Improving Broiler Chicken Intestinal Microbiota and Immune Function Using Black Cumin Seed Bioactive Peptides: A Comparative Study with Prebiotics and Organic Acids

Document Type : Original Paper

Authors

1 Department of Animal Science, University of Mohaghegh Ardabili, Ardabil, Iran

2 Department of Animal Science, Rodaki Higher Education Institute, Tonekabon, Mazandaran, Iran

3 Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Hamedan, Iran

10.22069/psj.2025.23324.2260

Abstract

This research investigated the impact of hydrolyzed black cumin seed protein (HBCP) as a potential source of bioactive peptides on the performance of broiler chickens. A total of 560 Ross 308 broiler chicks were randomly assigned to seven distinct dietary groups: Control, 0.05% HBCP, 0.1% HBCP, 0.15% HBCP, 0.2% HBCP, 0.2% Prebiotic and 0.2% Organic Acid. The experiment was conducted over a 42-day period. Birds were fasted for 8 hours before weighing, and feed intake was recorded daily to calculate feed conversion ratios. For microbial and enzyme analyses, two birds per replicate were selected and euthanized, with intestinal samples collected under sterile conditions. Serum samples were taken on day 42 to assess liver enzyme concentrations, and immune response was measured using a sheep red blood cell assay. The results indicated that including 0.1%, 0.15%, and 0.2% HBCP in the broiler diets significantly improved body weight gain and feed conversion ratio throughout the rearing period. Additionally, the inclusion of 0.2% prebiotic and organic acid in the diet notably increased body weight and decreased feed conversion ratio compared to the control group (P<0.05). These dietary interventions had a positive effect on the intestinal microbiota, decreasing Escherichia coli counts while boosting Lactobacillus populations (P<0.05). Additionally, supplementation with HBCP, prebiotics, and organic acids improved immune function, as indicated by elevated antibody titers and enhanced liver enzyme activity. (P<0.05). In conclusion, the study showed that bioactive peptides derived from black cumin hydrolysate had effects similar to those of prebiotics and organic acids, improving both productive traits and immune status in broilers at comparable dosage levels.

Keywords


Abdollahi MR, Zaefarian F, Gu Y, Xiao W, Jia J & Ravindran V. 2017. Influence of soybean bioactive peptides on growth performance, nutrient utilization, digestive tract development and intestinal histology in broilers. Journal of Applied Animal Nutrition, 5: e7. DOI: 10.1017/JAN.2017.6
Abdollahi MR, Zaefarian F, Gu Y, Xiao W, Jia J & Ravindran V. 2018. Influence of soybean bioactive peptides on performance, foot pad lesions and carcass characteristics in broilers. Journal of Applied Animal Nutrition, 6: e3. DOI:
Ağagündüz D, Cemali Ö & Çelik E. 2023. Interaction between natural products and gut microbiota. Current Pharmacology Reports, 9(1): 7–31. DOI: 10.1007/s40495-022-00309-5
Albakry Z, Karrar E, Ahmed IAM, Oz E, Proestos C, El Sheikha AF, Oz F, Wu G & Wang X. 2022. Nutritional composition and volatile compounds of black cumin (Nigella sativa L.) seed, fatty acid composition and tocopherols, polyphenols, and antioxidant activity of its essential oil. Horticulturae, 8(7): 575. DOI: 10.3390/horticulturae8070575
Aviagen. 2019. Ross 308: Broiler performance objectives and nutrition specifications. Newbridge, UK: Aviagen Ltd.
Chen X, Wang J & Ma H. 2009. Effects of oligopeptides on digestive enzyme secretion in poultry. Journal of Animal Science, 87(6): 1964–1972. DOI: 10.2527/jas.2008-1234
Choi SC, Ingale SL, Kim JS, Park YK, Kwon IK & Chae BJ. 2013. Effects of dietary supplementation with an antimicrobial peptide-P5 on growth performance, nutrient retention, excreta, intestinal microflora, and intestinal morphology of broilers. Animal Feed Science and Technology, 185(1–2): 78–84. DOI: 10.1016/j.anifeedsci.2013.01.021
Cruz-Casas DE, Aguilar CN, Ascacio-Valdés JA, Rodríguez-Herrera R, Chávez-González ML & Flores-Gallegos AC. 2021. Enzymatic hydrolysis and microbial fermentation: The most favorable biotechnological methods for the release of bioactive peptides. Food Chemistry: Molecular Sciences, 3: 100047. DOI: 10.1016/j.fochms.2021.100047
Eftekhari A, Rezaeipour V & Abdullahpour R. 2015. Effects of acidified drinking water on performance, carcass, immune response, jejunum morphology, and microbiota activity of broiler chickens fed diets containing graded levels of threonine. Livestock Science, 180: 158–163. DOI: 10.1016/j.livsci.2015.07.010
Feng J, Liu X, Xu ZR, Wang YZ & Liu JX. 2007. Effects of fermented soybean meal on digestive enzyme activities and intestinal morphology in broilers. Poultry Science, 86(6): 1149–1154. DOI: 10.1093/ps/86.6.1149
Gadde U, Kim WH, Oh ST & Lillehoj HS. 2017. Alternatives to antibiotics for maximizing growth performance and feed efficiency in poultry: A review. Animal Health Research Reviews, 18(1): 26–45. DOI: 10.1017/S1466252316000207
Gao D, Cao Y & Li H. 2010. Antioxidant activity of peptide fractions derived from cottonseed protein hydrolysate. Journal of the Science of Food and Agriculture, 90(11): 1855–1860. DOI: 10.1002/jsfa.4021
Ghadiri N, Javidan M, Sheikhi S, Taştan Ö, Parodi A, Liao Z & Ganjalikhani-Hakemi M. 2024. Bioactive peptides: An alternative therapeutic approach for cancer management. Frontiers in Immunology, 15: 1310443. DOI: 10.3389/fimmu.2024.1310443
Guo Z, Yi D, Hu B & Zhou Q. 2021. The alteration of gut microbiota by bioactive peptides: A review. Systems Microbiology and Biomanufacturing, 1(4): 363–377. DOI: 10.1007/s43393-021-00035-x
Hannan MA, Rahman MA, Sohag AAM, Uddin MJ, Dash R, Sikder MH, Rahman MS, Timalsina B, Munni YA, Sarker PP, Alam M, Mohibbullah M, Haque MN, Jahan I, Hossain MT, Afrin T, Rahman MM, Tahjib-ul-Arif M, Mitra S & Kim B. 2021. Black cumin (Nigella sativa L.): A comprehensive review on phytochemistry, health benefits, molecular pharmacology, and safety. Nutrients, 13(6): 1784. DOI: 10.3390/nu13061784
Hisham RI, Isono H & Miyata T. 2018. Potential antioxidant bioactive peptides from camel milk proteins. Animal Nutrition, 4(3): 273–280. DOI: 10.1016/j.aninu.2018.05.004
Hossain MS, Rahman MZ, Mozumder SN & Rahman J. 2024. Nutritional composition, fatty acids, bioactive compounds, and antioxidant activity of Nigella sativa seed grown in Bangladesh. Discover Food, 4: 52. DOI: 10.1007/s44187-024-00102-x
Karimzadeh S, Rezaei M & Teimouri Yansari A. 2016. Effects of canola bioactive peptides on performance, digestive enzyme activities, nutrient digestibility, intestinal morphology, and gut microflora in broiler chickens. Poultry Science Journal, 4(1): 27–36. DOI: 10.22069/psj.2016.2969
Karimzadeh S, Rezaei M & Teimouri Yansari A. 2017. Effects of different levels of canola meal peptides on growth performance and blood metabolites in broiler chickens. Livestock Science, 203: 37–40. DOI: 10.1016/j.livsci.2017.06.013
Kotzamanis YP, Gisbert E, Gatesoupe FJ, Zambonino Infante J & Cahu C. 2007. Effects of different dietary levels of fish protein hydrolysates on growth, digestive enzymes, gut microbiota, and resistance to Vibrio anguillarum in European sea bass larvae. Comparative Biochemistry and Physiology Part A: Molecular and Integrative Physiology, 147(1): 205–214. DOI: 10.1016/j.cbpa.2006.12.037
Mada SB, Ugwu CP & Abarshi MM. 2020. Health promoting effects of food-derived bioactive peptides: A review. International Journal of Peptide Research and Therapeutics, 26: 831–848. DOI: 10.1007/s10989-019-09890-8
Mokhtari R, Kazemi Fard M, Rezaei M & Dirandeh E. 2024. Effect of casein bioactive peptides on performance, nutrient digestibility, enzyme activity, and intestinal microbial population in broiler chickens. Journal of Animal Physiology and Animal Nutrition, 108(6): 1798–1806. DOI: 10.1111/jpn.14021
Mohammadrezaei M, Navidshad B, Gheisari A & Toghyani M. 2021. Cottonseed meal bioactive peptides as an alternative to antibiotic growth promoters in broiler chicks. International Journal of Peptide Research and Therapeutics, 27: 329–340. DOI: 10.1007/s10989-020-10086-8
Nasiri N, Ilaghi Nezhad M, Sharififar F, Khazaneha M, Najafzadeh MJ & Mohamadi N. 2022. The therapeutic effects of Nigella sativa on skin disease: A systematic review and meta-analysis of randomized controlled trials. Evidence-Based Complementary and Alternative Medicine, 2022: 7993579. DOI: 10.1155/2022/7993579
Osman A, Goda HA, Abdel-Hamid M, Badran SM & Otte J. 2016. Antibacterial peptides generated by alkaline hydrolysis of goat whey. LWT – Food Science and Technology, 65: 480–486. DOI: 10.1016/j.lwt.2015.08.058
Ovissipour M, Safari R, Motamedzadegan A & Shabanpou B. 2012. Use of hydrolysates from yellowfin tuna (Thunnus albacares) heads as a complex nitrogen source for lactic acid bacteria. Food and Bioprocess Technology, 5(1): 73–79. DOI: 10.1007/s11947-010-0481-1
Pasupuleti VK & Demain AL. 2010. Protein hydrolysates in biotechnology. Springer Science+Business Media. DOI: 10.1007/978-1-4020-6674-0
Paul SS, Rao SVR, Hegde N, Williams NJ, Chatterjee RN, Raju MVLN, Reddy GN, Kumar V, Kumar PSP, Mallick S & Gargi M. 2022. Effects of dietary antimicrobial growth promoters on performance parameters and abundance and diversity of broiler chicken gut microbiome and selection of antibiotic resistance genes. Frontiers in Microbiology, 13: 905050. DOI: 10.3389/fmicb.2022.905050
Power O, Jakeman P & Fitzgerald RJ. 2013. Antioxidative peptides: Enzymatic production, in vitro and in vivo antioxidant activity, and potential applications of milk-derived antioxidative peptides. Amino Acids, 44(3): 797–820. DOI: 10.1007/s00726-012-1393-9
Ryder K, Bekhit AED, McConnell M & Carne A. 2016. Towards generation of bioactive peptides from meat industry waste proteins: Generation of peptides using commercial microbial proteases. Food Chemistry, 208: 42–50. DOI: 10.1016/j.foodchem.2016.03.099
Sa’adoon WH & Abbas RJ. 2023. Effect of dietary supplementation of soybean bioactive peptides and vitamin E on productive performance and some carcass characteristics in broiler chickens. IOP Conference Series: Earth and Environmental Science, 1262: 062035. DOI: 10.1088/1755-1315/1262/6/062035
Saeid JM, Mohamed AB & Al-Baddy MA. 2013. Effect of garlic powder (Allium sativum) and black seed (Nigella sativa) on broiler growth performance and intestinal morphology. Iranian Journal of Applied Animal Science, 3(1): 123–130.
Salavati ME, Rezaeipour V, Abdullahpour R & Mousavi N. 2020. Effects of graded inclusion of bioactive peptides derived from sesame meal on the growth performance, internal organs, gut microbiota, and intestinal morphology of broiler chickens. International Journal of Peptide Research and Therapeutics, 26(3): 1541–1548. DOI: 10.1007/s10989-019-09947-8
Salavati ME, Rezaeipour V, Abdullahpour R & Mousavi SN. 2021. Bioactive peptides from sesame meal for broiler chickens: Its influence on the serum biochemical metabolites, immunity responses and nutrient digestibility. International Journal of Peptide Research and Therapeutics, 27: 1297–1303. DOI: 10.1007/s10989-021-10168-1
Seifi M, Rezaei M & Teimouri Yansari A. 2018. Effect of different levels of soybean meal peptides on performance, intestinal morphology, and intestinal bacterial population in broiler chicks. Journal of Animal Science, 9(22): 9–17. DOI: 10.29252/rap.9.22.9
Singh BP, Vij S & Hati S. 2014. Bioactive peptides from legumes and their bioavailability. Journal of Food Science and Technology, 51(9): 2176–2192. DOI: 10.1007/s13197-012-0674-5
Surolia R, Tyagi M & Singh A. 2024. A holistic approach: Exploring pre, pro, syn, post and paraprobiotics in sustainable diets. In: Thakur M (ed.), Sustainable Food Systems (Volume I). World Sustainability Series. Springer, Cham. DOI: 10.1007/978-3-031-47122-3_10
Tang Z, Yin Y, Zhang Y, Huang R, Sun Z, Li T & Wu G. 2012. Effects of dietary supplementation with amino acids on the growth performance and immune responses of broilers. British Journal of Nutrition, 108(1): 1–10. DOI: 10.1017/S0007114511005170
Tietz NW & Fiereck EA. 1966. A specific method for serum lipase determination. Journal of Clinical Chemistry and Laboratory Medicine, 13: 352–358.
Urban J, Kareem KY, Matuszewski A, Bień D, Ciborowska P, Lutostański K & Michalczuk M. 2024. Enhancing broiler chicken health and performance: The impact of phytobiotics on growth, gut microbiota, antioxidants, and immunity. Phytochemistry Reviews. DOI: 10.1007/s11101-024-09994-0
Wald M, Schwarz K, Rehbein H, Bußmann B & Beermann C. 2016. Detection of antibacterial activity of an enzymatic hydrolysate generated by processing rainbow trout by-products with trout pepsin. Food Chemistry, 205: 221–228. DOI: 10.1016/j.foodchem.2016.03.014
Wang G, Li X & Wang Z. 2016. APD3: The antimicrobial peptide database as a tool for research and education. Nucleic Acids Research, 44(D1): D1087–D1093. DOI: 10.1093/nar/gkv1278
Wang M, Feng J, Zhou D & Wang J. 2023. Bacterial lipopolysaccharide-induced endothelial activation and dysfunction: A new predictive and therapeutic paradigm for sepsis. European Journal of Medical Research, 28: 339. DOI: 10.1186/s40001-023-01301-5
Wei Y, Zhao X, Xu T, Liu Z, Zuo Y, Zhang M, Zhang Y & Yin H. 2024. Soybean bioactive peptide supplementation affects the intestinal immune antioxidant function, microbial diversity, and reproductive organ development in roosters. Animals, 14(13): 1954. DOI: 10.3390/ani14131954
Wijesekara T, Abeyrathne EDNS & Ahn DU. 2024. Effect of bioactive peptides on gut microbiota and their relations to human health. Foods, 13(12): 1853. DOI: 10.3390/foods13121853
Zaky AA, Simal-Gandara J, Eun JB, Shim JH & Abd El-Aty AM. 2022. Bioactivities, applications, safety, and health benefits of bioactive peptides from food and by-products: A review. Frontiers in Nutrition, 8: 815640. DOI: 10.3389/fnut.2021.815640
Zambrowicz A, Pokora M, Setner B, Dąbrowska A, Szołtysik M, Babij K, Szewczuk Z, Trziszka T, Lubec G & Chrzanowska J. 2015. Multifunctional peptides derived from an egg yolk protein hydrolysate: Isolation and characterization. Amino Acids, 47(2): 369–380. DOI: 10.1007/s00726-014-1869-x