اثرات 1-25 دی‌هیدروکسی کوله کلسیفرول (کلسیتریول) و عصاره هیدروالکلی ریشه گیاه دارویی بوزیدان (Withania somnifera) بر پاسخ ایمنی و ریخت شناسی روده جوجه‌های گوشتی

نوع مقاله : علمی پژوهشی- تغذیه طیور

نویسندگان

1 گروه علوم دامی، مجتمع آموزشی سراوان

2 گروه علوم دامی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران

3 گروه علوم دامی، مجتمع آموزش عالی سراوان

4 گروه علوم دامی، مرکز تحقیقات آموزش کشاورزی فارس، سازمان تحقیقات، آموزش و ترویج کشاورزی، شیراز، ایران.

چکیده

این آزمایش به منظور بررسی اثرات 1-25 دی‌هیدروکسی کوله کلسیفرول [1, 25 (OH)2 D3] (کلسیتریول) و عصاره هیدرو الکلی ریشه گیاه دارویی بوزیدان (Withania somnifera) بر پاسخ ایمنی و ریخت شناسی روده کوچک جوجه های گوشتی انجام شد. تیمارها در قالب فاکتوریل (2×3×2) شامل جیره کنترل مثبت با سطح کافی کلسیم و کنترل منفی (کاهش 30 درصدی سطح کلسیم)، 3 سطح عصاره بوزیدان (صفر، 75 و150 میلی‌گرم در کیلوگرم جیره) و 2 سطح 1-25 دی‌هیدروکسی کوله کلسیفرول (صفر و 5/0 میکروگرم در کیلوگرم جیره) بود. تعداد600 قطعه جوجه گوشتی یک روزه راس 308 به صورت تصادفی در 60 عدد پن و 10 پرنده در هر کدام توزیع گردید. هر تیمار دارای 5 تکرار (50 پرنده در هر تیمار) بود. جیره‌های آزمایشی به طور نا محدود در اختیار جوجه‌ها از 1 تا 42 روزگی قرار گرفت. برای بررسی ایمنی هومورال از تست SRBC استفاده گردید. لذا دو تزریق در روزهای 25 و 32 دوره آزمایش برای تعیین پاسخ آنتی‌بادی اولیه و ثانویه انجام شد. در 21 و 42 روزگی یک پرنده از هر تکرار کشتار و وزن اندام‌های لنفاوی اندازه‌گیری و 5/1 سانتی‌متر از بافت ژژونوم به منظور تعیین خصوصیات ریخت شناسی جدا گردید. آزمایشات پاسخ ایمنی هومورال نشان داد که کاهش 30 درصدی سطح کلسیم جیره باعث کاهش تیتر آنتی بادی کل می گردد. عصاره هیدروالکلی ریشه بوزیدان در سطح 150 میلیگرم فقط باعث افزایش تیتر ایمونوگلوبولین G در تست اولیه گردید. هیچ‌گونه اثرات اصلی از تیمارهای آزمایشی بر وزن اندام‌های لنفاوی مشاهده نگردید. اطلاعات ریخت شناسی روده نشان داد که مکمل‌سازی کلسیتریول در جیره باعث کاهش طول پرز در 21 روزگی و کاهش عرض آن در 42 روزگی می‌گردد. کاهش 30 درصدی سطح کلسیم جیره منجر به کاهش عمق کریپت در مقایسه با گروه کنترل مثبت گردید. این آزمایشات نشان داد که کاهش سطح کلسیم جیره باعث کاهش پاسخ ایمنی هومورال می‌گردد. مکمل سازی عصاره بوزیدان در جیره‌های آزمایشی تأثیری بر خصوصیات پرز نداشت.

کلیدواژه‌ها


عنوان مقاله [English]

The Effects of Dietary 1, 25-Dihydroxycholecalciferol (Calcitriol) and Root Hydroalcoholic Extract of Withania somnifera on Immune Response and Small Intestinal Morphology of Broiler Chickens

نویسندگان [English]

  • Mohammad Taher Mirakzehi 1
  • Hasan Kermanshahi 2
  • Abolghasem Golian 2
  • Hassan Saleh 3
  • Mohammad Javad Agah 4
1 Department of Animal Science, Saravan Educational Complex
2 Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran
3 Department of Animal Science , Higher Educational Complex of Saravan, Iran.
4 Animal Science Research Department, Fars Agricultural and Natural Resources Research and Education Center, Agricultural Research, Education and Extention Organization( AREEO), Shiraz, Iran
چکیده [English]

Introduction 1, 25-dihydroxycholecalciferol [1, 25 (OH)2 D3], the major biologically active metabolite of vitamin D3 is produced by two sequential hydroxylation reactions. The first hydroxylation occurred when vitamin D3 is transported to the liver, forming 25-hydroxycholecalciferol [25-(OH) D3] and the second in the kidney, by the enzyme 1-alpha-hydroxylase (VD3 1 hydroxylase) forming 1, 25 (OH)2 D3. Vitamin D3 may have a role in regulating the morphological and functional development of intestinal villus mucosa. Calcium ions have an essential role function in activation and maturation of lymphocytes. Previous studies have demonstrated that the intracellular concentration of free Ca2+ increases in several types of cells during stress. For example, acute restraint stress enhances Ca2+ mobilization in lymphocytes from mice. In addition, Ca2+ ions are essential for production of interleukin-2 by T cells (22). Withania somnifera (WS) is an annual herb and a rich source of bioactive compounds. Several pharmacological activities of the plant have been attributed to its roots. Oral administration of root extract of WS at 10, 30 and 300 mg/kg body weight for 15 days stimulated B and T cells proliferation, induced type 1 immunity, increased the activity of machrophage and levels of immunoglobulin which indicated the potent role of extract on humoral and cellular immunity. The present study aimed to evaluate the main effects and interaction of calcitriol and WS root extract in either low or adequate Ca diets on immune response and small intestinal morphology of broiler chickens.
Material and Methods Six hundred male day old Ross 308 broilers obtained from a commercial hatchery, and reared in 60 floor pens with wood shavings litter at a stocking rate of 10 birds per pen (1×1 m). Feed and water were provided ad libitum throughout the 6–wk experimental period. The experimental design was a 2×3×2 factorial comprising of two dietary concentration of Ca (negative and positive control), three concentrations of WS root extract (0, 75 and 150 mg/kg diet), two concentrations of 1, 25 (OH)2 D3 (0 and 0.5 g/kg diet). SRBC test was used to measure humoral immunity response. So, at 25 and 32 days of age chicks were injected to evaluate the primary and secondary antibody response. On d 21 and 42, one bird per replicate was killed to determine small intestinal morphology and lymphoid organ weights. Sample sections (2.5 cm in length) were taken from the middle region of the jejunum. The tissues were stained for measurement of villus length, villus width, crypth depth and thickness of muscle layer. The General Linear Models of SAS were used to analyse all the data.
Results and Discussion The results of humoral immune response showed that reduction of dietary Ca level by 30% resulted to depression of immunoglobulin M and total antibody levels. Supplementation of WS at 150 mg/kg significantly improved the level of immunoglobulin G only in primary test. Calcium ions have an essential role function in activation and maturation of lymphocytes (10). Previous studies have demonstrated that the intracellular concentration of free Ca2+ increases in several types of cells during stress. For example, acute restraint stress enhances Ca2+ mobilization in lymphocytes from mice. In addition, Ca2+ ions are essential for production of interleukin-2 by T cells. It was found that supplementation of calcitriol resulted in shorter villus length at 21 d and shorter villus width at 42 day of age, respectively. Reduction of dietary Ca level by 30% resulted to shorter crypth depth compared to positive control diet. A researcher reported that supplementation of 25-OH-D3 decreased the length and weight of small intestine. The duodenal putrescine content was enhanced in parallel with the increase in intestinal calcium absorption, suggesting that polyamines, in particular putrescine, may be involved somehow in the intestinal calcium transport mechanism. The aliphatic polyamines, putrescine, spermidine, and spermine, are considered to be essential cell constituents that play an important role in modulating cell proliferation and differentiation.
Conclusion The results of humoral immune response showed that decreasing of dietary Ca level to 30% resulted to reduction of total antibody response. The findings showed that supplementation of calcitriol did not exert beneficial effects on intestine morphology. Supplementation of WS root extract did not affect intestine characteristics.

کلیدواژه‌ها [English]

  • Broiler chicken
  • Calcitriol
  • Immune response
  • Intestine morphology
  • Withania somnifera
1- Aslam, S. M., J. D. Garlich, and M. A. Qureshi. 1998. Vitamin D deficiency alters the immune responses of broiler chicks. Poultry Science, 77(6): 842-9.
2- Beal, R. K., P. Wigley., C. Powers., S. D. Hulme., P. A. Barrow, and A. L. Smith. 2004. Age at primary infection with Salmonella enterica serovar Typhimurium in the chicken influences persistence of infection and subsequent immunity to re-challenge. Veterinary Immunology and Immunopathology, 100: 151-164.
3- Berridge, M. J. 1985. The molecular basis of communication within the cell. Scientific American, 253:142–152.
4- Berridge, M. J., M. D. Bootman, and H. L. Roderick. 2003. Calcium signalling: Dynamics, homeostasis and remodelling. Nature Reviews Molecular Cell Biology, 4: 517–529.
5- Cantorna, M. T., Y. Zhu., M. Froicu, and A.Wittke. 2004. Vitamin D status, 1,25-dihydroxyvitamin D3, and the immune system. The American Journal of Clinical Nutrition, 80(6): 1717S-1720S.
6- Chou, S. H., T. K. Chung, and B. Yu. 2009. Effects of supplemental 25-hydroxycholecalciferol on growth performance, small intestinal morphology, and immune response of broiler chickens. Poultry Science, 88: 2333–2341.
7- Davis, L, and G. Kuttan. 1998. Suppressive effect of cyclophosphamide-induced toxicity by Withania somnifera. Journal of Ethno pharmacology, 71: 193-200.
8- Davis, L, and G. Kuttan. 2000. Immunomodulatory activity of Withania somnifera”. Journal of Ethno pharmacology, 62: 209-214.
9- Eerola, E., T. Veromaa, and P. Toivanen. 1987. Special features in the structural organization of the avian lymphoid system. Pages 9–22 In Avian Immunology: Basis and Practice. Toivanen, A, and P. Toivanen, ed. CRC Press, Inc., Boca Raton, FL.
10- Feske, S. 2007. Calcium signalling in lymphocyte activation and disease. Nature Reviews Immunology, 7: 690-702.
11- Garlich, J. D., M. A. Qureshi., P. R. Ferket, and S. M. Aslam. 1992. Immune system modulation by dietary calcium. Pages 99–106 in: Proceedings of the 19th World’s Poultry Congress, Amsterdam, Netherlands.
12- Kazemi Fard, M., H. Kermanshahi., M. Rezaei, and A. Golian. Effect of fennel extract and vitamin D3 on hatchability and and qualitative characteristics of chicks in post molt broiler breeder performance. Iranian Journal of Animal Science Research, 6(3): 197-207. (In Persian)
13- Kenney, A. D. 1976. Vitamin D metabolism: Physiological regulation in egg-laying Japanese quail. American Journal of Physiology, 230: 1606– 1616.
14- Kimura, K., J. P. Goff, and M. E. Kehrli, J. 1999. Effects of the presence of the mammary gland on expression of neutrophil adhesion molecules and myeloperoxidase activity in periparturient dairy cows. Journal of Dairy Science, 82: 2385-92.
15- Kimura, K., T. A. Reinhardt, and J. P.Goff. 2006. Parturition and hypocalcemia blunts calcium signals in immune cells of dairy cattle. Journal of Dairy Science, 89: 2588-2595.
16- Linton, P. J. and N. R. Klinman. 1992. The generation of memory B cells. Seminars in Immunology, 4: 3-9.
17- Malik, F., J. Singh., A. Khajuria., K. Suri., N. Satti, and S. Singh. 2007. A standardized root extract of Withania somnifera and its major constituent withanolide-A elicit humoral and cell-mediated immune responses by up-regulation of Th1-dominant polarization in BALB/c mice. Life Science, 80: 525–1538.
18- McCrady, C. W., C. M. Ely., E. Westin, and R. A.Carchman. 1988. Coordination and reversibility of signals for proliferative activation and interleukin-2 mRNA production in resting human T lymphocytes by phorbol ester and calcium ionophore. The Journal of Biological Chemistry, 263: 18537-18544.
19- Mirakzehi, M. T., H. Kermanshahi., A. Golian, and A. R. Raji. 2014. The effects of dietary 1, 25-dihydroxycholecalciferol and hydroalcoholic extract of Withania somnifera root on bone mineralisation and strength in broiler chickens. Iranian Journal of Animal Science Research, 6(3): 313-324. (In Persian)
20- Montagne, L., J. R. Pluske, and D. J. Hampson. 2003. A review of interactions between DF and the intestinal mucosa, and their consequences on digestive health in young non-ruminant animals. Animal Feed Science and Technology, 108: 95-117.
21- Mu, H. H, and W. A. Sewell. 1994. Regulation of DTH and IgE responses by IL-4 and IFN-gamma in immunized mice given pertussis toxin. Immunology, 83: 639-45.
22- Negulescu, P. A., N. Shastri, and M. D. Cahalan. 1994. Intracellular calcium dependence of gene expression in single T lymphocytes. Proceedings of the National Academy of Sciences, 91(7): 2873-2877.
23- Qureshi, M. A, and G. B. Havenstein. 1994. A comparison of the immune performance of a 1991 commercial broiler with a 1957 randombred strain when fed ‘typical’ 1957 and 1991 broiler diets. Poultry Science, 73:1805–1812.
24- Reddy, G. S, and K. Y. Tserng.1989. Calcitroic acid, end product of renal metabolism of 1,25-dihydroxyvitamin D3 through C-24 oxidation pathway. Biochemistry, 28: 1763–1769.
25- Ross. 2007. Ross 308 Broiler: Nutrition Specification. Aviagen, Scotland, UK. Accessed May 25, 2009. Available at http://www.aviagen.com/.
26- SAS Institute. 2000. SAS User’s Guide: Statistics, Version 9.1 Edition. Cary, NC, USA.
27- Satoh, E., H. Edamatsu, and Y. Omata. 2006. Acute restraint stress enhances calcium mobilization and proliferative response in splenic lymphocytes from mice. Stress, 9(4): 223-30.
28- Sei, Y., T. McIntyre., P. Skolnick, and P. K. Arora. 1991. Stress modulates calcium mobilization in immune cells. Life Science, 49: 671-6.
29- Shinki, T., H. Tanaka., J. Takito., A. Yamaguchi., Y. Nakamura., S. Yoshiki, and T. Suda. 1991. Putrescine is involved in the vitamin D action in chick intestine. Gastroenterology, 100: 113–122.
30- Shinki, T., N. Takahashi., T. Kadofuku., T. Sato, and T. Suda. 1985. Induction of spermidine N1-acetyltransferase by 1α, 25-dihydroxyvitamin D3 as an early common event in the target tissues of vitamin D. The Journal of Biological Chemistry, 260: 2185–2190.
31- Shinki, T., T. Kadofuku., T. Sato, and T. Suda. 1986. Spermidine N1-acetyltransferase has a larger role than ornithine decarboxylase in 1 alpha, 25-dihydroxyvitamin D3-induced putrescine synthesis. The Journal of Biological Chemistry, 261: 11712-6.
32- Soares, J. H. Jr. 1984. Calcium metabolism and its control-a review. Poultry Science, 63: 2075–2083.
33- Spielvogel, A. M., R. D. Farley, and A. W. Norman. 1972. Studies on the mechanism of action of calciferol. V. Turnover time of chick intestinal epithelial cells in relation to the intestinal action of vitamin D. Experimental Cell Research, 74: 359-66.
34- Suvas, S., V. Singh., S. Sahdev., H. Vohra, and J. N. Agrewala. 2002. Distinct role of CD80 and CD86 in the regulation of the activation of B cell and B cell lymphoma. The Journal of Biological Chemistry, 277: 7766-75.
35- Toivanen, P., A. Naukkarinen, and O.Vainio. 1987. What is the function of bursa of Faricius? Pages 79–100 In Avian Immunology: Basis and Practice. Toivanen, A, and P. Toivanen, ed. CRC Press, Inc., Boca Raton, FL.
36- Uni, Z. and P. R. Ferket. 2004. Methods for early feeding and their potential. World Poultry Science Journal, 60: 101-111.
37- Uni, Z., A. Geyra., H. Ben-Hur, and D. Sklan. 2000. Small intestinal development in the young chick: crypt formation and enterocyte proliferation and migration. British Poultry Science, 41(5): 544-551.
38- Wang, Z. H., C. A.Vidair, and W. C. Dewey. 1991. Maintenance of intracellular free Ca2+ homeostasis following lethal heat shock. Radiation Research, 128: 104–107.
39- White, R. G., D. C. Henderson., M. B. Eslami, and K. H. Neilsen. 1975. Localization of a protein antigen in the chicken spleen. Effect of various manipulative procedures on the morphogenesis of the germinal centre. Immunology, 28: 1-21.
40- Yamamoto, Y, and B. Glick. 1982. A comparison of the immune response between two lines of chickens selected for differences in the weight of the bursa of Fabricius. Poultry Science, 61: 2129–2132.
41- Yamauchi, K., T. Buwjoom., K. Koge, and T. Ebashi. 2006. Histological alterations of the intestinal villi and epithelial cells in chickens fed dietary sugar cane extract. British Poultry Science, 47: 544-553.
42- Yang, S., C. Smith., J. M. Prahl., X. Luo, and H. F. DeLuca. 1993. Vitamin D deficiency suppresses cell-mediated immunity in vivo. Archives of Biochemistry and Biophysics, 303(1): 98-106.
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