_1689829435 20230720050355000 Rovedar Daryoushbabazadeh@gmail.com Rovedar Journal of Veterinary Physiology and Pathology J. Vet. Physiol. Pathol. 2821-0328 06 05 2023 2 2 Rahul Kumar Harmanjit Singh Banga Rajinder Singh Brar Introduction: Vitamin D3 is used as a supplement in the feeds of livestock, pets, and human infants. However, the presence of excessive Vitamin D3 has been shown to cause toxicity in humans and animals. This study investigated the clinicopathological aspects of Vitamin D3 toxicity in broiler chickens. Materials and methods: The median lethal dose (LD50) of Vitamin D3 was estimated by the up-and-down method. To determine long term (21 days) toxic effects of oral Vitamin D3 supplementation, 90 (14-day-old) IBL-80 unsexed chicks were randomly divided into three groups as group A (control, received basal diet), B (basal diet + Vitamin D3 at 16.67 mg/kg body weight daily), and C (basal diet + Vitamin D3 at 33.33 mg/kg body weight daily). Results: The findings indicated that broiler chickens tolerated a single oral dose of Vitamin D3 up to 550 mg/kg body weight (22,000,000 IU/kg) without mortality. The results of long-term (21 days) oral supplementation of divided doses of Vitamin D3 in broiler chickens (groups B and C) showed progressive emaciation, elevated hemoglobin, hypercalcemia, hypophosphatemia, and increased alkaline phosphatase activity. At necropsy, pale liver and kidneys, congestion and hardening of lungs, mild congestion in the brain, and soft bones were observed in Vitamin D3 treated chicks (groups B and C). Microscopically, degeneration and metastatic calcification in lung parenchyma and peribronchiolar epithelium, coagulative necrosis and calcification in kidneys, and calcification with fibroplasia in proventriculus was detected. Lungs and kidneys showed a significant difference in calcification score between groups B and C. Broiler chickens from Vitamin D3 treated groups (B and C) showed strong immunohistochemical expression of Calbindin D28K in the intestine and kidneys but weak expression in the lungs. Conclusion: This study demonstrates that broiler chicks can tolerate very high levels of a single oral dose of Vitamin D3. Toxic effects of prolonged exposure to Vitamin D3 are due to over-expression of Calbindin D28k in the intestine and kidneys, disturbing the calcium and phosphorous homeostasis and leading to metastatic calcification of vital organs. This study supports that prolonged over-supplementation of Vitamin D3 causes toxic effects; hence, appropriate dietary Vitamin D3 supplementation limits should be set. 06 25 2023 20 31 https://creativecommons.org/licenses/by/4.0 10.58803/jvpp.v2i2.21 https://jvpp.rovedar.com/index.php/JVPP/article/view/21 https://jvpp.rovedar.com/index.php/JVPP/article/download/21/31 https://jvpp.rovedar.com/index.php/JVPP/article/download/21/31 https://jvpp.rovedar.com/index.php/JVPP/article/download/21/35 https://jvpp.rovedar.com/index.php/JVPP/article/download/21/36 https://jvpp.rovedar.com/index.php/JVPP/article/download/21/37 10.1016/j.tem.2010.01.003 10.1258/ebm.2010.010014 10.58803/fahn.v1i1.7 Bronner F, Pansu D, and Stein WD. An analysis of intestinal calcium transport across the rat intestine. Am J Physiol Gastrointest Liver Physiol. 1986; 250(5): G561-G569. DOI: 10.1152/ajpgi.1986.250.5.G561 10.1080/00071660701302270 Kumar R, Brar RS, and Banga HS. Hypervitaminosis D3 in broiler chicks: Histopathological, immunomodulatory and immunohistochemical approach. Iran J Vet Res. 2017; 18(3): 170-176. PMCID: PMC5674439 Smith SE. Vitamins. In: Swenson M J, editor. Duke’s physiology of domestic animals. Cornell University Press, Ithaca and London: Comstock Publishing Associate; 1982. p. 380-382. National research council (NRC). Nutrient requirements of poultry. 9th rev ed. Washington, DC, USA: National Academy Press; 1994. Available at: https://nap.nationalacademies.org/catalog/2114/nutrient-requirements-of-poultry-ninth-revised-edition-1994 10.1093/jn/102.8.975 Long GG. Acute toxicosis in swine associated with excessive dietary intake of Vitamin D. JAVMA. 1984; 184(2), 164-170. PMID: 6321415 10.1292/jvms.57.831 10.2460/javma.2000.216.1115 10.1016/B0-72-160639-3/50039-3 10.3382/ps.2013-03278 10.1093/japr/14.1.156 Takeda M, Yamashita T, Sasaki N, Nakajima K, Kita T, Shinohara M, et al. Oral administration of an active form of Vitamin D3 (calcitriol) decreases atherosclerosis in mice by inducing regulatory T cells and immature dendritic cells with tolerogenic functions. 10.1161/ATVBAHA.110.215459 10.2141/jpsa.0120071 Mabelebele M, Ng’Ambi JW, and Norris D. Effect of dietary Vitamin D3 supplementation on meat quality of naked neck chickens. Afr J Biotechnol. 2013; 12(22): 3576-3582. Available at: https://www.ajol.info/index.php/ajb/article/view/132058 10.1080/00071660410001730941 10.1093/japr/15.4.493 10.1080/00071660903125826 10.1111/j.1439-0396.2009.00943.x 10.1017/S0043933913000597 10.1017/S0043933916001057 10.1016/j.cbpa.2007.08.023 10.1080/00071669888980 10.3382/ps.2013-03243 10.1016/0272-0590(85)90059-4 U. S. environmental protection agency (USEPA). Test guidelines/acute toxicity — Acute oral toxicity up-and-down procedure. 2003. Available at: https://www.epa.gov/pesticide-science-and-assessing-pesticide-risks/acute-oral-toxicity-and-down-procedure Benjamin MM. Outline of veterinary clinical pathology. 3rded. Ludhiana, India: Kalyani Publishers; 1985; p. 25, 48, 60. Luna LG. Manual of histological staining methods of armed forces institute of pathology. 3rd ed. New York, USA: Mc Graw Hill Book Company; 1968. p. 32-36. Available at: https://pesquisa.bvsalud.org/ portal/resource/pt/biblio-1081120 10.3382/ps.0741919 10.3382/japr.2011-00483 10.1080/03079459708419206 Machlin LJ. Handbook of vitamins. New York: Marcel Dekker, Inc; 1984. Meric SM. In: Ettiner SJ, Feldman EC, editors. Textbook of internal medicine. Philadelphia. Saunders; 1995. p. 159-163. Kaneko JJ, Harvey JW, and Bruss ML, editors. Clinical biochemistry of domestic animals. Academic press. 2008. Morrow C. Cholecalciferol poisoning. Vet Med. 2001; 96(12): 905-911. 10.1136/vr.146.6.161 Radostits OM, Gay CC, Blood DC, and Hinchcliff KW. Veterinary medicine- A textbook of the diseases of cattle, sheep, pigs, goats and horses. 9th Ed. (formerly ELST), UK: Book Power. 2000; p. 561, 563, 1543, Available at: https://www.cabdirect.org/cabdirect/abstract/ 19942214281 Sandhu HS, and Brar RS. Textbook of veterinary toxicology. 2nd ed. Ludhiana: Kalyani Publishers. 2009; p. 245. 10.1093/jn/107.6.1027 10.4103/0971-6580.75851 Harrington DD, and Page EH. Acute vitamin D3 toxicosis in horses: Case reports and experimental studies of the comparative toxicity of vitamins D2 and D3. J Am Vet Med Assoc, 1983; 182(12): 1358-1369. PMID: 6307954 Kumar R, Brar RS, Banga HS, and Sodhi S. Biochemical alterations in hypervitaminosis D3 in broiler chicks concomitantly challenged with endotoxin. J World Poult Res. 2018; 8(4): 100-104. Available at: https://jwpr.scienceline.com/attachments/article/47/J%20World%20Poult%20Res%208(4)%20100-104,%202018.pdf Abdi-Hachesoo B, Talebi A, and Asri-Rezaei S. Comparative study on blood profiles of indigenous and Ross-308 broiler breeders. Glob Vet. 2011; 7(3): 238-241. Available at: https://www.idosi.org/gv/GV7(3)11/6.pdf 10.1079/BJN19680081 Beasley VR. Veterinary toxicology. International Veterinary Information Service. Ithaca, New York. 1999. 10.1093/ps/77.5.666 10.1056/NEJM198904133201506 Lumeij JT. Avian medicine. Principles and Applications In: RitchieBW, Harrison GH, Harrison LR, editors. Lake Worth, FL: Wingers; 1994; p. 582-606. Norman AW. Vitamin D. Present knowledge of nutrition. 7th ed. Washington: 1996. 10.7326/0003-4819-122-7-199504010-00006 Scheftel J, Setzer S, Walser M, Pertile T, Hegstad RL, Felice LJ, et al. Elevated 25-hydroxy and normal 1, 25-dihydroxy cholecalciferol serum concentrations in a successfully treated case of Vitamin D3 toxicosis in a dog. Vet Hum Toxicol. 1991; 33(4): 345-348. PMID: 1654665 10.1093/jn/131.11.2910 10.1093/jn/115.7.929 10.1126/science.7025212 10.1152/ajpgi.1986.250.5.G561 10.1016/0300-9629(93)90510-B 10.1093/jn/99.2.217 10.1016/S1095-6433(03)00244-7 10.1590/rbz4720170015 10.1210/endo-127-2-580 10.1002/jbmr.5650050404 10.1016/0014-4827(92)90205-M 10.1111/j.1523-1755.2004.00858.x