اثر مصرف جیره های مکمل شده با سطوح مختلف ویتامین E، سیزامین و تیموکوئینون بر چالش پاسخ سیستم ایمنی سلولی، ریخت شناسی روده، جمعیت میکروبی روده و بیان ژن MUC2 در بلدرچین های ژاپنی تخم‌گذار

محورهای موضوعی : فصلنامه زیست شناسی جانوری

یاسر رحیمیان ¹ , فرشید خیری ² , مصطفی فغانی ³

1 - گروه علوم دامی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران
2 - گروه علوم دامی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران
3 - گروه علوم دامی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران

تاریخ دریافت : 1402/03/18 تاریخ پذیرش : 1402/05/05 تاریخ انتشار : 1402/12/01

کلید واژه: ویتامین E, سیزامین, تیموکوئینون, سیستم ایمنی سلولی, بیان ژن MUC2, بلدرچین ژاپنی تخم‌گذار,

چکیده مقاله :

به منظور بررسی اثر مصرف جیره های مکمل شده با سطوح مختلف ویتامین E، سیزامین و تیموکوئینون بر چالش پاسخ سیستم ایمنی سلولی ، ریخت شناسی، جمعیت میکروبی روده و بیان ژن MUC2 در بلدرچین های ژاپنی تخم‌گذار، تعداد210 قطعه بلدرچین ژاپنی ماده در سن 35-85 روزگی با 7 تیمار آزمایشی شامل گروه شاهد و مصرف کننده ویتامین E، تیموکوئنیون و سیزامین با سطوح 100 و 200 میلی‌گرم/کیلوگرم از خوراک استاندارد با 5 تکرار و 6 قطعه بلدرچین تخم‌گذار در هر تکرار استقاده گردید. در انتهای دوره (85 روزگی) پس از کشتار دو قطعه پرنده از هر تکرار وزن نسبی اندام های ایمنی (تیموس، طحال و بورس فابریسیوس) و فعالیت شاخص های کبدی نظیر آلبومین، آنزیم های آلکالین فسفاتاز (ALP)، آلانین ترانس آمیناز (ALT) و آسپارتات ترانس آمیناز (AST) بررسی شد. پاسخ های ایمنی سلولی نظیر تست چالش حساسیت به دی نیتروکلروبنزن و پاسخ به تزریق میتوژن فیتوهماگلوتینین به وب بال چپ مطالعه شد. برای ارزیابی مورفولوژی بافت روده و جمعیت فلور میکروبی روده باریک، نمونه برداری صورت گرفت. برای بررسی بیان ژن MUC2نمونه هایی از ژژنوم جمع آوری و از روش RT-PCR استفاده شد. نتایج نشان داد مصرف جیره های مکمل شده سبب افزایش وزن نسبی معنی دار بورس فابرسیوس (05/0 ≤ p) و کاهش غیرمعنی داری فعالیت آنزیم های کبدی گردید (05/0≥ p). افزایش معنی دار فلور میکروبی مفید (لاکتوباسیلوس ها) و در مقابل کاهش جمعیت کلونی باکتری های اشرشیاکولی و سالمونلا انتریکا در روده همراه با افزایش سطح ویلی ها و تراکم تعداد سلول‌های گابلت نشان دهنده اثرات مفید ترکیبات فعال استفاده شده در بهبود وضعیت سلامت بلدرچین ها بود (05/0 ≤ p). مصرف جیره های مکمل شده با سطوح ویتامین E، سیزامین و تیموکوئنیون سبب افزایش بیان mRNA ژن MUC2در روده شد. نتایج نشان دهنده ی سودمندی مصرف ویتامین E، سیزامین و تیموکوئنیون بر پاسخ سیستم ایمنی سلولی، مورفولوژی، فلور میکروبی روده و بیان mRNA ژن MUC2 در بلدرچین های ژاپنی تخم‌گذار بود.

چکیده انگلیسی:

In order to investigate the effect of consuming diets supplemented with different levels of vitamin E, sesamin and thymoquinone on cellular immune system challenges, intestinal morphology, intestinal microbial population and MUC2 gene expression in laying Japanese quails, 210 female Japanese quails aged 85-35 days with 7 experimental treatments including the control group and those consuming vitamin E, thymoquinone and sesamin at levels of 100 and 200 ml.gram/kg of standard feed was used with 5 repetitions and 6 pieces of laying quail in each repetition. At the end of the period (85 days), after killing two quails from each repetition, the relative weight of thymus, spleen and bursa of fabricius and the liver enzymes activity indices such as albumin, alkaline phosphatase (ALP), alanine transaminase (ALT) and aspartate transaminase (AST) were investigated. Dinitrochlorobenzene sensitization challenge test and response to the injection of mitogen phytohemagglutinin into the left wing web were studied. Sampling was done to evaluate the morphology of the intestinal tissue and the microbial flora population of the small intestine. To investigate the expression of MUC2 gene, samples from jejunum were collected and RT-PCR method was performed. The results showed that the consumption of supplemented diets caused a significant increase in the bursa.f relative weight (p ≤ 0.05) and a non-significant improvement of the liver enzyemes activity (p ≥ 0.05). The significant increase of beneficial microbial flora (Lactobacillus) and decrease of the colony population of (Escherichia coli and Salmonella enterica) bacteria in the intestine along with the increase in the surface of the villi and the density of the number of goblet cells indicated the beneficial effects of the active compounds used in improving the health status of quails (p ≤ 0.05). Consuming supplemented diets with levels of vitamin E, sesamin and thymoquinone caused an increase in MUC2 mRNA expression in the intestine. The results showed the benefits of vitamin E, sesamin and thymoquinone consumption on cell mediated mediated immunity, intestinal morphology, intestinal microbial flora and MUC2 mRNA expression in laying Japanese quails.

منابع و مأخذ:

1. Abbasnezhad A. 2015. Physiological effects of Nigella sativa seed on different body systems: A review study. The Horizon of Medical Sciences, 21:71-81.


2. 2005. Official methods of analysis, Association of official analytical chemists. AOAC Press, Gaithersburg, USA.


3. Al-Jishi S.A., Abuo Hozaifa B .2003. Effect of Nigella sativa on blood hemostatic function in rats. Journal of Ethnopharmacolgy, 85(1):7-14.


4. Aliakbarpour H.R., Chamani M., Rahimi M., Sadeghi A.A., Qujeq D. 2012. The bacillus subtilis and lactic acid bacteria probiotics influences intestinal mucin gene expression, histomorphology and growth performance in broilers. Asian-Australasian Journal of Animal Sciences, 25(9):1285-1293.


5. Ashayerizadeh O., Dastar B., Shams Shargh M. 2009. Use of garlic (Allium sativum), black cumin seeds (Nigella sativa L.) and wild mint (Mentha longifolia) in broiler chickens’ diets. Journal of Animal Veterinary Advance, 8(9):1860-1863.


6. Ait M.L., Ait M. H., Elabbadi N .2007. Anti-tumor properties of black seed (Nigella sativa) extracts. Brazilian Journal of Medical and Biological Research, 40(6):839-847.


7. Azzam M.M., Zou X.T., Dong X.Y., Xie P. 2011. Effect of supplemental L-threonine on mucin 2 gene expression and intestine mucosal immune and digestive enzymes activities of laying hens in environments with high temperature and humidity. Poultry Sciences, 90(10):2251-2256.


8. Azeem T., Zaib UrRehman, Umar S., Asif M., Arif M., Rahman A. 2014. Effect of Nigella Sativa on poultry health and production: A Review of Scientifics Letters, 2(2):76-82.


9. Badary O.A., Gamal El-Din A.M. 2001. Inhibitory effects of thymoquinone against 20-methylcholanthrene-induced fibrosarcoma tumorigenesis. Cancer Detection Prevention, 25:362 –328.


10. Barnett T.C., Bugrysheva J., VScott J.R. 2007. Role of mRNA stability in growth phase regulation of gene expression in the group A streptococcus. Journal of Bacteriology, 189:1866-1873.


11. Birchenough G.M., Nystrom E.E., Johansson M.E., Hansson G.C. 2016. A sentinel goblet cell guards the colonic crypt by triggering Nlrp6-dependent Muc2 secretion. Science, 352:1535-1542.


12. Cheadle C., Fan J., Cho-Chung Y.S., Werner T., Ray J.2005. Stability regulation of mRNA and the control of gene expression. Annual Academy Sciences, 1058:196-204.


13. Chen K.L., Kho W.L., You S.H. 2009. Effects of Bacillus subtilis var. natto and Saccharomyces cerevisiae mixed fermented feed on the enhanced growth performance of broilers. Poultry Science, 88:309-315.


14. Cooney R.V., Custer L.J., Okinaka L., Franke A.A. 2001. Effects of dietary sesame seeds on plasma tocopherol levels. Nutrition and Cancer, 39(1):66-71.


15. Duncan D.B. 1995. Multiple range test and F-test. Biometrics, 11:1-42.


16. Erf G.F. 2004. Cell-Mediated immunity in poultry. Poultry Science, 83:580-590.


17. Farrag A.R., Mahdy K.A., Abdel Rahman G.H., Osfor M.M. 2007. Protective effect of Nigella sativa seeds against lead-induced hepatorenal damage in male rats. Pakistan Journal of Biology Sciences, 10(17):2809-2816.


18. Flint J.F., Garner M.R. 2009. Feeding beneficial bacteria: A natural solution for increasing efficiency and decreasing pathogens in animal agriculture. Journal of Applied Poultry Reproduction, 18:367-378.


19. Forder R.E.A., Howarth G. S., Tivey D.R., Hughes. R.J. 2007. Bacterial modulation of small intestinal goblet cells and mucin composition during early post-hatch development of poultry. Poultry Science, 86:2396-2403.


20. Gilani A., Jabeen Q., Ullah Khan M. 2004. A review of medicinal uses and pharmacological activities of Nigella sativa. Pakistan Journal of Biology Sciences,7441–451.


21. Hernandez F., Madrid J., Garcia V., Orengo J., Megias M.D. 2004. Influence of two plant extracts on broiler performance digestibility and digestive organ size. Journal of Poultry Science, 83:169-174.


22. Hoan N., Khoa M. 2016. The Effect of different levels of sesame oil on productive performance, egg yolk and blood serum lipid profile in laying hens. Open Journal of Animal Sciences, 6:85-93.


23. Horn N.L., Donkin S.S., Applegate T.J., Adeola O. 2009. Intestinal mucin dynamics. Response of broiler chicks and white Pekin ducklings to dietary threonine. Poultry Sciences, 88:1906-1914.


24. Hosseinzadeh H., Fazl Bazzaz B.S., Haghi M.M. 2007. Antibacterial activity of total extracts and essential oil of Nigella sativa L seeds in mice. Pharmacology Online, 2:429-435.


25. Jamroz D., Wertelecki T., Houszka M., Kamel C. 2006. Influence of diet type on the inclusion of plant origin active substances on morphological and histochemical characteristics of the stomach and jejunum walls in chicken. Journal of Animal Physiology and Animal Nutrition, 90:255-268.


26. Johansson M.E., Sjovall H., Hansson G.C. 2013. The gastrointestinal mucus system in health and disease. Nat Rev Gastroenterol Hepatology, 10:352-361.


27. Kabir S.M.L. 2009. The role of probiotics in the poultry industry. International Journal of Molocular Sciences, 10:3531-3546.


28. Kamali-Sangani A., Masoudi A., Hosseini. S. 2014. The effects of herbal plants on Mucin 2 gene expression and performance in ascetic broilers. Iranian Journal of Veterinary Medicine, 8(1):47-52.


29. Kagnoff M.F. 1993. Immunology of the intestinal tract antibacterial and antifungal properties of essential oils. Current Medical Chemistry,10.813-829.


30. Kato J.M., Alex C., Laurence B. D., Norman G.L. 1988. Biosynthesis of antioxidant lignans in Sesamum indicum Photochemistry, 47:583-592.


31. Khan S.H., Anjum M.A., Parveen A., Khawaja T., Ashraf N.M. 2013. Effects of black cumin seed (Nigella sativa) on performance and immune system in newly evolved crossbred laying hens. Veterinary Quart, 33(1):13-19.


32. Law G.K., Bertolo R.F., Adjiri Awere A., Pencharz P.B., Ball R.O. 2007. Adequate oral threonine is critical for mucin production and gut function in neonatal piglets. American Journal of Physiology Gastrology, 292:1293-1301.


33. Lev R., Spicer S.S. 1964. Specific staining of sulphate groups with Alcian blue at low pH. Journal of Histochemical and Cytochemistry, 12:309-310.


34. NRC, National Research Council. 1994. Nutrient requirements of poultry.9^thEd., Washington, DC. National Academy Press.


35. Montagne L., Piel C., Lalles J.P. 2004. Effect of diet on mucin kinetics and composition: nutrition and health implications. Nutrition Reviews, 62:105–14.


36. Nikzad M., Mirdar S. 2020. Inhibitory effects of Nigella sativa nano-capsule on the expression of cyclin D1 in the lungs of wistar rats exposed to nicotine-derived nitrosamine ketone. Journal of Medicinal Plants,19(74):118-128.


37. Pahlavan-Afshari K., Sanaan H. 2022. Effect of dietary levels of lavender oil on Muc2 gene expression in broiler chickens', Journal of Animal Environment, 14(2):113-122.


38. Salim H., Kang H., Akter N., Kim D., Kim J., Kim M., Na J., Jong H., Choi H. and Suh O. 2013. Supplementation of direct fed microbials as an alternative to antibiotic on growth performance, immune response, cecal microbial population and ileal morphology of broiler chickens. Poultry Sciences, 92:2084-2090.


39. SAS Institute. 2001. SAS.STAT user’s guide for personal computer. Release 6.12 SAS Institute, Inc., Cary, N.C., USA.


40. Sepehri-Moghaddam H., Kermanshahi H., Heravi A., Raji A. 2010. The effect of vitamin A on mucin 2 gene expression, intestinal histology and performance   of   broiler   Global Veterinaria, 5:168-174.


41. A, Sklan. D, Uni Z. 2004. Mucin dynamics in the chick small intestine is altered by starvation. Journal of Nutrition,134:736-742.


42. Smirnov A., Perez R., Romach E.A, Sklan D., Uni Z. 2005. Mucin dynamics and microbial populations in chicken small intestine are changed by dietary probiotic and antibiotic growth promoter supplementation. American Society for Nutritional Sciences, 67:90-105.


43. Smirnov A., Tako E., Ferket., P.R, Uni Z. 2006. Mucin gene expression and mucin content in the chicken intestinal goblet cells are affected by In-ovo feeding of carbohydrates. Poultry Science Journal, 85:669-673.


44. Talebi , Maham M., Asri-Rezaei S. 2021. Effects of Nigella sativa on performance, blood profiles, and antibody titer against Newcastle disease in broilers. Evidence-Based Complementary and Alternative Medicine, 10:2-15.


45. Thompson K.L., Applegate T.J. 2006. Food withdrawal alters small-intestinal morphology and mucus of broilers. Poultry Sciences, 85:1535-1540.


46. Van Der Sluis M., Vincent A., Bouma J. 2008. Forkhead box transcription factors Foxa1 and Foxa2 are important regulators of Muc2 mucin expression in intestinal epithelial cells. Biochem Biophys Research Communication, 369:1108-1113.


47. Vali N. 2008. The Japanese quail. A Review. International Journal of Poultry Science, 7:925-931.


48. Verma J., Johri T.S., Swain B. 2004. Effect of graded levels of aflatoxin, ochratoxin and their combinations on the performance and immune response of broilers. British Poultry Sciences, 45:512-518.


49. Yamashita K., Nohara Y., Katayama Y., Namiki M. 1992. Sesame seed lignans and gamma tocopherol act synergistically to produce vitamin E activity in rats. Journal of Nutrition, 122(12):2440-2446.


50. Yesilbag D., Gezen S.S, Biricik H., Bulbul T. 2012. Effects of a rosemary and oregano volatile oil mixture on performance, lipid oxidation of meat and hematological parameters in Pharaoh quails. British Poultry Science, 53:89-97.