Veterinary World

     Open access and peer reviewed journal  

ISSN (Online): 2231-0916


Home l Editorial board l Instructions for authors l Reviewer guideline l Open access policy l Archives l FAQ

Open Access

Copyright: The authors. This article is an open access article licensed under the terms of the Creative Commons Attribution License

( which permits unrestricted use, distribution and reproduction in any medium, provided the work is properly cited.

Research (Published online: 17-06-2015)

9.  Evaluation of subacute bisphenol – A toxicity on male reproductive system - S. S. Karnam, R. C. Ghosh, S. Mondal and M. Mondal

Veterinary World, 8(6): 738-744



   doi: 10.14202/vetworld.2015.738-744


S. S. Karnam: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Chhattisgarh Kamdhenu

Vishwavidyalaya, Durg, Chhattisgarh, India;

R. C. Ghosh: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Chhattisgarh Kamdhenu

Vishwavidyalaya, Durg, Chhattisgarh, India;

S. Mondal: Department of Veterinary Pathology, West Bengal University of Animal and Fishery Sciences, Kolkata, West Bengal, India;

M. Mondal: Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University Rewa, Madhya Pradesh, India;


Received: 19-01-2015, Revised: 05-05-2015, Accepted: 13-05-2015, Published online: 17-06-2015


Corresponding author: S. S. Karnam, e-mail:

Citation: Karnam SS, Ghosh RC, Mondal S, Mondal M (2015) Evaluation of subacute bisphenol - A toxicity on male reproductive system, Veterinary World 8(6): 738-744.

Aim: The aim was to evaluate the effect of multiple oral administration of bisphenol A (BPA) for 28 days on seminal characteristic on mammal using Wistar rat as a model.

Materials and Methods: Rats were randomly divided into five different groups having 6 male rats in each group. The doses chosen were 50, 200, and 600 mg/kg body weight for Groups III, IV and V, respectively, based on preliminary dose range finding study and Group II served as vehicle control and Group I was negative control.

Results: Reproductive study in the BPA-treated rats on day 28 revealed that there was significant (p0.05) reduction in the epididymal sperm count of rats of Group IV and significant (p0.01) decrease in Group V. Sperm motility percentage, dead count percentage, head and tail abnormality percentage were found to be significantly (p0.01) increased in rats of BPA-treated groups as compared to rats of control groups. Testes showed necrosis of germinal layer and spermatogonial cells in the seminiferous tubules. Hematological examination revealed significant (p0.01) decrease in the mean values of total erythrocyte count (TEC), total leukocyte count (TLC), hemoglobin, packed cell volume, and there was also significant (p0.05) lymphocytopenia in treated animals.

Conclusion: It can be concluded from this study that subacute toxicity of BPA caused a reduction in the epididymal sperm count, sperm motility, dead count, head and tail abnormality, as well as hematological indices such as TLC, TEC etc. Hence, it appears that BPA affects the germ cells leading to impairment in the spermatogenesis, and thus having its property as reproductive toxicant and it also suppresses bone marrow functioning, which leads to normocytic hypochromic anemia in rats.

Keywords: bisphenol A, hematology, rat, spermatozoa, testes.

1. Staples, C.A., Dorn, P.B., Klecka, G.M., O'Block, S.T. and Harris, L.R. (1998) A review of the environmental fate, effects and exposures of bisphenol A. Chemosphere, 36: 2149-2173.
2. Flint, S., Markle, T., Thompson, S. and Wallace, E. (2012) Bisphenol A exposure, effects, and policy: A wildlife perspective. J. Environ. Manage., 104: 19-34.
3. Roberts, R. (2012) BPA exposure and health effects: Educating physicians and patients. Am. Fam. Physician, 85(11): 1040-1044.
4. Schierow, L.J. and Lister, L.A. (2010) Bisphenol A (BPA) in plastics and possible human health effects. Congr. Res. Serv., 7-5700:1-10.
5. Fernandez, S.V. and Russo, J. (2010) Estrogen and xenoestrogens in breast cancer. Toxicol. Pathol., 38: 110-122.
PMid:19933552 PMCid:PMC2907875
6. Howdeshell, K.L., Peterman, P.H., Judy, B.M., Taylor, J.A., Orazio, C.E., Ruhlen, R.L., Vomsaal, F.S. and Welshons, W.V. (2003) Bisphenol A is released from used polycarbonate animal cages into water at room temperature. Environ. Health Perspect., 111: 1180-1187.
PMid:12842771 PMCid:PMC1241572
7. Joseph, M.B., Bruce, P.L., Antonia, M.C., Sirad Deria, J.K., Chanelle, J.H. and Scott, A.V. (2014) Early-life bisphenol a exposure and child body mass index: A prospective cohort study. Environ. Health Perspect., 122(11): 1239-1245.
8. Del Val, G.M. and Robledano, P.M. (2013) In vivo serial sampling of epididymal sperm in mice. Lab. Anim., 47: 168-174.
9. Saalu, L.C., Kpela, T., Benebo, A.S., Oyewopo, A.O., Anifowope, E.O. and Oguntola, J.A. (2010) The dose-dependent testiculoprotective and testiculotoxic potentials of telfairia occidentalis hook f. Leaves extract in rat. Int. J. Appl. Res. Nat. Prod., 3: 27-38.
10. Luke, L., Vicens, A., Tourmente, M. and Roldan, E.R.S. (2014) Evolution of protamine genes and changes in sperm head phenotype in rodents. Biol. Reprod., 90: 67.
11. Correia, S., Oliveira, P.F., Guerreiro, P.M., Lopes, G., Alves, M.G., Canário, A.V.M., Cavaco, J.E. and Socorro, S. (2013) Sperm parameters and epididymis function in transgenic rats overexpressing the Ca2+-binding protein regucalcin: A hidden role for Ca2+ in sperm maturation? Mol. Hum. Reprod., 19: 581-589.
12. Longo, U.G.F., Ruzzini, L., Rabitti, C., Morini, S., Maffulli, N. and Denaro, V. (2009) Characteristics at haematoxylin and eosin staining of ruptures of the long head of the bicepstendon. Br. J. Sports Med., 43: 603-607.
13. Kourouma, A., Peng, D., Chao, Q., Yaima, M.L.T., Changjiang, L., Chengmin, W., Wenjuan, F., Suqin, Q., Tingting, Y. and Kedi, Y. (2014) Bisphenol A induced reactive oxygen species (ROS) in the liver and affect epididymal semen quality in adults sprague-dawley rats. J. Toxicol. Environ. Health Sci., 6(4): 103-112.
14. Manfo, F.P., Jubendradass, R., Nantia, E.A., Moundipa, P.F. and Mathur, P.P. (2014) Adverse effects of bisphenol A on male reproductive function. Rev. Environ. Contam. Toxicol., 228: 57-82.
15. Meeker, J.D., Ehrlich, S., Toth, T.L., Wright, D.L., Calafat, A.M., Trisini, A.T., Ye, X. and Hauser, R. (2010) Semen quality and sperm DNA damage in relation to urinary bisphenol A among men from an infertility clinic. Reprod. Toxicol., 30(4): 532-539.
PMid:20656017 PMCid:PMC2993767
16. Li, D.K., Zhou, Z., Miao, M., He, Y., Wang, J., Ferber, J., Herrinton, L.J., Gao, E. and Yuan, W.(2011) Urine bisphenol A (BPA) level in relation to semen quality. Fertil. Steril., 95(2): 625-630.
17. Jin, P., Wang, X., Chang, F., Bai, Y., Li, Y., Zhou, R. and Chen, L. (2013) Low dose bisphenol A aimpairs spermatogenesis by suppressing reproductive hormone production and promoting germ cell apoptosis in adult rats. J. Biomed. Res., 27(2): 135-144.
PMid:23554804 PMCid:PMC3602871
18. Salian, S., Doshi, T. and Vanage, G. (2009) Neonatal exposure of male rats to bisphenol A impairsfertility and expression of sertoli cell junctional proteins in the testis. Toxicology, 265(1-2): 56-67.
19. Salian, S., Doshi, T. and Vanage, G. (2011) Perinatal exposure of rats to bisphenol A affects fertility of male offspring-an overview. Reprod. Toxicol., 31(3): 359-62.
20. Qiu, L.L., Wang, X., Zhang, X.H., Zhang, Z., Gu, J., Liu, L., Wang, Y., Wang, X. and Wang, S.L. (2013) Decreased androgen receptor expression may contribute to spermatogenesis failure in rats exposed to low concentration of bisphenol A. Toxicol. Lett., 219(2): 116-124.
21. Isling, L.K., Boberg, J., Jacobsen, P.R., Mandrup, K.R., Axelstad, M., Christiansen, S., Vinggaard, A.M., Taxvig, C., Kortenkamp, A. and Hass, U. (2014) Late-Life effects on rat reproductive system after development exposure to mixture of endocrine disruptors. Reproduction, 147(4): 465-476.
22. Takahashi, O. and Oishi, S. (2003) Testicular toxicity of dietarily or parenterally administered bisphenol A in rats and mice. Food Chem. Toxicol., 41: 1035-1044.
23. Liu, C., Duan, W., Li, R., Xu, S., Zhang, L., Chen, C., He, M., Lu, Y., Wu, H., Pi, H., Luo, X., Zhang, Y., Zhong, M., Yu, Z. and Zhou, Z. (2013) Exposure to bisphenol A disrupts meiotic progression during spermatogenesis in adult rats through estrogen-like activity. Cell Death Dis., 4: 676.
PMid:23788033 PMCid:PMC3702305
24. Benjamin, L.L.T., Normadiah, M.K. and Mustafa, A.M. (2003) Assessment of pubertal development in juvenile male rats after sub-acute exposure to bisphenol A and nonylphenol. Toxicol. Lett., 143(3): 261-270.
25. Jurewicz, J., Hanke, W., Radwan, M. and Bonde, J.P. (2009) Environmental factors and semenquality. Int. J. Occup. Med. Environ. Health, 22(4): 305-329.
26. Vilela, J., Hartmann, A., Silva, E.F., Cardoso, T., Corcini, C.D., Varela-Junior, A.S., Martinez, P.E. and Colares, E.P. (2014) Sperm impairments in adult vesper mice (Calomys laucha) caused by in uteroexposure to bisphenol A. Andrologia, 46(9): 971-978.
27. Hulak, M., Gazo, I., Shaliutina, A. and Linhartova, P. (2013) In vitro effects of bisphenol A on thequality parameters, oxidative stress, DNA integrity and adenosine triphosphate content in starlet (Acipenser ruthenus) spermatozoa. Comp. Biochem. Physiol. C. Toxicol. Pharmacol., 158(2): 64-71.
28. Lizarraga, D., Vinardell, M.P., Noé, V., van Delft, J.H., Alcarraz-Vizán, G., van Breda, S.G., Staal, Y., Günther, U.L., Reed, M.A., Ciudad, C.J., Torres, J.L. and Cascante, M. (2011) A lyophilized red grape pomace containing proanthocyanidin-rich dietary fiber induces genetic and metabolic alterations in colon mucosa of female C57BL/6J mice. J. Nutr., 141: 1597-1604.
29. Clegg, E.D., Perreault, S.D. and Klinefelter, G.R. (2001) Assessment of male reproductive toxicity. In: Hayes, A.W., editor. Principles and Methods of Toxicology. 4th ed. Taylor & Francis, Philadelphia, PA. p11263-1300.
30. Smith, M.C. (1986) Infertility in the buck. In: Marrow, D.A., editor. Current Therapy in Theriogenology. 2nd ed. W.B. Saunders, Philadelphia. p622-624.
31. Sangai, N.P., Verma, R.J. and Trivedi, M.H. (2014) Testing the efficacy of quercetin in mitigating bisphenol A toxicity in liver and kidney of mice. Toxicol. Ind. Health, 30: 581-597.
32. ACT 2011 Annual Meeting Abstracts. (2012) Int. J. Toxicol., 31: 97-126.
33. Lemasters, G.K. and Selevan, S.G. (1993) Toxic exposures and reproduction: A view of epidemiology and surveillance. In: Scialli, A.R. and Zinaman, M.J., editors. Reproductive Toxicology and Infertility. McGraw Hill, New York. p307-321.
34. Kim, S.T., Omurtag, K. and Moley, K.H. (2012) Decreased spermatogenesis, fertility, and altered Slc2A expression in Akt1–/– and Akt2–/– testes and sperm. Reprod. Sci., 19: 31-42.
PMid:22228739 PMCid:PMC3343127
35. Takzare, N., Hosseini, M.J., Mortazavi, S.H., Safaie, S. and Moradi, R. (2011) The effect of Achillea millefolium extract on spermatogenesis of male Wistar rats. Hum. Exp. Toxicol., 30: 328-334.
36. Zhou, D., Zhang, J. and Wang, H. (2011) Assessment of the potential reproductive toxicity of long-term exposure of adult male rats to low-dose formaldehyde. Toxicol. Ind. Health, 27: 591-598.
37. Awal, M.A., Kurohmaru, M., Mizukami, T., Bibin, B.A., Kanai, Y. and Hayashi, Y. (2004) Effects of bisphenol A on testes in prepubrtal male wistar rats: In vitro study. J. Med. Sci., 4(3): 225-231.
38. Delclos, K.B., Camacho, L., Lewis, S.M., Vanlandingham, M.M., Latendresse, J.R., Greg, R., Olson, G.R., Kelly, J., Davis, K.J., Ralph, E., Patton, R.E., Costa, G.G.D., Woodling, K.A., Bryant, M.S., Chidambaram, M., Trbojevich, R., Juliar, B.E., Felton, R.P. and Thorn, B.T. (2014) Toxicity evaluation of bisphenol A administered by gavage to sprague dawley rats from gestation day 6 through postnatal day 90. Toxicol. Sci., 139: 174-197.
PMid:24496637 PMCid:PMC4038785
39. Krishnan, A.V., Stathis, P., Permuth, S.F., Tokes, L. and Feldman, D. (1993) Bisphenol A: Anestrogenic substance is released from polycarbonate flasks during autoclaving. Endocrinology, 132: 2279-2286.
40. Furukawa, F., Nishikawa, A., Mitsui, M., Sato, M., Suzuki, J., Imazawa, T. and Takahashi, M. (1994) A 13-week subchronic toxicity study of bisphenol A in B6C3F1 mice. Bull. Natl. Instur. Hyg. Sci., ???: ???. 112:89-96
41. Matsumoto, K., Ochiai, T., Sekita, K., Uchida, O., Furuya, T. and Kurokawa, Y. (1991) Chronictoxicity of 2,4,6-tri-tert-butylphenol in rats. J. Toxicol. Sci., 16(4): 167-179.
42. Tilak, K.S., Veeraiah, K. and Butchiram, M.S. (2007) Effect of phenol on haematological components of Indian major carps Catla catla, Labeo rohita and Cirrhinus mrigala. J. Environ. Biol., 28(2): 177-179.
43. Woo, G.H., Shibutani, M., Ichiki, T., Hamamura, M., Lee, K.Y., Inoue, K. and Hirose, M. (2006) A repeated 28-day oral dose toxicity study of nonylphenol in rats, based on the 'Enhanced OECD Test Guideline 407' for screening of endocrine-disrupting chemicals. J. Arch. Toxicol., 81(2): 77-88.
44. Santa, N., Bhagyabati, S., Jeetenkumar, T., Motu, T., Lokeshwar, K. and Kulachandra, M.S. (2003) Phenol-induced haemolytic anaemia. J. Indian Acade. Clin. Med., 4(2): 174-175.
45. Benjamin, M.M. (2001) Outline of Veterinary Clinical Pathology. 3rd ed. Colorado State University, U.S.A.
46. Hall, R.L. (2001) Principles of clinical pathology for toxicology studies. In: Hayes, A.W., editor. Principles and Methods of Toxicology. 4th ed. Taylor & Francis, Philadelphia. p1301-1381.