The health of broiler chickens’ intestines when using a phytogenic composition in their diet

Relevance. The reduction in the use of feed antibiotics in poultry farming necessitates the development and study of alternative feed additives that positively affect gut health. Such additives include phytobiotics, which exhibit anti-inflammatory, antioxidant, antibacterial, and immunomodulatory properties. To identify the effect of different variants of a phytobiotic composition on gut health, an experiment was conducted on broiler chickens.

Methods. Three groups were formed, two of which were fed two variants of the phytobiotic composition with different ratios of chicory extract (40/35%), St. John’s wort (10/5%), safflower leuzea (35/40%), and creeping thyme (15/20%) in their diets. The impact on gut health was assessed through morphological characteristics of the cecum tissues, relative expression of genes related to resistance, and levels of opportunistic pathogenic microbiota.

Results. The results indicated a decrease in the expression of genes associated with inflammation and pathogen resistance in the gut tissues of the experimental groups: AvBD9 activity decreased 7.0-fold (p ≤ 0.05), AvBD10 by 12.7 to 140.0-fold (p ≤ 0.01), IL8L2 by 154.0-fold (p ≤ 0.01), and IRF7 by 47.0-fold (p ≤ 0.05). Histological studies showed reductions in epithelial height by 24.7 to 49.1% (p ≤ 0.001) and submucosal thickness by 2.8-2.9 times (p ≤ 0.001), an increase in crypt depth by 80.5% (p ≤ 0.001), muscularis mucosa thickness by 6.5% (p ≤ 0.05), and muscular layer thickness by 14.8% (p ≤ 0.001). No signs of degradation or suppression of epithelial cells were observed in the treated groups compared to control. Escherichia coli strains harboring virulence factors such as ybtA, iroE, fliA, epaQ, ycbV, ykgK/ecpR, fliP, espL4, cheB, stgC, among others, accounted for 11.98% of the cecal microbiome in the control group, whereas their abundance decreased to 3.74% in the 2nd experimental group and to 0.71% in the 3rd experimental group (p ≤ 0.05). These findings allow considering the developed phytogenic compositions as feed biopreparations aimed at improving gut health in broiler chickens.

1. Fisinin V. Global and domestic poultry farming: realities and challenges of the future. Animal Husbandry of Russia. 2025; (1): 8‒13 (in Russian). EDN GEOATN

2. Shatskikh E.V., Korolkova-Subbotkina D.E. The state of the intestines of broiler chickens when a phytobiotic supplement is included in the diet. Agrarian Bulletin of the Urals. 2022; (S14): 81‒91 (in Russian). https://doi.org/10.32417/1997-4868-2022-229-14-81-91

3. Sevastyanova T.V., Usha B.V., Noskov S.B., Alshevskaya A.A. The effect of functional targeted probiotics on the indicators of nonspecific resistance in young farm animals. Agrarian science. 2025; (11): 22‒32 (in Russian). https://doi.org/10.32634/0869-8155-2025-400-11-22-32

4. Vertiprakhov V.G. Physiology of intestinal digestion in chickens (Experimental approach). Moscow: Russian State Agrarian University - Moscow Timiryazev Agricultural Academy. 2022; 175 (in Russian). ISBN 978-5-9675-1887-4. EDN: VCFFIY

5. Oni A.I., Oke O.E. Gut health modulation through phytogenics in poultry: mechanisms, benefits, and applications. Frontiers in Veterinary Science. 2025; 12: 1616734. https://doi.org/10.3389/fvets.2025.1616734

6. Oni A.I., Adeleye O.O., Adebowale T.O., Oke O.E. The role of phytogenic feed additives in stress mitigation in broiler chickens. Journal of Animal Physiology and Animal Nutrition. 2024; 108(1): 81‒98. https://doi.org/10.1111/jpn.13869

7. Obianwuna U.E. et al. Phytobiotics in poultry: revolutionizing broiler chicken nutrition with plant-derived gut health enhancers. Journal of Animal Science and Biotechnology. 2024; 15: 169. https://doi.org/10.1186/s40104-024-01101-9

8. Gou H.-Z., Zhang Y.-L., Ren L.-F., Li Z.-J., Zhang Z. How do intestinal probiotics restore the intestinal barrier?. Frontiers in Microbiology. 2022; 13: 929346. https://doi.org/10.3389/fmicb.2022.929346

9. Zainuddin, Darmawan A., Sumiati, Wiryawan K.G., Nahrowi. Effects of Dietary Bacillus coagulans D3372 Supplementation as Probiotics on Broiler Performance, Ileal Microflora, Meat Quality, Nutrient Retention, and Metabolizable Energy. Advances in Animal and Veterinary Sciences. 2020; 8(1): 115‒123. https://doi.org/10.17582/journal.aavs/2020/8.1.115.123

10. Laptev G.Yu. et al. Microbiome of farm animals: relationship with health and productivity. St. Petersburg: Prospekt nauki. 2020; 334 (in Russian). ISBN 978-5-906109-99-6. EDN ONQNKC

11. Selionova M.I., Trukhachev V.I., Zagarin A.Y., Kulikov E.I., Belyaeva N.P. Effects of Dietary Supplementation Using Phytobiotics with Different Functional Properties on Expression of Immunity Genes, Intestinal Histology, Growth, and Meat Productivity of Broiler Chickens. Veterinary Sciences. 2025; 12(4): 302. https://doi.org/10.3390/vetsci12040302

12. Livak K.J., Schmittgen T.D. Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔC T Method. Methods. 2001; 25(4): 402‒408. https://doi.org/10.1006/meth.2001.1262

13. Laptev G.Yu. et al. Expression of genes of immune response and adaptation and cecal microbiome composition in males and females of chickens (Gallus gallus L.) in CM5 and CM9 preparental lines of Smena 9 cross. Agricultural Biology. 2023; 58(2): 313‒332. https://doi.org/10.15389/agrobiology.2023.2.313eng

14. Tyurina D.G. et al. Influence of antibiotics, glyphosate and a Bacillus sp. strain on productivity performance and gene expression in cross Ross 308 broiler chickens (Gallus gallus L.). Agricultural Biology. 2022; 57(6): 1147‒1165. https://doi.org/10.15389/agrobiology.2022.6.1147eng

15. Yildirim E.A. et al. Expression of key genes in the caecum in CM5 and CM9 chickens of the meat cross «Smena 9» against the background of the replacement of fishmeal. Agrarian science. 2023; (9): 52‒58 (in Russian). https://doi.org/10.32634/0869-8155-2023-374-9-52-58

16. Fancher C.A., Zhang L., Kiess A.S., Adhikari P.A., Dinh T.T.N., Sukumaran A.T. Avian Pathogenic Escherichia coli and Clostridium perfringens: Challenges in No Antibiotics Ever Broiler Production and Potential Solutions. Microorganisms. 2020; 8(10): 1533. https://doi.org/10.3390/microorganisms8101533

17. Waliaula P.K. Kiarie E.G., Diarra M.S. Predisposition factors and control strategies of avian pathogenic Escherichia coli in laying hens. Frontiers in Veterinary Science. 2024; 11: 1474549. https://doi.org/10.3389/fvets.2024.1474549

18. Panin A.N., Motorygin A.V., Abrosimova N.S., Mezhieva Z.Kh., Prasolova O.V., Manoyan M.G. Definition of the species Escherichia coli: problems of taxonomy and phylogeny. Agrarian science. 2025; (12): 22‒27 (in Russian). https://doi.org/10.32634/0869-8155-2025-401-12-22-27

19. Smati M. et al. Strain-specific impact of the high-pathogenicity island on virulence in extra-intestinal pathogenic Escherichia coli. International Journal of Medical Microbiology. 2017; 307(1): 44‒56. https://doi.org/10.1016/j.ijmm.2016.11.004

20. Johnson T.J., Wannemuehler Y., Doetkott C., Johnson S.J., Rosenberger S.C., Nolan L.K. Identification of Minimal Predictors of Avian Pathogenic Escherichia coli Virulence for Use as a Rapid Diagnostic Tool. Journal of Clinical Microbiology. 2008; 46(12): 3987‒3996. https://doi.org/10.1128/JCM.00816-08

21. Claret L., Miquel S., Vieille N., Ryjenkov D.A., Gomelsky M., DarfeuilleMichaud A. The Flagellar Sigma Factor FliA Regulates Adhesion and Invasion of Crohn Disease-associated Escherichia coli via a Cyclic Dimeric GMP-dependent Pathway. Journal of Biological Chemistry. 2007; 282(46): 33275‒33283. https://doi.org/10.1074/jbc.M702800200

22. Yildirim E.A. et al. Effect of different forms of lysine and methionine in broiler diets on the key genes transcriptional profile. Agrarian science. 2025; (2): 87‒94 (in Russian). https://doi.org/10.32634/0869-8155-2025-391-02-87-94

23. Mirakzehi M.T., Agah M.J., Baranzehi T., Saleh H. The Effects of Saccharomyces cerevisiae and Citric Acid on Productive Performance, Egg Quality Parameters, Small Intestinal Morphology, and Immune-Related Gene Expression in Laying Japanese Quails. Brazilian Journal of Poultry Science. 2022; 24(4): 1‒12. https://doi.org/10.1590/1806-9061-2022-1678

24. Shimizu M., Nii T., Isobe N., Yoshimura Y. Effects of avian infectious bronchitis with Newcastle disease and Marek’s disease vaccinations on the expression of toll-like receptors and avian β-defensins in the kidneys of broiler chicks. Poultry Science. 2020; 99(12): 7092‒7100. https://doi.org/10.1016/j.psj.2020.08.071

25. Yildirim E.A. et al. L-carnitine-induced changes in the expression profile of key genes in broiler chickens of the “Smena 9” cross, taking into account sex differences. Agrarian science. 2025; (10): 81‒90 (in Russian). https://doi.org/10.32634/0869-8155-2025-399-10-81-90

26. Kim T.H. Zhou H. Overexpression of Chicken IRF7 Increased Viral Replication and Programmed Cell Death to the Avian Influenza Virus Infection Through TGFBeta/FoxO Signaling Axis in DF-1. Frontiers in Genetics. 2018; 9: 415. https://doi.org/10.3389/fgene.2018.00415

27. Sarkar D. The anti-inflammatory activity of plant derived ingredients: an analytical review. International Journal of Pharmaceutical Sciences and Research. 2020; 11(2): 496‒506. https://doi.org/10.13040/IJPSR.0975-8232.11(2).496-06

28. Saleh H.A. Yousef M.H., Abdelnaser A. The Anti-Inflammatory Properties of Phytochemicals and Their Effects on Epigenetic Mechanisms Involved in TLR4/ NF-κB-Mediated Inflammation. Frontiers in Immunology. 2021; 12: 606069. https://doi.org/10.3389/fimmu.2021.606069

29. Pandey S. et al. Cutting-edge knowledge on the roles of phytobiotics and their proposed modes of action in swine. Frontiers in Veterinary Science. 2023; 10: 1265689. https://doi.org/10.3389/fvets.2023.1265689

30. Belote B.L. et al. Applying different morphometric intestinal mucosa methods and the correlation with broilers performance under Eimeria challenge. Poultry Science. 2023; 102(9): 102849. https://doi.org/10.1016/j.psj.2023.102849

31. Li L., Kubasová T., Rychlik I., Hoerr F.J., Rautenschlein S. Infectious bursal disease virus infection leads to changes in the gut associated-lymphoid tissue and the microbiota composition. PLOS One. 2018; 13(2): e0192066. https://doi.org/10.1371/journal.pone.0192066

32. Kitagawa H., Imagawa T., Uehara M. The apical caecal diverticulum of the chicken identified as a lymphoid organ. Journal of Anatomy. 1996; 189(3): 667‒672.

33. Selionova M., Zagarin A., Lysenko Yu. Impact of a phytogenic composition on feed conversion and certain digestive mechanisms in broiler chickens. BIO Web of Conferences. 2025; 194: 01039. https://doi.org/10.1051/bioconf/202519401039