Effect of Molybdenum Induced Copper Deficiency on Peripheral Blood Cells and Bone Marrow in Buffalo Calves

  • Randhawa, C.S. (Department of Clinical Veterinary Medicine, Ethics and Jurisprudence, Punjab Agricultural University) ;
  • Randhawa, S.S. (Department of Clinical Veterinary Medicine, Ethics and Jurisprudence, Punjab Agricultural University) ;
  • Sood, N.K. (Department of Veterinary Pathology, Punjab Agricultural University)
  • Received : 2001.07.05
  • Accepted : 2001.11.15
  • Published : 2002.04.01


Copper deficiency was induced in eight male buffalo calves by adding molybdenum (30 ppm wet basis) to their diet. Copper status was monitored from the liver copper concentration and a level below 30 ppm (DM basis) was considered as deficient. Haemoglobin, haematocrit, total and differential leucocyte numbers were determined. The functions of peripheral neutrophils were assessed by in vitro phagocytosis and killing of Staphylococcus aureus. The effect of molybdenum induced copper deficiency on bone marrow was monitored. The mean total leucocyte count was unaffected whereas a significant fall in neutrophil count coincided with the fall in hepatic copper level to $23.9{\pm}2.69$ ppm. Reduced blood neutrophil numbers was not accompanied by any change in the proportion of different neutrophil precursor cells in bone marrow. It was hypothesised that buffalo calves were more tolerant to dietary molybdenum excess than cattle. It was concluded that neutropenia in molybdenum induced copper deficiency occurred without any effect on their synthesis and maturation process. Bone marrow studies in healthy calves revealed higher percentage of neutrophilic myelocytes and metamyelocytes as compared to cattle.


  1. Das, M. N. and N. C. Giri. 1986. Design and Analysis of Experiments. John Wiley and Sons Inc. New York, USA.
  2. Fujita, M., T. Itakura, Y. Takagi and A. Okada. 1989. Copper deficiency during total parenteral nutrition : clinical analysis of three cases. J. Parenter. Enteral Nutr. 13:421-425.
  3. Holtenius, P. 1961. Cytological puncture. A new method for study of bovine hepatic disease. Cornell Vet. 51:56-63.
  4. Jain, N. C. and D. E. Jasper. 1967. Phagocytosis and destruction of Aerobacter aerogenes by leucocytes from bovine milk. Am. J. Vet. Res. 28:405-411.
  5. Phillip, M., A. Singer and A. Tal. 1990. Copper deficiency in an infant with giardiasis fed with cow' milk. Isr. J. Med. Sci. 26:289-290.
  6. Radostits, O. M., D. C. Blood and C. C. Gay. 1994. Veterinary Medicine. 9th Ed. Bailliere Tindall, London.
  7. Soodan, J. S. 1996. Clinico-biochemical, immunological and therapeutic studies in experimentally induced copper deficiency in animals. Ph.D. Thesis, Punjab Agricultural University, Ludhiana, India.
  8. Sriram, K., J. A. O'ara, J. R. Strunk and J. K. Peterson. 1986. Neutropenia due to copper deficiency in total parenteral nutrition. J.Parenter. Enteral. Nutr. 13:530-532.
  9. Tizzard, I. 1996. Veterinary Immunology: An Introduction. $5^{st}$ Ed. W. B. Saunders Company, Philadelphia, USA.
  10. Wilde, J. K. H. 1964. The cellular elements of the bone marrow. Res.Vet. Sci. 5:213-227.
  11. AOAC. 1970. Official Method of Analysis. Association of agricultural Chemists, Washington, DC., USA.
  12. Babu, U. and M. L. Failla. 1990a. Respiratory burst and candidacidal activity of peritoneal macrophages are impaired in copper deficient rats. J. Nutr. 120:1692-1699.
  13. Randhawa, S. S. 1993. Clinico-biochemical studies on phosphorus, copper and molybdenum status in dairy animals with haemoglobinuria, pica and leucoderma and their interaction with agro-ecological conditions. Ph.D. Thesis, C.S.Azad University of Agriculture and Techmnology, Kanpur, India.
  14. Roth, J. A., M. L. Kaeberle and W. H. Hsu. 1982. Effect of ACTH administration on bovine polymorphonucleat function and lymphocyte blastogenesis. Am. J. Vet. Res. 43:412-416.
  15. Badwey, J. A. and M. L. Karnovsky. 1980. Active oxygen species and the functions of phagocytic leucocytes. Annu. Rev. Biochem. 49:695-726.
  16. Jones, D. G. and N. F. Suttle. 1981. Some effects of copper deficiency on leucocyte function in sheep and cattle. Res. Vet. Sci. 31:151-156.
  17. Xin, Z., D. F. Watermann, R. W. Hemken and R. J. Harmon. 1991. Effects of copper status on neutrophil function, superoxide dismutase and copper distribution in steers. J. Dairy Sci. 74:3078-3085.
  18. Gordon, P. A., J. Sturt, T. R. Lee, G. R. Breeze and R. N. H. Pugh. 1975. The cytocentrifuge NBT test. J. Clin. Path. 28:674-679.
  19. Goyens, P., D. Brasseur and S. Cadranel. 1985. Copper deficiency in infants with active celiac disease. J.Paediatr. Gastroenterol. Nutr. 4:677-680.
  20. Schalm, O. W. and J. Lasmanis. 1976. Cytological features of bone marrow in normal and mastitic cows. Am. J. Vet. Res. 37:359-363.
  21. Van Soest, P. J. and R. M. Wine. 1967. Use of detergents in the analysis of fibrous material. IV. Determination of plant cell wall constituents. J. Assoc. Agric. Chem. 50:50-55.
  22. Dunlap, W. M., J. S. W. Jones and D. M. Hume. 1974. Anaemia and neutropenia caused by deficiency. Ann. Intern. Med. 80:470-476.
  23. Higuchi, S., A. Higashi, T. Nakamura and I. Matasuda. 1988. Nutritional copper deficiency in severely handicapped patients on a low enteral diet for a prolonged period : Estimation of the required dose of dietary copper. J. Paediatr. Gastroenterol. Nutr. 7:583-587.
  24. Stabel, J. R., J. W. Spears and Jr. T. T. Brown. 1993. Effect of copper deficiency on tissue characteristics and immune function of calves challenged with infectious bovine rhinotracheitis virus and and Pasteurella haemolytica. J. Anim. Sci. 71:1247-1255.
  25. Jain, N. C. 1986. Schalm'Veterinary Haematology.$4^{th}$ Ed. Lea and Febiger, Philadelphia, USA.
  26. Boyne, R. and J. R. Arthur. 1986. Effects of molybdenum or iron induced copper deficiency on viability and function of neutrophils from cattle. Res. Vet. Sci. 41:417-419.
  27. Becton, D. L., W. H. Schultz and T. R. Kinney. 1986. Severe neutropenia caused by copper deficiency in a child receiving continuous ambulatory peritoneal dialysis. J. Paediatr. 108:735-737.
  28. Cerone, S. I., A. S. Sansinanea, S. A. Streitenberge, M. C. Garcia and N. J. Auza. 1998. The effect of copper deficiency on the peripheral blood cells of cattle. Vet. Res. Comm. 22:47-57.
  29. Winquist, G. 1954. Morphology of the blood and the hemopoietic organs in cattle under normal and some experimental conditions. Acta. Anat. Suppl. 21:1 (original not seen. Cited froim N. C. Jain. 1986. Schalm' Veterinary Hematology $4^{th}$ edn. pp. 178-207.
  30. Zidar, B. L., R. K. Shadduck, Z. Zeiglar and Winkelstein. 1977. Observations in the anaemia and neutropenia of human copper deficiency. Am. J. Haemtol. 37:177-185.
  31. Kellaway, R. C., P. Sitorus and J. M. L. Leibholz. 1978. The use of copper levels in hair to diagnose hypocuprosis. Res. Vet. Sci. 24:352-357.
  32. Babu, U. and M. L. Failla. 1990b. Copper status and function of neutrophil are reversible depressed in marginal and severly copper deficient rats. J. Nutr. 120:1700-1709.
  33. Percival, S. S. 1995. Neutropenia caused by copper deficiency : possible mechanisms. Nutr. Rev. 53:59-66.
  34. Arthington, J. D., L. R. Corah and F. Blecha. 1996a. The effect of molybdenum-induced on acute-phase protein concentrations' superoxide dismutase activity, leucocyte numbers and lymphocyte proliferation in beef heifers inoculated with Bovine Herpesvirus-1. J. Anim. Sci. 74:211-217.
  35. Arthington, J. D., A. R. Spell, L. R. Corah and F. Blecha. 1996b. Effect of Molybdenum induced copper deficiency on in vivo and in vitro measures of neutrophil chemotaxis both before and following an inflammatory stressor. J. Anim. Sci. 74:2759-2764.
  36. Hirase, N., Y. Abe and S. Sadamura et al. 1992. Anaemia and neutropenia in a case of copper deficiency : Role of copper in normal haematopoiesis. Acta Haematol. 87:193-197 Original not seen. (Ed. S. S. Percival). Nutr. Rev. 53:59-66.
  37. Bala, S., M. Failla and J. K. Lunney. 1991. Alterations in splenic lyjphiod cell subsets and activation of antigens in copper deficient rats. J. Nutr. 121:745-753.
  38. Bhuyan, U. N. 1992. Evaluation of neutrophil function. In: A Hand Book of Practical and Clinical Immunology $1^{st}$ Ed. (Ed. G. P. Talwar and S. K. Gupta). CBS Publishers and Distributors, New Delhi, India. pp. 301-307.
  39. Boyne, R. and J. R. Arthur. 1981. Effects of selenium and copper deficiency on neutrophil function in cattle. J. Comp. Path. 91:271-278.
  40. Humphries, W. R., M. Phillippo, B. W. Young and I. Bremner. 1983. The influence of dietary iron and molybdenum on copper metabolism in calves. Br. J. Nutr. 49:77-86.
  41. Gangelbach, G. P., J. D. Ward, J. W. Spears and T. T. Jr. Brown. 1997. Effects of copper deficiency and copper deficiency coupled with high dietary iron or molybdenum phagocytic cell function and response of calves to a respiratory disease challenge. J. Anim. Sci. 75:1112-1118.
  42. Yeo, S. P., C. R. Stokes, F. G. R. Taylor and F. J. Bourne. 1993. Maturation of celluar defence in the respiratory tract of young calves. Res. Vet. Sci. 55:292-297.
  43. Carlson, G. P. and J. J. Kaneko 1973. Isolation of leucocytes from bovine peripheral blood. Proc. Soc. Exp. Biol. Med. 142:853-860.
  44. Jones, Jr. J. B. 1977. Elemental analysis of soil extracts and plant tissue ash by plasma emmision spectroscopy. Comm. Soil Sci. Pl. Anal. 8:349-365.