DDT in Eggs and Tissues of Chickens Fed Varying Levels of DDT

DDT in Eggs and Tissues of Chickens Fed Varying Levels of DDT MELVIN J. BRYSON , C. I. DRAPER, JOSEPH R. HARRIS, CLYDE BIDDULPH, D. A. G R E E N W O O D , L. E. HARRIS, WAYNE BINNS, M. L. MINER, and L. L. MADSEN 1

Utah Agricultural Experiment Station, Logan, Utah

Downloaded by UNIV LAVAL on May 4, 2016 | http://pubs.acs.org Publication Date: January 1, 1950 | doi: 10.1021/ba-1950-0001.ch043

The concentration of DDT in eggs and tissues of chickens fed a mash made with DDT-treated alfalfa hay is reported. The amount of DDT in the fat, muscle, and eggs was correlated with the DDT intake.

I he use of D D T to control lygus bugs (Lygus elisus V a n D ; Lygus hesperus K n i g h t ) and alfalfa weevil (Hypera postica Gyll.) i n the production of seed alfalfa has been demonstrated by Lieberman (10), Smith and Michelbacher (17), and Lieberman and Hare (11). Alfalfa hay is also markedly improved b y the use of this insecticide (7). This has resulted in widespread use of the compound. W i t h the dissemination of D D T dusts and sprays over large forage crop areas, i t is important to know the effect of the insecticide on livestock and poultry. A series of studies has been conducted in the authors laboratories to gain information on this problem. When D D T is fed to animals, even i n small quantities, there is an accumulation of the compound i n the tissues, particularly the fat. Telford and Guthrie (18), Orr and M o t t (13), Woodward et al. (20, 21), and Laug and Fitzhugh (9) have demonstrated that D D T will accumulate i n certain tissues and i n milk fat of domestic and laboratory animals. Marsden and B i r d (12) found that D D T was toxic to turkeys i n concentrations above 0.075% of the diet, and that turkeys fed the insecticide for 7 to 8 weeks stored D D T i n their fat at concentrations ranging from 4 to 8 times that i n the diet. R u b i n et al. (14) reported that hens fed 0.062% D D T in their diet for 12 weeks showed reduced egg production with lowered hatchability. A t one half this concentration there was a detrimental effect on egg production, but hatchability was not seriously affected. The hens were killed by doses of 0.125% D D T . The insecticide was found i n the eggs i n quantities much smaller than i n the body fat. Harris et al. (8) have shown that D D T will accumulate i n considerable quantities i n the fat of lambs fed D D T - t r e a t e d hay. Small amounts of the insecticide were found i n other tissues. Carter and Fitzhugh (2, 5) have summarized the results of experiments on domestic 1

Table I.

Basal Mash %

Ground wheat Bran Ground barley Fish meal Soybean oil meal Salt Ground limestone Bone meal Fish liver oil Vitamin concentrate

25 24.25 25 10 10 1 1.75 2.25 0.25 0.5

Present address, Department of Biochemistry and Nutrition, Agricultural and Mechanical College of Texas, College Station, Tex. 1

232

AGRICULTURAL CONTROL CHEMICALS Advances in Chemistry; American Chemical Society: Washington, DC, 1950.

BRYSON ef al.—DDT IN EGGS AND TISSUES OF CHICKENS

233

and laboratory animals i n which D D T was found to have accumulated in small amounts in the tissues. Carter et al. have shown (3) that cooking meat from D D T - t r e a t e d animals did not seriously change its D D T content and (4) that the insecticide accumulated in the fat of pigs fed beef that contained D D T . The purpose of this work was to determine the extent of accumulation of D D T i n eggs and the tissues of chickens during and after a period of feeding a mash made with D D T - t r e a t e d alfalfa hay.


Design of the Experiment The alfalfa hay used i n making the mashes i n this experiment was from the same field as that described b y Harris et al. (1, 8). This alfalfa was dusted with 0, 1, 2, and 4 pounds of technical D D T per acre, which was approximately the same amount used b y hay producers. The harvested hay was ground into meal and a mash was made for l a y ing hens, containing 1 5 % alfalfa meal. D D T was also added to mash made from u n treated alfalfa meal at levels of 0, 50, 100, and 200 p.p.m. to determine a possible margin of safety for feeding D D T - t r e a t e d alfalfa. A total of 48 White Leghorn pullets approximately 6 months of age were used i n the experiment. Thirty-two pullets were selected at random from the original 48 birds, and divided into four groups of 8 birds each. One group was then assigned to each mash made with the hay dusted with 0, 1, 2, and 4 pounds of technical D D T per acre. The remaining 16 pullets were divided into four groups of 4 birds each, and were assigned to mashes containing 0, 50, 100, and 200 p.p.m. of D D T . This mash was made by thoroughly mixing an ether solution of D D T with 1 pound of mash and then mixing this with 19 pounds of mash (see Table I ) . The mash fed was made by mixing 1 5 % alfalfa meal with 8 5 % of the basal mash. This was available to the hens at all times. I n addition, the hens were fed a grain mixture once daily. This was made up of 5 0 % wheat, 4 0 % barley, and 10% oats. A l l hens were i n excellent laying condition at the start of the experiment. E a c h hen w as assigned at random to a compartment of a laying battery. The eggs were collected daily and were kept under refrigeration until they were ana lyzed for their D D T content. Feed consumption and production records were kept for each hen. The data on weight gains, food consumption, and egg production will be pub lished at the end of a 3-year feeding period. A t the end of one year of feeding D D T - t r e a t e d mash, 16 of the hens were killed and their leg and breast muscle and fatty tissues were analyzed for D D T . During this time 8 hens had died of leukemia complex, and their death was not attributable to the D D T . A t the beginning of the second year 24 of the original 48 hens were still on the experiment. I n order to replace those hens that were killed and those that had died, 24 more 6-monthold pullets were assigned at random to the eight groups of the experiment, making a total of 48 hens at the beginning of the second year. r

The D D T residue of the hay was determined b y the total chloride method of U m hoefer (19) as described b y Harris et al. (8). The eggs and tissues were analyzed b y the method of Schechter.ei at. (16) with the modifications described b y Harris et al. (7). T h e eggs were sampled b y mixing the yolks and the whites of a l l the eggs from one hen for one month i n a Waring Blendor. A 50-gram sample of the eggs was placed i n a 250-ml. centri fuge bottle and 75 m l . of 9 5 % ethyl alcohol and 50 m l . of Skellysolve Β were added. The Table II. D

D

DDT Content of Eggs of Chickens Consuming Mash Containing DDT-Treated Hay (1947-1948)°

T

Applied D D T Residue, P . P . M . per Acre, L b . On hay In mash6 0 0 0 1 15 2.3 2 22 3.3 4 42 6.3 Each value is average * S.D. of 6 to 8 hens. b D D T content of mash by calculation.

Months after Beginning D D T Feeding 9 10 D D T in Eggs, P . P . M . 1.7*2.0 1.7*2.0 0.9±1 2 2.0*2.0 2.1*1.6 1.8*0.8 1.6*1.3 3.7*4.9 2.4*2.5 3.3*1.6 4.1*1.0 2.5*1.2 8

a


ADVANCES IN CHEMISTRY SERIES

234

addition of the ethyl alcohol served to precipitate the protein and to prevent the for mation of emulsions with the Skellysolve B . The D D T was extracted from the egg m i x ture b y the Skellysolve B . After centrifuging, the Skellysolve Β was easily separated from the mixture. F r o m this point the analysis was carried out as with milk i n the method of Schechter et al. (15). A l l colorimetric readings were made on the Beckman quartz spectrophotometer.


Results and Discussions None of the chickens showed i l l effects from the ingestion of D D T . The mortality of the 48 hens used was 15% over a period of a year, which is below the average mortality for laying hens in this area. The deaths were not associated with D D T feeding. There was no significant difference in live weight gain between any of the hens on any of the D D T treatments or between any of the D D T levels and the control chickens. The amount of D D T present on the treated hay and in the eggs for the first year of the experiment is shown in Table I I . The D D T residue on the alfalfa hay varied from 0 to 42 p.p.m. The undusted hay contained no D D T and the dusted hay contained amounts that increased progressively with the quantity of D D T applied. Small amounts of D D T that appeared i n the eggs of the control chickens might be explained by t h e hens scattering the mash during feeding to other compartments i n the laying battery. The presence of D D T i n small amounts does not detract from the significance of the other data. W i t h an increase of D D T in the feed there is generally an increase of the com pound i n the eggs. The D D T content of the eggs from chickens fed 0, 50, 100, and 200 p.p.m. i n the mash during the first year is presented in Table I I I . The eggs of the 12 control chickens were analyzed as one group; therefore, the results in both Tables I I and I I I are the same for the control chickens. The amount of D D T found i n the eggs of the chickens fed mash containing D D T i n most cases is roughly proportional to the amount of D D T ingested.. Table III.

DDT Content of Eggs of Chickens Consuming Mash Containing DDT (1947-1948)°

. ... Months after Beginning D D T Feeding D D T Applied r to Mash, . „ , P.P.M. D D T in Eggs, P . P . M . 0 1.7*2.0 1.7*2.0 50 7.5*8.7 7.7*6.9 100 13.3*5.8 18.7*8.4 200 65.6*25.2 57.1 * 5 . 9 ° Each value is average * S.D. of 2 to 4 hens. l> Value of one hen consistently high during 3-month period. 8

y

Ώ

Ώ

τ

τγ. ι

υ

0.9*0.8 23.3±0& 18.3*10.6 43.5*12.8

In Table I V are tabulated the concentrations of D D T found i n the·tissues of the chickens killed after one year of feeding. The concentration of D D T was markedly higher in the fat than in the diet. Only small amounts of the insecticide were found in the breast and leg muscle tissues. I n each case the fatty tissues contained D D T at markedly higher levels than the eggs. Table IV.

DDT Content of Chickens Killed at End of 1 2 Months of Feeding DDT (1947-1948)

D D T Applied per Acre, Lb.

D D T Residue, P . P . M . On hay In mash 0 15 22 42

0 1 2 4 D D T Added to Mash, P.p.m.

0 2.3 3.3 6.3 50 100 200

No. of Chickens

Leg, P.P.M.

3 2 3 4

0.3 0.5 0.7 1.0

Breast, P.P.M. 0.2 0.1 0.8 0.4

1 1 2

1.8 11.6

0.2 0.5 4.0

Fat, P.P.M 30.0 29.8 44.1 103.6 502.6

The concentration of D D T in the eggs during the second year is shown i n Tables V and V I . During the first month of the second year D D T was not fed. However, 24 of the hens that had been on the experiment during the first year still had D D T present i n their AGRICULTURAL CONTROL CHEMICALS Advances in Chemistry; American Chemical Society: Washington, DC, 1950.

235

BRYSON et al.—DDT IN EGGS AND TISSUES OF CHICKENS

eggs. The 24 new pullets did not have D D T i n their eggs during the first month, but thereafter the level of D D T was about the same as that in the eggs during the first year. Table V.

DDT Content of Eggs of Chickens Consuming Mash Containing DDT-Treated Hay (1948-1949) a

DDT DDT6 Applied Residue per in Acre, Mash, Lb. P.P.M.

I

e

2

Months after Beginning D D T Feeding 3 4 5 6 7 D D T in Eggs, P . P . M .

0 1 2 4

0 0.3±0.6 0.3±0.5 0.3±0.6 0.2±0.4 0.7±0.8 0.2±0.3 0.3±0.8 0.4±0.5 0.1±0.4 2.3±0.6 2.2±0.6 3.6±1.0 2.6±0.8 2.9±0.8 2.3 3.3 1.0±0.4 2.1±0.6 0.8±0.3 3.4±0.5 3.4±0.7 3.1±0.9 6.3 6.3 0 . 2 ± 0 . 3 0 . 5 ± 0 . 4 4 . 3 ± 1 . 5 2.7*1.5 8 . 2 ± 1 . 9 7 . 9 ± 1 . 9 8 . 3 ± 2 . 8 Each value is average ± S.D. of 6 to 8 hens. b D D T content of mash by calculation. D D T not fed during first month. 16 hens were from experiment the preceding year D D T in their eggs. 16 new pullets did not have D D T in their eggs during first month.

8

9

0.8±0.7 1.2±0.4 2.6±0.8 3.1±0.9 2.7±0.9 3.1±1.3 7.6±2.0 6.9±2.2

a


c

Table VI.

and still contained

DDT Content of Eggs of Chickens Consuming Mash Containing DDT (19481 949)*

DDT in Months after Mash, lb 2 3 4 P.P.M. DDT 0 0.3±0.6 0.3±0.5 0.3±0.6 0.2±0.4 50 1.1 ± 1 . 1 15.9 ± 3 . 3 1 0 . 5 ± 8 . 8 100 3 . 2 ± 5 . 5 1 1 . 2 * 8 . 9 2 9 . 0 ± 1 . 0 2 9 . 3 ± 4 . 7 200 6 . 6 ± 9 . 3 7 . 7 ± 3 . 0 2 8 . 9 ± 4 . 5 4 8 . 4 ± 1 6 . 4

Beginning D D T Feeding 5 6" 7 8 9 in Eggs, P . P . M . 0 . 7 ± 0 . 8 0 . 2 ± 0 . 3 0.3*0.8 0 . 8 ± 0 . 7 1.2±0.4 23.0 ± 5 . 6 19.8 ± 0 . 8 22.6 ± 1 . 8 18.5 ± 2 . 1 21.0 ± 0 . 9 56.1 ± 2 . 2 5 0 . 5 ± 6 . 8 4 6 . 7 ± 6 . 9 3 2 . 2 ± 2 . 9 4 7 . 3 ± 3 . 3 6 6 . 5 ± 2 . 7 64.1 ± 8 . 1 5 1 . 9 ± 7 . 6 4 2 . 9 ± 3 . 7 5 8 . 2 ± 4 . 1

Each value is average * S.D. of 3 to 4 hens. 6 D D T was not fed during first month. 8 hens were from experiment of preceding year and still contained D D T in their eggs. 8 new pullets did not have D D T in their eggs during first month. α

Rubin et al. have shown that 1250 p.p.m. of D D T in the diet of laying hens for 12 weeks produced toxic symptoms and death. I n the work reported here the highest level of D D T used was 200 p.p.m. The authors did not observe toxic symptoms at this or lower levels. The results of this study indicate that when alfalfa is dusted with D D T and the har vested hay is used in making a mash for chickens, D D T will appear i n considerable quan tities in the tissues and eggs of chickens consuming such mash. The daily intake of D D T in this study was not sufficient to produce toxic symptoms. However, if the same level of D D T is consumed for a long period of time, D D T may accumulate i n sufficient concentra tion to produce harmful effects in the chickens, or in a human consuming the meat or eggs. Fitzhugh and Nelson (6) have shown that the withdrawal of food from animals fed a high level of D D T produced characteristic D D T tremors; D D T - t r e a t e d animals made sick by infection also showed characteristic D D T tremors. They attributed these effects to the fact that the animals were metabolizing their body fat, which contained large amounts of D D T , at an increased rate during starvation or infection. This effect could be impor tant in cases of human infection or starvation if there was a large storage of D D T i n the fat tissue.

Summary Alfalfa was dusted with 0, 1, 2, and 4 pounds of technical D D T per acre as an insecti cide and the harvested hay was ground into meal. A mash was made for laying hens containing 1 5 % alfalfa meal. D D T was also added to mash made with untreated alfalfa hay at 0, 50, 100, and 200 p.p.m. Four groups of 8 hens each were fed the mash made with the field-dusted hay and four groups of 4 hens each were fed the mash containing 0, 50, 100, and 200 p.p.m. of D D T . The D D T residue of the field-treated hay was 0.0, 15, 22, and 42 p.p.m. for the 0, 1, 2, and 4 pounds of D D T per acre treatment, respectively. During the first year of feeding the mash, D D T was present i n the eggs at a maxi mum concentration of 1.7, 2.1, 3.7, and 4.1 p.p.m. from the hens consuming the mashes AGRICULTURAL CONTROL CHEMICALS Advances in Chemistry; American Chemical Society: Washington, DC, 1950.

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ADVANCES IN CHEMISTRY SERIES

containing alfalfa treated with 0, 1, 2, and 4 pounds of D D T per acre, respectively. T h e eggs from the hens consuming mashes containing 0, 50, 100, and 200 p.p.m. of D D T c o n tained maximum amounts of 1.7, 23.3, 18.7, and 65.6 p.p.m. of D D T , respectively. The highest concentration of D D T was found i n the fat of the chickens killed after being fed D D T for 1 year. The average amount present was 30.0, 29.8, 44.1, 103.6, and 502.6 p.p.m. when hay treated with 1, 2, and 4 pounds of D D T per acre and 100 and 200 p.p.m. of D D T , respectively, was fed. M u s c l e tissues varied from 0.1 to 11.6 p.p.m. The concentrations of D D T i n the eggs during the second year of feeding the D D T treated mash were generally slightly higher than the first year.

Literature Cited (1) Biddulph, C., Bateman, G. Q., Bryson, M. J., Harris, J. R., Greenwood, D. Α., Binns, W.,


Miner, M . L . , Harris, L . E . , and Madsen, L . L . , A D V A N C E S I N CHEMISTRY SERIES, 1, 237

(2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16) (17) (18) (19) (20) (21)

(1950). Carter, R. H . , Ind. Eng. Chem., 40, 716 (1948). Carter, R. H . , Hubanks, P. E . , Mann, H . D., Alexander, L . M . , and Schopmeyer, G . H., Science, 107, 347 (1948). Carter, R. H . , Hubanks, P. E., Mann, H . D., Zeller, J . H . , and Hankins, O. G., J. Animal Sci., 7, 509 (1948). Fitzhugh, O. G., Ind. Eng. Chem., 40, 704 (1948). Fitzhugh, O. G., and Nelson, Α. Α., J. Pharmacol. Exptl. Therap., 89, 18 (1947). Harris, L . E., Biddulph, C., Greenwood, D . Α., Binns, W., Miner, M . L., and Madsen, L . L . , unpublished data. Harris, J . R., Biddulph, C., Greenwood, D. Α., Harris, L . E., Bryson, M . J., Binns, W., Miner, M . L., and Madsen, L . L., Arch. Biochem., 21, 370 (1949). Laug, E. P., and Fitzhugh, O. G., J. Pharmacol. Exptl. Therap., 87, 18 (1946). Lieberman, F . V., U . S. Dept. Agr., Bur. Entomol. Plant Quarantine, Bull. E-658 (1945). Lieberman, F . V., and Hare, Q. Α., Ibid., Bull. E-697 (1946). Marsden, S. J . , and Bird, H . R., Poultry Sci., 26, 3 (1947). Orr, L . W., and Mott, L . O., J. Econ. Entomol., 38, 428 (1945). Rubin, M . , Bird, H . R., Green, N . , and Carter, R. H . , Poultry Sci., 26, 410 (1947). Schechter, M . S., Pogorelskin, Μ. Α., and Haller, H . L., Anal. Chem., 19, 51 (1947). Schechter, M . S., Soloway, S. B., Hayes, R. Α., and Haller, H . L., Ind. Eng. Chem., Anal. Ed., 17, 704 (1945). Smith, R. F., and Michelbacher, A. E., J. Econ. Entomol., 39, 638 (1946). Telford, H . S., and Guthrie, J . E., Science, 102, 647 (1945). Umhoefer, R. R., Ind. Eng. Chem., Anal. Ed., 15, 383 (1943). Woodward, G., and Ofner, R. R., Federation Am. Soc. Exptl. Biol., Federation Proc., 5, 215 (1946). Woodward, G., Ofner, R. R., and Montgomery, C. M . , Science, 102, 177 (1945).

R E S E A R C H supported in part by a research grant from the Division of Research Grants and Fellowships, N a tional Institutes of Health, U . S. Public Health Service, and published with the approval of the director of the Utah Agricultural Experiment Station.


DDT in Eggs and Tissues of Chickens Fed Varying Levels of DDT

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