Effect of Prophylactic Supplementation of Vitamin E and Se on Antioxidant Enzymes during Endotoxic Shock in Buffalo Calves

  • Sandhu, T.S. (Veterinary Officer, Department of Animal Husbandry, Government of Punjab) ;
  • Singha, S.P.S (Veterinary Officer, Department of Animal Husbandry, Government of Punjab)
  • Received : 2002.09.17
  • Accepted : 2003.06.23
  • Published : 2003.11.01


This study was conducted to determine the effects of prophylactic supplementation of vitamin E and Se on oxidative damage and antioxidant status. Fifteen healthy male buffalo (Bubalus bubalis) calves between the age of 6 to12 months were divided into three groups of five animals each: Group I-control, group II-endotoxic shock group infused with lyophilized E coli endotoxin @ 5 ${\mu}g$/kg body wt, and group III-supplemented with vitamin E @ 250 mg and Se @ 7.5 mg, one month prior to induction of endotoxic shock. All the animals in group II and group III exhibited signs of endotoxic shock. When the endotoxic shock was induced, there was significant (p<0.05) increase in the circulating levels of malonyl dialdehyde MDA (an indicator of lipid peroxidation). In the supplemented group III the magnitude of formation of MDA was also less as compared to group II at every stage of study. There was significant (p<0.05) decrease in circulating levels of SOD, GSH-Px, Catalase and G-6-PD activity from the normal (0 h) value with passage of time. As a result of endotoxic shock, these values reached a lowest value, and then showed a tendency towards the 0 h value. Prophylactic supplementation with vitamin E and Se was successful in reducing the quantum of oxidative damage due to formation of free radicals because of endotoxic shock.


Buffalo Calves;Vitamin E;$\alpha$-tocopherol;Se Lipid Peroxidation;Endotoxic Shock;SOD;GSH-Px;G-6-PD;Catalase


  1. Barja, G., S. S. Kadenas, C. Rojar, R. Perez-campo, M. Lopez-Torres, J. Trat and R. Pamploma. 1996. Effect of dietary vitamin E levels on fatty acid profiles and non-enzymatic lipid peroxidation in the guinea pig liver. Lipids. 9:963-17.
  2. Beckman, J. S., R. L. Minor and B. A. Freeman. 1986. Augmentation of antioxidant enzymes in vascular endothelium. J. Free Radic. Biol. Med. 2(5-6):359-65.
  3. Cadenas, S., C. Rojas, R. Perez-campo, M. Lopaz-Torres and G. Barja. 1995. Vitamin E protects guinea pig liver from lipid peroxidation without depressing levels of antioxidants. Int J. Biochem. Cell Biol. 11:1175-81.
  4. Halliwell, B., M. A. Murica, S. Chirico and O. I. Aruoma. 1995. Free radicals and antioxidants in food and in vivo: what they do and how they work. Crit. Rev. Food Sci. Nutr. 35:7-20.
  5. Kadota, T., H. Ono, T. Aihara, H. Mochizuki and S. Tamakuma. 1989. The role of oxygen free radical and free radical scavenger in septic and hemorrhagic shock. Nippon Geka Gakkai Zasshi. 9:1366-9.
  6. Mueller, S., H. D. Riedel and W. Stremmel. 1997. Direct evidence for catalase as the predominant $H_2O_2$ removing enzyme in human erythrocytes. Blood 90(12):4973-8.
  7. Murray, R. K., D. K. Granner, P. A. Mayes and V. W. Rodwell. 1996. In 'Harper's Biochemistry' 24th edn. Prentice Hall International, USA.
  8. Osame, S., H. Yamaguchi and S. Ichijo. 1992. Changes in blood tocopherol, selenium and lipid peroxide levels in pregnant cows injected with tocopherol and selenium. J. Japan Vet. Med. Assoc. 45:543-46.
  9. Kayden, H. J., C. K. Chow and L. K. Bjornson. 1973. Spectrophotometric method for determination of tocopherol in red blood cells. J. Lipid Res. 14:533-540.
  10. Sakaguchi, S., N. Kanda, C. C. Hsu and O. Sakaguchi. 1981. Lipid peroxide formation and membrane damage in endotoxinpoisoned mice. Microbiol. Immunol. 25(3):229-44.
  11. Portoles, M. T., M. Catala, A. Anton and R. Pagani. 1996. Hepatic response to the oxidative stress induced by E. coli endotoxin; glutathione as an index of the acute phase during the endotoxic shock. Mol. Cell Biochem. 159(2):115-121.
  12. Hugo, E. A. 1984. Catalases in vitro. Methods Enzymol. 105:270.
  13. Pamela, C. C. and A. H. Richard. 1994. Lippincott’s Illustrated Reviews: Biochemistry 2nd ed. Lippincott Raven Publishers, Pennsylvania.
  14. Chow, C. K. 1991. Nutritional influence on cellular antioxidant defense system. Am. J. Clin. Nutr. 32:1066-1081.
  15. Sakaguchi, O., N. Kanda, S. Sakaguchi, C. C. Hsu and H. Abe. 1981. Effect of alpha-tocopherol on endotoxicosis. Microbiol Immunol. 25(8):787-99.
  16. Hafeman, D. G., R. A. Sunde and W. G. Hoekstra. 1974. Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat. J. Nutr. 104:580-587.
  17. Halliwell, B. 1999. Antioxidant defence mechanism: from the beginning to the end (of the beginning). Free Radic res. 4:261-72.
  18. Nishikimi, M., N. A. Rao and K. Yagi. 1972. The occurrence of superoxide anion in the reaction of reduced phenazine methosufate and molecular oxygen. Biochem. Biophys. Res. Commun. 46:849-854.
  19. Yoshida, A. 1973. Hemolytic anemia and G6PD deficiency. Science 179:532-537.
  20. Kayden, H. J., C. K. Chow and L. K. Bjornson. 1973. Spectrophotometric method for determination of tocopherol in red blood cells. J. Lipid Res. 14:533-540.
  21. Burton, G. W. and M. G. Traber. 1990. Vitamin E: antioxidant activity, biokinetics and bioavailability. Ann. Rev. Nutr. 10:357-382.
  22. Mayes, P. A. 1996. Structure and function of lipid soluble vitamins in: (Ed. R. K. Murray, D. K. Granner, P. A. Mayes and V. M. Rodwell) Harper's Biochemistry. 34th edn. pp. 614-624.
  23. Turrens, J. F., J. D. Crapo and B. A. Freeman. 1984. Protection against oxygen toxicity by intravenous injection of liposome entrapped catalase and superoxide dismutase J. Clin. Invest. 73(1):87-95.
  24. Harvey, J. W. 1989. Erythrocyte metabolism in ruminants: (Ed. J. Kaneko) Clinical Biochemistry of Domestic Animals. 4th edn, pp. 185-234. Academic Press, New York.
  25. Pironi, L., L. L. Cushman and B. C. Johnson. 1996. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal. Biochem. 16:359-64.
  26. Nagaraja, T. G., E. E. Bartley, H. D. Anthony, H. W. Leipold and L. R. Fina. 1979. Endotoxin shock in calves from intravenous injection of Rumen bacterial endotoxin. J. Anim. Sci. 49(2):567-571.
  27. Prentice hall International Limited, London. Mills, G. C. 1958. The purification and properties of glutathione peroxidase of erythrocytes. J. Biol. Chem. 234:502-506.
  28. Kovacheva, S. and S. Ribarov. 1996. Lipid peroxidation in the lung of rats exposed to endotoxin: effect of vitamin E supplementation. Pharmacol Toxicol. 79 (4):177-82.
  29. Placer, Z. A., L. L. Cushman and B. C. Johnson. 1966. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal. Biochem. 16:359-365.
  30. Kurt, A. Z., O. P. Jones and S. D. Perreault. 1997. Glutathione oxidation is associated with altered microtubule function in mature Hamster oocytes. Biology reproduction 57:1413-19.
  31. Deutsch, J. 1978. Maleimide as an inhibitor in measurement of erythrocyte glucose-6-phosphate dehydrogenase activity. Clin Chem. 24:885-889.