 42.3 28.5 Dendrinos & Thorpe, 1987
Chaetoceros gracilis 32 20.1 44.9 Ben-Amotz et al., 1987
Chlorella stigmatophora 28.1 7.4 10 Ben-Amotz et al., 1987
Isochrysis galbana 29.1 11.1 7 Ben-Amotz et al., 1987
Nannochloropsis salina 28 16.4 24.1 Ben-Amotz et al., 1987
Phaeodactylum tricomutum 54.4 8.9 13.5 Ben-Amotz et al., 1987
Yeast 55.4 4.5 28 Ben-Amotz et al., 1987
Yeast 65-67 15-21 10 Watanabe etal., 1983
Chlorella minutissima 58-72 21-31 4-9 Watanabe et al., 1983
Chlorella + yeast 63-67 20-22 4-6 Watanabe et al., 1983
Nannochloropsis oculata 50-53 9-13 Minkoff, 1987
Isochrysis galbana 50-54 10-12 Minkoff, 1987
Yeast 53-57 6-9 Minkoff, 1987

(HUFA) of the n-3 series. These fatty acids most cial yeast enriched with (n-3)HUFA (Kitajima
likely form a part of the larvae's cellular mem- et aI., 1980). (3) emulsions which are based on
branes and therefore are crucial in determining (n-3)HUFA rich oil from marine fish or cuttlefish
the rates of enzymatic processes taking place at extracts (Watanabe et al., 1983). (4) a dry diet for
these sites. It is well established that in rotifers, rotifers which both supports their culture and
the fatty acid profile is chiefly determined by diet improves their quality as food for sea bass larvae
(Scott & Middleton, 1979; Watanabe et aI., 1983) (Gatesoupe & Luquet, 1981 ; Gatesoupe & Robin,
and that only minor alterations in the fatty acid 1982). (5) microcapsules (Walford & Lam, 1987).
composition can be achieved through de novo The dietary HUFA requirements for most
synthesis by the rotifer itself (Lubzens et al., marine fish larvae have not yet been determined
1985). These lipids are digested by the rotifer and in a way that will differentiate between a need for
incorporated into its cellular phospholipids either 20: 5 n-3 or 22: 6 n-3 or both. Red sea
(Lubzens et aI., 1985). Rotifers which are raised bream larvae have very similar growth rates when
on baker's yeast alone are inadequate as food for fed on rotifers containing the C : 20 HUF A incor-
larvae due to their lack ofthese fatty acids. There- porated from Chlorella, as on rotifers containing
fore, rotifer enrichment procedures were devel- both the C: 20 and C: 22 HUF A incorporated
oped to insure the ready supply of nutrients from yeasts cultured in the presence of marine
required by the developing larva. The enrichment lipids (Kitajima et al., 1980). The ayu, Plecoglos-
regimes are based on the immersion of rotifers for sus altivelis, which spends most of its early life
8-24 hours in a medium rich in essential nutrients, history in fresh water, has a dietary requirement
especially n-3 HUFA. The enrichment medium for 18: 3 n-3 (Watanabe et al., 1983) which is very
may contain one of the following constituents: similar to what is known for salmonids
(1) different algal species (Kitajima et al., 1979), (Kanazawa, 1985). However, the ayu can benefit
for example: Isochrysis galbana (Howell, 1979; from the input of both the C: 20 and C : 22 FA
Tandler & Helps, 1985); Nannochloropsis sp. and into its diet (Oka et al., 1980). A recent attempt to
Tetraselmis sp. (Fukusho et al., 1984); (2) a spe- differentiate between the requirements for C: 20
396

and C: 22 FA in a range of larvae suggests that Culture of crustacean larvae important in aqua-
only turbot have an essential fatty acid (EF A) culture with B. plicatilis
requirement for both C: 20 and C: 22 HUFA,
while herring and plaice thrive equally well when Several reports show that rotifers may fill the gap
only the C : 20 HUFA or both the C : 20 and the between the period phytoplankton and Artemia
C : 22 are present in the diet (Minkoff, 1987). are offered to developing shrimp larvae (Mock
et a/., 1980). Rotifers are consumed by zoea
stages 2-3 of Penaeus japonicus (Hirata et al.,
Can rotifers be replaced by inert diets? 1985), P. kerathurus (Yufera et a/., 1984), and
P. semisulcatus (Watanabe, 1980; Samocha et aI.,
At present the aquaculturist views the rotifer 1988). P. indicus has been shown to consume
mainly as a 'living food capsule' through which rotifers as early as Z 1 (Emmerson, 1984). It has
nutrients essential to the fish larva can be trans- been recently challenged by Samocha et al. (1988)
ferred (Watanabe et al., 1983). Special rotifer that since rotifers are less adequate and more
feeding techniques were developed for their expensive than Artemia (Tandler, 1984/85), they
'enrichment' with various essential nutrients. are not essential in culturing commercially impor-
Lately, larvae are being offered rotifers which tant crustaceans, but if available may replace
have been enriched with antibiotics. Gatesoupe Artemia for short periods (Yamasaki & Hirata,
(1982), for example, found that larval turbot given 1982).
rotifers which were previously immersed in a
48 mg 1- 1 Tribrissen for 0.5 hr grew faster and
survived better than the untreated control. Summary and conclusions
The general dependence on live feeds, mainly
rotifers, is a constraint as far as dietary manipu- Rotifers of the species Brachionus plicatilis were
lation of feed body composition is concerned. found to be primarily an essential live food source
Therefore, efforts were recently made by various for larvae of several species of marine fish and to
laboratories to develop inert microdiets in which a lesser extent for larvae of shrimp or crabs.
the composition is easy to alter. At present, how- Rotifers possess several characteristics that make
ever, microdiets cannot completely replace them attractive as live food in mariculture:
rotifers in first feeding marine fish larvae. In most (1) They are relatively small, ranging in size from
of the reports, microdiets are offered to marine 0.06-1.00 mm, depending on zoogeographical
fish larvae as a partial replacement only 8-10 days strain and stage of development; (2) They are
after hatching (Kanazawa et a!., 1982; Kana- slow swimmers, maintaining their position in the
zawa, 1985). Kissil (1984/85) correlated the larval water column; (3) They can be cultured at high
size offrrst feeding larvae of 10 species and found densities; (4) They reproduce rapidly, making
that the minimum length of fish reported to feed them available in large quantities within a rela-
on formulated diets is 7 mm. This suggests that tively short period; (5) They can be enriched with
the minimum age/size recommended for success- fatty acids or antibiotics which are required for
ful use of inert diets is associated with the stage growth and survival of larvae.
of development of the digestive system. A similar The development of standardized mass culture
conclusion was reached by Dabrowski (1984), techniques insures, to some extent, an adequate
who proposed that the advantage of live feeds supply of the large numbers of rotifers required to
results from the presence of digestive enzymes, raise commercially important fish species. The
and the activation of zymogens in the developing number of rotifers required depends on the size of
larval digestive tract. the rotifer strain and the duration it is supplied in
the fish larval tanks. The temperature prevailing
in the culture site, the salinity of the water, and the
 397

food offered to rotifers will determine to a large Berghahn, R., S. Euteneuer & E. Lubzens, 1989. High density
extent the rotifer strain chosen for cultivation. storage of rotifers (Brachionus plicatilis) in cooled and
undercooled water. Spec. Publ. Europ. Aquacult. Soc. (in
Choosing the appropriate strain is important for press).
obtaining high reproductive rates, thus reducing Beyer,1. E., 1980. Feeding success of c1upeoid fish larvae and
the rotifer 'standing stock' in the hatchery. stochastic thinking. Dana 1: 65-91.
In order to ensure adequate amounts of essen- Beyer, J. E. & G. C. Laurence, 1981. Aspects ofstochasticity
tiallipids, rotifers must be enriched with either an in modelling growth and survival of c1upeoid fish larvae.
Rapp. P.-v. Reun. Cons. Int. Explor. Mer 178: 17-23.
appropriate alga, emulsified fresh oils or inert par-
Blaxter, J. H. S., 1968. Rearing herring larvae to meta-
ticles such as microcapsules. The nutritional morphosis and beyond. 1. mar. bioI. Ass. UK. 48: 17-28.
quality of the rotifers, which depends on their Blaxter,1. H. S., 1975. The eyes of larval fish. In: M. A. Ali
state of satiation, is maintained in the presence of (ed), Vision in Fishes. Plenum Press, New York: 427-443.
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retinomotor responses in larval teleosts. 1. mar. bioI. Ass.
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with the concomitant development of inert foods. Boehlert, G. W. & M. M. Yoklavich, 1984. Carbon assimi-
The possible future development of preserved live lation as a function of ingestion rate in larval Pacific
rotifers may improve the management of supply herring, Clupea harengus pallasi Valenciennes. 1. expo mar.
and demand of rotifers in the hatchery and may BioI. Ecol. 79: 251-262.
Brett, J. R. & T. D. D. Groves, 1979. Physiological
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Buckley, L. 1. & D. W. Dillmann, 1982. Nitrogen utilization
Acknowledgements of larval summer flounder, Parallichthys dentatus
(Linnaeus).1. expo mar. BioI. Ecol. 59: 243-256.
Castell, 1. D., D. E. Conklin, 1. S. Craigie, S. P. Lall & K.
We would like to thank the U.S. Agricultural Norman-Boudreau, 1986. Aquaculture nutrition. In M.
Research and Development Fund (BARD) and Bilio, H. Rosenthal & c. J. Sindermann (eds), Realism in
the National Council for Research and Develop- Aquaculture; Achievements, Constraints, Perspectives.
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251-308.
respectively) supporting our research.
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Nitrogen flow through a Brachionus / Chlorella mass culture system

Warren D. Nagata
Present address: Aquaculture Department, Malaspina College, 900 Fifth Street, Nanaimo, B.C., Canada,
V9R 5S5

Key words: mariculture, rotifers, batch culture, nitrogen, recycling

Abstract

A model of nitrogen flow is presented through the Brachionus plicatilis/ Chiorella saccharophila mass (batch)
culture system, from the initial input of inorganic nitrogen to the algal culture medium to the final
production of rotifers, dissolved nitrogen and particulate nitrogen.
A nitrogen budget was first formulated for B. plicatilis relating ingestion, excretion, egestion, somatic
growth and reproductive growth. Measurements were made on rotifers from 20° and 10° cultures.
The calculated model of nitrogen flow through the rotifer/algal batch culture system estimates the
percentages of the original input nitrogen which will be incorporated into algal nitrogen, rotifer nitrogen
and the particulate and dissolved nitrogen pools. It is suggested that the dissolved nitrogen pool could
be recycled directly for use in subsequent algal culture.

Introduction development of more efficient modes for this

species.
Since its initial application as a live food organism The object of this study was to examine the
for the larval sweet smelt, the euryhaline rotifer dynamics of nitrogen cycling within the
Brachionus plicatilis Milller has become a widely- B. plicatilis mass culture system in the batch cul-
used food organism in the larval culture of over 60 ture mode of operation. Initially, nitrogen budgets
marine finfish and 18 crustacean species were formulated for this species. These budgets
throughout the world (Fujita, 1983; Lubzens, were then expanded into general models of nitro-
1987). Although mass culture techniques for this gen flow within the culture system.
rotifer have been greatly improved over the last 25
years (Hirata, 1980; Lubzens, 1987), unexplained
decline or inhibited growth in the cultured popu- Materials and methods
lations is still periodically encountered using the
standard batch culture mode. Self-pollution of the Cultures of Brachionus plicatilis and its food
culture environment through accumulation of organism Chlorella saccharophila var. saccharo-
waste products has been deemed a major con- phila Kruger were obtained from the same sources
tributing factor to this culture 'crash' phenome- and cultured as previously described (Nagata,
non (Hirata, 1980; Hirayama, 1987). A more 1985a).
thorough understanding of the nutrient dynamics The following formula modified from Conover
of this culture system may lead to improvement of (1978) was used to formulate the nitrogen budgets
its present modes of operation and the future for B. plicatilis.
402

R = G + U + E, (1) Algal cell counts were carried out by the method

of Nagata (1985b), while clearance and ingestion
where R = ration, food consumed per unit time;
rates were calculated by the equation of Schlosser
G = growth or production, the change in weight
& Anger (1982). Nutrient analyses for ammo-
per unit time; U = excretion, material released as
nium, urea and total dissolved nitrogen were
urine or through the body surface per unit time;
carried out by the methods of Solorzano (1969),
and E = egestion, the amount of feces produced
McCarthy (1970), and Solorzano & Sharp (1980)
per unit time. The growth term (G) was also sub-
and by automated modifications of these methods
divided into somatic growth (GB) and reproduc-
adapted to run on the Technicon Autoanalyzer
tive growth (G R ).
IIc system. Intracellular ammonium pools of the
Assimilation efficiency (A '), the percentage of
algae were also measured by the C-2 method of
the ingested food digested and absorbed into the
Thoreson et al. (1982) and later used to correct
body was calculated as
the rotifer excretion data.
A' = (G + M)/R. (2)

Since B. plicatilis does not produce coherent Results and discussion

fecal pellets collection of feces was not feasible.
Egestion was estimated by difference A mean dry weight of 5.8 ± 1.2 pg cell- 1 was
(E = R - G - U). obtained for C. saccharophila (n = 38) from cul-
Carbon and nitrogen contents of dried algal tures ranging in density from 1.5 to
and rotifer samples were analyzed by a Hitachi 17.5 x 106 cells ml- 1. Mean carbon and nitrogen
Model 026 CHN Analyzer, coupled to ~ Takeda composition of this alga were found to be
Riken TR-2217 integrator and were calibrated 54.58 ± 8.38 and 5.34 ± 1.64%, respectively.
against both caffeine and acetanilide standards. Dry weight data for eggs, neonates and adult
Both rotifer ingestion and excretion were rotifers at 10 0 and 20 ° C, are presented in
measured concurrently. Algae were centrifuged Table 1. The neonate mean dry weights at the two
then resuspended in ASP low nitrogen medium temperatures were 0.32 ± 0.06 Jig ind - 1 (20 0C)
(Hirata & Nagata, 1982) and allowed to acclimate and 0.38 ± 0.02 Jig ind -1 (10 0C). Mean dry
in the dark. Rotifers were filtered from their cul- weights of egg-bearing and non-egg-bearing
ture medium, rinsed several times with ASP females at the two temperatures were not signifi-
medium and resuspended into the algal/ASP sus- cantly different, so pooled means of 0.82 (1 egg)
pension. They were maintained at a density of and 0.55 (no egg) Jig ind - 1 were used in sub-
about 100 ind ml- 1, and were never concentrated sequent calculations.
to densities greater than their stock cultures. The Mean carbon and nitrogen contents of
combined rotifer/algal culture was then accli- B. plicatilis were: 6.56 ± 0.61 % N and
mated for an additional 2 h period, transferred to 39.57 ± 2.24% Cat 20 °c, and 6.23 ± 0.21% N
experimental flasks and tightly stoppered. The and 42.68 ± 1.65% C at 10 °C. Carbon and
flasks were attached to a plankton rotor within an nitrogen contents of eggs and neonates at 20 ° C
incubator and rotated over their longitudinal axes were determined to be 5.60% N and 37.09% C,
through a radius of 10 cm at 5 rpm for an incuba- and 2.69% N and 25.47% C, respectively.
tion period of 3 h at 10 or 20 ± 0.2 ° C. Two From the mean calculated dry weights
control flasks correcting for nutrient uptake and (Table 1), nitrogen and carbon content ofindivid-
cell increase by the algae, and changes in the ual eggless adult rotifers were: 28.54 ng N ind - 1
nutrient levels due to bacterial and physical and 172.13 ng C ind - 1 at 20 ° C, and 28.97 ng N
processes were also prepared. Prior to and follow- ind -1 and 198.46 ng C ind - 1 at 10 °c (Table 2).
ing the incubation period samples were taken for Those of neonates and amictic eggs were 8.61 ng
algal cell counts and dissolved nutrient analyses. N and 81.50 ng C ind -1, and 14.56 ng N and
 403

Table 1. Mean dry weight of B. plicatilis at 20 ° and 10 ° and the results of variance ratio testing between data from the two
temperatures. Each sample was comprised of 50 individuals. Egg dry weight was determined by difference.

Stage Temp. n Dry weight (Jlg) Variance ratio test results

Neonate 20 8 0.32 ± 0.06 Fo = 6.86> FO.05(2)7,6 = 5.70

Neonate 10 7 0.38 ± 0.02 0.02 < P < 0.05
Adult 20 20 0.55 ± 0.08 Fo = 2.29> FO.05(2)19,13 = 2.96
(no egg) 10 14 0.55 ± 0.05 0.01 < P < 0.02
Adult 20 15 0.81 ± 0.05 Fo = 1.71 > FO.05(2)14,7 = 4.60
(1 egg) 10 8 0.83 ± 0.04 0.20 < P < 0.50
Egg 20 0.26
10 0.28

96.43 ng C egg - I, respectively at 20 ° C. At 10 ° C A summary ofthe clearance and ingestion rates

these values were slightly higher. Droop (1976) of this rotifer is presented in Table 3. Clearance
obtained a similar carbon content of this species rates ranged from 0.90 to 2.44 III ind - I h - I at
of 180 ng C ind - I, while Schlosser & Anger 20°C and 0.99 to 1.99 III ind - I h -I at 10 DC.
(1982) found values of 9.3 % N or 26 ng N ind - I Ingestion rates, on the other hand ranged from 45
and 41.5% Cor 116ng C ind- ' . to 345 cells ind -I min -I at 20°C, and 35 to 77
In terms of elemental composition the growth cells ind - I min - I at 10 ° C. Clearance rates
from neonate to adult at 20 ° C results in a gain of appeared to be little affected by algal concen-
27.47 ng N and 136.14 ng C ind - I (Table 2). At tration and temperature within the tested ranges,
10°C, similar values were 24.05 ng N and while ingestion rates seem to be greatly affected by
137.95 ng C ind - I. As for total reproduction these parameters. Mean clearance rates obtained
(22.5 eggs ind- ' lifetime-I, from lifetable in: in the present study were within the range of 1 to
Nagata, 1985a) 327.60 ng Nand 2169.68 ng Care 10 III ind - I h - I obtained for other rotifers at
partitioned into eggs at 20°C, while 276.50 ng N 20-25 °C (Starkweather, 1980) and 0.38 to
and 1809.94 ng C at 10 °C (16.1 eggs ind- ' 10.00 III ind -I h - I for B. plicatilis at 20-28 °C
lifetime - I). Thus, about 12 times as much nitro- (Hirayama, 1983).
gen and 16 times the carbon are diverted into As a conservative mean estimate of daily con-
reproduction as opposed to somatic growth over sumption, an adult rotifer weighing 0.44 Ilg dry
the lifetime of an amictic female of this species at weight at 20 ° C, or 0.46 Ilg at 10 ° C, could ingest
20°C. The same values for 10 °C females were 174960 cells or 1.0 Illg dry weight of C. saccharo-
10.5 x Nand 12 x C. phila per day at 20 ° C; and 98640 cells or 0.57 Ilg

Table 2. Elemental composition (C,N) of B. plicatilis from egg to adult at 20°C and 10 0c.
Temp. (oq Stage Dry weight N (ng ind- I ) C (ng ind- I )
(ng ind - I )

20 egg 260 14.56 96.43

20 neonate 320 8.61 81.50
20 adult 550 36.08 217.64
10 *egg 280 15.68 103.85
10 *neonate 380 10.22 96.79
10 adult 550 34.27 234.74
X 20°C = 435 28.54 172.13
X 10 °C = 465 28.97 198.46

* Assuming same carbon and nitrogen percentage composition as 20 ° C.

404

Table 3. Summary of clearance and ingestion rates of B. plicatiiis at 20°C and 10 °C and various algal densities.

Temp. n Algal density Clearance rate Ingestion rate

CC) (104 cells ind -I) (/ll ind - I h - I) (cells ind - I h - I)

20 6 16.72 1.46 ± 0.31 290 ± 49

20 8 4.21 1.78 ± 0.53 91 ± 29
10 5 2.50 1.60 ± 0.37 52 ± 16
Total X= 1.63 ± 0.47 143 ± 108

dry weight per day at 10 ° C. In terms of carbon those of this study are seen in the studies on
and nitrogen: 54 ng N and 550 ng C at 20 ° C, and B. calyciflorus of Galkovskaya & Ejsmont-
30 ng Nand 310 ng Cat 10 °C. Thus the amount Karabin (1981) and Ejsmont-Karabin (1983), in
of food required per rotifer at 10 0 C is about 56 % which only the rates of ammonium excretion were
that required at 20 ° C. measured. Excretion rates of 0.26 x 10 - 3 f..lg N
A summary of the derived excretion rates of ind -I h - lor 16.91 f..lg N mg dry wt -I day-I at
B. plicatilis is also presented (Table 4). Nitro- 20 °C were obtained in the former, while a rate of
genous excretion in this species was comprised 24 f..lg N mg dry wt - 1 day - 1 was obtained in the
mainly of ammonium and urea-N as previously latter at 20 ° C.
observed by Hirata & Nagata (1982). The pro- Nitrogen budgets were calculated for three
portions of urea excreted ranged from 27.27 to experimental groups of B. plicatilis which varied in
77.78%, with a mean of 48.59% of the total per- acclimation temperature and food concentration
sulfate nitrogen (TPN). Proportions of am- (Table 5). The values for excretion labeled (a) are
monium ranged from 16.33 to 72.53 % with a the initial uncorrected values, while those labeled
mean of 43.34 %. Despite the great variation in (b) and (c) are those corrected for 5 and 10%
proportions of these two nutrients, they always leakage (Dagg, 1976) from the intracellular ammo-
comprised together the majority of the TPN ex- nium pools of the algae, respectively.
cretion (90.24 ± 9.45%). From Table 5, at 20 °C and low algal density
Mean TPN excretion rates at 20 ° C of (1.20 f..lg N ml- I) about 83 % of the ingested algal
3.94 ± 0.76 and 0.67 ± 0.15 ng N ind -I h- I nitrogen was assimilated. Ofthis, 66 %was chan-
were obtained for the high and low algal trials, neled into growth, while 34 %was excreted. At the
respectively, indicating a dependence of excretion higher algal density (4.35 f..lg N ml- I ) 85% of the
rates of B. plicatilis on algal density. The excretion ingested algal nitrogen was assimilated, of which
rates of B. plicatilis at 10 °C of NH4-N, urea-N, 28 % was used for growth and 72 % excreted. At
and TPN at 10 °C were considerably lower than 10 0 C, assimilation was decreased to 61 %, of
those at 20 ° C under similar food regimes. which 52% was channeled into growth and 49%
The only comparable rotifer excretion rates to was excreted.

Table 4. Summary of nitrogenous excretion in B. plicatilis at 20°C and 10 0c.

Temp. Algal density Rotifer dry wt Rotifer N Excretion (/lg N mg dry wt- I day-I)
°C (/lg dry wt ml- I ) (ngind- I)
NH4-N Urea-N Total-N

20 81.4 0.44 28.54 82.64 68.48 151.19

20 22.5 0.44 28.54 15.66 13.89 31.42
10 15.5 0.47 28.97 5.24 11.97 19.41
 405

Table5. Daily nitrogen budget of B.plicatilis at 20°C and 10 0c. Figures in brackets indicate percentage of assimilated algal
ration. GB • GR' U. E, and R expressed as: ng N ind -I day-I. (a)-uncorrected excretion values. (b)-excretion values corrected
for 5 % spillage. (c)-excretion values corrected for 10% spillage. Refer to text for further details of correction method.

Temp. Algal density Somatic Reproductive Excretion Egestion Ingestion Assimilation

(0C) (JLg N ml- I) growth (GB ) growth (GR ) (U) (E) (R) (%)

20 1.20 2.39(5.1) 28.49(60.7) (a)16.08(34.2) 9.12 56.08 83.74

20 1.20 2.39(5.1) 28.49(61.3 ) (b )15.62(33.6) 9.58 56.08 82.92
20 1.20 2.39(5.2) 28.49(61.9) (c)15.15(32.9) 10.05 56.08 82.08
20 4.35 2.39( 1.9) 28.49(22.7) (a)94.56(75.4) 3.18 128.62 97.53
20 4.35 2.39(2.2) 28.49(25.9) (b)79.08(71.6) 18.66 128.62 85.49
20 4.35 2.39(2.5) 28.49(30.0) (c)63.60(67.0) 34.14 128.62 73.46
10 0.83 0.91(4.4) 9.53(45.9) (a) 10.32(49.7) 12.42 33.18 62.57
10 0.83 0.91(4.5) 9.53(47.0) (b) 9.86(48.6) 12.88 33.18 61.18
10 0.83 0.91(4.6) 9.53(47.9) (c) 9.44(47.5) 13.30 33.18 59.92

The assimilation efficiencies from this study are Chlorella cellular nitrogen. This in turn will yield
comparable to values of 64.4 and 74.1 % for this 2.70 g of rotifer nitrogen. A rotifer mortality rate
species (Droop & Scott, 1978), and of 78 % for of 1.62 % day - 1 would result in 0.24 g N being
B. calyciflorus (Galkovskaya, 1971). lost to the particulate pool. Thus the net output of
In Table 6 the above terms are expressed as rotifer biomass will contain 45.05 % of the input
gross (K 1) and net (K2 ) growth efficiencies. These nitrogen, while 29.85 and 25.09% of the remain-
efficiencies were calculated as: K 1 = GB+ GR/R ing nitrogen will enter the particulate and dis-
and K2 = GB + GR/GB + GR + U. solved nitrogen pools, respectively.
The K 1 value of 55.06 % for this rotifer is high, but A similar model was constructed for this cul-
comparable to those of other zooplankton, (i.e. ture system at 10°C (Fig. 2). The major dif-
Artemia sp., Kl = 53% (Gibor, 1952); Rhino- ferences between the models at the two tempera-
calanus nasutus Kl = 55% (Mullin & Brooks, tures was seen in the lower final yield of rotifer
1970). nitrogen (1.15 g N), which contained only 21.06%
The K2 value of 66.41 % is comparable to those of the original input nitrogen, and increase in the
of other brachionid rotifers, i.e. B. calyciflorus, particulate nitrogen pool (2.85 g N) at 10°C. The
K2 = 69% (Galkovskaya, 1963); and K2 = 74% latter comprised 52.2 % of the original input nitro-
(Galkovskaya & Ejsmont-Karabin, 1981). gen. Of this particulate nitrogen pool, rotifer fecal
From the preceding information, the nitrogen material was the major contributor (1. 91 g) while
flow through the culture system as a whole can be 0.39 g N was contained in dead rotifer nitrogen.
estimated. A model of nitrogen flow through the The dissolved nitrogen pool, however, was very
B. plicatilis/C. saccharophila culture system oper- similar to that at 20°C comprising 26.74% of the
ating in the batch culture mode at 20 ° C is illus- original input nitrogen. Thus it can be seen that
trated in Fig. 1. Transfer efficiencies were the overall efficiency of the culture system at
assumed to be at maximum levels, such that: 10°C can be expected to be less than half that at
assimilation of medium N to Chlorella 20°C.
N = 100 %; rotifer ingestion efficiency = 90 % ; Although the nitrogen contained in the particu-
rotifer assimilation efficiency = 83.0%, and late N pool cannot be readily reutilized, the
rotifer net growth efficiency = 66.4 %. The origi- amount of nitrogen contained in the dissolved
nal input of nitrogen contained in the Yashima nitrogen pool is almost entirely in remineralized
I-A medium of 5.46 g per m 3 will under ideal forms which can be utilized directly as N sources
growth conditions yield an equivalent amount of by Chiarella sp. Nevertheless, this amount of
406

Table 6. Gross (Kd and net (K2 ) growth efficiencies of B. plicatilis at 20 °C and 10 °C.

Temp. Algal density Assimilation KI K2

( 0 C) (JLg dry wt ml- I) (%) (G B + GR/R) (GB + GR/GB + GR + U)

20 22.5 83.74 55.06 65.76

20 22.5 82.92 55.06 66.41
20 22.5 82.08 55.06 67.09
20 81.4 97.53 24.01 24.62
20 81.4 85.49 24.01 28.08
20 81.4 73.46 24.01 29.60
10 15.5 62.57 31.46 50.29
10 15.5 61.18 31.46 51.43
10 15.5 59.92 31.46 52.52

Input Yashima medium

5.46 9 N

1
C. saccharophila culture
Uptake: 5.46 9 N
Assimilation: 5.46 9 N
Growth: 5.46 9 N
-17.73xl0 6cells ml- l

5.46 9 Chlorella N
~
Ingestion loss ~. plicatilis culture
0.55 9 N Ingestion: 4.91 9 N
Feces
Assimilation: 4.07 9 N
0.84 9 N
Growth: 2.70 9 N
-94.6 rotifers ml- l
Excretion
Rotifer 2.46 9 Rltifer N Spillage
mortality (45.05%)
0.24 9 N
!
Larval culture
Particulate N pool Dissolved N pool
1. 63 9 N 1. 37 9 N
(29.85%) (25.09%)
Fig. 1. Model of nitrogen flow through the B. plicatilis/ C. saccharophila mass culture system at 20 °C. Figures in brackets indicate
the percentage of the input nitrogen. All nitrogen values are expressed as N per m3 •
 407

Input Yashima medium

5.46 g N

C. saccharophila culture
Uptake: 5.46 g N
Assimilation: 5.46 g N
Growth: 5.46 g N
-17.73xl0 6 cells ml- l

5.46 g Chlorella N
l
Ingestion loss B. plicatilis culture
0.55 g N Ingestion: 4.91 g N
Feces Assimilation: 3.00 g N
1. 91 g N Growth: 1.54 g N
-53.2 rotifers ml- l
Excretion
Rotifer 1.15 g Rltifer N Spillage
mortality (21.06%)
0.39 g N
1
Larval culture
Particulate N pool Dissolved N pool
2.85 g N 1. 46 g N
(52.20%) (26.74%)
Fig. 2. Model of nitrogen flow through the B. plicatilis/C. saccharophila mass culture system at 10 C. Figures in brackets indicate
0

the percentage of the input nitrogen. All nitrogen values are expressed as N per m 3 •

nitrogen can only account for about 25 % of the Hokkaido University for his encouragement and
nitrogen requirement for suitable algal biomass guidance throughout this study. The author is also
production. indebted to the Ministry of Education of the
Although these simplified models overlook Japanese Government for financial support.
several potential fluxes of nitrogen, they may
facilitate the development of more efficient mass
culture systems for B. plicatilis.
References

Conover, R. J., 1978. Transformation of organic matter., In:

Acknowledgements O. Kinne (ed.), Marine Ecology Vol. IV Dynamics, Wiley,
London: 221-499.
Dagg, M. J., 1976. Complete carbon and nitrogen budgets for
The author wishes to express his deepest gratitude the carnivorous amphipod, Calliopius laeviusculus (Kroyer).
to Prof. T. Minoda of the Plankton Laboratory, Int. Revue ges. Hydrobiol. 61: 297-357.
408

Droop, M. R, 1976. The chemostat in mariculture., In: C. Hirayama, K., 1987. A consideration of why mass culture of
Persoone & E. Jaspers (eds.), Proc. 10th Europ. Symp. the rotifer Brachionus plicatilis with baker's yeast is
Mar. Bio!. 1: 71-93. unstable. Hydrobiologia 147: 269-270.
Droop, M. R & 1. M. Scott, 1978. Steady-state energetics of Lubzens, E., 1987. Raising rotifers for use in aquaculture.
a planktonic herbivore. 1. mar. bio!. Ass. U.K. 58: 749-772. Hydrobiologia 147: 245-255.
Ejsmont-Karabin, 1.,1983. Ammonia nitrogen and inorganic MacCarthy,1. 1., 1970. A urease method for urea in seawater.
phosphorus excretion by the planktonic rotifers. Limnol. Oceanogr. 15: 309-313.
Hydrobiologia 104: 231-236. Mullin, M. M. & E. R Brooks, 1970. Growth and metabo-
Fujita, S., 1983. The rotifer as living food for seedling produc- lism of two planktonic, marine copepods as influenced by
tion., In: Jpn. Soc. Sci. Fish. (ed.) The Rotifer Brachionus temperature and type offood. In: J. H. Steele (ed.) Marine
plicatilis, Biology and Mass Culture. Koseisha-Koseikaku, Food Chains. Oliver & Boyd, Edinburgh: 74-95.
Tokyo, Vo!' 44: 9-21, (In Japanese). Nagata, W. D., 1985a. Long-term acclimation of a partheno-
Galkovskaya, G. A., 1963. On the utilization of food for genetic strain of Brachionus plicatilis to subnormal tem-
growth and conditions for the maximum production of the peratures. I. Influence on size, growth and reproduction.
rotifer Brachionus calyciflorus PALLAS. Zoo!. zh. Mosk. Bull. Mar. Sci. 37: 716-725.
42: 506-512, (In Russian), Fish. Res. Bd Can. Trans!. no. Nagata, W. D., 1985b. Long-term acclimation of a partheno-
997. genetic strain of Brachionus plicatilis Miiller to subnormal
Galkovskaya, G. A., 1971. The production of planktonic temperatures II. Effect on clearance and ingestion rates.
Rotatoria. In: G. G. Winberg (ed.), Methods for the Esti- Bull. Fac. Fish. Hokkaido Univ. 36: 1-11.
mation of Production of Aquatic Animals, Academic Schlosser, H. 1. & K. Anger, 1982. The significance of some
Press, London: 123-128. methodological effects of filtration and ingestion rates of
Galkovskaya, G. A. & J. Ejsmont-Karabin, 1981. The the rotifer, Brachionus plicatilis. He1go1ander Meeresunters.
metabolic 0: N ratio in rotifers. Dok. Akad. N auk. 35: 215-225.
belorussk. SSR 25: 472-474, (In Russian), Ca. Trans!. Solorzano, L., 1969. The determination of ammonium in
Fish. aquat. Sci. no. 4811. natural waters by the phenol-hypochlorite method. Limnol.
Gibor, A., 1957. Conversion of phytoplankton to zoo- Oceanogr. 14: 799-801.
plankton. Nature, Lond. 179: l304. Solorzano, L. & J. H. Sharp, 1980. Determination of total
Hirata, H., 1980. Culture methods of the marine rotifer, dissolved nitrogen in natural waters. Limnol. Oceanogr.
Brachionus plicatilis. Min. Rev. Data File Fish. Res., 1: 25: 751-754.
27-46. Starkweather, P. L. 1980. Aspects of the feeding behavior
Hirata, H. & W. D. Nagata, 1982. Excretion rates and and trophic ecology of suspension-feeding rotifers. Hydro-
excreted components of the rotifer Brachionus plicatilis biologia 73: 63-72.
O. F. MOLLER in culture. Mem. Fac. Fish. Kagoshima Thoreson, S. S., Q. Dortch & S. I. Ahmed, 1982. Compari-
Univ. 31: 161-174. son of methods for extracting intracellular pools of
Hirayama, K., 1983. Physiology of population growth. In: inorganic nitrogen from marine phytoplankton. J. Plankton
Jpn. Soc. Sci. Fish. (ed.), The Rotifer Brachionusplicatilis, Res. 4: 695-704.
Biology and Mass Culture. Koseikosha-Koseikaku,
Tokyo, Vol. 44: 52-68 (in Japanese).
Hydrobiologia 186/187: 409-413, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 409
© 1989 Kluwer Academic Publishers.

Kinetics of n-3 fatty acids in Brachionus plicatilis and changes in the

food supply

Yngvar Olsen, Jose Rodriguez Rainuzzo 1, Olav Vadstein & Arne Jensen 1
SINTEF, Division of Applied Chemistry, Aquaculture Group, N-7034 Trondheim, Norway; 1 Laboratory of
Biotechnology, Norwegian Institute of Technology, University of Trondheim, N-7034 Trondheim, Norway

Key words: Brachionus plicatilis, live feed, enrichment, n-3 fatty acids

Abstract

Moderately starved rotifers exhibited a two-phased increase in n-3 fatty acids when they were fed a diet
rich in these fatty acids. The fIrst 20-30 min of enrichment, the increase in n-3 fatty acids was primarily
due to increased gut content. The subsequent slow increase was due to an incorporation of n-3 fatty acids
into rotifers tissues. Saturation was achieved before 24 h of exposure and the saturation level was
independent of the initial content of n-3 fatty acids in the rotifers.
Starvation and limited feeding of the enriched rotifers for additional 4-8 h at 10-20 °C did not affect
the accumulated fatty acids signillcantly. This was found for rotifers with high and low initial content
of n-3 fatty acids. The n-3 fatty acids were assimilated with high efficiency from the feed and were not
metabolized faster than other groups of fatty acids.
Enriched rotifers retained their nutritional value for a sufficient period after enrichment to serve well
as live feed for marine fIsh larvae.

Introduction lism of the long-chain fatty acids could dominate

over assimilation if the ingestion rate of the fatty
The use of the rotifer Brachionus plicatilis as live acids was reduced. The goal of this work was to
feed for marine fIsh larvae has been successful study the kinetics of the n-3 fatty acids in
(Ito, 1960; Lubzens, 1987). This is primarily B. plicatilis after shifts in the ingestion rate of
because it can be efficiently cultured and enriched these fatty acids.
with essential fatty acids (Watanabe et al., 1983)
to satisfy the high demand of most marine larvae
for polyunsaturated, long-chain n-3 fatty acids Materials and methods
(Owen et aI., 1972; Cowey et al., 1976).
When enriched rotifers are transferred to larval The rotifer Brachionus plicatilis was grown in 250 I
rearing tanks with particle-free seawater, the conical, fIberglass vessels in 20%0 seawater at
rotifers experience a sudden decrease in food con- 20 ° C. Each 200 I culture was aerated and fed
centration. If the enriched fatty acids were pri- 20-50 g bakers yeast (wet weight) plus 2-5 g
marily stored in the gut, a rapid decrease in n-3 capelin oil (10% by weight) once per day. Oil and
fatty acids would result because of reduced yeast were mixed with 0.5 I freshwater in a blender
feeding rate and defecation. In addition, metabo- for 10-15 s. The cultures were used in the experi-
410

ments when they had reached a density of Table 1. Summary of the experimental conditions of the
200-250 ind ml- 1. feeding experiments in 20%0 seawater
Two feeding experiments were undertaken, Exp. 1 Exp. 2
each with two phases. At time zero, moderately
starved rotifers were fed a diet rich in n-3 fatty Cultivation feed Baker's yeast Baker's yeast
acids, obtained from 25 /lm micronized herring + capelin oil
roe powder. After 4-8 h the animals were Feeding phase
removed and starved or fed a low concentration Density. ind ml- 1 200 200
(5 mg 1- 1) of the roe powder, respectively. Temp,.oC 20 20*
In each feeding experiment two replicate 100 1 Duration. h 4 8
cultures ofrotifers at density of200 ind ml- 1 were Postfeeding phase
given 0.1 g 1- 1 of blended roe powder under Density. ind ml- 1 10 10
Time from start of
standard growth conditions. Five liter samples for
feedingphase, h 6 24
lipid analysis were removed from one culture just Temp,.oC 20 10
before and just after addition of the feed and Duration. h 4 8
thereafter 5-7 times through the following 4-8 h.
The second culture was used for post-feeding * Cooled from 20 to 10 °C between 8 and 24 h after start of
feeding.
analysis. One liter was washed and transferred to
a 20 1 plastic vessel. Five such vessels contained
19 liter 5 /lm filtered, 20%0 seawater and five (1959) method. The fatty acids were methylated
vessels contained 19 liter 5 /lm filtered seawater with 12% BF3 in methanol (Metcalfe et aI., 1966)
plus 5 mg roe powder 1- 1. The final concentration after alkaline hydrolysis (0.5 N NaOH), and the
of rotifers through post-feeding was approxi- methyl esters were determined by capillary gas
mately 10 ind ml - 1 and the added food was suf- chromatography (Carlo Erba HRGC 5160 series
ficient to keep rotifer guts filled for several hours. equipped with a SP-2330 glass capillary column,
Samples for lipid analysis were collected at dif- splitless injection and flame ionization detector)
ferent times through the following 4-8 h by har- fitted with a Shimadzu-Chromatopac C-R3A
vesting two entire vessels. In addition, the culture computing integrator. Fatty acid identifications
was sampled at start of the post-feeding phase. were based on standard references (Nu-Chek
The two feeding experiments differed in various Prep., Elysium Minn.). Total lipids and total
ways summarized in Table 1. In Exp. 1, the methylated fatty acids were determined gravi-
capelin oil was excluded from the diet through 5 d metrically in separate samples.
before start of the experiment. Moreover, the
post-feeding phase of Exp. 1 was initiated 6 h
after the addition of the roe powder diet, whereas Results
it was initiated after 24 h in Exp. 2. The rotifers
were cooled from 20 to 10 0 C between 8 and 24 h The content of total lipid and the lipid com-
in Exp. 2. This cooling procedure is routinely position of the rotifers are shown in Table 2. The
applied for preparation of enriched live feed in our concentration of n-3 fatty acids was higher in
laboratory. animals fed bakers yeast + capelin oil (yeast-oil
Samples of rotifers (0.1-2 g dry weight) were rotifers) than in those which were given bakers'
collected on 70 /lm screens, washed with 20%0 yeast through the five days preceding the experi-
seawater, frozen on dry ice, transferred to test ment (yeast rotifers). The latter animals still
tubes, freeze dried, and stored at - 80 0 C in the contained a significant amount of 20 : 5 n-3 from
dark under a nitrogen atmosphere. earlier feeding on yeast + oil. The content of total
Total lipids were extracted from 25 mg freeze lipids was the same in both groups. The fatty acid
dried samples using a modified Bligh & Dyer fraction of total lipids was not dependent on
 411

Table 2. Content of n-3 fatty acids and polyunsaturated fatty acids (PUFA)* in rotifers cultured on yeast (yeast rotifers) or
yeast + oil (yeast-oil rotifers) and in the rotifer feeds.

Fatty acids, Rotifers Feeds

mg (g dry wW I
Yeast Yeast + oil Yeast Capelin oil Roe powder

20: 5 n-3 2.2 3.3 54 12

22: 6 n-3 0.63 2.3 42 35
Sum n-3 4.3 S.4 0.2 153 52
Sum PUFA 7.0 15 2.0 164 54

Total lipid,
mg (g dry wt)-I 100 94 21 150

* Sum of fatty acids with more than one double bond.

rotifer feed in the present experiments and in total lipid in the rotifers as also insignificant
averaged 0.57 ± 0.02 % (mean ± SE, n = 20) of (P < 0.01) through post-feeding, which is not sur-
total lipids. prising because this phase was short and tempera-
Lipid composition of the feeds was rather dif- ture was low.
ferent. Baker's yeast was practically free from n-3 The average concentration of some n-3 fatty
fatty acids, whereas both capelin oil and herring acids through the post-feeding period are shown
roe powder were very rich. The concentration of in Table 3. The concentration of n-3 fatty acids
22 : 6 n-3 in the roe powder is notable. The con- were slightly, but significantly (P < 0.05), higher
tent of lipids was also much higher in the roe in the yeast-oil rotifers than in the yeast rotifers,
powder than in baker's yeast. whereas the content of polyunsaturated fatty
The fatty acid composition of both yeast-oil acids remained lower. These differences are
rotifers and yeast rotifers changed rapidly when regarded as accidental because some variations
the animals were fed the roe powder. The concen- occurred in fatty acid content from one experi-
tration of 22 : 6 n-3 and the sum of n-3 fatty acids ment to another.
are shown in Fig. 1A. The rate of increase in the
n-3 fatty acids was high through the first half hour,
whereafter it leveled off and continued increasing Discussion
at low rate for at least 4 h. The concentration of
22: 6 n-3 and the total n-3 fatty acids remained The main conclusion of the present study is that
uniformly higher in the yeast-oil rotifers than in the increased content of highly unsaturated n-3
the yeast rotifers, but became equal within start of fatty acids obtained by feeding rotifers for short
the post-feeding phase of the experiments
(Table 1 and Fig. 1B). Table 3. Average content (± SE) of n-3 fatty acids and
The concentrations of 22 : 6 n-3 and n-3 fatty PUF A in the rotifers through the post-feeding phase. Data
for rotifers with and without food added are pooled (n = 11).
acids in the rotifers through post-feeding are
shown as a function of time in Fig. 1B. The con- Fatty acids, Yeast rotifers Yeast-oil rotifers
tents remained constant in both yeast-oil rotifers mg (g dry Wt)-I
and yeast rotifers. The addition of roe powder at
a concentration sufficient to maintain maximum 20: 5 n-3 4.S0 ±0.08 5.33 ± O.OS
22: 6 n-3 6.60 ± 0.14 7.04 ±O.OS
ingestion rate and filled guts (Korstad et aI., 1989)
Sum n-3 13.3 ± 0.26 14.1 ± O.1S
did not have any significant effect upon their fatty PUFA 18.3 ±0.37 15.5 ± 0.29
acid content through post-feeding. The decrease
 412

A B
.... Yeast rotiters tJ. ... O. fed
tJ. 0 Yeast-Oil rotifers ~)I(~ not fed

?;
15
tJ. ~ H ...
.....
:A:

•
-0 .A.{( ~ ~
:{j: ~
Ol X tJ.

---EOl ...
L. w3 ... tJ.

~ 10
0
-... d
LI:
i «i )I( a ..• li
of ~ g...

5
•

o
o 0.5 1 5 10 0 0.5 5 10

Time, h

Fig. 1. The kinetics of some n-3 (w3)fatty acids of yeast fed and yeast + oil fed rotifers. A: Feeding phase. Variation in 22: 3
n-3 and the sum of n-3 fatty acids upon addition of a diet rich in n-3 fatty acids at time zero. B: Post-feeding phase. Variation
in 22 : 6 n-3 and the sum of n-3 fatty acids after transferring the rotifers to low food concentration. Crossed values represent
starved groups and the remaining values represent groups fed 5 mg roe powder 1- 1.

time (6-24 h) with a feed rich in n-3 fatty acids The accumulation rate of 22 : 6 n-3 and other
(enrichment) is stable for hours after transferring n-3 fatty acids in the rotifers tissue must neces-
the rotifers to conditions of very low food concen- sarily level off at some level dependent on the
tration. This was found in rotifers which exhibited composition of the fed and the metabolic
different initial contents of n-3 fatty acids in their processes in the rotifers. In the present experi-
tissues and at different temperatures through ments, an apparent saturation level was obtained
post-feeding (10-20 QC). This conclusion sug- within 24 h. Other feeding experiments have indi-
gests that enriched B. plicatilis will maintain their cated that only minor changes take place in the
improved nutritional value for several hours after fatty acid distribution of rotifers after 24 h of
application as live food for marine fish larvae feeding on a given diet. These findings are also in
The kinetics of fatty acids through the feeding agreement with the fact that the same saturation
period indicate a two-step process of fatty acid level of n-3 fatty acids was obtained for rotifers
accumulation. The rotifers filled their empty guts with different initial content of n-3 fatty acids in
in about 20-30 min and this resulted in a rapid the present experiments (cf. Table 2). However,
initial increase in the principal n-3 fatty acids of further additions of roe powder could theoreti-
the roe powder (cf. Table 2). The slower increase cally have resulted in significantly higher content
which followed probably represented an accumu- of n-3 fatty acids after 24 h, because the rotifers
lation of the actual n-3 fatty acids in the body probably reduced the food level below saturation
tissues of the rotifers. over that time in Exp. 2.
 413

The stability of the fatty acids in the rotifers in References

the post-feeding period was surprising. The guts
of the rotifers were filled when they were trans- Bligh, E. G. & W. 1. Dyer, 1959. A rapid method of total lipid
extraction and purification. Can. 1. Biochem. Physiol. 37:
ferred to post-feeding in both experiments, even in
911-917.
Exp. 2 where the concentration of food was low Cowey, C. B., J. M. Owen, 1. W. Adron & C. Midde1ton,
after 24 h. The groups of rotifers which were fed 1976. Studies on the nutrition of marine flatfish. The effect
low concentrations of roe powder through post- of different diatary fatty acids on the growth and fatty acid
feeding were expected to maintain their guts filled composition of turbot (Scophthalmus maximus). Br. J. Nutr.
36: 479-486.
and to have a constant content on n-3 fatty acids
Ito, T., 1960. On the culture of the mixohaline rotifer
(cf. Fig. 1B). The fact that the starved groups also Brachionus plicatilis O. F. Muller in the sea water. Rep. Fac.
kept a constant and high level of n-3 fatty acids Fish Pref. Univ. Mie. 3: 708-740.
implies that the assimilation efficiency of the Korstad, 1. E., O. Vadstein & Y. Olsen, 1988. Feeding
present fatty acids were high and that insignificant kinetics of Brachionus plicatilis fed Isochrysis galbana. This
volume.
amounts were released through defecation. It also
Lubzens, E., 1987. Raising rotifers for use in aquaculture.
follows that the assimilated n-3 fatty acids were Hydrobiologia. 147: 245-255.
not metabolized at higher rates than other fatty Metcalfe, L. D., A. A. Schimtz & J. R. Pelka, 1966. Rapid
acids in the post-feeding phase. preparation of fatty acid esters from lipids for gas chroma-
tography. Analyt. Chern. 38: 514-515.
Owen,1. M., 1. W. Adron, J. R. Sargent & C B. Cowey, 1972.
Studies on the nutrition of marine flatfish. The effect of
Acknowledgements different diatary fatty acids on the tissue fatty acids of the
plaice Pleuronectes platessa. Mar. BioI. 13: 160-166.
This study forms part of a research project on live Watanabe, T., C. Kitajima & S. Fujita, 1983. Nutritional
feed for marine fish larvae (HB4925.18213) values of live food organisms used in Japan for mass
propagation of fish: a review. Aquaculture. 34: 115-143.
financed by The Royal Norwegian Council for
Scientific and Industrial Research. We thank dr.
Grethe Rosenlund for access to the herring roe
powder and Marte Morkved for technical
assistance.
Hydrobiologia 186/187: 415-421, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 415
© 1989 Kluwer Academic Publishers.

Effect of incubation and preservation on resting egg hatching and mixis

in the derived clones of the rotifer Brachionus plicatilis

Atsushi Hagiwara 1, Akinori Hin0 2

IGraduate School of Marine Science & Engineering, Nagasaki University, Bunkyo 1-14, Nagasaki 852,
Japan; 2Department of Fisheries, Faculty of Agriculture, The University of Tokyo, Yayoi, Bunkyo, Tokyo
113, Japan

Key words: Rotifera, resting eggs, light, temperature, hatching, mixis

Abstract

The marine rotifer Brachionus plicatilis typicus (Clone 8105A, Univ. of Tokyo) was cultured in 500 ml
beakers to form resting eggs. Tetraselmis tetrathele was used as a culture food. Just after formation, resting
eggs were exposed to various temperature (5-25 0c) and light regimes (24L: OD and OL: 24D). When
eggs were exposed to light just after formation, the eggs hatched sporadically over a month. No hatching
was observed for six months when eggs were preserved under dark conditions regardless of the tem-
perature. These eggs hatched simultaneously after being exposed to light and eggs preserved at 5 ° C
showed twice as high hatching rate (40%) as that of eggs preserved at 15-25 °C (24%). Clones from
resting eggs that were kept under different temperature and light regimes were reared individually to the
third generation. Incubation at 25°C with lighting produced the highest (5.4% and 5.2%) rate ofmictic
females during their 2nd and 3rd generations, respectively. The lowest rates (0 and 1.5%) were found
when the eggs were kept at 5 ° C in total darkness for six months. A lower rate of amictic female
production was found in clones with higher rates of mixis.

Introduction noticeable (Gilbert, 1977; Hino & Hirano, 1977).

Thus, genetic differences among stocks are the
Sexual reproduction in monogonont rotifer is primary determinants of sexual reproduction in
affected by external and internal factors (reviewed B. plicatilis. The age of the parental female is also
by Gilbert, 1974, 1977; Pourriot & Snell, 1983; a significant internal factor that affects the pro-
Hino, 1983). Of these, internal factors have been duction of mictic daughters (Pourriot & Rougier,
less investigated. Brachionus plicatilis stocks col- 1976; Clement et aI., 1976: Rougier & Pourriot,
lected from different sites show variable charac- 1977). Lately, Hino & Hirano (1985, 1988)
teristics in the sexual reproduction (Snell & Hoff, reported that external factors such as temperature
1985; Hagiwara et aI., 1988). These stocks some- and salinity during the resting egg formation affect
times show distinctive genetic differences among the internal factors such as the mictic repro-
each other (Snell & Winkler, 1984; Fu et aI., duction in derivative clones, although the physio-
1989). The differences of mictic reproduction logical mechanism of this action is unknown.
among clones in a stock may be smaller than that Gilbert (1974) and Pourriot & Snell (1983)
among stocks, but these differences are still reviewed the hatching of rotifer resting eggs. For
416

B. plicatilis, it is known that incubation tempera- condition EB (RC) and 4) incubation in condition
ture and salinity significantly affect the hatching EC (RD).
rate and incubation time (Ito, 1960; Minkoff Five to ten clones were randomly selected from
et al., 1983; Hagiwara et aI., 1985). Culture con- four groups and reared individually (1 individ-
ditions during the resting egg formation affect the ual *0.2 ml- 1) from the first to the third gen-
optimum salinity for hatching (Ito, 1960; Gilbert, eration according to the method of Hino &
1980). Hirano (1977). Culture salinity was 14.5 ppt and
In this report, we examined the effect of exter- the medium was changed daily. Tetraselmis tetra-
nal conditions during the resting stage and incuba- thele and Chlamydomonas sp. grown in Miquel
tion period of resting eggs on the hatching pattern medium (Allen & Nelson, 1910)was suspended in
(sporadic or simultaneous), hatching rate and fresh medium at a density of 5 x 105 cells *ml - 1
sexual and asexual reproductive characteristics in and fed to rotifers. The female type (mictic or
the hatched clones. amictic) was determined and the life-span and
fecundity monitored for all the females of the 1st
and 2nd generation. As an index of amictic female
Materials and methods viability, the number of offspring per day of repro-
ductive life was calculated (Snell & King, 1977).
Brachionus plicatilis(Clone 8105A, Univ. ofTokyo ) A Chi square test was used to evaluate the
was mass cultured in 500 ml beakers to produce effects of incubation and preservation conditions
resting eggs. The temperature was 25 ± 1 ° C, on the hatching rate of resting eggs and the rate
salinity was 14.5 ppt obtained by diluting natural of mixis in the derivative clones. Analysis of
sea water with distilled water and the food was variance was calculated to compare reproductive
Tetraselmis tetrathele. Resting eggs were removed characteristics among experimental clones.
from the culture every day. At the late-exponential Regression analysis was applied to determine if
growth stage when mixis rate reached to the the rate of mixis was related to the viability of a
highest level, the resting eggs deposited in the last clone.
24 hours were collected for the hatching experi-
ment. Just after the collection, samples of 100
eggs were exposed to three different conditions of Results
temperature and lighting: 1) 25°C and 24L: OD
since formation (condition EA), 2) 25°C and Cumulative percentages of resting egg hatching in
OL: 24D for the first 15 days after the formation three different conditions are presented in Fig. 1.
and 25°C and 24L: OD from day 16 (condition When resting eggs were exposed to light just after
EB), 3) 5 °C and OL: 24D for the first month and their formation at 25 ° C (EA), they started hatch-
25°C and 24L: OD thereafter (EC) and 4) 5 °C ing on Day 11. These eggs continued to hatch
and OL: 24D for the first six months and 25°C sporadically for 40 days from the first hatch. The
and 24 L : 0 D thereafter (ED). Light intensity was resting eggs kept in the dark for the first 15 days
2000 lux with white fluorescence bulbs. The (EB) hatched simultaneously 2-3 days after being
hatching of resting eggs was observed directly and exposed to light. The final hatching rates of rest-
number of hatched eggs was monitored every day ing eggs incubated in conditions EA or EB, how-
until no more hatching occurred. ever, remained 26 and';2%, respectively. On the
The hatched clones were classified into four other hand, when resting eggs were kept at 5 ° C
groups according to the conditions of resting egg for 1 and 6 months (condition EC or ED), hatch-
incubation and their hatching patterns. These ing rates were 38 and 42%, respectively. The
were: 1) incubation in condition EA and hatching hatching rates of the resting eggs incubated in
on day 11-14 (RA), 2) incubation in condition EA conditions EC or ED were significantly higher
and hatching on day 26-48 (RB), 3) incubation in than those incubated in EA or EB (Chi-square
 417

/6
40
--.
;!. / /
-...
C!J
0
/ /
j
0'1
c
£.
. --ad
oJ--4
J
I
·
U

20 or-~-d
j)
0
£.

;l-r
C!J
>
0
::J
E
::J
u

0 20 f 40 60 Jf1soi Light
200
Li ght Light
Ti me(days)

Fig. 1. Effect of conditions of the preservation and incubation on the hatching of rotifer resting eggs. 0: Condition EA. . : EB.
L: EC. . : ED. See text for explanation of conditions.

test, P < 0.05). The resting eggs kept at 5 0 C for cant differences in the lifespan, reproductive
one month hatched sporadically, whereas those period, fecundity and number of offspring among
kept at 5 0 C for six months hatched simultane- clones from the four different incubation and
ously. hatching conditions.
The reproductive results of the 1st and 2nd The rate of mictic female production in the 2nd
generations of clones in the four different groups generation of the stem mothers is shown in Fig. 2.
are presented in Table 1. There were no signifi- The rats were: RA- 5.03%, n = 199; RB- 5.75%,

c c
o 0
o
Q;1O
c
~lO
QJ
QJ c
QJ
01
01
"0 -
....,...
"0
C
N
QJ
£. 5 - - QJ
.c 5
c c

t
~ ~

~
~
u.:
~ 0 '---'-....I....-....L..-....L.....Jn----'_ __
RA R B R C RD
~
u

0
RA RB
n
RD

Fig. 2. Percent mictic females produced in the 2nd and 3rd generations hatched from resting eggs preserved and incubated in
different conditions. See text for explanation of RA, RB, RC and RD.
418

Table 1. Reproductive characteristics of the I st and 2nd generation of B. plicatilis clones hatched from resting eggs which are
exposed to the different conditions of preservation and incubation.

Condition Lifespan * Fecundity** Viability Mic. F.

index**'" production
Reproductive days No. of offspring (%)

1st generation
RA 11.3 ± 4.0 23.2 ± 3.0 4.1 ± 1.0 5.0
5.8 ± 1.1 22.7 ± 3.3
RB 12.4 ± 3.9 23.4 ± 0.9 3.7 ± 0.2 5.8
6.2 ± 0.4 22.8 ± 0.4
RC 12.8 ± 4.1 21.2 ± 3.6 3.3 ± 0.9 2.0
6.2 ± 0.8 20.2 ± 4.0
RD 10.3 ± 3.2 25.7 ± 3.1 4.6 ± 0.6 0.0
5.7 ± 0.6 25.7 ± 3.1
2nd generation
RA 10.5 ± 1.0 27.9 ± 0.6 4.3 ± 0.5 2.6
6.3 ± 0.7 26.6 ± 1.2
RB 11.0 ± 0.8 26.6 ± 0.5 3.1 ± 0.5 8.0
6.7 ± 0.9 20.5 ± 1.8
RC 9.8 ± 0.4 26.3 ± 0.6 3.9 ± 0.2
6.2 ± 0.5 22.6 ± 1.7
RD 9.0 ± 0.8 28.3 ± 0.4 5.0 ± 0.0 0.7
5.4 ± 0.1 27.1 ± 0.7

* Days; ** No. of offspring plus No. of unhatched egg; *** Offspring per day of reproductive life. Each value shows
average ± standard error. See text for the explanation of RA, RB, RC and RD.

0 0

-
;t.
.........
u.:
10

u
~
......
0
~ 5
§L-
I::.
a 1::.1::.
~ Do
a. 0
<t
.1::.
1::.1::. °0
0
2 3 4
'" I::.
~
Offspring/day of reproductive life of 2nd generation
Fig. 3. Relation between the viability index of amictic females in the 2nd generation and the percent mictic female produced
by clones hatched from resting eggs preserved and incubated in different conditions. 0: Hatched after preservation at 5 0 C for
I month. /::,.: Hatched after preservation at 25 C for 15 days. 0: Hatched without preservation.
0
 419

n = 226; RC- 2.04%, n = 98; RD- 0%, n = 136. nal viability in the 2nd generation is presented in
Differences in the rate of mictic female production Fig. 3. Regression analysis did not reveal a signifi-
was significant among all groups except RA and cant relation between the viability index and mixis
RB (Chi-square, P < 0.05 between RC and RD, rate. But highest rate of the mictic female produc-
P < 0.01 among other groups). In the offspring of tion was observed in the RD group (1.6-12.2%,5
the 2nd generation, the rate was 2.57% clones), followed by RA (0-7.46 %, 10 clones) and
(n = 4483) and 7.96% (n = 4348) in RA and RB, RB (0-1.2 %, 3 clones).
respectively. These are significantly different with When clones were fed Chlamydomonas, there
P < 0.01. The mixis rate of the RD group was was a more noticeable trend (Fig. 4). The results
1.45% (n = 2746). The rate was highest in the of regression analysis on RD (3 clones) and RB
group RB, followed by RA and RD in both 2nd group (5 clones) indicate a highly significant nega-
and 3rd generations. In groups RB and RD, the tive relationship (P < 0.01, y = - 8.6 x + 29.9,
rates in the 3rd generation were significantly r = - 0.93).
higher than in the 2nd generation. In group RA,
however, the rate was higher in the 2nd gen-
eration. Discussion
The relation between the conditions of
preservation and incubation and the index of clo- Environmental conditions during preservation
and incubation significantly affected the hatching
rates and hatching patterns of B. plicatilis resting
eggs. Resting eggs exposed to lower temperature
showed higher hatching rates (EC, ED). The
highest hatching rate of the eggs used for this
0 experiment was 40 %. We have conducted pre-
liminary experiments with resting eggs from a dif-
....... 10
c;e ferent clone (clone 8401, University of Tokyo)
........ and obtained 90 %hatching rate without exposing
u.: y= -B.60x+29.B7 them to low temperature. This illustrates the varia-
u
~
( r=-0.93) bility among clones in storage requirements and
'+- hatching rates.
0
<!J Two hatching patterns, sporadic and simul-
u
c taneous, were observed depending on the condi-
cI- 5 tions of preservation and incubation. The control
C 0
<!J mechanism of hatching pattern seems to be
Q.
0.. species specific in monogonont rotifers (Pourriot
<{
& Snell 1983). When resting eggs of B. plicatilis
were incubated just after their formation (EA), or
when the preservation period was comparatively
o 2 3
shorter (EC), resting eggs hatched sporadically.
This suggests that each resting egg has a different
Offspring/day of reproductive diapause period. Accordingly, it seems to be con-
Iife of 2nd generat ion tradictory that one group (EA; no preservation)
showed sporadic hatching for over 50 days while
Fig. 4. Relation between the viability index of amictic
the other group (EB; preserved in darkness for 15
females in the 2nd generation and the percent mictic female
produced by clones hatched from resting eggs preserved and days at 25 0 C) finished hatching in almost 2 days.
incubated in different conditions. 0: Hatched after preserva- One possible explanation is that each resting egg
tion at 5 C for 1 month. 0: Hatched without preservation.
0 has a different level of sensitivity to the environ-
420

ment. Changes of environmental conditions, such exposed to inhibitory conditions, they hatched
as from darkness to light, or lower to higher tem- into clones with higher mixis rates and lower
perature, seems to stimulate hatching of resting population growth rates. Brachionus plicatilis
eggs. Resting eggs hatched simultaneously when populations in nature may be controlled by such
preserved for 15 days at 25°C (EB) and for 6 mechanisms. When resting eggs in the sediments
months at 5 ° C (ED). This indicates that the hatch after a winter season of diapause, the hatch-
diapause period of resting eggs increases with a ing clones will probably show fast partheno-
decrease of temperature. We incubated resting genetic growth. In contrast, if resting eggs hatch
eggs just after the formation at various tempera- without experiencing inhibition, the clones will
tures (5-30 0c) and salinity levels (7.2-28.9 ppt), show high mixis rates and produce a large number
and observed that they started to hatch earlier of resting eggs. These resting eggs can be the seeds
when they were incubated at higher temperature for initiating population next season.
or lower salinity (Hagiwara & Hino, in prepa- It has been suggested that sexual reproduction
ration). This suggests that the completion of of B. plicatilis occurs when parthenogenetic repro-
diapause by rotifer resting eggs depends on the duction is vigorous in the moderate environmental
environmental conditions. Development of conditions (Lubzens et al., 1985; Snell, 1986;
embryos in resting eggs probably stops at some Hagiwara et a!., 1988). The highest mixis rate in
point if resting eggs are kept in darkness. Light B. plicatilis populations is observed when they are
triggers the resumption of egg development, fol- in exponential growth. Current results suggest
lowed by synchronous hatching. Study of rotifer that clones with lower viability have higher rates
embryological development will be of interest to of mixis. These clones are less vigorous and the
examine these hypotheses. population growth of these clones is probably
No mictic females appeared in stem females slower. But even these clones show the highest
hatched from resting eggs. The rate of mictic mixis during exponential growth phase.
female appearance in the 2nd and 3rd generation Current results are also of interest for aqua-
varies depending on conditions during the period culture. B. plicatilis is the most commonly used
from the egg formation to hatching. The percent live food for larval rearing of marine fish. Resting
of mixis in the 3rd generation (groups RB and eggs can be seeded similar to brine shrimp cysts
RD) was higher than that in the 2nd generation (Hino & Hirano, 1976; Lubzens, 1981; Snell &
and this is consistent with the result of Hino & Hoff, 1985). The current study suggests that it will
Hirano (1977). Current results showed that the be possible to select favourable clones for the
higher mixis rate was observed in clones hatched purpose of mass culture and resting egg produc-
from resting eggs which were incubated just after tion.
their formation (RA, RB). Moreover, resting eggs
requiring longer incubation time hatched into
clones with the highest mixis rate (RB). When Acknowledgements
eggs were moderately preserved at 25 ° C and
darkness, they hatched into clones with moderate Portion of this research was supported by a Grant
mixis rate. Resting eggs experiencing extreme in Aid to Scientific Research from the Ministry of
preservation conditions had the lowest mixis rates Education, Japan. Authors wish to thank review-
upon hatching (RD). ers for improving the manuscript.
The viability index was inversely related to
mixis rates. The viability of a clone tended to be
References
high when its mixis rate is low. This suggests that
rotifer clones experiencing inhibitory conditions Allen, E. & E. Nelson, 1910. On the artificial culture of
for hatching will show higher population growth marine plankton organisms. J. mar. bioI. Ass. UK. 8:
rates and less mixis. If resting eggs were not 421-474.
 421

Clement, P., R. Pourriot & c. Rougier, 1976. Les facteurs rotifer Brachionus plicatilis. Nippon Suisan Gakkaishi. 51:
exogenes et endogenes qui controlent i'apparition des 511-514.
males chez les Rotiferes. Bull. Soc. zool. Fr. 101: 86-95. Hino, A. & R. Hirano, 1988. Relationship between water
Fu, Y., Y. Natsukari & K. Hirayama, 1989. A preliminary chlorinity and bisexual reproduction rate in the rotifer
study on genetics of two types of the rotifer Brachionus Brachionus plicatilis. Nippon Suisan Gakkaishi. 54:
plicatilis. NOAA Technical Report NMFS, in press. 1329-1332.
Gilbert, 1. J., 1974. Dormancy in Rotifers. Trans. am. Ito, T., 1960. On the culture of the mixohaline rotifer
microsc. Soc. 93: 490-513. Brachionus plicatilis o. F. Muller in the sea water. Rep. Fac.
Gilbert,1. 1.,1977. Mictic female production in monogonont Fish. Prefect. Univ. Mie 3: 708-740.
rotifers. Arch. Hydrobiol. Beih. 8: 142-155. Lubzens, E., 1981. Rotifer resting eggs and their application
Gilbert, J. 1., 1980. Some effects of diet on the biology of the to marine aquaculture. European Maric. Soc., Spec. Publ.
Rotifer Asplanchna and Brachionus. In: Nutrition in the 6: 163-179.
lower Metazoa. Pergamon Press, Oxford: 57-71. Lubzens, E., G. Minkoff & S. Marom, 1985. Salinity depend-
Hagiwara, A., A. Hino & R. Hirano, 1985. Combined effects ence of sexual and asexual reproduction in the rotifer
of environmental conditions on the hatching of fertilized Brachionus plicatilis. Mar. BioI. 85: 123-126.
eggs of the rotifer Brachionus plicatilis collected from an Minkoff, G., E. Lubzens & D. Kahan, 1983. Environmental
outdoor pond. Nippon Suisan Gakkaishi. 51: 755-758. factors affecting hatching of rotifer (Brachionus plicatilis)
Hagiwara, A., A. Hino & R. Hirano, 1988. Comparison of resting eggs. Hydrobiologia, 104: 61-69.
resting egg formation among five Japanese stocks of the Pourriot, R. & R. Rougier, 1976. Influence de i'age des
rotifer Brachionus plicatilis. Nippon Suisan Gakkaishi. 54: parents sur la production de femelles mictiques chez
577-580. Brachionus calyciflorus (Pallas) et B. rubens Ehr. (Roti-
Hino,A. & R. Hirano, 1976. Ecological studies on the mecha- feres). C.R. Acad. Sc. Paris 283: 1497-1500.
nism of bisexual reproduction in the rotifer Brachionus Pourriot, R. & T. Snell, 1983. Resting eggs in rotifers. Hydro-
plicatilis-1. General aspects of bisexual reproduction. biologia, 104: 213-224.
Nippon Suisan Gakkaishi. 43: 1147-1155. Rougier, C. & R. Pourriot, 1977. Aging and control of the
Hino, A. & R. Hirano, 1977. Ecological studies on the mecha- reproduction in Brachionus calyciflorus (Pallas) (Rotatoria).
nism of bisexual reproduction in the rotifer Brachionus Exp. Gerontol. 12: 137-151.
plicatilis-II. Effects of cumulative parthenogenetic gen- Snell, T., 1986. Effect of temperature, salinity and food level
eration. Nippon Suisan Gakkaishi. 43: 1147-1155. on sexual and asexual reproduction in Brachionus plicatilis
Hino, A., 1983. Life cycle - especially on the bisexual repro- (Rotifers). Mar. BioI., 92: 157-162.
duction inducing factors. In: The rotifer Brachionus pli- Snell, T. & F. Hoff, 1985. The effect of environmental factors
catilis - Biology and mass culture, pp 22-34. Ed. by on resting egg production in the rotifer Brachionus plicatilis.
Nippon Suisan Gakkai. Tokyo: Koseisha Koseikaku. (In J. World Maricul. Soc. 16: 484-497.
Japanese). Snell, T. & c. King, 1977. Lifespan and fecundity patterns in
Hino, A. & R. Hirano, 1985. Relationship between the tem- rotifers: the cost of reproduction. Evolution 31: 882-890.
perature given at the time of fertilized egg formation and Snell, T. & B. Winkler, 1984. Isozyme analysis of rotifer
bisexual reproduction pattern in the deriving strain of the proteins. Biochem. System. Ecol. 12: 199-202.
Hydrobiologia 186/187: 423-430, 1989.
C. Ricci, T. W. Snell and C. E. King (eds), Rotifer Symposium V. 423
© 1989 Kluwer Academic Publishers.

Intensive rotifer cultures using chemostats *

Charles M. James & T. Abu Rezeq

Kuwait Institute/or Scientific Research, Mariculture and Fisheries Department, P.O. Box: 1638,22017
Salmiya, Kuwait

Key words: Brachionus plicatilis, production, Chlorella, Nannochloropsis, fatty acids

Abstract

Continuous production of the rotifer Brachionus plicatilis rotundiformis (S-type) in an intensive chemostat
culture system has been investigated. The production dynamics of rotifers in relation to different flow rates
and feed regimes show that the growth rate and production depends on the type of algal feed and flow
rate utilized in the culture system. It was possible to achieve a mean production of up to 318.84 X 106
rotifers m - 3 d - 1 at a flow rate of 6 I h - 1 in 100 I chemostats and up to 261.21 x 106 rotifers m - 3 d - 1
at a flow rate of 40 I h - 1 while using 1 m 3 capacity rotifer chemostats as production units. The w 3 fatty
acid composition of rotifers while using Chlorella and Nannochloropsis in the culture system has been
described. The results of this investigation show that the rotifer productivity in the continuous culture
system is considerably higher than in any of the conventional culture systems described to date for
aquacultural purposes.

Introduction The usual technique for producing microalgae

such as Chlorella and rotifers consists of a
One of the problems in aquaculture is the ade- multistep backup batch culture system involving
quate production of nutritionally enriched live several tanks that occupy a great deal of hatchery
microorganisms such as rotifers at the lowest pos- space and is labour intensive (Lubzens, 1987).
sible cost for feeding the larval stages of fin fishes Synchronized continuous culture systems have
and crustaceans. The success of a marine fish been given importance in recent years for the cul-
hatchery, especially during early larval rearing ture of planktonic rotifers, and many investigators
stages, exclusively depends on the high quality live have discussed the production dynamics of roti-
microorganisms such as micro algae (eg. Chlorella) fers under small scale experimental conditions
and rotifers (Scott & Middleton, 1979; Hirata, (Droop, 1976; Droop & Scott, 1982; Boraas,
1980; Gatesoupe & Luquet, 1981; Watanabe 1983; Trotta, 1980; Walz, 1983; Rothhaupt,
eta!., 1983; James eta!., 1983, 1987; James & 1985). Taub (1980) stated that continuous cul-
Rezeq, 1989). In recent years, many attempts tures would not only provide constant yield, but
have been made to improve rotifer culture condi- also control the nutritional quality of the cultured
tions and the nutritional quality of the rotifers organisms. However, most of the work done to
produced for aquaculture. date on continuous culture systems has been

*This research was financed by the Kuwait Foundation for the Advancement of Sciences (KFAS), Kuwait, under a contract
research project code 86-04-02.
424

largely restricted to small-scale experiments to pacity and two 1 m3 capacity chemostats pro-
study the population dynamics of various species vided each with 50 I and 500 I capacity mixing
of micro algae and rotifers. The present investi- reactors. The desired food level in the rotifer
gation was aimed at developing an automated, chemostats were synchronized by metering
intensive continuous culture system using the pumps from the mixing reactors. The washout
chemostat principle for producing rotifers for from the rotifer chemostats was collected using
aquaculture. The study examines the production 100 I and 1 m 3 capacity rotifer concentration
dynamics and fatty acid composition of rotifers tanks to facilitate daily harvest. The rotifer
under different feed regimes and culture condi- chemostats were kept in a temperature controlled
tions. room provided with aeration and temperature
controllers to maintain the temperature at 25 0 c.
The experiments were carried out in two parts.
Materials and methods The first part of the experimental program investi-
gated the effect of different flow rates and cell
The investigations were carried out using the densities of two species of micro algae (Chlorella
S-type rotifer Brachionus plicatilis rotundiformis strain MFD-l and Nannochloropsis strain
(size 95-160 Jlm) in a two-stage intensive con- MFD-2) isolated from local sea water for pro-
tinuous culture system (Fig. 1). The stage-l con- ducing S-type rotifers using 100 I capacity
tinuous algal culture (using filtered and diluted sea chemostats. The algal flow rates (from the mixing
water enriched with nutrients at 30%0 salinity) has reactor) at desired algal cell densities (10 and
been described by James et al. (l988). The stage-2 20 x 106 cells ml- 1) were maintained at 2, 4, 6,
rotifer culture system consisted of three 100 I ca- 8 and 10 I h - 1 to determine the optimum food

Stage 1: Algal Chemostat Stage2: Rotifer Chemostat

AI r ---------.-----------
•
I
!.------j

pump ~ !
pH control 0 0' I
0::: _----:
0:::

---
I
0::: ) I
Controls

~ O)-----!
0::: :

0:::
~ 8)-----: I

o---j
Light

~
0::
0::
I
:

~ 0)-----[---------------------------,----
Fig. 1. Schematic diagram of the two stage rotifer chemostat system. R-Algal reservoir; TR-Temperature monitor and regulator.
 425

level requirements. Baker's yeast was used four cells ml- 1, the Nannochloropsis fed rotifers give
times a day, as a feed supplement which varied significantly higher (P < 0.001) rotifer production
from 38 to 144 mg 10 - 6 rotifers d - 1. The dilution at a flow rate of 6 1 h - 1 compared to rotifers fed
rates were synchronized with the population with Chlorella at 10 X 106 cells ml- 1. Further-
growth rate using a three channelled metering more, the rotifer productivity significantly in-
pump and a timer. During the observation period, creased (P < 0.01) by feeding Nannochloropsis at
Chlorella strain MFD-1 and Nannochloropsis 20 x 106 cells ml- 1 as compared to rotifers fed
strain MFD-2 from the algal chemostats were lOx 106 cells ml- 1. The results of these experi-
diluted with 30%0 sea water in the mixing reactor ments show that rotifer productivity increase with
to make up the desired cell densities of 10 and increasing cell density and flow rates of up to 6 1
20 x 106 cells ml- 1 for different treatments. h - 1 used in the culture system. The dilution rate
Amictic rotifers acclimated in each algal species varied up to 0.6 d -1 (60% of culture volume per
were used to seed the cultures to obtain an initial day). Rotifer production declined significantly
density of about 50 rotifers ml- 1. The population (P < 0.001) at flow rates above 61 h -1 (Nan-
counts, pH and DO were monitored every 24 h. nochloropsis at 10 and 20 x 106 cells ml- 1). Roti-
The production dynamics of rotifers were calcu- fer population growth rate and doubling time were
lated according to the equation discussed by similarly reduced. Rotifer production at flow rates
James et al. (1986). of 4 and 6 1 h - 1, was not significantly different
Based on the results obtained in 1001 capacity (P> 0.05).
rotifer chemostats, the second part of the experi-
mental program investigated the use of Chlorella
strain MFD-1 and Nannochloropsis strain MFD-2 Production dynamics of rotifers in 1 m3 chemostats
at 10 and 20 x 106 cells ml- 1 using 1 m3 capacity
rotifer chemostats. The algal flow rate was Table 2 shows the production dynamics of roti-
maintained at 40 1 h - 1. The dilution rates were fers in 1 m3 chemostats using Chlorella and Nan-
adjusted once in 24 h according to the growth rate nochloropsis at 10 and 20 x 106 cells ml- 1. The
of rotifers in the culture system. Dry weights of flow rate was constant at 40 1h - 1. The results are
rotifers were calculated from freeze-dried and similar to those of the 1001 rotifer chemostats.
oven dried samples and averaged 0.22 jlg per Rotifer production significantly increased
rotifer. Feeding efficiency equaled the total (P < 0.001) using Chlorella at 20 x 106 cells ml- 1
biomass offood provided d - 1 divided by biomass as compared to lOx 106 cells ml- 1. Further-
(same units) of rotifers produced d - 1. The fatty more, rotifer productivity significantly increased
acid composition in rotifers were analyzed as de- (P < 0.001) using Nannochloropsis at 10 x 106
scribed by James etal. (1987, 1988). cells ml- 1 as compared to Chlorella at lOx 106
cells ml- 1. However, this difference disappeared
at 20 x 106 cells ml- 1.
Results

Production dynamics of rotifers in 100 I chemostats Fatty acid composition of rotifers

Table 1 shows the production dynamics of roti- Studies carried out using different cell densities
fers in relation to different flow rates and algal and flow rates of Nannochloropsis in 100 1 rotifer
feeds in 100 1 capacity chemostats. While using chemostats show that the fatty acid content in
different flow rates from 2 to 10 1h - 1, a flow rate rotifers depends on algal flow rate and cell
of 6 1 h - 1 appears to be most conducive for pro- density. Rotifer w 3 PUFA content significantly
ducing rotifers in the culture system. Also while increases (P < 0.01) using Nannochloropsis at
using Chlorella and Nannochloropsis at 10 X 106 20 x 106 cells ml - I as compared to 10 X 106 cells
 .j::..
tv
0\

Table 1. Production dynamics of rotifers in relation to different flow rates and algal feeds in 100 I capacity chemostats
(B.Y. = Bakers' yeast, n = 14)

Algal strain Flow rate Feed amount Growth rate Doubling time Egg ratio Birth rate Production Feed
(x10 6 cells ml- 1) (lh - 1) (gd- 1 drywt.) (ml d - 1) (d) (E) (ml d - 1) (x 106 m - 3 d - 1) efficiency

Algae B.Y.
Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D.

MFD-1, at 10 x 106 2 3.7 ± 0.9 14.5 ± 5.8 1.0 ± 0.4 0.8 ± 0.3 0.4 ± 0.2 0.7 ± 0.3 87.4 ± 44.4 12.2 ± 4.0
4 3.7 ± 0.9 15.6 ± 5.7 1.3 ± 0.3 0.6 ± 0.2 0.5 ± 0.1 0.8 ± 0.1 108.8 ± 41.3 9.4 ± 2.6
6 3.6 ± 0.9 16.2 ± 5.3 1.3 ± 0.3 0.6 ± 0.2 0.4 ± 0.1 0.7 ± 0.1 114.5 ± 47.2 9.1 ± 2.6
8 3.3 ± 0.9 18.9 ± 5.6 1.3 ± 0.6 0.9 ± 0.9 0.4 ± 0.1 0.6 ± 0.2 94.0 ± 35.4 13.8 ± 6.7
10 3.2 ± 0.9 18.6 ± 5.4 1.3 ± 0.4 0.7 ± 0.5 0.4 ± 0.1 0.7 ± 0.1 75.3 ± 28.6 17.0 ± 9.1

MFD-2, at 10 X 106 2 3.4 ± 1.6 20.1 ± 7.5 1.0 ± 0.5 0.4 ± 1.2 0.4 ± 0.1 0.7 ± 0.2 148.3 ± 61.9 9.3 ± 5.0
4 4.2 ± 1.5 22.6 ± 7.0 1.6 ± 0.4 0.5 ± 0.4 0.4 ± 0.1 0.6 ± 0.3 231.0 ± 20.9 5.7 ± 0.5
6 4.2 ± 1.5 21.6 ± 6.5 1.8 ± 0.4 0.5 ± 0.4 0.4 ± 0.1 0.7 ± 0.2 239.0 ± 36.5 5.4 ± 0.6
8 4.2 ± 1.7 11.5 ± 5.2 0.3 ± 0.6 -1.9±8.1 0.3 ± 0.1 0.5 ± 0.2 189.6 ± 118.6 6.0 ± 4.4
10 3.7 ± 1.5 12.5 ± 5.0 0.6 ± 0.3 2.5 ± 3.7 0.3 ± 0.1 0.5 ± 0.2 140.5 ± 59.8 6.4 ± 2.4

MFD-2, at 20 X 106 2 6.5 ± 2.0 13.9 ± 6.0 1.1 ± 0.9 0.8 ± 5.5 0.4 ± 0.1 0.7 ± 0.1 224.5 ± 74.2 4.7 ± 0.9
4 9.0 ± 3.0 11.2 ± 4.3 0.6 ± 1.0 1.3 ± 0.4 0.3 ± 0.1 0.6 ± 1.0 259.7 ± 67.8 3.1 ± 1.0
6 8.2 ± 2.7 12.3 ± 4.8 0.8 ± 0.2 1.0 ± 0.4 0.4 ± 0.2 0.6 ± 0.3 318.8 ± 80.4 3.2 ± 0.7
8 8.1 ± 2.8 11.0 ± 4.8 0.6 ± 0.1 1.2 ± 0.3 0.4 ± 0.2 0.6 ± 0.3 225.6 ± 67.3 4.4 ± 1.4
10 8.1 ± 2.8 10.7 ± 4.4 0.6 ± 0.2 0.6 ± 1.8 0.4 ± 0.2 0.7 ± 0.3 182.4 ± 35.2 5.0 ± 1.0
Table 2. Production dynamics of rotifers in relation to different algal feeds in 1 m 3 capacity chemostats (B.Y. = Bakers' yeast,
n = 14)

Algal strain Flow rate Feed amount Growth rate Doubling time Egg ratio Birth rate Production Feed
(x10 6 cells ml- I) (lh- 1 ) (gd- 1 drywt.) (ml d - I) (d) (E) (ml d - I) (x 106 m- 3 d- 1 ) efficiency

Algae B.Y.
Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D. Mean ± S.D.

MFD-l, at 10 X 106 40 31.9 ± 10.0 140.3 ± 68.4 1.1 ± 0.3 0.7 ± 0.2 0.4 ± 0.1 0.7 ± 0.1 128.4 ± 38.6 7.7 ± 2.1
MFD-l, at 20 x 106 40 67.3 ± 18.1 206.3 ± 77.7 1.4 ± 0.2 0.5±0.1 0.4 ± 0.0 0.7 ± 0.1 246.9 ± 77.3 6.1 ± 1.6
MFD-2, at 10 X 106 40 43.5 ± 14.9 177.5 ± 54.5 0.6 ± 0.2 1.2 ± 0.3 0.4 ± 0.1 0.7 ± 0.1 255.9 ± 59.5 4.5 ± 1.0
MFD-2, at 20 x 106 40 85.5 ± 27.3 154.3 ± 75.5 1.0 ± 0.1 0.9 ± 0.4 0.4 ± 0.1 0.7 ± 0.1 261.2 ± 23.5 4.9 ± 1.2

~
N
-..J
428

ml- 1. A highly significant increase (P < 0.001) in Discussion

EFA 20 : 5 w 3 was also observed on this feeding
regime and at a flow rate of 6 1 h - 1. The rotifer production achieved during this inves-
Rotifers produced in 1 m 3 rotifer chemostats tigation (318 x 106 m - 3 d - 1 in 1001 units and
show that the fatty acid composition depends on 261 x 106 m - 3 d - 1 in 1 m 3 tanks) is considerably
the cell density and the type of micro algae used in higher than any of the conventional rotifer pro-
the culture system (Fig. 2). A cell density of duction systems reported to date for aquacultural
20 x 106 cells ml- 1 of Chlorella yielded signifi- purposes. Lubzens (1987), while reviewing the
cantly higher (P < O.OI)rotifer w3 PUFA content conventional rotifer production systems, stated
than a density of 10 x 106 cells ml- 1. However, that low yields were mostly associated with large
the total rotifer w 3 PUF A content on Nan- culture tanks (10-20 m 3 ). The highest rotifer pro-
nochloropsis at 10 x 106 cells ml- 1 was signifi- duction achieved was in 60 1containers and only
cantly higher (P < 0.01) than Chlorella at 20 x 106 122 rotifers ml- 1 d - 1 were recorded (Gatesoupe
cells ml- 1. Total w 3 PUFA content of rotifers & Luquet, 1981). This is considerably lower than
fed Nannochloropsis at 20 x 106 cells ml- 1 our mean rotifer production of 318 rotifers ml- 1
(18.9 ± 3.62%) was significantly higher d - 1. Furthermore, no significant difference
(P < 0.01) than a feeding level of 10 x 106 cells (P < 0.05) in rotifer production was observed
ml- I . using 100 1capacity and 1000 1(1 m 3 ) chemostats.

c::::
Q)

...
u
Q)
D..

Fatty Acids

Fig. 2. Fatty acid composition in rotifers in relation to different cell densities of Chlorella strain MFD-l and Nannochloropsis strain
MFD-2.
 429

This illustrates that 1 m3 capacity rotifer as compared to the 100 I capacity chemostats
chemostats could be adapted as suitable units for where maximum rotifer w 3 PUF A content was
the large scale production of rotifers in aquacul- observed at a flow rate of 6 I h - 1 ( = 60 I h - 1 in
ture. 1 m 3 tank).
Boraas (1983) cultured B. calyciflorus in a James et al. (1989) while investigating the
two-stage chemostat system and obtained a dry growth and w 3 fatty acid composition of marine
weight biomass of 0.0202 g 1- 1 d - 1 at a dilution algal species under different temperature regimes
rate of 0.83 d - 1 and 0.0252 g 1- 1 d - 1 at a dilution concluded that fatty acid composition, especially
rate of 1.08 d - 1. In our investigation, a dilution the EPA (eicosapentaenoic acid, 20: 5 w 3)
rate of 0.6 d - 1 yielded a dry-weight rotifer profile, is species dependent and varies according
biomass of 0.0699g 1-1 d- 1 (318 X 106 m- 3 to different temperatures prevailing in the culture
d - 1) in 100 I chemostats and up to 0.0574 g 1- 1 system. In our investigation the temperature was
d- 1 (261 X 106 m- 3 d- 1 ) in 1m3 capacity controlled at 25 0 C for all the treatments. Hence,
chemostats. This is higher rotifer production than variations in EF A content observed in rotifers
any previously described culture systems (Hirata, while using the two species of micro algae appears
1979; Groeneweg & Schluter, 1981; James et a!., to be due to the type of algal feed utilized (James
1987). & Rezeq, 1988, James et al., 1989) in the chemo-
It is generally known that different algal foods stat culture system. Moreover, adequate quan-
can yield substantially different reproductive rates tities of EF A are present in rotifers fed with Nan-
(Hirayama etal. 1979; Yufera eta!' 1983; Snell nochloropsis at 20 x 106 cells ml- 1, so that further
et al. 1983; James & Rezeq, 1988). Furthermore, nutritional enrichment is not required for feeding
rotifer growth and productivity also depends on marine fish larvae (Watanabe et al., 1983).
the cell densities of Chlorella utilized in the culture
system (Hirayama et al., 1973; Halbach &
Halbach-Keup, 1974; Pilarska, 1977a, b; Endo & Acknowledgement
Mochizuki, 1979; Yamasaki & Hirata, 1985;
Rezeq & James, 1987) as does nutritional quality This research was financially supported by
(J ames et a!., 1986; 1987; 1988). The results of Kuwait Foundation for the Advancement of
our investigation are in accordance with the above Sciences, Kuwait, under Project Code 86-04-02.
observations showing increased rotifer produc- Our sincere thanks are due to Dr. Ziad Shehadeh,
tivity with higher cell densities of micro algae. Also Manager, Aquaculture, Kuwait Institute for
the studies show that increased rotifer productivi- Scientific Research who provided invaluable
ty could be achieved while using Nannochloropsis insight and guidance through all phases of this
strain MFD-2 as compared to Chlorella strain study.
MFD-l. This is in accordance with the observa-
tions of James & Rezeq (1988) who used a differ-
ent species of Chlorella. References
The presence of w 3 PUF A in rotifers, which
Boraas, M. E., 1983. Population dynamics of food limited
are indispensable for rearing marine fish larvae
rotifers in two-stage chemostat culture. Limno!. Oceanogr.
(Watanabe et aI., 1983), shows that the level of 28: 546-563.
these essential fatty acids in rotifers depends on Droop, M. R., 1976. The chemostat in mariculture. In: A.
the cell density and species of micro algae used in Persoone & E. Jaspers (ed.). Proceedings of the 10th
the culture system. Also, the flow rate of algal feed European Symposium on Marine Biology. Belgium
University Press, Ostend, pp. 71-93.
in the culture system has an impact on the availa-
Droop, M. R. & J. M. Scott, 1982. A steady state approach
bility of EF A in rotifers. The slight reduction in to some micro-plankton problems. Annales de l'Institut
the w 3 PUF A content observed in the 1 m3 Oceanographique 58: 47-54.
chemostat may be due to the flow rate (40 I h - 1 ) Endo, E. & T. Mochizuki, 1979. Relation between specific
430

growth rate of rotifer, Brachionus plicatilis, and concen- James, C. M., A. M. AI-Khars & P. Chorbani, 1988. pH
tration of food, marine species of Chlorella. J. Fermen- dependent growth of Chlorella in a continuous culture
tation Techno!., Osaka 57: 372-374. system. Journal of the World Aquaculture Society 19:
Gatesoupe, F. J. & P. Luquet, 1981. Practical diet for mass 27-35.
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Hydrobiologia 186/187: 431,1989.
C. Ricci, T.W. Snell and CEo King (eds), Rotifer Symposium V. 431

Author Index
Abu-Rezeq T. 423 Hirayama K. 39 Pascual E. 371
Amsellem J. 255 Hollowday E.D. 319 Pauli H.R 355
Berzins B. 137 Hussey C. 319 Pejler B. 137
Bogaert G. 167 JamesC.423 Pontin RM. 229
Bolzern A.M. 145 Jensen A. 409 Popiolek B. 235
Brett M.T. 181 King C.E. 375 Radwan S. 235
Carmona MJ. 325 Korstad J. 43,51 Ricci C. 145
Carrillo P. 103,215 Koste W. 129, 191,239,279 Rodgriguez Rainuzzo J. 409
Chengalath R 191 Kutikova L.A. 285 Rooth J. 29, 347
Clement P. 255 Litt A.H. 109 Ruttner-Kolisko A. 291
Colburn RA. 311 Lubzens E. 387 Saunders-Davies A.P. 153
Cruz-Pizarro L. 103,215 Maeda T. 39 Serra M. 81,325,331,381
Dumont HJ. 167 Markevich G.I. 285 Serrano L. (Valencia) 381
Duncan A. 11 Maruyama I. 39 Serrano Martin L. (Sevilla) 339
Edmondson W.T. 1, 109 Matsumoto T. 247 Shiel RJ. 239, 279
Ejsmont-Karabin J. 29,347 Matveeva L.K. 69 Siewertsen K. 347
Esparcia A. 331 MayL. 59 Snell T.W. 299
Farabegoli A. 201 Mazzoni R 201 Starkweather P.L. 35
Ferrari I. 201 Mikschi E. 209 Sudzuki M. 247
Galkovskaya G .A. 119 Minkoff G. 387 TanL.W. 239
Gilbert J J. 75 Miracle M.R 81,325,331,381 Tandler A. 387
Gschloessl T. 363 Mityanina I.F. 119 Vadstein O. 43,51,409
Guisande C. 339 Morales-Baquero R 103,215 Wallace RL. 311
Gulati RD. 29,347 Nagata W.D. 401 Walsh EJ. 35, 157
Hagiwara A. 415 Narita K. 247 Wa1zN. 363
Hartmann U. 363 Nogrady T. 223 YUfera M. 371
Herzig A. 129 Olsen Y. 43,51,409 Zoufal W. 163
Hino A. 415 Pagani M. 145
Hydrobiologia 186/187: 433, 1989.
C. Ricci, T.W. Snell and CEo King (eds), Rotifer Symposiwn V. 433

SUbject Index
aging 325 ingestion rate 51
algae 29,39,43,51 interference 59, 69, 75
arctica 109, 191
light 103, 153,415
behavior 157,255 life cycle 11,43,81
biogeography 109, 167, 191,239,279,319 life table 39, 43, 81
biometry 355, 381
body size 11,347,355 mating 299
metabolic rate 11,363,371
Cladocera 11,69,75 migration 167
cladogram 311 diurnal vertical 103, 119
clearance rate 51 muscles 255
communities 181, 191,201,209,215,229,235,247
competition 69, 75 nitrogen 347, 371,401
conductivity 215
Crustacea 11,69,75,167, 181 oxygen 209, 215,331
cyanobacteria 35
parasites 59
development time 11, 119, 163 pH 181
DNA 375 phylogeny 255, 285
polymorphism 109,279,291
electrophoresis 145, 325, 375 predation 35, 69
epizoic 59, 69
evolution 311 resting eggs 291, 381,415
excretion 347
salinity 81, 129,381
fatty acids 409, 423 sexuality 299
fecundity 11,39,43,81 systematics 279, 291, 299, 311, 319
food 11,29,35,39,43,51,363,387 substrate selection 137
(rotifers as .. ) 387,409
temperature 81, 129, 163,209,363,381,415
genetics 145,291,375,381
ultrastructure 255, 285
hatching 415
zooplankton 75, 109, 181,201,235