Effect of Bacteriophage Cocktail on Growth Performance, Diarrhea Score, and Gut Bacteria of Broilers in Comparison with Bacillus subtilis

Subject Areas :

H. Dm ¹ , G. Akib ² , A. Dutta ³ , S. Mahfuz ⁴ , C. Wang ⁵ , Z. Qq ⁶ , Z. Jy ^{7 *}

1 - College of Life Science, Linyi University, Linyi City 276000, Linyi, China
2 - Department of Animal Nutrition, Sylhet Agricultural University, Sylhet 3100, Sylhet, Bangladesh
3 - Department of Animal Nutrition, Sylhet Agricultural University, Sylhet 3100, Sylhet, Bangladesh
4 - Department of Animal Nutrition, Sylhet Agricultural University, Sylhet 3100, Sylhet, Bangladesh
5 - College of Life Science, Linyi University, Linyi City 276000, Linyi, China
6 - College of Life Science, Linyi University, Linyi City 276000, Linyi, China
7 - College of Life Science, Linyi University, Linyi City 276000, Linyi, China

Received: 2025-05-23 Accepted : 2025-06-26 Published : 2025-06-30

Keywords: Bacillus subtilis, bacteriophage cocktail, broiler, growth performance, health condition,

Abstract :

This study investigated the efficacy of a bacteriophage cocktail on growth parameters, diarrhea score, and ileal and cloacal bacterial counts of broilers, compared to Bacillus subtilis. A total of 4131 mixed-sex broiler day old chicks (DOCs) were procured from a commercial company and reared for 42 days in cages within a controlled broiler shed. Birds were assigned to three treatment groups: T1 (supplemented with B. subtilis at 200 g/kg of feed), T2 (bacteriophage added to basal feed at 300 g/kg of feed), and T3 (feed sup-plemented with bacteriophage at 500 g/ton of feed). Each treatment was replicated into 81 cages, with 17 broilers per cage. Feed conversion ratio (FCR) was lowest, and the survival rate and European production efficiency factor (EPEF) were highest in the T3 group, with T2 showing intermediate performance. Bacte-riophage-supplemented groups exhibited lower diarrhea scores, as feces appeared normal. Escherichia coli and Salmonella counts were lowest in the T3 group throughout the experiment, followed by T2, while these groups had higher counts of Lactobacillus and Bifidobacterium. The T3 group demonstrated superior over-all performance in comparison with the other two experimental groups. In essence, supplementing a bacte-riophage cocktail at a rate of 500 g/ton of feed could serve as an effective feed additive for enhancing growth performance, overall welfare, and modulation of gut bacteria by increasing beneficial bacteria and reducing targeted pathogens.

References:

Abbas R.Z., Alsayeqh A.F. and Aqib A.I. (2022). Role of bacte-riophages for optimized health and production of poultry. Animals. 12(23), 3378-3387.
Abd-El Wahab A., Basiouni S., El-Seedi H.R., Ahmed M.F.E., Bielke L.R., Hargis B., Tellez-Isaias G., Eisenreich W., Lehnherr H., Kittler S., Shehata A.A. and Visscher C. (2023). An overview of the use of bacteriophages in the poultry indus-try: Successes, challenges, and possibilities for overcoming breakdowns. Front. Microbiol. 14, 1-10.
Abreu R., Semedo-Lemsaddek T., Cunha E., Tavares L. and Oliveira M. (2023). Antimicrobial drug resistance in poultry production: Current status and innovative strategies for bacte-rial control. Microorganisms. 11(4), 1-8.
Aljuhaishi D.A. and Albawi F.H.K. (2024). Using bacteriophage in poultry diet as therapeutics against Escherichia coli in broiler chickens. J. Anim. Health Prod. 12(1), 23-31.
Asghar A., Khalid A., Baqar Z., Hussain N., Saleem M.Z., Sairash and Rizwan K. (2024). An insights into emerging trends to control the threats of antimicrobial resistance (AMR): An ad-dress to public health risks. Arch. Microbiol. 206(2), 72-81.
Castanon J.I.R. (2007). History of the use of antibiotic as growth promoters in European poultry feeds. Poult. Sci. 86(11), 2466-2471.
Clavijo V. and Flórez M.J.V. (2018). The gastrointestinal micro-biome and its association with the control of pathogens in broiler chicken production: A review. Poult. Sci. 97(3), 1006-1021.
Curea C.D., Usturoi M.G., Custură I., Radu-Rusu R.M., Rațu R.N., Prisăcaru M. and Usturoi A. (2023). Efficiency of grow-ing of chicken broilers under conditions of compliance with EU rules of welfare. Sci. Papers Ser. D. Anim. Sci. 66(1), 273-278.
Ducatelle R., Goossens E., Eeckhaut V. and Van Immerseel F. (2023). Poultry gut health and beyond. Anim. Nutr. 13, 240-248.
European Food Safety Authority (EFSA) and European Centre for Disease Prevention and Control (ECDC). (2024). The Euro-pean Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2021-2022. EFSA J. 22(2), e8583.
Fang Q., Yin X., He Y., Feng Y., Zhang L., Luo H., Yin G., McNally A. and Zong Z. (2024). Safety and efficacy of phage application in bacterial decolonisation: A systematic review. Lancet Microbe. 5(5), 489-499.
Gindin M., Febvre H.P., Rao S., Wallace T.C. and Weir T.L. (2019). Bacteriophage for gastrointestinal health (PHAGE) study: Evaluating the safety and tolerability of supplemental bacteriophage consumption. J. Am. Coll Nutr. 38(1), 68-75.
Gómez-Gómez C., Blanco-Picazo P., Brown-Jaque M., Quirós P., Rodríguez-Rubio L., Cerdà-Cuellar M. and Muniesa M. (2019). Infectious phage particles packaging antibiotic resis-tance genes found in meat products and chicken feces. Sci. Rep. 9(1), 13281-13292.
Habteweld H.A. and Asfaw T. (2023). Novel dietary approach with probiotics, prebiotics, and synbiotics to mitigate antim-icrobial resistance and subsequent out marketplace of antim-icrobial agents: A review. Infect. Drug Resist. 16, 3191-3211.
Hashem M.A., Hassan A.E.A., Abou-Elnaga H.M.M., Abdo W., Dahran N., Alghamdi A.H. and Elmahallawy E.K. (2022). Modulatory effect of dietary probiotic and prebiotic supple-mentation on growth, immuno-biochemical alterations, DNA damage, and pathological changes in E. coli-infected broiler chicks. Front. Vet. Sci. 9, 964738-964747.
Huff W.E., Huff G.R., Rath N.C., Balog J.M. and Donoghue A.M. (2003). Bacteriophage treatment of a severe Escherichia coli respiratory infection in broiler chickens. Avian Dis. 47(4), 1399-1405.
Jayaraman S., Das P.P., Saini P.C., Roy B. and Chatterjee P.N. (2017). Use of Bacillus subtilis PB6 as a potential antibiotic growth promoter replacement in improving performance of broiler birds. Poult. Sci. 96(8), 2614-2622.
Kim K.H., Ingale S.L., Kim J.S., Lee S.H., Lee J.H., Kwon I.K. and Chae B.J. (2014). Bacteriophage and probiotics both en-hance the performance of growing pigs but bacteriophage are more effective. Anim. Feed Sci. Technol. 196, 88-95.
Korver D.R. (2023). Review: Current challenges in poultry nutri-tion, health, and welfare. Animal. 17, 100755-100768.
Ling H., Lou X., Luo Q., He Z., Sun M. and Sun J. (2022). Recent advances in bacteriophage-based therapeutics: Insight into the post-antibiotic era. Acta Pharm. Sin. B. 12(12), 4348-4364.
Liu Y.L., Yan T., Li X.Y., Duan Y.L., Yang X. and Yang X.J. (2020). Effects of Bacillus subtilis and antibiotic growth pro-moters on the growth performance, intestinal function and gut microbiota of pullets from 0 to 6 weeks. Animal. 14(8), 1619-1628.
Miller R.W., Skinner J., Sulakvelidze A., Mathis G.F. and Hofacre C.L. (2010). Bacteriophage therapy for control of necrotic en-teritis of broiler chickens experimentally infected with Clos-tridium perfringens. Avian Dis. 54(1), 33-40.
Mosimann S., Desiree K. and Ebner P. (2021). Efficacy of phage therapy in poultry: a systematic review and meta-analysis. Poult. Sci. 100(12), 101472-101483.
Ningsih N., Respati A.N., Astuti D., Triswanto T., Purnamayanti L., Yano A.A., Putra R.P., Jayanegara A., Ratriyanto A. and Irawan A. (2023). Efficacy of Bacillus subtilis to replace in-feed antibiotics of broiler chickens under necrotic enteritis-challenged experiments: A systematic review and meta-analysis. Poult. Sci. 102(10), 102923-102933.
Noor M., Runa N., Husna A., Rahman M., Rajib D.M., Mahbub-e-Elahi A.T. and Rahman M.M. (2020). Evaluation of the effect of dietary supplementation of bacteriophage on production performance and excreta microflora of commercial broiler and layer chickens in Bangladesh. MOJ Proteom. Bioinform. 9(2), 27-31.
NRC. (1994). Nutrient Requirements of Poultry, 9th Rev. Ed. Na-tional Academy Press, Washington, DC., USA.
Pelyuntha W., Yafa A., Ngasaman R., Yingkajorn M., Chukiatsiri K., Champoochana N. and Vongkamjan K. (2022). Oral ad-ministration of a phage cocktail to reduce Salmonella Coloni-zation in broiler gastrointestinal tract-a pilot study. Animals. 12(22), 24-31.
Pickard J.M., Zeng M.Y., Caruso R. and Núñez G. (2017). Gut microbiota: Role in pathogen colonization, immune responses, and inflammatory disease. Immunol. Rev. 279(1), 70-89.
Qiu K., Li C., Wang J., Qi G., Gao J., Zhang H. and Wu S. (2021). Effects of dietary supplementation with Bacillus subtilis, as an alternative to antibiotics, on growth performance, serum im-munity, and intestinal health in broiler chickens. Front. Nutr. 8, 1-9.
Ramlucken U., Lalloo R., Roets Y., Moonsamy G., van Rensburg C.J. and Thantsha M.S. (2020). Advantages of Bacillus-based probiotics in poultry production. Livest. Sci. 241, 104215-104226.
Sarrami Z., Sedghi M., Mohammadi I., Bedford M., Miranzadeh H. and Ghasemi R. (2023). Effects of bacteriophage on Sal-monella Enteritidis infection in broilers. Sci. Rep. 13(1), 12198-12207.
Sarrami Z., Sedghi M., Mohammadi I., Kim W.K. and Mahdavi A.H. (2022). Effects of bacteriophage supplement on the growth performance, microbial population, and PGC-1α and TLR4 gene expressions of broiler chickens. Sci. Rep. 12(1), 14391-14401.
SAS Institute. (1996). SAS®/STAT Software, Release 6.11. SAS Institute, Inc., Cary, NC. USA.
Tariq S., Samad A., Hamza M., Ahmer A., Muazzam A., Ahmad S. and Amhabj A.M.A. (2022). Salmonella in poultry; an overview. Int. J. Multidiscip. Sci. Arts. 1(1), 80-84.
Tian R., Xu S., Li P., Li M., Liu Y., Wang K., Liu G., Li Y., Dai L. and Zhang W. (2023). Characterization of G-type Clostrid-ium perfringens bacteriophages and their disinfection effect on chicken meat. Anaerobe. 81, 102736-102745.
Upadhaya S.D., Ahn J.M., Cho J.H., Kim J.Y., Kang D.K., Kim S.W., Kim H.B. and Kim I.H. (2021). Bacteriophage cocktail supplementation improves growth performance, gut micro-biome and production traits in broiler chickens. J. Anim. Sci. Biotechnol. 12(1), 49-54.
Wang J.P., Yan L., Lee J.H. and Kim I.H. (2013). Evaluation of bacteriophage supplementation on growth performance, blood characteristics, relative organ weight, breast muscle character-istics and excreta microbial shedding in broilers. Asian-Australasian J. Anim. Sci. 26(4), 573-578.
Wernicki A., Nowaczek A. and Urban-Chmiel R. (2017). Bacte-riophage therapy to combat bacterial infections in poultry. Vi-rol. J. 14, 1-13.