Aquatic Invertebrates as Indicators

Of Stream Pollution
By ARDEN R. GAUFIN, Ph. D., and CLARENCE M. TARZWELL, Ph. D.
Year-round field studies of the biology of various sections of the stream and diurnal
stream sanitation were initiated by the biology changes in environmental conditions are dis-
section of the Public Health Service's Environ- cussed here.
mental Health Center on Lytle Creek in Octo- Lytle Creek, which was especially selected
ber 1949. The aims of these investigations for the study, is about 45 miles northeast of
were: Cincinnati, Ohio, and is a tributary of Todds
1. To develop or devise and field test pro- Fork, a part of the Little Miami River system.
cedures and equipment for biological surveys It is a small stream approximately 11 miles
and investigations of polluted streams. long, 3 to 35 feet wide during non-floodstage,
2. To investigate seasonal and diurnal en- and a few inches up to 6 feet deep. The prin-
vironmental changes in a stream polluted with cipal natural source of water in the stream is
oxygen-depleting wastes. surface drainage from the surrounding area.
3. To determine how the physical-chemical Some 7 miles above its mouth, the creek receives
environment in the various pollutional zones the effluent from the primary sewage treatment
affects the qualitative and quantitative com- plant of Wilmington, Ohio, a city of about
position of aquatic populations and how these 10,000 people. Lytle Creek is particularly fa-
populations in turn affect or change physical- vorable for studies of the pollutional effects of
chemical conditions. oxygen-depleting wastes because it has only
4. To relate various qualitative and quantita- one source and type of pollution (domestic
tive compositions of aquatic populations to en- wastes from Wilmington) ; it has no permanent
vironmental conditions found in streams pol- tributaries below the source of pollution; and
luted with organic wastes and to test the value it has all degrees of pollution from a definite
and use of aquatic organisms as indicators of septic zone through recovery and back to clean
clean water and various degrees of pollution at water.'
all seasons of the year.
5. To determine the rate of satisfaction of
biochemical oxygen demand (BOD), the area Procedures
in which most of the demand is satisfied, and One of the early objectives of the Lytle Creek
how seasonal and other environmental condi- investigations was to evaluate the reliability
tions affect the process. and possible use of aquatic organisms (chiefly
6. To determine the value of fishes as indica- benthic forms) as indicators of the extent and
tors of pollution and their use in evaluating the severity of pollution and the degree of stream
effects of pollution. recovery. Plans were made for a comprehen-
The composition of the aquatic fauna in
'Zones of pollution, as herein considered for Lytle
Creek, are based principally on the amount of dissolved
Dr. Tarzwell is chief and Dr. Gaufin is biolo- oxygen occurring throughout the stream. The septic
ist of the biology section of the Public Health zone is the area of stream showing oxygen depletion
during at least some portion of the year. It grades
Service Environmental Health Center, Cincnn- gradually into the recovery zone where the minimum
nati, Ohio. oxygen concentrations range from 0.1 to 2 ppm. In the
clean-water zone minimal oxygen concentrations may
be as low as 2 ppm for short periods of time.
Vol. 67, No. 1, January 1952 57
 sive study. A detailed survey was made of the These data were then correlated with those ob-
stream and a map prepared showing the loca- tained by the quantitative and qualitative
tion and extent of pools, runs, and riffles, and studies of the benthic, marginal, and surface
the stream profile and gradient. A sharp
crested rectangular contracted weir and contin-
uous float level r ecorder were placed in the
stream about 3 miles above its mouth for the
continuous recording of flows.
During the winter and spring, flows average
about 6 to 7 cubic feet per second (cfs) with S
frequent floods of 40 cfs or more. In late 2.
"9.0
summer and fall, flows are generally small, 0
0~8.0
about 0.1 cfs above the sewage plant outfall and Io 7.0
1 cfs below it. During periods of low summer ~0
flow, the sewagre plant effluent overwhelms the 0~6.0 Minimui
stream with oxygen-depleting wastes and causes .15.0
a septic zone to be produced immediately below 4.0
tlle outfall. 3.0
Ten stations were selected along the stream 2.0
course for periodic sampling in all zones. These 1.0
stations were desiginated by their distance in 0.0 --- -T -T-
miles upstream fiom the mouth, as stations: Miles 8.7 7.6 7.2 6.5 5.2 4.2 2.8 1.0 0.O
0, 1, 2.8, 4.2, 5.2, 6.5, 7.2, 7.3 (sewage outfall),
Actual distances between stations
7.6, and 8.7. Monthly, or more frequently, sam-
ples were taken at these stations for the deter- Range in dissolved oxygen, Lytle Creek, August 22-23, 1951.
mination of dissolved oxygen, pH, CO2, methyl
orange, and phenolphthalein alkalinity, and fauna to provide information on (a) aquatic
temperatures. Quantitative bottom samples populations which can be expected to develop
were taken at monthly intervals at each station under certain ecologic conditions, and con-
in pools, runs, and riffles. A Surber square- versely (b) environmental conditions or varia-
foot samnpler was used in riffle areas, while a tions in environmental conditions which are in-
Petersen or Ekman dredge was used in other dicated by various compositions and densities of
areas. Marginal samples were also taken, and aquatic organisms.
reconnaissance surveys of bottom and surface
fauna were made along the entire stream.
(BOD studies have been made periodically at Results
all stations, as well as plankton, microbottom The chemical and physical data collected dur-
fauna, and fish population studies.) ing 6 sampling runs for the 10 stations are
In the Lytle Creek investigations emphasis enumerated in table 1. These data present the
was placed on year-round studies to determine maximum and minimum oxygen concentrations
seasonal variations in aquatic populations and and water temperatures occurring in the stream
ecologic conditions. Attention was also di-
rected to diurnal variations in physical and during the early spring and summer months.
chemical conditions, especially duringr the Since a wide difference in the amounts of
spring and summer when they are more pro- oxygen present in the stream during day and
nounced. Physical-chemical variations were night was noted, special attention was paid to
determined by taking hourly samples at each of lhourly variations in the dissolved oxygen con-
the stations for a 24-hour period. By these and tent. The range of these variations for the
other studies, information was obtained con- period 9: 00 a. m. August 22 to 8:00 a. m. Au-
cerning ecologic conditions and their diurnal gust 23 is presented in the chart. During the
and seasonal variations in each of the zones. reconnaissance surveys, conducted in connec-

s8 Public Health Reports

 tloll vitl the samiipliing rulns, 68 genera and 79 sp., were abunidant. Of the 23 species of may-
species of animals were collected and identi- flies, stoneflies, and caddisflies taken, only one
fied. Of these species, 29 percent were Diptera, species, Callibaetis sp., occurred outside of the
or fly larvae, a group which is adapted to live clean-water zone. The organisms collected at
un(der a wide variety of environmental condi- five stations representative of each of the zones
tions. Fifteen species of organisms were taken are listed in table 2. Their comparative abun-
in the septic zone, but only six of them, Chi- dance by month and zone is also indicated.
ronomfUs tentans, Cule pipiens, Eristalis sp., Among the primary requisites for animal
Physa integra, Li?nnodrilus sp., and Tubifex existence are oxygen and food. When estab-

Table 1. Summary of physical and chemical conditions in Lytle Creek, 1951

Dates of hourly sampling over 24-hour period
Station Range
4/24/51 5/1/51 5/9, 10/51 6/28,29/51 8/15/51 8/22, 23/51

- | S]
D. 0 ---- 15 ppm--- 13.9
pppm.
-
8.7 | aximinum1Tenp - 63-. - - - |82.6|||450pp.
-M n m m
Te p 450- --
--- -- 590.
7D. 0 6----- ppm 8. - - - - - 4.5 ppm.
Miaximum{TemP-----|
D.O
62
14.7 ppm = --- =- m-- - 770I
8.8 ppm.
7.65 Mininimum{Teinp-
D. 0 ------
470
7.2 ppm-- -- - - -
---
0
81°-------- ppm----0--61p.
ppm.

5Maximum{TemP 590 -
710.pm---------------------- 710.
7.2Al D. O 12.1 ppm | 0.8 m | O---
----|---7]° 0.2pp,.
7.2ir m

ND.
em
----6.0 ppm
480 -
-- 680.
-------------- -- -----------0.0 ppm.
6.5MaY ~fTemp- - -60g.. - 700.... 810.... 790 --- 790.
4 2 M0taximum D 0 ------11.2 ppm. 14.
-------| p
1 pm 1. ppm------| -40.2ppm | 0.8 ppm.
_ 5 -lin.i.5um T -5-ppm --
-- --- 65°--- 710 ------- 64.50 - 610. -----

ND. O 4.7 ppm --- 0.6 ppm... 0.0 ppm... 0.0 ppm.... 0.0 ppm.
Mfaximrum{Temp- 60.50 ----760-----700 -----80.50---
D. 0 ----12.4 19.1 19.4 11.6
800 -----750.
7.3 ppm..-.-.. 6.9 ppm.
5.2 ppm- ppm- ppm..- ppm-
Ali nimurlm JTemp...... 540 ----- 650 -----590------770 710----- 610.
ND. O 5.4 ppm... 0.8 ppm---- 2.9 ppm.... 0.2 ppm-.. 0.0 ppm... 0.2 ppm.
4.2 ai km em 660 -- 760 770 -- 71.50.
4.2 ND.e 0 16.4 ppm..- 18.1 ppm--------------- 5.4 ppm....- 5.5 ppm.
M iir[ M LD. Op 540. -- 650. - --- 720 -------- 550.
---- 6.5 ppm... 1.0 ppm.----..---- --------1.4 ppm- 2.0 ppm.
Temp -----
Maximum1ID.O. - 690 - - - 790 870 870 . 800.
2.8 13.8 ppm.... 14.3 ppm...... 10.1 ppm... 12.3 ppm.... 14.7 ppm.
Minimum Temp....-. 8.1 540 650 720.pp 72-. 590.
ND. 0.--- -1-01 ppm. -.... I- 1.6 ppm..-.-. 3.6 ppm....- 1.6 ppm.... 3.2 ppm.
I.
Temp- - - - 70 .
Alaximum ND. - 700..... 730.
1 -0-.-. - 13.5 ppm 14.2 ppm.. 9.6 ppm.
Minimum' Tem. 5406.
p 580 ._ 65.50.
6.9 ppm......- 5.2 ppm...... 5.3 ppm.
II I.I,
1 Temp......
maximum ND. ------------ 750.
0 - ---- 11.6 ppm.
Temp.......
Minimum'ND. 63.50.
0.--- 5.0 ppm.
_
_1
II - I___ _I
Temp.-
Todds Maximlum ND. ---- ------------
700.
Fork 0.----- ------------
10.7 ppm.
Ternp......
Mfinimum'ND. 62.50.
0 ---- 5.1 ppm.

Vol. 67, No. 1, January 1952

59
 Table 2. Distribution of invertebrates in Lytle Creek, May and August 1951
Stations I
Organisms i 8.7 6.5 5.2 4.2 _ 2 -_
__ -_~~~~~~~~~~~~~
M
May , A ug. May Aug. May Aug. May Aug. May Aug.
.1
DIPTERA

Chironomus flavus 2p P p
Chironomus tentans - - --- p A P p
Chironomus flavicingula P P ------ p
---I-- P
Chironomus quadripunctatum P P--
Chironomus sp. A P
Chironomus sp. B P
Pentaneura flavifrons P - -

Dictya sp
Anopheles punctipennis P
Culex pipiens- p A A p A
Eristalis sp A A
Stratiomyia sp P P P
Nemotelus sp P -P
Tabanus sp -- P- -P p p P
P
Palpomyia sp P- -P -P
Stilobezzia sp- P P p
Brachydeutera argentata --I--- p
Simulium vittatum P P .

Hemerodromia sp- . PP [ P
Pilaria sp - - ---i-- P
Tipula sp - --
-ii- 1- P
Eriocera sp - -ii--P
Paradixa sp P -I- -
p --i---
p
COLEOPTERA
p p - -i ----
Stenelmis crenata_______ p
Stenelmis sp
Simsonia quadrinotata P
Bidessus sp. A
Laccophilus sp
Berosus sp-
Tropisternus lateralis- p
-I----p P
Tropisternus sp p p 7-- p p P
Laccobius sp- p p p
Peltodytes sp P P
Gyrinus sp -- -------
Dineutes sp-
EPHEMEROPTERA Pp
Baetis cingulatus- p P
Baetis parvus ---- -- p P
Caenis sp p ---I--
p p
Callibaetis sp-- p ------ p p P
Isonychia albomanicata-
Stenonema femoratum _ -_ p p pI
Stenonema ohioense _ p p
I
See footnotes at end of table.

lishlinlg biological indices of pollution, such stream. From the standpoint of dissolved
requisites must be taken into consideration. In oxygen, the stream was satisfactory for most
Lytle Creek from November 15,1950, to April 1, types of bottom organisms throughout its
1951, the volume of flow was more than 6 cfs length during the entire period. Chemically,
on all except three occasions. The time of flow the water appeared to be relatively free from
from the sewage outfall to the mouth at Todds harmful pollution, but the fauna that was pres-
Fork never exceeded 48 hours. Water tempera- ent still reflected the critically low oxygen
tures were under 600 F., and oxygen concentra- concentrations occurring in the past.
tions approached saturation throughout the From April 1 to October 1, the volume of flow
60 Public Health Reports
 Table 2. Distribution of invertebrates in Lytle Creek, May and August 1951-Continued
Stations I

Organisms

TRICHOPTERA

Brachycentrus americanus P
Cheumatopsyche sp p pP P
Hydropsyche betteni pP P
HIydropsyche bifida p P P
Chimarra obscura p p~~~~~~~~~~~P P
Dolophilus shawnee
.--

P
p
Phyacophila lobifera _ p _--

Ochrotrichia sp p _--

Hydroptila consimilis p p _--

Hydroptila sp
PLECOPTERA

Acroneuria evoluta p
Allocapnia viviparia P
Nemoura venosa P
Perlesta placida P _-
Neoperla clymene p
Isoperla minuta
ODONATA
Plathemis sp p p P
Paltothemis sp p
Argia sp --P-
P- p P
Agrion sp -- -I---
p
Enallagma sp p P
NEUROPTERA P
Corydalis cornutus-
Sialis sp- p P-P
HEMIPTERA
P
Belostoma sp p p p p p P
Corixidae p p p p P
Gerris sp p p p P
Microvelia sp ---------------- p P
Notonecta sp _ p
Ranatra sp .- p
CRUSTACEA
Asellus sp P p
Cambarus rusticus Pp p p
- -- -- -

p
Hyalella sp --------- p ---

MOLLUSCA p
Physa integra ------------
p p p p A p A p P
Sphaerium solidulum - - p
p A-
ANNELIDA
Limnodrilus sp- p A p A p A p p
Tubifex sp p A p A p A p p
(l108Tphonia spee p----------------- p p
L1- P
Total species per station_ -- 40 29 ", 9 10 10 10 19I 37
1 10
I
2
The stations are classified into zones of pollution as follows: clean water, 8.7 and 2.8; septic zone, 6.5 and 5.2; zone of recovery, 4.2.
A=abundant; P=present.

Vol.67, No. 1,January 1952 61

977220-52
 dropped to an average of 1 cfs, the time of flow floods, and abundant food, prevailing through.
extended to 7 or 8 days, water temperatures rose out the summer of 1951, a surprisingly large
to as high as 870 F., and oxygen concentrations and varied macrobottom fauna developed ill
became critically low in the 4-mile section of the stream. This condition was observed dur.
stream below the sewage treatment plant out- ing May and again in August when special col.
fall. Definite pollutional zones were estab- lections of aquatic organisms were made at all
lished in May and persisted throughout the sampling points along the stream, and their dis-
summer. An abundant food supply and a tribution correlated with the diurnal physical
variable oxygen supply below the treatment and chemical conditions prevailing at the vari-
plant outfall resulted in a distinctive fauna in ous stations.
each of these zones. In evaluating aquatic organisms as indicators
Critically low oxygen concentrations were of pollutional conditions, great caution must be
found in the stream during the night at station used because of several complicating ecologic
6.5 as early as May 9. Concomitant with the conditions. First, many organisms which occur
low nocturnal values, remarkably high oxygen in large numbers in extremely polluted water
concentrations were recorded during the after- may also be found in limited numbers in cleaner
noon at the same station. Peak concentrations situations. Several species of invertebrates,
of oxygen for the season occurred before the such as the mosquito, (u1ea pipi ; beetle,
deciduous stream-side trees began to exercise a Tropi8ternmw sp.; and sludgeworms, Limrnod-
shading influence. A dissolved oxygen value of riWm and Tubifeac spp., which occurred in
19.4 ppm was found at station 5.2 at 3 p. m. on abundance at stations 6.5 and 5.2, also occurred
May 9. This peak was produced by a heavy in the clean-water zones. Second, many species
plankton bloom. When marginal vegetation listed in table 2 occurred in such small numbers
shaded the stream, blooms became less marked as to discourage their individual use as indi-
and daytime oxygen values were not as high. cators. Third, several ecologic factors other
As the summer advanced, oxygen depletion than the presence of a pollutant may limit the
became of progressively greater duration and distribution of certain species; for example,
extent. The result was a well-defined oxygen- erosion, floods, the size of the stream, the type
less zone extending from the sewage treatment of bottom, the flight range of the insect, and
plant outfall to a point 2 miles downstream, as the portion of the stream under study. It is
indicated by studies carried out on the night of believed that the moderate abundance of single
August 22. This depletion persisted through- species should not be considered as biological
out the day at stations 7.2 and 6.5. The range indicators of pollution because organisms such
in hourly dissolved oxygen values found late in as Tubl3fex usuaRly associated with polluted
August at the various sampling stations along areas are also found in clean waters. It is the
the stream is shown in the chart. complex or association of organisms which is
important for indicating clean or polluted
Discussion water. All organisms present and their rela-
tive abundance must be considered.
One of the primary objectives of the Lytle In satisfactorily using associations of aquatic
Creek studies has been to arrive at a better defi- organisms as indicators of pollution, the ab-
nition of indicator organisms. The finding of sence or much reduced numbers of formerly
certain organisms, such as rat-tail maggots, present clean-water species in an area may be
Eristalis sp.; sludgeworms, Tiubifex tubifex; as important, or more so, as numbers of known
and bloodworms, Chironamus tentanm, presents pollutional forms (2). In this connection, table
important evidence of the pollutional condition 2 shows that such insects as mayflies, caddis-
of a stream (1), while the absence of cleaner flies, stoneflies, and hellgramites were almost
water forms is likewise a valuable measure of entirely limited to the clean water occurring at
existing conditions. stations 8.7 and 2.8. The only exception was a
With favorable ecologic conditions, such as mayfly, Callibaeti sp. This form was collected
high temperatures, uniform flow, freedom from in a marginal surface sample and evidently had
62 Public Health Reports
 been able to obtain sufficient oxygen from the these forms to attain a great abundance in
surface film to survive. polluted areas.
The absence of organisms, which are indica- When conditions are favorable for organisms
tive of clean-water conditions, from a section of which can adapt to pollution, they thrive and
stream should not always be taken as definite build high populations (4). For this reason,
indication of pollution. A knowledge of the the society of organisms found in zones of pol-
life histories of the various groups of aquatic lution is highly significant in offering clues
insects often is helpful in interpreting the mean- to the intensity of pollution and the degree of
ing of their distribution. For example, in the recovery. If changing conditions brought
collection of invertebrates made in August, only about by pollution are unfavorable, organisms
one species of stonefly and five species of cad- must resist these changes, migrate, or be de-
disflies were represented. In the May collection stroyed. Since many of the species of insects
five species of stoneflies and nine species of found in Lytle Creek are found in both the
caddisflies were present. Their absence later on lower and upper sections of the stream but are
was due to their emergence as adults during the missing in the pollutional zones, and since such
intervening months. Similarly, the absence of organisms are continuous in other nonpolluted
some beetles, dragonflies, and damselflies in the streams in the area, it seems probable that pol-
earlier collections was partly due to their pres- lution is responsible for their discontinuous
ence in immature stages so that they were distribution in Lytle Creek.
missed or overlooked. In a stream polluted with organic wastes, a
Apart from the fact that pollutional organ- certain reduction of the supply of dissolved
isms must be found and clean-water species oxygen may occur in a certain section through-
must be missing in delineating the zones of out the year, during the summer months, or
pollution in a stream, the mode of occurrence for only a few days in the year. Because pol-
of the organisms must be taken into account lution surveys frequently cannot be made dur-
(3). For example, the assemblage of organ- ing the period of critical conditions, and
isms found in the polluted zones of Lytle Creek chemical studies give information on physical-
presented the following characteristics: (a) chemical conditions only at the time of sam-
very large numbers of individuals; (b) few pling, there is need for additional methods
species represented in the fauna; (c) princi- which can be used throughout the year for de-
pally scavenger types present; and (d) the termining over-all conditions and the extent
presence of forms having low oxygen require- and severity of brief critical or limiting en-
ments or special adaptations for obtaining their vironmental factors. The qualitative and quan-
oxygen supply. Not over 10 different species titative composition of an aquatic population
of macroinvertebrates were taken from sta- is determined by recurring critical conditions,
tions 6.5 and 5.2 in the septic zone at any one even though of short duration, as well as the
time, while as many as 40 species were found more stable or long-time environmental fac-
in the clean-water zones (table 2). However, as tors. Therefore, the complex of organisms
many as 3,000 Culew, pipien. or 20,000 Tubificids which develops in a given area is in turn in-
were collected per square foot sample in the dicative of environmental conditions which
septic zone, while no species exceeded 500 in- have occurred during its development. Or-
dividuals per square foot in the clean-water ganisms having life histories of a year or more
area. All the insects occurring in the septic will thus serve to indicate unfavorable or lim-
zone have special adaptations for obtaining iting conditions that have occurred several
oxygen, such as the caudal respiratory tubes of months previously. Because aquatic popula-
the mosquito and rattail maggot or the air tions are a result of past environmental condi-
space under the elytra of the beetles. With tions, they serve as a means for determining
such conditions in a
such adaptations, depletion of oxygen in the cially valuable because they can They stream. are espe-
be used during
water does not serve as a barrier to their dis- fall, winter, or spring months, when flows are
tribution. An abundant food supply enables large, dilution is at a maximum, dissolved oxy-
Vol. 67, No. 1, January 1952
63
 gen is near saturation, and visual evidence of able index in delineating zones of pollution in a
pollution at a minimum, to delineate former stream.
septic areas or to indicate critical conditions of 4. In interpreting the distribution of orgal-
short duration. isms in a stream as an index of pollutional con-
In order to utilize aquatic populations in this ditions, all environmental factors should be con-
way, however, it is essential to have a knowledge sidered.
of the species composition and abundance of the 5. Pollutional associations are characterized
various organisms in the population under the by few species but large numbers of individuals.
various ecologic conditions which prevail in 6. The association of organisms normally
clean and in polluted waters. Further, consid- present under most septic conditions is charac-
eration must be given to the fact that pollution terized by the absence of plant and animal eaters
with organic wastes is only one of the several and the presence of scavengers.
environmental factors determining the compo- 7. Organisms characteristic of septic zones
sition of aquatic populations. Other environ- are those which can exist under conditions of
mental factors, such as type, size and character very low oxygen or have adaptations for breath-
of the stream, variations in flow, severity of silt- ing atmospheric oxygen.
ing, floods and erosion, water temperatures and 8. Nocturnal deficiencies in dissolved oxygen
the character of the watershed, are all important are often the critical environmental factor
in determining the make-up of aquatic popula- which determines the distribution of organisms
tions. Occasionally, there is a sparsity of bot- in a stream.
tom organisms which makes the drawing of any 9. Under conditions of organic pollution
conclusions in regard to organic pollution diffi- diurnal-nocturnal deficiencies in oxygen are
cult. However, one who has knowledge of and sometimes concomitant with supersaturation of
experience with aquatic organisms can, by a dissolved oxygen.
reconnaissance survey, learn a great deal about
the severity and extent of pollution and the ACKNOWLEDGMENTS
extent of stream recovery even though the sur- An investigation of this type requires the services of
vey is made at a time of year when the physical- a considerable number of people. The entire staff of
the biology section has assisted in certain phases of
chemical indicators of pollution are least evi- the study, and their contributions are acknowledged.
dent or largely absent. The authors express their appreciation for extensive
assistance to Dr. 0. M. Palmer, George Paine, Harold
Walter, Jack Hammond, Dr. Max Katz, and Thomas
Conclusions Maloney.
1. Single species of organisms such as Tubi-
fex tubifex, or Clhiroinowm tentanm cannot be REFERENCES
used safely as indicators of pollution unless (1) Bartsch, A. F.: Biological aspects of stream pollu-
their relative abundance is considered. tion. Sewage Works Journal 20: 292-302 (1948).
2. The absence or much reduced numbers of (2) Richardson, R. E.: The bottom fauna of the Mid-
dle Illinois River, 1913-1925. Ill. Nat. Hist.
formerly present clean-water species constitutes Surv. Bull. 17: 387-475 (1928).
an important index in evaluating the degree of (3) Whipple, G. S.: The microscopy of drinking water.
pollution. Ed. 4. New York, John Wiley and Sons, 1948.
3. The quantitative and qualitative composi- (4) Lackey, James B.: Stream microbiology. In
Stream sanitation, by Earle B. Phelps. New
tion of an aquatic population constitutes a valu- York, John Wiley and Sons, 1944.

Public Health Reports