Journal of Toxicology and Environmental Health

ISSN: 0098-4108 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/uteh19

Lack of effect of dietary diethylstilbestrol on reproductive performance S. L. Liu , W. T. Allaben & G. H. Gass To cite this article: S. L. Liu , W. T. Allaben & G. H. Gass (1976) Lack of effect of dietary diethylstilbestrol on reproductive performance, Journal of Toxicology and Environmental Health, 1:5, 817-821, DOI: 10.1080/15287397609529380 To link to this article: http://dx.doi.org/10.1080/15287397609529380

Published online: 20 Oct 2009.

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Date: 15 November 2015, At: 04:32

LACK OF EFFECT OF DIETARY DIETHYLSTILBESTROL ON REPRODUCTIVE PERFORMANCE S. L. Liu, W. T. Allaben Department of Physiology, Southern Illinois University, Carbondale, Illinois G. H. Gass

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National Center for Toxicological Research, Jefferson, Arkansas

Groups of male or female mice were pretreated for 2 wk and 1 wk, respectively, with flesh (liver or muscle) diets prepared from steers. In one experiment diethylstilbestrol (DES) was added to the diet at 0.5 or 5.0 ppb. In a second experiment diets prepared from DBS-implanted steer flesh (liver or muscle) were fed. Tissues used in the control diet and DES-added diets were from DES-free steers. The animals were allowed to mate and diets continued until the first litter was delivered. Increasing DES levels in either liver or muscle diets or flesh from DES-implanted steers resulted in no significant differences either in litter size or in the number of fertile male or female mice between the control group and experimental groups. The offspring from each litter were mated and showed no significant difference in their reproductive performance. No abnormalities were noted in any offspring.

INTRODUCTION Natural and synthetic estrogens have long been recognized to increase feeding efficiency and to promote growth in cattle, whether administered orally or by subcutaneous implants. The Department of Agriculture estimates that diethylstilbestrol (DES) implants reduce the length of the feeding period by 15% and may save about 225 kg feed per animal (Hearing before Subcommittee of House Committee on Government Operations, 1971). Although this synthetic estrogen has been widely used as an implant or an additive in the diets of cattle raised for human consumption, oral DES has long been known to induce mammary carcinoma in certain strains of mice (Gass et al., 1964, 1974; Okey and Gass, 1968; Shimkin and Grady, 1940). Therefore, the use of DES was banned because DES or DES conjugates were detected in the livers of test steers (Hearing before Subcommitte of House Committee on Government Operations, 1971). This work was supported in part by the Fraternal Order of Eagles. W. T. Allaben's present address is National Center for Toxicological Research, Jefferson, Arkansas 72079. Requests for reprints should be sent to G. H. Gass, National Center for Toxicological Research, Jefferson, Arkansas 72079. 817 Journal of Toxicology and Environmental Health, 1:817-821, 1976 Copyright © 1976 by Hemisphere Publishing Corporation

818

S. L. LIUETAL.

A recent report by a French researcher (Ferrando et al., 1971) has purportedly shown that female mice and rats that had consumed rations of 20% lyophilized meat from DES-implanted cattle were incapable of reproducing. The fact that DES has antifertility properties is not disputed. The problem that exists revolves around a single issue: edible tissues of cattle that have been implanted with estrogenic compounds may have a detrimental effect on the consumer. This particular study examines the effect that diets of calf muscle or liver incorporated with DES and a diet of muscle or liver from DES-implanted cattle had on the reproductive performance of Swiss mice.

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MATERIALS AND METHODS Male and female mice for this experiment were obtained from our breeding colony. To assure that all animals used were fertile, they were mated and produced at least one litter prior to their use in the study. The animals were housed in polycarbonate shoe box cages with wood chips for bedding. Water and diets were administered ad libitum. Groups of male and groups of female mice were pretreated for 2 wk and for 1 wk, respectively, with test diets as follows: control diet (80% liver or muscle plus 20% chow); DES diet A (80% liver or muscle plus 20% chow plus 0.5 ppb DES); DES diet B (80% liver or muscle plus 20% chow plus 5.0 ppb DES); and DES-implanted meat diet (80% liver or muscle from 30 mg DES-implanted calf plus 20% chow). Meat for DES-implanted meat diet was obtained from a calf slaughtered 15 days following the implant of 30 mg DES pellets in the ear. Control diet, DES diet A, and DES diet B were prepared using muscle or liver obtained from DES-free calves. Diets containing DES were prepared as previously described by Umberger and co-workers (1958; Gass et al., 1974). Each pretreated male was placed into a cage with one pretreated female of the corresponding diet group. The animals were allowed to mate and the diets continued until the first litter was delivered. At that time the males were removed and the diets discontinued. The females were observed for 25 days, and the number of litters and litter size were recorded. All pups were examined for any gross abnormality. RESULTS Animals that were either on liver/chow/DES or muscle/chow/DES diets were found to have no significant difference in either the number of litters produced or in the average size of the litter born to each female. Summaries of the data obtained are illustrated in Table 1. Even when the animals were fed diets containing 5.0 ppb, which is 10 times the maximum dose thus far recovered from the livers of DES-implanted cattle (Rumsey et al., 1974),

DES AND REPRODUCTIVE PERFORMANCE

TABLE 1. Effect of Diethylstilbestrol Muscle Diet Diet amount of DES (ppb)

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Control 0.5 ppb DES liver diet 5.0 ppb DES liver diet 0.5 ppb DES muscle diet 5.0 ppb DES muscle diet

819

(DES) in 80% Liver or

No. of mice"

No. of litters

No. of offspring per litter (average)

20

10

10.5

20

9

10.0

20

10

9.1

20

10

11.7

20

10

10.7

"One male and one female per cage.

there was no significant difference in the reproductive performance of animals. Table 2 presents the feeding of muscle or liver from the calf given one 30-mg DES implant and slaughtered 15 days after implantation. The feeding did not significantly influence either litter size or fertility of male and female mice. The Fj generation from DES-treated parents is shown in Table 3. The offspring from both control and treated groups showed no significant difference in their ability to reproduce or in the number of offspring produced per litter. DISCUSSION In a study carried out at Beltsville, Maryland, the maximum amount of DES thus far identified from DES-implanted cattle (30-mg pellets in ear) was TABLE 2. Effect of 80% Liver or Muscle from Diethylstilbestrol (DES)-lmplanted Calf

Diet Control Liver from implanted calf Muscle from implanted calf

No. of mice 0

No. of offspring per litter (average)

No. of litters

20

9

9.0

20

10

8.5

20

8

"One male and one female per cage.

,

11.5

820

S. L. LIUETAL.

TABLE 3. Fertility of F, Generation of Diethylstilbestrol (DES)-Treated Parents

Parents' diet (ppb DES) Control 0.5 ppb 5.0 ppb

No. of mice0

No. of litters

No. of offspring per litter (average)

20 20 20

9 10 10

9.1 10.7 9.1

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"One male and one female per cage.

0.5 ppb in the liver with none detected in the muscle tissue (Rumsey et al., 1974). Animals were killed at 30, 60, 90, or 120 days following implanting. However, in our study we killed 15 days following implantation. Based on USDA findings, the muscle and liver from the implanted calf in our study should contain either the maximum detectable amount of DES or higher than that. In addition, the dose levels of DES incorporated into the diets fed to Swiss mice in this study were (1) 0.5 ppb, the maximum found by the USDA and (2) 5.0 ppb, 10 times that found in cattle so treated. Data obtained from this experiment do not support the hypothesis that edible tissues from DES-implanted cattle could have an unfavorable influence on the reproductive function of mice.

SUMMARY Groups of male and groups of female mice were pretreated for 2 wk and 1 wk, respectively, with DES diets as follows: control (80% liver or muscle + 20% chow); DES diet A (80% liver or muscle + 20% chow +0.5 ppb DES); DES diet B (80% liver or muscle + 20% chow + 5.0 ppb DES); and DES-implanted meat diet (80% liver or muscle from a 30 mg DES-implanted calf + 20% chow). Tissues used in control diet and DES diet A and B were from DES-free steers. Each pretreated male was placed into a cage with one pretreated female. The animals were allowed to mate and diets continued until the first litter was delivered. The number of litters and litter size were observed, as were any abnormalities of the pups. Increasing DES levels in either liver or muscle diets resulted in no significant differences either in litter size or in the number of fertile male or female mice between the control group and experimental groups. The offspring from each litter were mated and showed no significant difference in their reproductive performance. REFERENCES Ferrando, M. R., Henry, M. N., Valette, J. P. and Parodi, A. 1971. Rural economy-Effect of a diet containing meat from diethylstilbestrol-implanted calves on rat testicular histology. C.R. Acad. Sci. [3] (Paris) 275:587.

DES AND REPRODUCTIVE PERFORMANCE

Gass,

G.

H.,

Coats, D.

and

Graham,

N.

1964. Carcinogenic

821

dose response curve to oral

diethylstilbestrol. J. Natl. Cancer Inst. 33:971. Gass, G. H., Brown, J. and Okey, A. B. 1974. Carcinogenic effects of oral diethylstilbestrol on C3H male mice with and without the mammary tumor virus. J. Natl. Cancer Inst. 53:1369. Hearing before Subcommittee of House Committee on Government Operations. Regulation of feed additives in medicated animal feeds. 1971. 92nd Congress, 1st Session at 446-58. Okey, A. B. and Gass, G. H. 1968. Continuous versus cyclic estrogen administration: Mammary carcinoma in C3H mice. J. Natl. Cancer Inst. 40:225. Rumsey, T. S., Oltjen, R. R. and Kozak, A. S. 1974. Implant absorption, performance and tissue analysis for beef steers implanted with diethylstilbestrol and fed an all-concentrate diet. J. Anim. Sci. 39:1193. Shimkin, M. B. and Grady, H. B. 1940. Mammary carcinomas in mice following oral administration of stilbestrol. Proc. Soc. Exp. Biol. Med. 45:246. Umberger, E. J., Gass, G. H. and Curtis, J. M. 1958. Design of a biological assay method for the detection and estimation of estrogenic residues in the edible tissues of domestic animals treated with

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estrogen. Endocrinology

63:806. Received December 15, 1975 Accepted December 16, 1975

Lack of effect of dietary diethylstilbestrol on reproductive performance.

Groups of male or female mice were pretreated for 2 wk and 1 wk, respectively, with flesh (liver or muscle) diets prepared from steers. In one experim...
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