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Research (Published online: 30-09-2015)

20. Replacement of inorganic zinc with lower levels of organic zinc (zinc nicotinate) on performance, hematological and serum biochemical constituents, antioxidants status, and immune responses in rats - D. Nagalakshmi, K. Sridhar and S. Parashuramulu

Veterinary World, 8(9): 1156-1162



   doi: 10.14202/vetworld.2015.1156-1162


D. Nagalakshmi: Department of Animal Nutrition, College of Veterinary Science, Korutla, Karimnagar - 505 326, Telangana, India;

K. Sridhar: Department of Animal Nutrition, College of Veterinary Science, Hyderabad - 500 030, Telangana, India;

S. Parashuramulu: Department of Animal Nutrition, College of Veterinary Science, Hyderabad - 500 030, Telangana, India;


Received: 01-04-2015, Revised: 22-08-2015, Accepted: 31-08-2015, Published online: 30-09-2015


Corresponding author: D. Nagalakshmi, e-mail:

Citation: Nagalakshmi D, Sridhar K, Parashuramulu S (2015) Replacement of inorganic zinc with lower levels of organic zinc (zinc nicotinate) on performance, hematological and serum biochemical constituents, antioxidants status and immune responses in rats, Veterinary World 8(9):1156-1162.

Aim: A study was undertaken to investigate the effect of organic zinc (zinc nicotinate, Zn-nic) supplementation (6, 9, and 12 ppm) compared to inorganic zinc (12 ppm) on growth performance, hematology, serum biochemical constituents oxidative stress, and immunity in weaned female Sprague–Dawley rats.

Material and Methods: A 48 weaned rats (285.20±1.95 g) were randomly distributed to 4 dietary treatments with 6 replicates in each and reared in polypropylene cages for 10 weeks. Basal diet (BD) was formulated with purified ingredients without zinc (Zn). Four dietary treatments were prepared by adding 12 ppm Zn from ZnCO3 (control) and 6, 9, and 12 ppm Zn from Zn-nic to the BD. On 42nd day, blood was collected by retro-orbital puncture for analyzing hematological constituents, glucose, cholesterol, alkaline phosphatase, total protein, albumin, and globulin and antioxidant enzyme activities. At 43rd day, rats were antigenically challenged with sheep red blood cell (RBC) to assess humoral immune response and on 70th day cell-mediated immune response.

Results: Weekly body weight gains, daily feed intake, blood hematological constituents (white blood cell, RBC, hemoglobin concentration, packed cell volume, mean corpuscular volume, lymphocyte, monocyte, and granulocyte concentration) and serum glucose, total protein levels were comparable among the rats feed Zn from ZnCO3 and Zn-nic (6, 9, and 12 ppm). Serum cholesterol reduced with organic Zn supplementation at either concentration (6-12 ppm). Serum globulin concentration reduced (p<0.05) with 6 ppm Zn-nic supplementation compared to other dietary treatments. Lipid peroxidation lowered (p<0.05) reduced with 12 ppm organic Zn; thiobarbituric acid reacting substances and protein carbonyls concentrations in liver reduced (p<0.05) with 9 and 12 ppm levels of organic Zn supplementation compared to 12 ppm Zn supplementation from inorganic source. RBC catalase and glutathione peroxidase enzymes activities were highest (p<0.05) in rats supplemented with 12 ppm Zn-nic, followed by 9 ppm. Comparable immune response (humoral and cell-mediated) was observed between 12 ppm inorganic Zn and 9 ppm organic Zn and higher (p<0.05) immune response was noticed at 12 ppm Zn-nic supplementation.

Conclusion: Based on the results, it is concluded that dietary Zn concentration can be reduced by 50% (6 ppm) as Zn nicotinate without affecting growth performance, hemato-biochemical constituents, antioxidant status, and immunity. In addition, replacement of 12 ppm inorganic Zn with 12 ppm organic Zn significantly improved antioxidant status and immune response.

Keywords: antioxidants status, hematological and serum biochemical constituents, immune responses, performance, rats, zinc nicotinate.

1. Chasapis, C.T., Loutsidou, A.C., Spiliopoulou, C.A. and Stefanidou, M.E. (2012) Zinc and human health: An update. Arch. Toxicol., 86(4): 521-534.
2. Gruber, K., Rink, L. (2013) The role of zinc in immunity and inflammation In: Calder, P.C. and Yaqoob, P. editor. Diet. Immun. Inflam. 1st edition, Cambridge, U.K. 123-156.
3. Oteiza, P.I. (2012) Zinc and the modulation of redox homeostasis. Free Radic. Biol. Med., 53(9): 1748-1759.
PMid:22960578 PMCid:PMC3506432
4. Świątkiewicz, S., Arczewska-włosek, A. and Jozefiak, D. (2014) The efficacy of organic minerals in poultry nutrition: Review and implications of recent studies. World's Poult. Sci. J., 70(3): 475-486.
5. Moghaddam, H.N. and Jahanian, R. (2009) Immunological responses of broiler chicks can be modulated by dietary supplementation of zinc-methionine in place of inorganic zinc sources. Asian – Aust. J. Anim. Sci., 22(3): 396-403.
6. Feng, J., Ma, W.Q., Niu, H.H., Wu, X.M., Wang, Y. and Feng, J. (2010) Effects of zinc glycine chelate on growth, hematological, and immunological characteristics in broilers. Biol. Trace. Elem. Res., 133: 203-211.
7. Ao, T., Pierce, J.L., Power, R., Pescatore, A.J., Cantor, A.H., Dawson, K.A., Ford, M.J. and Paul, M. (2011) Effects of feeding different concentration and forms of zinc on the performance and tissue mineral status of broiler chicks. Br. Poult. Sci., 52: 466-471.
8. Swiatkiewicz, S., Arczewska-włosek, A. and Jozefiak, D. (2014) The efficacy of organic minerals in poultry nutrition: Review and implications of recent studies. World's Poult. Sci. J., 70(03): 475-486.
9. Reinhold, J.G. (1953) In: Rynner, M., editor. Standard Methods of Clinical Chemistry C. Academic Press, New York. p88.
10. Gustafsson, E.J. (1976) Improved specificity of serum albumin determination and estimation of acute phase of reactants by use of the bromocresol green. Quinchemisty, 22(5): 616-622.
11. Cooper, G.R. and Mc Daniel, V. (1970) Assay methods. In: Mc Donald, R.P., editor. Standard Methods for Clinical Chemistry. John Wiley and Sons, New York. p159-170.
12. Wybenga, D.R., Pileggi, V.J., Dirstine, P.H. and Di Giorgio, J. (1970) Direct manual determination of serum total cholesterol with a single stable reagent. Clin. Chem., 16: 980-984.
13. Kind, P.R. and King, E.J. (1954) Estimation of plasma phosphatase by determination of hydrolyzed phenol with amino-antipyrine. J. Clin. Pathol., 7: 322-326.
PMid:13286357 PMCid:PMC1023845
14. Bergmeyer, H.U. (1983) Catalase. Methods of Enzymatic Analysis. Verlag Chemie, Weinheim. p165-166.
15. Placer, Z.A., Cushman, L.L. and Johnson, B.C. (1966) Estimation of product of lipid peroxidation (Malonyl Dialdehyde) in biochemical systems. Anal. Biochem., 16: 359-364.
16. Paglia, D.E. and Valentine, W.N. (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J. Lab. Clin. Med., 70(1): 158-169.
17. Carlberg, I. and Mannervik, B. (1985) Glutathione reductase. Methods Enzymol., 113: 484-490.
18. Cannan, R.K. (1958) Laboratory methods-proposal for a certified standard for use in hemoglobinometry second and final report. J. Lab. Clin. Med., 52(3): 471-476.
19. Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Boil. Chem., 193(1): 265-275.
20. Balasubramanian, K.A., Manohar, M. and Mathan, V.I. (1988) An unidentified inhibitor of lipid peroxidation in intestinal mucosa. Biochim. Biophy. Acta, 962(1): 51-58.
21. Moron, M.J., Diperre, J.W. and Mannerv, K.B. (1979) Levels of glutathione, glutathione reductase and glutathiones transferase activities in rat lungs and liver. Biochim. Biophys. Acta, 582: 67-71.
22. Levine, R.L., Garland, D., Oliver, C.N., Amici, A., Climent, I., Lenz, A.G., Ahn, B.W., Shaltiel, S. and Stadtman, E.R. (1990) Determination of carbonyl content in oxdatively modified proteins. Methods Enzymol., 186: 464-478.
23. Liew, F.Y. (1977) Regulation of delayed type hypersensitivity, T suppressor cells for delayed type hypersensitivity to sheep erythrocytes in mice. Eur. J. Immunol., 7: 714-718.
24. Snedecor, G.W. and Cochran, W.G. (1994) Statistical Methods. 8th ed. Iowa State University Press, Ames, Iowa, USA.
25. Duncan, D.B. (1955) Multiple range and multiple F tests. Biometrics, 11(1): 1-42.
26. Deshpande, J.D., Joshi, M.M. and Giri, P.A. (2013) Zinc: The trace element of major importance in human nutrition and health. Int. J. Med. Sci. Public Health, (1): 1-6.
27. Rossi, P., Rutz, F., Amciuti, M.A., Rech, J.L. and Zauk, N.H.F (2007) Influence of graded levels of organic zinc on growth performance and carcass traits of broilers. J. Appl. Poult. Res., 16: 219-225.
28. Wang, Y., Tang, J.W., Ma, W.Q., Feng, J. and Feng, J. (2010) Dietary zinc glycine chelate on growth performance, tissue mineral concentration, and serum enzyme activity in weanling piglets. Biol. Trace Elem. Res., 133: 325-334.
29. El Hendy, H.A., Yousef, M.I. and El-Naga, N.I.A. (2001) Effect of dietary zinc deficiency on hematological and biochemical parameters and concentrations of zinc, copper, and iron in growing rats. Toxicology, 167(2): 163-170.
30. Dardenne, M. (2002) Zinc and immune function. Eur. J. Clin. Nutr., 56: S20-S23.
31. Someya, Y., Ichinose, T., Nomura, S., Kawashima, Y.U., Sugiyama, M., Tachiyashiki, K. and Imaizumi, K. (2007) Effects of zinc deficiency on the number of white blood cells in rats. FASEB J., 21: 697.2.
32. Akbari, M.R., Kermanshahi, H., Moghaddam, H.N., Moussavi, A.R.H. and Afshari, J.T. (2008) Effects of wheat-soybean meal based diet supplementation with vitamin A, vitamin E and zinc on blood cells, organ weights and humoral immune response in broiler chickens. J. Anim. Vet. Adv., 7(3): 297-304.
33. Ao, T., Pierce, J.L., Power, R., Pescatore, A.J., Cantor, A.H., Dawson, K.A. and Ford, M.J. (2009) Effect of different forms of zinc and copper on the performance and tissue mineral content of chicks. Poult. Sci., 88: 2171-2175.
34. Al-Daraji, H.J. and Amen, M.H.M. (2011) Effect of dietary zinc on certain blood traits of broiler breeder chickens. Int. J. Poult. Res., 10(10): 807-813.
35. Beattie, J.H., Gordon, M.J., Duthie, S.J., McNeil, C.J., Horgan, G.W., Nixon, G.F., Feldmann, J. and Kwun, I.S. (2012) Suboptimal dietary zinc intake promotes vascular inflammation and atherogenesis in a mouse model of atherosclerosis. Mol. Nutr. Food Res., 56(7): 1097-1105.
36. Reiterer, G., Macdonald, R., Browning, J.D., Morrow, J., Matveev, S.V., Daugherty, A., Smart, E., Toborek, M. and Hennig, B. (2005) Zinc deficiency increases plasma lipids and atherosclerotic markers in LDL-receptor-deficient mice. J. Nutr., 135: 2114-2118.
37. Bolkent, S., Yanardag, R., Bolkent, S., Mutlu, O., Yildirim, S., Kangawa, K., Minegishi, Y. and Suzuki, H. (2006) The effect of zinc supplementation on ghrelin-immunoreactive cells and lipid parameters in gastrointestinal tissue of streptozotocin-induced female diabetic rats. Mol. Cell. Biochem., 286: 77-85.
38. Parák., T. and Straková, E. (2011) Zinc as a feed supplement and its impact on plasma cholesterol concentrations in breeding cocks. Acta Vet. Brno., 80: 281-285.
39. Sahin, K., Smith, M.O., Onderci, M., Sahin, N., Gursu, M.F. and Kucuk, O. (2005) Supplementation of zinc from organic or inorganic source improves performance and antioxidant status of heat-distressed quail. Poult. Sci., 84: 882-887.
40. Sies, H., editor. (2013) Oxidative Stress. Elsevier, San Diego.
41. Osaretin, A.T.E. and Gabriel, A.A. (2009) Effect of zinc deficiency on memory, oxidative stress and blood chemistry in rats. Int. J. Biol. Chem. Sci., 3(3): 513-523.
42. Prasad, A.S. (2014) Zinc: An antioxidant and anti-inflammatory agent: Role of zinc in degenerative disorders of aging. J. Trace Elem. Med. Biol., 28(4): 364-371.
43. Flohe, L. (2009) Glutathione peroxidase: Fact and fiction. Oxygen Free Radicals and Tissue Damage. Oxford University Press, New York. p95-120.
44. Peerapatdit, T. and Sriratanasathavorn, C. (2010) Lipid peroxidation and antioxidant enzyme activities in erythrocytes of type 2 diabetic patients. J. Med. Assoc. Thai, 93(6): 682-693.
45. Bun, S.D., Guo, Y.M., Guo, F.C., Ji, F.J. and Cao, H. (2011) Influence of organic zinc supplementation on the antioxidant status and immune responses of broilers challenged with Eimeria tenella. Poult. Sci., 90: 1220-1226.
46. Ma, W., Niu, H., Feng, J., Wang, Y. and Feng, J. (2011) Effect of zinc glycine on oxidative stress, contents of trace elements, and intestinal morphology in broilers. Biol. Trace Elem. Res., 142: 546-556.
47. Gajula, S.S., Chelasani, V.K., Panda, A.K., Mantena, V.L.N. and Rama Rao, S. (2011) Effect of supplemental inorganic Zn and Mn and their interactions on the performance of broiler chicken, mineral bioavailability, and immune response. Biol. Trace Elem. Res., 139: 177-187.
48. Sajadifar, S., Miranzadeh, H. and Moazeni, M. (2011) Immune responses of broiler chicks supplemented with high levels of zinc. Online J. Anim. Feed Res., 6(2): 493-496.
49. Soni, N., Mishra, S.K., Swain, R., Das, A., Chichilichi, B. and Sethy, K. (2013) Bioavailability and immunity response in broiler breeders on organically complexed zinc supplementation. Food Nutr. Sci., 4(12): 1293-1300.
50. Ezzati, M.S., Bozorgmehrifard, M.H., Bijanzad, P., Rasoulinezhad, S., Moomivand, H., Faramarzi, S., Ghaedi, A., Ghabel, H. and Stabraghi, E. (2013) Effects of different levels of zinc supplementation on broilers performance and immunity response to Newcastle disease vaccine. Eur. J. Exp. Biol., 3(5): 497-501.
51. Hudson, B.P., Dozier 3rd, W.A., Wilson, J.L., Sander, J.E. and Ward, T.L. (2004) Reproductive performance and immune status of caged broiler breeder hens provided diets supplemented with either inorganic or organic sources of zinc from hatching to 65 wk of age. J. Appl. Poult. Res., 13: 349-359.