Effects of DDT and Charcoal on Performance of

White Leghorn Hens1'2

G. P. WAIBEL, G. M. SPEERS3 AND P. E. WAIBEL 3
Department of Animal Science, University of Minnesota, St. Paul, Minnesota 55101

(Received for publication March 6, 1972)

ABSTRACT White Leghorn hens were fed diets containing DDT (0 or 300 p.p.m.) and
activated charcoal (0 or i%) in factorial arrangement for a 30 day period to study the
effects on hen performance, egg quality, embryo and progeny growth. The presence of
charcoal in the diet reduced the DDT deposition in the hens' abdominal fat and in the egg
yolk. Charcoal also reduced yolk pigmentation. Neither DDT nor charcoal affected the egg
weight or shell thickness. Embryo development from hens fed D D T and/or charcoal appeared
normal. There were no differences in hatchability, fertility, or progeny growth related to the
treatments.
POULTRY SCIENCE 51: 1963-1967, 1972

INTRODUCTION greater during the first part of a clutch be-

D DT has been a highly valuable insec- cause the calcium had a chance to restore
ticide for many years. Its uses in- itself after being depleted during the previ-
clude control of mosquitoes, grasshoppers, ous clutch.
and other insects harmful to crops and There have been numerous instances
man. More recently, questions have been where DDT or related compounds were
raised about the overall effect DDT will blamed for the death or reduced level of re-
have on the ecosystem including man and production in wild birds. Heath et al.
his domestic animals. (1969) found that 40 p.p.m. DDE in the
Technical DDT includes a series of diet of Mallard ducks caused a 13 percent
closely related compounds (the three pres- decrease in shell thickness as compared
ent in greatest amounts are p,p' DDT, o,p with the control group. As a result, 25% of
DDT and DDE). These compounds are the eggs laid in the DDE group were
relatively stable and characteristically be- cracked or broken compared with only 5%
come deposited in the fatty tissues of ani- of the eggs laid by the control ducks. In the
mals, including man (Liska et al., 1964; second year, there were 38 hatchlings out
Rubin et al., 1947). of 228 eggs incubated from the DDE
Using quail, Bitman et al. (1969) found group, as compared with the control group
that DDT caused thinner egg shells of re- where there were 318 hatchlings from 607
duced calcium content. Their treatments eggs. Only 26 of the 38 hatchlings from the
utilized a low calcium level (0.56%), DDE group lived to 14 days of age,
which posed a stress condition. The p,p' whereas 274 of the 318 hatchlings from the
DDT group produced fewer eggs and had control group lived to 14 days. Similar re-
more broken eggs than either the control or sults were shown with 25 p.p.m. DDT in
o,p DDT groups. Shell thickness was the diet.
Rubin et al. (1947) reported that hens
1
Published as Paper No. 7908 Scientific Journal fed DDT at 0, 300, 620, 1250, and 2500
Series of the Minnesota Agricultural Experiment
p.p.m. produced eggs at daily percentage
Station.
2
Supported in part by funds of the North Cen- rates of 55, 36, 24, 0 and 0, respectively,
tral Regional Project NCM-40. during the third lunar month of the study.
3
Department of Animal Science. During this time mortality was 11, 11, 10,
1963
1964 G. P. WAIBEL, G. M. SPEERS AND P. E. WAIBEL

TABLE 1.—Composition of the chicken breeder ration, MATERIALS AND METHODS

Single Comb White Leghorn hens, nine
Ingredient %
months of age and in good egg production,
Corn, ground yellow1 63.5
Soybean meal, 44% protein 16.5 were assigned on the basis of body weight
Meat and bone meal, 50% protein 2.5 to eight experimental groups of 4 hens
Fish solubles, 100% equiv. dried on soy-
bean meal 2.0 each. The hens in each group were housed
Whey, dried whole 2.0 in adjacent individual cages sharing a com-
Alfalfa meal, 17% protein, dehydrated 2.5
Dicalcium phosphate 1.25 mon feeder and waterer in a heated and
Calcium carbonate 5.5 ventilated building with 16 hours of artifi-
Salt, iodized 0.25
Manganese sulfate, 27% Mn 0.025 cial light per day. The two replicate groups
Zinc oxide, 72% Zn 0.0025 per treatment were housed in separate
Vitamin premix2 1.0
blocks on either side of the hen battery, all
Total 100.0275 on one cage level.
1
4 % corn omitted for inclusion of sand, charcoal The diet, shown in Table 1, was of the
or DDT premix ( 1 % corn carrier).
2
corn-soybean meal type with 2 times the
Supplied per kilogram of diet: vitamin A, 11,000
I.U.; vitamin D 3 , 3520 I.C.U.; vitamin E, 11 I.U.; usual vitamin supplementation. It was
vitamin K, 4.4 mg.; riboflavin, 8.8 mg.; calcium available ad libitum. The four experimental
pantothenate, 15.4 mg.; niacin, 39.6 mg.; vitamin
B12, 17.6 meg.; folic acid, 1.1 mg.; corn carrier. treatments listed below were in factorial ar-
rangement.
Treatment A. Control (3% sand (puri-
90 and 100 percent, respectively. Egg pro-
fied) replacing corn)
duction during the first lunar month was
57, 46, 34, 36 and 17 percent respectively. Treatment B. Control diet plus 300
p.p.m. of DDT and 3 % sand
The aspects of DDT intake and elimi-
Treatment C. Control diet with 3% acti-
nation from animals are also important.
vated charcoal replacing sand
Wesley et al. (1969) reported that the level
Treatment D. Control diet with 300
of protein in the diet, injection of androgen
p.p.m. of DDT and 3 % activated
hormone (15 mg. per chick), or a starva-
charcoal replacing sand
tion period reduced the amount of DDT in
the fatty tissues and egg yolks. Activated After a pre-experimental period of one
charcoal has been shown to reduce the week during which all birds received the
availability of vitamins and pigment to the control diet, the above treatments were ad-
chicken (Ewing, 1963). Charcoal might ministered for a 30 day period. Egg pro-
have the same effect on DDT, since DDT duction and egg weights were recorded
is a lipid soluble compound, and provide a throughout the experiment. Body weights
means of counteracting a dietary exposure were taken at weekly intervals. At the ter-
to DDT. mination of the experiment the hens were
This experiment was designed to study killed by an injection of sodium pentobar-
the influence of a substantial level of di- bital for necropsy examination and abdomi-
etary DDT on the performance of the hen, nal (leaf) fat samples were collected.
egg quality characteristics, deposition of The hens were artificially inseminated at
DDT in abdominal fat and egg yolk, em- weekly intervals using pooled semen from
bryonic development, and progeny perfor- New Hampshire roosters. The eggs laid
mance. The effect of activated charcoal on during the 10th-13th days of the experi-
the biological availability of DDT was also ment were incubated and the embryos were
investigated. examined by gross visual observation at 3,
 DDT AND CHARCOAL FOR HENS 1965

7, and IS days of incubation. Eggs laid treatment group were housed together (16
from the 16th-25th days of the experiment pens total) with 7-12 chicks per pen. They
were incubated for measurement of fertility were weighed and observed twice weekly.
and hatchability. The chicks hatching from The data were subjected to an analysis
these eggs were used in a progeny growth of variance according to procedures of Steel
and livability study. The eggs laid on the and Torrie (1960). Treatment means were
14th and 15th days were used for shell differentiated using Duncan's new multiple
thickness and yolk color measurements. range test.
Shell thickness and yolk color were mea-
sured again in eggs laid on the 26th-28th RESULTS AND DISCUSSION
days. The results are presented in Table 2.
The concentration of DDT in egg yolk There was no significant change in hen
was determined on pooled samples from the weights during the pre-experimental period
four hens of each replicate group from the or the 30 days of the experiment. Egg pro-
14th and 26th days of the experiment. Egg duction did not differ significantly between
yolk and abdominal fat samples were pre- treatments, and the variations observed
pared for analysis using the procedure of during the experiment were due mainly to
Stemp et al. (1964). Florisil was placed one or two birds going out of production
into a chromatographic column and egg rather than to a change in the intensity of
yolk or abdominal fat was ground into an- lay of those remaining in production. In
other portion of florisil. This was placed on view of the variation between hens and the
top of the florisil in the column and eluted recognition that hens will occasionally go
with 20% methylene chloride in petroleum out of production, it is concluded that no
ether. The solvent was evaporated, and the differences attributable to the treatments
residue was taken up in hexane. The DDT were demonstrated. This is in contrast to
compounds were determined by gas-liquid the findings of Rubin et al. (1947) that a
chromatography.4 similar level of DDT decreased egg produc-
A Roche Yolk Colour Fan5 was used to tion.
estimate the yolk color by visual compari- Egg weights increased slightly during the
son after separating the yolk from the egg experiment, however, there was no signifi-
white. The scale ranged from 1 to IS with cant treatment difference. There was a
the highest score representing the deepest marked difference in egg yolk color; yolks
color. Shell thickness was measured on two from hens fed charcoal were significantly
pieces of shell (without membrane) taken lighter (P < .01) in color than those from
from opposite sides at the equator of the hens not receiving charcoal. There was no
egg using a Starrett anvil jawed microme- difference in yolk color due to DDT treat-
ter.6 ment in the presence or absence of char-
In the two week progeny growth study coal. Xanthophyll pigments, which give the
the chicks were wing banded and fed a yolk its color, are normally absorbed from
standard chick starting feed. Chicks from the digestive tract and deposited in the
pairs of hens from the same experimental fatty tissues and egg yolk. Since charcoal is
not absorbed from the digestive tract, it is
4 assumed that charcoal adsorbed the xan-
Beckman Model GC-4.
5
Hoffmann-LaRoche, Inc., NutJey, New Jersey. thophyll pigments within the digestive
"Starrett No. 25-136, L. S. Starrett and Com- tract and reduced their absorption into the
pany, Athol, Massachusetts. body.
1966 G. P. WAIBEI., G. M. SPEEES AND P. E. WAIBEL

TABLE 2.—Effects of DDT and charcoal on hen performance, reproduction, egg quality,
progeny growth, and DDT residues in tissue and egg

Treatment
A B C D
Characteristic
DDT, p.p.m. 0 300 0 300
Sand,% 3 3 0 0
Charcoal, % 0 0 3 3
Average hen weight, gm.1 1849 1838 1828 1777
Egg production, % 77.7 76.3 77.1 65.2
Egg weight, gm. 59.2 E3 57.6 B 57.2 A 58.9
Yolk color2 9.80 9.73 6.43 6.43 A
Shell thickness, mm. .317 .314 .322 .336
Fertility, % 74 70 72 66
Hatchability, % 92 94 94 90
Chick weight, gm. (14 day) 155.9 150.0 152.3 152.9
D D T Concentration, p.p.m.
Egg yolk, 14th day DDE 0.0» 26.8" 0.5* 23.9 b
o, p D D T O.O 68.1= 0.0* 49.9b
p, p ' D D T 0.0" 241.2" 0.1* 177.3"
Egg yolk, 26th day DDE 0.0" 45.8° 0.0* 34.7 b
o, p D D T 0.0* 101.5" 0.0* 68.0 b
p, p ' D D T 0.0" 332.6" 0.0* 244.6 b
Abdominal fat, 30th day DDE 0.5" IS.2* 0.5* 89.2b
o, p D D T 0.2" 256.8" 2.0* 208.8 b
p, p ' D D T 0.0» 790.9" 2.0* 636.6 b
1
Average of the weights taken on the 7th, 14th, 21st and 28th days.
2
Average of data collected on the 14th, 15th, 26th and 28th days of the experimental period.
* Values followed by the same superscript are not significantly different at P < . 0 1 , capital letters; and
at P < . 0 5 . small letters.

The shell thickness of none of the groups servation) or progeny growth and liabil-
changed significantly during the experi- ity. Necropsy examination of hens revealed
ment. Except for treatment D, DDT and no gross pathological changes associated
charcoal, the average shell thickness was with the treatments.
similar among the treatments. Three hens Analysis of abdominal fat and egg yolks
of one replicate of treatment D laid eggs demonstrated the presence of a high con-
with thicker shells during both the pre-ex- centration of p,p' DDT (l,l,l-trichloro-2,2-
perimental and experimental periods. In bis (p-chlorophenyl) ethane; o,p DDT
contrast, Heath et al. (1969) using Mal- (l,l,l-trichloro-2-(o-chlorophenyl)-2-(p-
lards and Bitman et al. (1969) using quail chlorophenyl) ethane; and DDE (1,1-
found that DDT compounds did reduce dichloro-2,2-bis (p-chlorophenyl) ethylene).
shell thickness, indicating perhaps specie as The DDT content in the egg yolk increased
well as possible dietary differences between from the 14th to the 26th day of the experi-
these studies. ment. The inclusion of 3 % charcoal in the
There was little difference in fertility in diet significantly reduced the deposition of
treatments A, B, and C, however, the fertil- all DDT constituents in the egg yolk.
ity of hens on treatment D (DDT and The concentrations of o,p DDT and p,p'
charcoal) was 8% lower than that of the DDT in the body fat were also reduced by
controls. The hatchability of fertile eggs the dietary charcoal. DDE concentration in
was excellent for all treatments. Neither egg yolk but not in body fat was reduced
DDT nor charcoal in the hens' diet seemed by dietary charcoal; no explanation can be
to affect embryo development (by gross ob- given for this inconsistency. Ragland
 DDT AND CHARCOAL FOE HENS 1967

(1969) fed much lower doses of DDT (0.1 Cutkomp, Entomology Department, for
and 0.5 p.p.m.), and found that 3 or 6 per- providing the DDT used in this study, Dr.
cent dietary charcoal decreased residues in R. S. Adams, Soils Department, for assis-
abdominal fat when chicks were exposed to tance with the DDT analysis, and Dr.
0.5 p.p.m., but not 0.1 p.p.m. The DDT D. M. Barnes, Veterinary Diagnostic Lab-
concentration was reduced more when char- oratory, for participating in the necropsy
coal was mixed into the diet than when ad- examination. Appreciation is extended to
ministered by capsule. Three percent char- Merck and Company, Rahway, New Jer-
coal had a greater effect on DDT retention sey, Hoffmann-LaRoche Inc., Nutley, New
than did six percent. In their study the rate Jersey, Commercial Solvents Corporation,
of DDT depletion from contaminated chicks Terre Haute, Indiana and Distillation
was not influenced by feeding charcoal for Products Corp., Rochester, New York, for
21 days. Our data, as well as those of Rag- providing vitamins.
land (1969), suggest that the lower DDT
REFERENCES
content observed in the egg yolk and fatty
Bitman, J., H. C. Cecil, S. J. Harris and G. F.
tissue of hens receiving charcoal in their diet Fries, 1969. DDT induces a decrease in calcium.
along with DDT was due to the adsorption Nature, 224: 44-46.
of DDT by charcoal in the digestive tract Ewing, W. R., 1963. Poultry Nutrition, Sth edi-
of the hen, thereby preventing its total ab- tion, Ray Ewing Co., Pasadena, California, pp.
702-703.
sorption from the gut.
Heath, R. G., J. W. Spann and J. F. Kreitzer,
The presence of small quantities of DDT 1969. Marked DDE impairment of Mallard re-
constituents in the body fat in treatments production in controlled studies. Nature, 224:
A and C demonstrated the ease of contami- 47-48.
nation in the environment. In spite of pre- Liska, B. J., B. E. Langlois, G. C. Mostert and
W. J. Stadelman, 1964. Residues in eggs and
cautions taken to prevent cross consump-
tissues of chickens in rations containing low
tion between treatments (e.g., use of sepa- levels of DDT. Poultry Sci. 4 3 : 982-984.
rate feed scoops, cages on one level of the Ragland, W. W., 1969. Retention of DDT in tis-
battery only, and separate feeders and wa- sues of chickens. Unpublished Thesis. Purdue
terers for each group) DDT was found in University, Lafayette, Indiana.
Rubin, M., H. R. Bird, N. Green and R. H. Car-
fatty tissues of these birds. These birds
ter, 1947. Toxicity of DDT to laying hens.
could possibly have been contaminated be- Poultry Sci. 26:410-413.
fore the project began. Steel, R. D. G. and J. H. Torrie, 1960. Principles
By all criteria observed, DDT at the and Procedures of Statistics, McGraw-Hill
level of 300 p.p.m. was nontoxic to the lay- Book Co., N. Y.
Stemp, A. R., B. J. Liska, B. E. Langlois and
ing hens during the 30 day feeding period.
W. J. Stadelman, 1964. Analysis of egg yolk and
Over a longer period this level may have poultry tissues for chlorinated insecticide resi-
resulted in a toxic accumulation as reported dues. Poultry Sci. 43 : 273-27S.
by other workers. Wesley, R. L., A. R. Stemp, R. B. Harrington,
B. J. Liska, R. L. Adams and W. J. Stadelman,
1969. Further studies on depletion of DDT
ACKNOWLEDGMENTS
residues from laying hens. Poultry Sci. 48:
The authors wish to thank Dr. L. K. 1269-1275.

MARCH 14-16, PACIFIC EGG AND POULTRY SUPERSHOW, TOWN AND

COUNTRY HOTEL, SAN DIEGO, CALIFORNIA