Studies on the Reproductive Performance and Progeny Performance of Caged White Leghorns Fed Malathion and Carbaryl

Studies on the Reproductive Performance and Progeny Performance of Caged White Leghorns Fed Malathion and Carbaryl ROBERT J.

LILLIE

United States Department of Agriculture1 (Received for publication May 1, 1972)

ABSTRACT Caged White Leghorn pullets were fed a breeder diet supplemented with 0, 250 or 500 p.p.m. malathion and/or carbaryl. The traits studied in a 36-week period were body weight changes, egg production, egg weights, specific gravity of eggs, feed consumption, mortality, fertility, hatchability, embryonic abnormalities and progeny performance. In the progeny performance studies, progeny from hens fed 0 and 500 p.p.m. malathion and/or carbaryl were fed a broiler diet supplemented with 0 or 500 p.p.m. malathion or carbaryl for a 4-week period in batteries. The only significant differences resulting from malathion and/or carbaryl supplementation in the feed were pullet weights and 4week progeny weights. The pullet weight gains were significantly reduced at the 1% level by carbaryl with or without malathion. A significant growth depression was observed with the progeny fed carbaryl, irrespective of maternal diet. In a separate 4-week study, the incorporation of 500 p.p.m. malathion and/or carbaryl in the caged Leghorn male diet exerted no significant changes in the fertility pattern or the incidence of sperm and embryonic abnormalities. POULTRY SCIENCE 52:

G

266-272,

1973

HADIRI et al. (1967) showed that a Leghorn pullets and that the two higher combination of malathion and carba- levels significantly reduced the hatchability ryl fed to Single Comb White Leghorn hens of fertile eggs after 10 weeks of pesticide and males in a three-week period resulted feeding, as compared with the control in a significant decrease in hatchability. group. Both pesticides fed singly or in combinaSince both malathion and carbaryl have tion produced a significant increase in been regarded by the poultry industry as pathological changes in embryos and de- the safest pesticides to use, studies were formities in progeny. The decrease in conducted to test the effect of these two hatchability and increase in embryonic and pesticides on the performance of layers. progeny deformities were directly propor- The results of these studies are presented tional to the pesticide level (0, 75, ISO, herein. 300, 600 p.p.m.). Embryonic deformities have been described by Ghadiri and GreenEXPERIMENTAL PROCEDURE wood (1966), Dunachie and Fletcher Experiment 1. Single Comb White Leghorn (1969) and Greenberg and LaHam pullets were obtained from a commercial (1970), all of whom employed the egg-in- hatchery at 20 weeks of age and housed in jection technique with malathion and/or individual cages in a gravity-ventilated, carbaryl in preincubated fertilized eggs. open-front building equipped with winSauter and Steele (1972) reported that 0.1, dows. Supplementary heat was supplied 1 and 10 p.p.m. of malathion significantly during the cold weather to maintain room reduced egg production of caged White temperature at approximately 10°C. A 1 Animal Science Research Division, A.R.S., Ag- stock breeder diet containing 3.5% calcium ricultural Research Center, Beltsville, Maryland was fed ad libitum to the birds exposed to a 14-hour light regime. At 32 weeks of age 20705. 266

PESTICIDES AND CHICKEN PERFORMANCE

the pullets were individually weighed and randomly distributed into seven groups of 20 each on the basis of body weight and sexual maturity. The mean body weight of all groups was 1737 grams at the initiation of the study. The seven groups were continued on the breeder diet supplemented with 0, 250 or 500 p.p.m. malathion,2 250 or 500 p.p.m. carbaryl,3 250 p.p.m. each of malathion and carbaryl or 500 p.p.m. each of both pesticides. Diets containing the pesticides were prepared every two weeks. The traits studied in a 36-week period were body weight changes, egg production, egg weights, egg shell thickness as measured by specific gravity, feed consumption, mortality, fertility, hatchability, embryonic abnormalities and progeny performance. In the fertility studies, a composite sample of undiluted semen obtained from caged White Leghorn males fed the same diet as the control pullets was used to inseminate the pullets once a month at the rate of 0.05 cc. per bird. Two days after insemination, eggs were saved for a 5-day period and set according to hen and group number. All embryos that did not hatch by 21 days were examined for evidence of abnormalities that might be attributed to pesticide feeding. In progeny performance studies, the progeny from the group fed no pesticides and from those three groups fed 500 p.p.m. malathion or carbaryl singly and in combination were housed in chick brooder batteries and fed a standard broiler diet supplemented with 0, 500 p.p.m. malathion or 500 p.p.m. carbaryl for a 4-week period. In 2 95% of 0,0-dimethyl phosphorodithioate of diethyl mercaptosuccinate and 5% inert material (Cythion, American Cyanamid Co.). 5 Sevin, 97.5% aerosol grade (Union Carbide Chemicals Co.).

267

all cases, each dietary regime contained progeny from each of the four maternal groups to eliminate position effect. In the 4-bimonthly progeny tests growth and mortality were the criteria used to determine pesticide effects. Experiment 2. At the termination of Experiment 1, 20 males were retained for a 4week feeding trial to determine the effect of pesticide supplementation on the subsequent fertilizing capacity of the male. The males were randomly distributed into four groups of five each on the basis of semen volume and sperm motility. The four groups were fed the same breeder diet supplemented with 0 or 500 p.p.m. of malathion or carbaryl singly and in combination. Females used in this study were those that had been fed either 0 or 500 p.p.m. of malathion and/or carbaryl in Experiment 1 and were continued on their respective diets in Experiment 2. One week after the initiation of pesticide supplementation in the male diet, a composite sample of undiluted semen was obtained from each group of males and used to inseminate both nonsupplemented and pesticide-supplemented females at the rate of 0.05 cc. per bird. This procedure was carried out a total of three times on a weekly basis. The criteria used in the male fertility studies was percent fertility, percent hatchability, semen production and embryonic abnormalities. Data on all traits except feed consumption, adult and progeny mortality, and embryonic and sperm abnormalities were analyzed by least squares analysis of variance. Whenever appropriate, the significant differences were determined by the least significance difference test (LeClerg, 1957). RESULTS AND DISCUSSION

Experiment 1. The data obtained on adjusted body weight gains, egg production,

268

R. J. LlLLIE TABLE 1.—Effect of malathion and/or carbaryl supplementation on body weight gains, egg production, egg weights, and specific gravity of eggs of caged S.C.W.L. pullets in a 36-week period

Pesticide supplementation

None Malathion (M)

Level

Adjusted body weight gains

Egg production (hen-day)

Egg Weights

Specific gravity of eggs 1

Average feed consumption (hen-day)

p.p.m. 0

(grams) 312 + 48.9a 2

76.8±2.3

%

(grams) 60.8 + 0.78

1.085 + 0.001

(grams) 120

289±47.7 236±47.7

77.3±2.3 75.0+2.3

58.9 + 0.76 60.9+0.76

1.085 + 0.001 1.082+0.001

118 120

263+33.7a

76.1+1.6

59.9 + 0.53

1.083+0.001

119

129±52.1 104±51.9

71.7+2.5 75.7+2.5

58.4+0.83 60.1 + 0.82

1.082 + 0.001 1.082 + 0.001

118 108

117±36.8b

73.7+1.8

59.2 + 0.58

1.082 + 0.001

113

91 + 50.4 153 + 48.9

75.9 + 2 . 4 73.2 + 2 . 3

60.9 + 0.80 58.8 + 0.78

1.085 + 0.001 1.084±0.001

112 110

122 ± 3 5 . 2 b

74.5+1.7

59.9 + 0.56

1.085 + 0.001

111

250 500

Means Carbaryl (C)

250 500

Means M plus C, each

250 500

Means

Source Total Reduction MU Pesticide Level ( L ) / M L/C L/M+C BW(1) Error 1

!

Specific gravity = -

Least Squares Analysis of Variance Adjusted body weight gains Egg Production

df 123 8 1 3 1 1 1 1

<0.01 N.S. N.S. N.S. N.S.

N.S. N.S. N.S. N.S. N.S.

Egg Weights

Specific gravity

N.S. N.S. N.S. N.S. <0.01

N.S. N.S. N.S. N.S. N.O.

115 egg weight in air

egg weight in air—egg weight in water Means with different superscripts are significantly different at the 1% level of probability.

egg weights, specific gravity of whole eggs, 22 weeks of a 29-week period and 1000 and average feed consumption per hen per p.p.m. malathion in an 8-week period, reday in a 36-week period are tabulated in spectively. Furthermore, our data with carTable 1, the data on fertility, hatchability baryl substantiate the work of Nir et al. and embryonic abnormalities in Table 2, (1966) who reported decreases in body and the data on progeny performance in weight gains when layers were given 540 Table 3. The only significant differences re- p.p.m. carbaryl administered orally in gelasulting from malathion and/or carbaryl tin capsules in a 60-day period, whereas, a supplementation in the feed were adult lower level (180 p.p.m.) was ineffective. body weight gains (Table 1) and 4-week Progeny body weight gains: The incorbody weights of progeny (Table 3). poration of 500 p.p.m. carbaryl in the progAdult body weight gains: The adult eny diet significantly reduced the 4-week body weight gains were significantly re- body weight gains, as compared with that duced by carbaryl with or without mala- of progeny fed the control or malathionthion. No differences were observed be- supplemented diets (Table 3). In all cases, tween the control group and those groups pesticide supplementation in the maternal fed malathion alone. These results with ma- diet exerted no effects on the subsequent lathion are in agreement with those of Ross growth or livability of progeny, irrespective and Sherman (1960) and Toepfer and of progeny diet. The significant growth deMorgareidge (1971) who observed no sig- pression observed with progeny fed 500 nificant body weight changes as a result of p.p.m. carbaryl was not in accordance with feeding 1100 p.p.m. malathion in the last the data of Sherman and Ross (1961) who

PESTICIDES AND CHICKEN PERFORMANCE

found no changes in the growth rate as a result of feeding 440 p.p.m. carbaryl in a 2week period. Higher levels of carbaryl depressed growth and increased mortality of young chickens (Acciarri et al., 1969). The finding that 500 p.p.m. malathion exerted no adverse effects on the 4-week body weight gains of progeny (Table 3) differed from the results of Ross and Sherman (1960), who observed a significant decrease in body weight gains at 4 weeks of age only when chicks were fed 220 p.p.m. malathion during the grower period. However, Rehfeld et al. (1969) reported no growth-depressing effects of 1000 p.p.m. malathion but did report a toxic effect of higher levels (2500 and 5000 p.p.m.) on growth and livability of young chickens. Likewise, Acciarri et al. (1969) showed a TABLE 2.—Effect

None Malathion (M)

Level

Carbaryl (C)

Fertility

461

93.2 + 1.31

89.1 + 1.7

507 501

93.1 + 1.31 94.6±1.31

92.3±1.7 89.5 + 1.7

1008

93.8 + 0.09

90.9 + 1.2

403 451

93.1 + 1.31 95.5 + 1.31

88.8 + 1.7 87.5 + 1.7

854

94.3±0.09

88.2 + 1.2

449 469

92.5 + 1.31 94.6+1.31

85.9+1.7 88.1 + 1.7

918

93.6+0.09

87.0+1.2

250 500

250 500

Totals and Means 250 500

Totals and Means Source Total Reduction MU Pesticide L/M L/C L/M+C Error 2

Hatchability of fertile eggs

Eggs set

p.p.m.

Totals and Means

1

depressing effect on growth and livability of chicks fed 5000 p.p.m. malathion supplied by a pesticide compound containing 50% malathion and 20% parathion. Production traits and feed consumption: The feeding of malathion and/or carbaryl resulted in no significant changes in egg production, egg weights, specific gravity of eggs and average feed consumption (Table 1). The data of Sauter and Steele (1972) indicated that egg production was significantly reduced by low levels of malathion (10 p.p.m. or less). Conversely, egg production was not affected by high levels of malathion (1000 p.p.m. or more) in studies conducted by Ross and Sherman (1960) and Toepfer and Morgareidge (1971) and by low levels of carbaryl (200 p.p.m. or less) in studies by McCay and Arthur

of malathion and/or carbaryl supplementation on fertility, hatchability and embryonic abnormalities of eggs laid by caged S.C.W.L. pullets (8 monthly studies)

Pesticide supplementation

M plus C, each

269

From the 8th and final hatch. From the 6th hatch.

%

Presence of macroscopic embryonic abnormalities

%

Least Squares Analysis of Variance df Fertility 63 7 1 3 N.S. 1 N.S. 1 N.S. 1 N.S. 56

One 16-day embryo with cranial hernia, 4 legs and deformed beak 1

one 17-day embryo with no beak2 two 18-day embryos with cranial hernia one of which had a smaller upper beak2

Hatchability

N.S. N.S. N.S. N.S.

270

R. J. LILLIE TABLE 3.—Effect

of malathion and/or carbaryl in the maternal diet upon 4-week body weight means of progeny fed nonsupplemented and pesticide-supplemented broiler diets (4 bi-monthly progeny studies) Pesticide supplementation in progeny diet

Pesticide supplementation in • maternal diet

None 500 p.p.m, malathion (M) 500 p.p.m. carbaryl (C) 500 p.p.m. each of M and C Means and (totals)

vr«„„ None

500 p.p.m. malathion

500 p.p.m. carbaryl

Means and totals

(grams) 290 + 9.8 (70)%' 294 + 9.8 (75)a 293 + 9.8(67)a 292 + 9.8 (70)a 292 + 4.9 (282)a

(grams) 289±9.8(69)a 297±9.8(75)a 291±9.8(65)a 289±9.8(66)a 292±4.9(275)a

(grams) 258 + 9.8 (75)b 267 + 9.8 (74)b 263 + 9.8 (65)b 257 + 9.8 (64)b 261 + 4.9(278)b

(grams) 279 + 5.6(214) 286 + 5.6(224) 282 + 5.6(197) 280 + 5.6(200) 282 + 2.8(835)

Least Squares of Analysis of Variance df Mean 4-week body weights of progeny Total 48 Reduction 12 MU 1 Maternal Diet (M) 3 N.S. Progeny Diet (P) 2 <0.01 MXP 6 N.S. Error 36 Source

1 Number in parenthesis represents survivors at the end of the 4-week study. The mortality rate was less than 1% in all cases. 2 Means with different superscripts are significantly different at the 1% level of probability.

(1962) and Nir et al. (1966). However, a higher level of carbaryl (S40 p.p.m. via gelatin capsule) did cause a cessation of egg production and resulted in 100% mortality by the 35th day of the study (Nir et al., 1966). Hence, the mode of administration could be responsible for the degree of toxicity on egg production and mortality, since 500 p.p.m. carbaryl fed in the diet for a 36week period was not toxic for both egg production and mortality in our studies. Higher levels of malathion (1100 p.p.m.) for the last 22 out of 29 weeks resulted in 25% mortality when administered in the feed (Ross and Sherman, 1960). The regression of initial body weight [BW(1)] on egg weights (Table 1), significant at the 1% level, uncovered a source of variation which could affect the group means; differences between the resulting adjusted group means could then be assessed free of this influence. Accordingly, no significant differences due to pesticide feeding were observed for egg weights,

Reproduction traits: Pesticide supplementation in the breeder diet exerted no effects on the fertility, hatchability and incidence of embryonic abnormalities (Table 2). Out of 3,241 eggs set in the 36-week period, only four embryos were found to possess abnormalities—three in the 6th hatch and one in the 8th and final hatch. These results do not support those of Ghadiri et al. (1967) for hatchability and embryonic abnormalities in eggs laid by hens fed the pesticides and those of Ghadiri and Greenwood (1966), Dunachie and Fletcher (1969) and Greenberg and LaHam (1970) for embryonic abnormalities observed in eggs injected with either or both pesticides. Experiment 2. As indicated in Table 4, the incorporation of 500 p.p.m. malathion or carbaryl singly or in combination in the male diet for a 4-week period exerted no significant changes in the fertilizing capacity of the male. Furthermore, neither the incidence of sperm abnormalities nor the

271

PESTICIDES AND CHICKEN PERFORMANCE

incidence of embryonic abnormalities was increased by pesticide feeding. Sosnierz et al. (1970) reported that rats fed malathion at the level of 16 p.p.m. via oral administration exhibited inhibition of spermatogenesis in addition to other toxic symptoms. GENERAL DISCUSSION

The data clearly indicate that malathion or carbaryl fed singly or in combination at the 250 or 500 p.p.m. concentrations are not toxic to chickens with respect to reproduction, including embryonic development. Several possibilities may account for the discrepancies between our data and those reported by Ghadiri et al. (1967) and by

TABLE 4.—Effect

Sauter and Steele (1972). One possibility is that the birds used by Ghadiri et al. (1967) were maintained in floor pens equipped with litter, where coprophagy is common, whereas our birds were deprived of coprophagy in wire cages. A second possibility is the mode of pesticide administration in the feed. Dunachie and Fletcher (1969) reported that the toxicity is much greater if malathion is dissolved in corn oil than in acetone and that carbaryl is insoluble in corn oil. In our studies, both malathion in liquid form and cabaryl in powdered form were incorporated into the feed without any dilution. The method of pesticide incorporation in the feed by Ghadiri et al. (1967) was not reported whereas malathion used by Sauter

of malathion (M) and/or carbaryl (C) in the male diet upon the subsequent fertilizing capacity of the male (3 weekly inseminations)

Pesticide supplementation In the male diet (M.D.) None (MD-1)

Eggs set

Fertility

In the female diet (F.D.)

Hatchability of fertile eggs

None 500 p.p.m. each of M + C

48 43

% 95.2 + 3.1 89.9 + 3.1

% 86.5 + 10.3 8 8 . 4 ± 10.3

91

92.5 + 2.2

8 7 . 5 ± 7.3

55 52

96.1 + 3.1 88.6 + 3.1

79.2 + 10.3 62.0 + 10.3

107

92.3±2.2

70.6+ 7.3

59 55

94.1 + 3.1 95.5 + 3.1

96.6+10.3 88.5±10.3

114

94.8±2.2

9 2 . 5 + 7.3

64 64

96.4±3.1 99.9 + 3.1

80.8±10.3 95.8+10.3

128

98.2±2.2

8 8 . 3 ± 7.3

Totals and Means 500 p.p.m. M (MD-2)

None 500 p.p.m. M Totals and Means

500 p.p.m. C (MD-3)

None 500 p.p.m. C Totals and Means

500 p.p.m. each of M + C (MD-4)

None 500 p.p.m. each of M + C Totals and Means

Least Squares Analysis of Variance df Fertility Total 24 Reduction 8 MU 1 Male Diet (MD) 3 N.S. FD/MD-1 1 N.S. FD/MD-2 1 N.S. FD/MD-3 1 N.S. N.S. FD/MD-4 1 Error 16 Source

Hatchability

N.S. N.S. N.S. N.S. N.S.

272

R . J . LlLLIE

and Steele (1972) was in powdered form. A third possibility would be the source of malathion which might account for the discrepancies between our data and those of Ghadiri et al. (1967) and of Sauter and Steel (1972). Oregon and New York were the sources of malathion used by Sauter and Steele (1972) and in our studies, respectively, whereas the source was not reported in the data of Ghadiri et al. (1967). A fourth possibility may be the difference in the tryptophan and niacin content of the diets used. According to Greenberg and LaHam (1970), tryptophan, niacin, nicotinamide or quinolinic acid via the egg-injection technique counteracted the adverse effects of injected malathion on embryonic malformations. Similar results were obtained with nicotinamide via the egg-injection technique by Walker (1971). In this respect, the stock breeder diet used in our studies was properly balanced in all known nutrients, including tryptophan and niacin. No information is available on the composition of the diet used by Ghadiri et al. (1967), and Sauter and Steele (1972). ACKNOWLEDGEMENT

We wish to express our gratitude for the valuable services of Dr. B. T. Weinland of the Biometrical Services Staff, A.R.S., Beltsville, Maryland. REFERENCES Acciarri, C , A. Restuccia and F. Gramenzi, 1969. Toxicity tests for some pesticides (organic phosphoesters, chlorinated insecticides, carbamates) in experimentally treated chickens. Veterinaria Italiana, 20: 609-630. Dunachie, J. F., and W. W. Fletcher, 1969. An investigation of the toxicity of insecticides to birds' eggs using the egg-injection technique. Ann. Appl. Biol. 64: 409-423.

Ghadiri, M., and D. A. Greenwood, 1966. Toxicity and biologic effects of malathion, phosdrin and sevin on the chick embryo. Toxicol. App. Pharmacol. 8:342. Ghadiri, M., D. A. Greenwood and W. Binns, 1967. Feeding of malathion and carbaryl to laying hens and roosters. Toxicol. Appl. Pharmacol. 10:392. Greenberg, J., and Q. N. LaHam, 1970. Reversal of malathion-induced teratisms and its biochemical implications in the developing chick. Canad. J. Zool. 48: 1047-1053. LeCIerg, E. L., 1957. Mean separation by the functional analysis of variance and multiple comparisons. Agric. Res. Serv. A.R.S.—20-3 (May). McCay, C. F., and B. W. Arthur, 1962. Sevin residues in poultry products. J. Econ. Entomol. 55: 936-938. Nir, I., E. Weisenberg, A. Hadani and M. Egyed, 1966. Studies of the toxicity, excretion and residues of Sevin in poultry. Poultry Sci. 4 5 : 720728. Rehfeld, B. M., D. E. Pratt and M. L. Sunde, 1969. Effect of various levels of dietary malathion on performance of chicks. Poultry Sci. 48: 1718-1723. Ross, E., and M. Sherman, 1960. The effect of selected insecticides on growth and egg production when administered continuously in the feed. Poultry Sci. 39: 1203-1211. Sauter, E. A., and E. E. Steele, 1972. The effect of low level pesticide feeding on the fertility and hatchability of chicken eggs. Poultry Sci. 5 1 : 71-76. Sherman, M., and E. Ross, 1961. Acute and subacute toxicity of insecticides to chicks. Toxicol. Appl. Pharmacol. 3 : 521-533. Sosnierz, M., Z. Szczurek, R. Knapek and A. Kolodziejczyk, 1970. Pathomorphologic experiments on accelerated chronic toxicity of Sadofos (malathion). Patol. Pol. 2 1 : 737-744. Toepfer, E. W., and K. Morgareidge, 1971. Effect of malathion on nutrient composition of eggs and flavor of meat from laying hens. J. Food Sci. 36: 511-514. Walker, N. E., 1971. The effect of malathion and malaoxon on esterases and gross development of the chick embryo. Toxicol. Appl. Pharmacol. 19: 590-601.