Effects of single injection of vitamin D3 on some immune and oxidative stress characteristics in transition dairy cows

Document Type : Research Articles


1 Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran &Center of Excellence in Ruminant Abortion and Neonatal Mortality, Ferdowsi University of Mashhad, Mashhad, Iran.

3 Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran & Center of Excellence in Ruminant Abortion and Neonatal Mortality, Ferdowsi University of Mashhad, Mashhad, Iran.


Recent studies suggest that vitamin D may have preventive and therapeutic effects on autoimmune disease, cancer, and diabetes type 1 and 2 beyond the skeletal condition and calcium metabolism. To demonstrate the effects of an over-supplemented single 8 million I.U. vitamin D3 IM injection on the modulation of immune responses and oxidative/antioxidative variables in transition dairy cows, this study was conducted on a commercial dairy farm with about 1500 lactating cows in the Tehran province, Iran. Twenty-four multiparous Holstein cows were randomly categorized into control and treatment groups. In the treatment group, 12 cows received a single dose of 8,000,000 IU vitamin D intramuscularly. In the control group, a placebo (distilled water) was injected into 12 cows 2 to 8 days before the expected calving time. Blood samples were collected on 21 and 7 days before calving and 1,3,7,15, and 30 days after calving. 25(OH)vitamin D3, tumor necrosis factor- α (TNF-α), interferon-γ (INF-γ), haptoglobin, interleukin 6 (IL-6), ferric reducing the ability of plasma (FRAP), glutathione peroxidase (GPx), superoxide dismutase (SOD), and hemolysate GPx were measured. This study showed that the treatment group had significantly higher amounts of 25(OH) vitamin D3, hemolysate GPx, and IL-6 values than the control group. According to our results, vitamin D3 injection increased the amounts of IL-6 and hemolysate GPx activity and tended to affect serum GPx activity. 


1.    Mathieu RC, Gysemans C, Giulietti A, Bouillon, R. Vitamin D and diabetes. Diabetologia. 2005; 48:1247–1257.
2.    Adams JS , Hewison M. Unexpected actions of vitamin D : new perspectives on the regulation of innate and adaptive immunity. Nat. Clin. Pract. Endoc 2008; 4(2):80–90.
3.    Hoenderop JG, Nilius B, Bindels RJ. Calcium Absorption Across Epithelia. Physiol Rev. 2005; 85(1):373–422.
4.    Goff JP, Horst RL, Littledik ET, Beitz DC. Use 24-F-1,25-Dihydroxyvitamin D3 to prevent parturient paresis in dairy cows. Int. J. Dairy Sci 1988; 71(5):1211–1219.
5.    Horst RL, Goff JP, Reinhardt TA. Calcium and vitamin D metabolism in the dairy cow. Int. J. Dairy Sci 1994; 77(7):1936–1951.
6.    De Garis PJ, Lean IJ. Milk fever in dairy cows: A review of pathophysiology and control principles. Vet. J 2008; 176(1): 58–69.
7.    Holick MF. Vitamin D : importance in the prevention of cancers, type 1 diabetes, heart disease and osteoprosis. Am. J. Clin. Nutr 2004; 79(3):362-71.
8.    Nelson CD, Reinhardt TA, Lippolis JD, Sacco RE, Nonnecke BJ. Vitamin D signaling in the bovine immune system: a model for understanding human  vitamin D requirements. Nutrients. 2012; 4(3): 181–196.
9.    O’Brien MA, Jackson MW. Vitamin D and the immune system: Beyond rickets. Vet. J. 2012; 194(1): 27–33.
10.    Pittas AG, Lau J, Hu FB, Dawson-Hughes B. The role of vitamin D and calcium in type 2 diabetes. A systematic review and meta-analysis. J. Clin. Endocrinol. Metab. 2007; 92(6): 2017–2029
11.    Chiu KC, Chu A, Go VL,  Saad MF. Hypovitaminosis D is associated with insulin resistance and beta cell dysfunction. Am. J. Clin. Nutr 2004; 25(4): 820–825.
12.    Nelson CD, Lippolis JD, Reinhardt TA, Sacco RE, Powell JL, Drewnoski ME, et.al. Vitamin D status of dairy cattle: outcomes of current practices in the dairy industry. Int. J. Dairy Sci 2016; 99(12): 10150–10160.
13.    Lean IJ, Saun RV, Degaris PJ. Mineral an antioxidant management of transition dairy cows. Vet. Clin. North. Am. Food. Anim. Pract. 2013; 29: 367–386.
14.    Holcombe SJ, Wisnieski L, Gandy J,  Norby B, Sordillo LM. Reduced serum vitamin D concentrations in healthy early-lactation dairy cattle. Int. J. Dairy Sci  2018; 3: 1–7.
15.    Contreras GA, Clarissa Strieder-Barboza C, Raphael W. Adipose tissue lipolysis and remodeling during the transition period of dairy cows. J. Anim. Sci. Biotechnol 2017; 8: 41-53.
16.    Querfeld U. Vitamin D and inflammation. Pediatr. Nephrol., 2013; 28: 605–610.
17.    Dimitrov V, White JH. Molecular and Cellular Endocrinology Vitamin D signaling in intestinal innate immunity and homeostasis. Mol. Cell. Endocrinol. 2017; 453: 68–78.
18.    Borges MC, Martinin LA, Rogero MM. Current perspectives on vitamin D, immune system, and chronic diseases. Nutr. J., 2011; 27(4): 399–404.
19.    Das M, Tomar N, SreenivasV, Gupta1 N, Goswami1 R. Effect of vitamin D supplementation on cathelicidin, INF-γ, IL-4 and Th1/Th2 transcription factors in young healthy female. Eur. j. Clin. Nutr. 2014; 68: 338-343.
20.    Jorde R, Sneve M, Torjesen PA, Figenschau Y, Goransson LG, Omdal R. No effect of supplementation with cholecalciferol on cytokines and markers of inflammation in overweight and obese subjects. Cytokine, 2010; 50: 175-180.
21.    Ohtsuka H, Koiwa M, Hatsugaya A, Kudo K, Hoshi F, Itoh N, et.al. Relationship between serum TNF activity and insulin resistance in dairy cows affected with naturally occurring fatty liver. J. Vet. Med. Sci. 2001; 63(9): 1021–1025.
22.    Dandona P, Aljada A, Bandyopadhyay A. Inflammation: the link between insulin resistance, obesity and diabetes. Trends. Immunol. 2004; 25(1): 4–7.
23.    Bertoni G; Minuti A, Trevisi, E. Immune system, inflammation and nutrition in dairy cattle. Anim. Prod. Sci. 2015; 55(7): 943–948.
24.    De Koster JD, Opsomer G. Insulin Resistance in Dairy Cows. Vet. Clin. North. Am. Food. Anim. Pract. 2013; 29(2): 299–322.
25.    Giulietti A, van Etten E, Overbergh L. Monocytes from type 2 diabetic patients have a pro-inflammatory profile.1,25-Dihydroxyvitamin D3 works as anti-inflammatory. Diabetes. Res. Clin. Pract. 2007; 77:47–57.
26.    Di Rosa M,  Malaguarnera G,  De Gregorio C, Palumbo M, Nunnari G,  Malaguarnera L. Immuno-modulatory effect of vitamin D3 in human monocyte and macrophage. Cell. Immunol., 2012; 280: 36-43.
27.    Naghavi Gargari B, Behmanesh M, Shirvani Farsani Z, Pahlevan Kakhki M, Azimi AR. Vitamin D supplementation up-regulates IL-6 and IL-17A gene expression in multiple sclerosis patients. Int. Immunopharmacol., 2015; 28: 414-419
28.    Wöbke TK, Sorg BL, Steinhilber D. Vitamin D in inflammatory diseases. Front Physiol., 2014; 5: 244.
29.    Pires JA, Pescara JB, Grummer RR. Reduction of plasma NEFA concentration by nicotinic acid enhances the response to insulin in feed-restricted holstein cows. Int. J. Dairy Sci. 2007; 90(10): 4635–4642.
30.    Sordillo LM, Aitken SL. Impact of oxidative stress on the health and immune function of dairy cattle. Vet. Immunol. Immunopathol 2009; 128: 104–109.
31.    Benedito JL, Abuelo A, Hernández J, Castillo C. The importance of the oxidative status of dairy cattle in the periparturient period : revisiting antioxidant supplementation. J. Anim. Physiol. An. N 2015; 99(6): 1003–1016.
32.    Littledike ET, Horst RL.Vitamin D3 toxicity in dairy cows. Int. J. Dairy Sci. 1982; 65: 749-759.
33.    Julien WE, Conard HR, Hibbs JW, Crist, WL, Milk fever in dairy cows: effect of injected vitamin D3 and calcium and phosphorus intake on incidence. Int. J. Dairy Sci. 1977; 60: 431-436.
34.    Edmonson AJ, Lean IJ, Weaver LD, Farver T, Webster G. A body condition score chart for Holstein dairy cows. Int. J. Dairy Sci. 1989; 72: 68-78
35.    Duffield TF, Leslie KE, Sandals D,  Lissemore K,  McBride BW,  Lumsden JH, et.al. Effect of a monensin-controlled release capsule on cow health and reproductive performance. Int. J. Dairy Sci., 1999; 82(11): 2377–2384.