Abstract
Background
The rapid emergence of multidrug-resistantSalmonellain poultry demands alternative control strategies beyond conventional antibiotics. In this study, we evaluated a combination of lyticSalmonella-infecting bacteriophages (SLAM_phiST45 and SLAM_phiST56) and a probiotic bacteriumLimosilactobacillus reuteri(SLAM_LAR11) in a chick model challenged withSalmonella entericaserovar Typhimurium infection.
Results
Co-administration with two-phage cocktail and a probiotic showed markedly reducedSalmonellacolonization in the gut and systemic organs of chicks, comparable to the effect of phage-only treatment. In contrast with phage-only treatment, the combined therapy significantly improved the rate of body-weight change from the day of infection to necropsy (P< 0.0001) and alleviated infection-associated splenomegaly (P= 0.028) and hepatomegaly (P= 0.011). In the ileum, the villus height-to-crypt depth ratio (VH/CD) increased significantly (P= 0.044). In the colon, expression of tight-junction genesOCLN(P= 0.014),TJP1(P< 0.0001), andMUC2(P= 0.011) was elevated, whereas the pro-inflammatory cytokineIL6was reduced (P= 0.018). These improvements were accompanied, in the cecum, by trends toward decreases inEscherichia–Shigella(P= 0.09) andClostridium(P= 0.16) and a trend toward an increase inBlautia(P= 0.11); additionally, in the ileum,Lactobacillus(P= 0.037) andBlautia(P= 0.016) increased significantly, yielding a more balanced microbiota than with phage-only treatment. Consistently, levels of functional metabolites, including acetic acid (LDA = 3.32) and lactic acid (LDA = 5.29), were increased.
Conclusion
Taken together, these findings demonstrate that phage–probiotic co-administration not only enhances the clearance of multidrug-resistantSalmonellamore effectively than phage treatment alone but also promotes intestinal health, highlighting its potential as an antibiotic-alternatives strategy to improve intestinal health and ensure food safety in poultry production systems.
Keywords
Data Availability
WGS data have been deposited in NCBI GenBank (phage accession numbers PP948674.1 and PP948675.1; the accession number for LAR11 is CP196337). The sequencing data generated in this study have been deposited in the NCBI Sequence Read Archive (SRA) under BioProject accession PRJNA1291055. This includes 16S rRNA gene sequencing reads from cecal content samples of all 5 groups (BL, SA, PR, PC, PP) and from ileal and jejunal content samples of the PC and PP groups. These datasets are publicly available from NCBI. All data needed to evaluate the conclusions in the paper are present in the manuscript. Additional data are available from the authors upon request.
Abbreviations
- ADFI:: Average daily feed intake
- ARGs:: Antibiotic resistance genes
- CLDN1 :: Claudin-1 (tight junction protein gene)
- CLDN4 :: Claudin-4 (tight junction protein gene)
- FCR:: Feed conversion ratio
- GAPDH :: Glyceraldehyde-3-phosphate dehydrogenase (housekeeping gene)
- GPR41 (FFAR3):: G protein-coupled receptor 41 (free fatty acid receptor 3)
- GPR43 (FFAR2):: G protein-coupled receptor 43 (free fatty acid receptor 2)
- H&E:: Hematoxylin and eosin (stain)
- IAA:: Indole-3-acetic acid
- IFNG :: Interferon-gamma (Th1 cytokine gene)
- IL1B :: Interleukin-1 beta (pro-inflammatory cytokine gene)
- IL6 :: Interleukin-6 (pro-inflammatory cytokine gene)
- IL8 :: Interleukin-8 (CXCLi2, pro-inflammatory cytokine gene)
- IL10 :: Interleukin-10 (anti-inflammatory cytokine gene)
- MDR:: Multidrug-resistant
- MUC2 :: Mucin 2 (intestinal mucus glycoprotein gene)
- OCLN :: Occludin (tight junction protein gene)
- SCFA:: Short-chain fatty acid
- TGFB3 :: Transforming growth factor beta 3 (anti-inflammatory cytokine gene)
- TJP1 :: Tight junction protein 1 (ZO-1 gene)
- WGS:: Whole-genome sequencing
References
- 1.Raza MA, Kim E, Shakeel M, Fiaz M, Ma L, Kim H, et al. Evaluation of zinc oxide and copper oxide nanoparticles as potential alternatives to antibiotics for managing fowl typhoid in broilers. J Anim Sci Technol. 2024;66(5).(2024)2023.e91.: 962.
- 2.Kim H, Lee J, Kim S, Kim B, Chang S, Song D, et al. Supplemental illite or in combination with probiotics improves performance and gut health in broilers challenged withSalmonellaTyphimurium. J Anim Sci Technol. 2024.https.(2024)//doi. org/10.5187/jast.
- 3.Chang CH, Teng PY, Lee TT, Yu B. Effects of multi-strain probiotic supplementation on intestinal microbiota, tight junctions, and inflammation in young broiler chickens challenged withSalmonella entericasubsp.enterica. Asian-Australas J Anim Sci. 2019;33(11).(2019)1797.: 1797.
- 4.Ibrahim D, Abdelfattah-Hassan A, Badawi M, Ismail TA, Bendary MM, Abdelaziz AM, et al. Thymol nanoemulsion promoted broiler chicken’s growth, gastrointestinal barrier and bacterial community and conferred protection againstSalmonellaTyphimurium. Sci Rep. 2021;11.(2021)7742.: 7742.
- 5.Zhang S, Wang Y, Ye J, Fan Q, Lin X, Gou Z, et al. Dietary supplementation of bilberry anthocyanin on growth performance, intestinal mucosal barrier and cecal microbes of chickens challenged withSalmonellaTyphimurium. J Anim Sci Biotechnol. 2023;14.(2023)org/10.1186/s40104-022-00799-9.: 15.
- 6.Oh JY, Jeong JH, Kwak SM, Chae J-C. Complete genome of blaCTX-M-65-carryingSalmonellaInfantis strain Z01323CSL0015 isolated from broiler chicken. J Anim Sci Technol. 2024.https.(2024)//doi. org/10.5187/jast.
- 7.Pandey S, Doo H, Keum GB, Kim ES, Kwak J, Ryu S, et al. Antibiotic resistance in livestock, environment and humans.(2024)one Health perspective.J Anim Sci Technol.: 266.
- 8.Jo H, Han G, Kim EB, Kong C, Kim BG. Effects of supplemental bacteriophage on the gut microbiota and nutrient digestibility of ileal-cannulated pigs. J Anim Sci Technol. 2024;66(2).(2024)2023.e96.: 340.
- 9.Torkashvand N, Kamyab H, Aarabi P, Shahverdi AR, Torshizi MAK, Khoshayand MR, et al. Evaluating the effectiveness and safety of a novel phage cocktail as a biocontrol ofSalmonellain biofilm, food products, and broiler chicken. Front Microbiol. 2024.https.(2024)//doi. org/10.3389/fmicb.
- 10.Hosny RA, Gaber AF, Sorour HK. Bacteriophage mediated control of necrotic enteritis caused byC. perfringensin broiler chickens. Vet Res Commun. 2021;45(4).(2021)org/10.1007/s11259-021-09821-3.: 409.
- 11.Jhandai P, Mittal D, Gupta R, Kumar M, Khurana R. Therapeutics and prophylactic efficacy of novel lyticEscherichiaphage vB_EcoS_PJ16 against multidrug-resistant avian pathogenicE. coliusing in vivo study. Int Microbiol. 2024;27(3).(2024)org/10.1007/s10123-023-00420-7.: 673.
- 12.Egido JE, Costa AR, Aparicio-Maldonado C, Haas P-J, Brouns SJJ. Mechanisms and clinical importance of bacteriophage resistance. FEMS Microbiol Rev. 2021.https.(2021)org/10.1093/femsre/fuab048.
- 13.Esteves NC, Scharf BE. Flagellotropic bacteriophages.(2022)opportunities and challenges for antimicrobial applications.Int J Mol Sci.
- 14.Yoo S, Lee KM, Kim N, Vu TN, Abadie R, Yong D. Designing phage cocktails to combat the emergence of bacteriophage-resistant mutants in multidrug-resistantKlebsiella pneumoniae. Microbiol Spectr. 2024;12(1).(2024)1128/spectrum.01258-23.
- 15.Chen H, Liu H, Gong Y, Dunstan RA, Ma Z, Zhou C, et al. AKlebsiella-phage cocktail to broaden the host range and delay bacteriophage resistance both in vitro and in vivo. NPJ Biofilms Microbiomes. 2024;10(1).(2024)org/10.1038/s41522-024-00603-8.: 127.
- 16.Tagliaferri TL, Jansen M, Horz H-P. Fighting pathogenic bacteria on two fronts.(2019)phages and antibiotics as combined strategy.Front Cell Infect Microbiol.
- 17.Mu A, McDonald D, Jarmusch AK, Martino C, Brennan C, Bryant M, et al. Assessment of the microbiome during bacteriophage therapy in combination with systemic antibiotics to treat a case of staphylococcal device infection. Microbiome. 2021;9.(2021)org/10.1186/s40168-021-01026-9.: 92.
- 18.Kurilovich E, Geva-Zatorsky N. Effects of bacteriophages on gut microbiome functionality. Gut Microbes. 2025;17(1).(2025)2481178. https://doi. org/10.1080/19490976.: 2481178.
- 19.Naeem M, Bourassa D. Probiotics in poultry.(2025)unlocking productivity through microbiome modulation and gut health.Microorganisms.: 257.
- 20.Prado-Rebolledo OF, Delgado-Machuca JJ, Macedo-Barragan RJ, Garcia-Márquez LJ, Morales-Barrera JE, Latorre JD, et al. Evaluation of a selected lactic acid bacteria-based probiotic onSalmonella entericaserovar Enteritidis colonization and intestinal permeability in broiler chickens. Avian Pathol. 2017;46(1).(2017)2016.1222808.: 90.
- 21.Guan L, Hu A, Ma S, Liu J, Yao X, Ye T, et al.Lactiplantibacillus plantarumpostbiotic protects againstSalmonellainfection in broilers via modulating NLRP3 inflammasome and gut microbiota. Poult Sci. 2024;103(4).(2024)103483. https://doi. org/10. 1016/j.psj.: 103483.
- 22.Kim K, Lee J, Kang S-J, Oh S. Whole genome sequence analysis ofBifidobacterium animalissubsp.lactisKL101 and comparative genomics with BB12. J Anim Sci Technol. 2024.https.(2024)//doi. org/10.5187/jast.
- 23.Kang J, Doo H, Kwak J, Kim ES, Keum GB, Choi Y, et al. Complete genome sequence ofEnterococcus faeciumstrain GB_C_05 with potential characteristics applicable as a bacteriocin-producing probiotic feed additive. J Anim Sci Technol. 2025.https.(2025)//doi. org/10.5187/jast.
- 24.Park S, Son S, Park MA, Kim D-H, Kim Y. Complete genome sequence ofLatilactobacillus curvatusCACC879 and its functional probiotic properties. J Anim Sci Technol. 2024;66(3).(2024)2023.e50.: 630.
- 25.Soffritti I, D’Accolti M, Bini F, Mazziga E, Volta A, Bisi M, et al. Harnessing probiotics and bacteriophages to fightSalmonellaand limit the use of antibiotics in broilers.(2025)a study in commercial conditions.Poult Sci.: 105595.
- 26.Shaufi MAM, Sieo CC, Chong CW, Geok Hun T, Omar AR, Han Ming G, et al. Effects of phage cocktail, probiotics, and their combination on growth performance and gut microbiota of broiler chickens. Animals. 2023;13(8).(2023)org/10.3390/ani13081328.: 1328.
- 27.Wang X, Ji Y, Qiu C, Zhang H, Bi L, Xi H, et al. A phage cocktail combined with the enteric probioticLactobacillus reuteriameliorated mouse colitis caused byS. Typhimurium. Food Funct. 2022;13(16).(2022)org/10.1039/d2fo00699e.: 8509.
- 28.Nii T, Kakuya H, Isobe N, Yoshimura Y.Lactobacillus reuterienhances the mucosal barrier function against heat-killedSalmonellaTyphimurium in the intestine of broiler chicks. J Poult Sci. 2020;57(2).(2020)2141/jpsa.0190044.: 148.
- 29.Karaffová V, Kiššová Z, Tóthová C, Tráj P, Mackei M, Mátis G.Limosilactobacillus reuteriB1/1 modulated the intestinal immune response in preventingSalmonellaEnteritidis PT4 infection in a chicken ileal explant model. Vet Res Commun. 2024;49.(2024)org/10.1007/s11259-024-10609-4.: 32.
- 30.Choi Y, Kang A, Kang M-G, Lee DJ, Kyoung D, Kwak M-J, et al. Exploring synergistic effect of bacteriophages with probiotics against multidrug resistantSalmonellaTyphimurium in a simulated chicken gastrointestinal system using metagenomic- and culturomic approaches. J Anim Sci Technol. 2025.https.(2025)//doi. org/10.5187/jast.
- 31.Lu H, Giordano F, Ning Z. Oxford nanopore MinION sequencing and genome assembly. Genomics Proteomics Bioinformatics. 2016;14(5).(2016)265–79. https://doi. org/10. 1016/j.gpb.: 265.
- 32.Kolmogorov M, Yuan J, Lin Y, Pevzner PA. Assembly of long, error-prone reads using repeat graphs. Nat Biotechnol. 2019;37(5).(2019)540–6.: 540.
- 33.Seemann T. Prokka.(2014)rapid prokaryotic genome annotation.Bioinformatics.: 2068.
- 34.Thompson JD, Gibson TJ, Higgins DG. Multiple sequence alignment using ClustalW and ClustalX. Curr Protoc Bioinform. 2003;(1).(2003)3.22.: 2.
- 35.Stothard P, Grant JR, Van Domselaar G. Visualizing and comparing circular genomes using the CGView family of tools. Brief Bioinform. 2019;20(4).(2019)1576–82.: 1576.
- 36.Heidari M, Wang D, Delekta P, Sun S. Marek’s disease virus immunosuppression alters host cellular responses and immune gene expression in the skin of infected chickens. Vet Immunol Immunopathol. 2016;180.(2016)21–8.: 21.
- 37.Heymans R, Vila A, van Heerwaarden CA, Jansen CC, Castelijn GA, van der Voort M, et al. Rapid detection and differentiation ofSalmonellaspecies,SalmonellaTyphimurium andSalmonellaEnteritidis by multiplex quantitative PCR. PLoS ONE. 2018;13(10).(2018)e0206316.
- 38.Feldman AT, Wolfe D. Tissue processing and hematoxylin and eosin staining. In.(2014)Day C, editors. Histopathology. Methods in molecular biology, vol 1180.New York, NY: Humana Press;.
- 39.Song B, Li H, Wu Y, Zhen W, Wang Z, Xia Z, et al. Effect of microencapsulated sodium butyrate dietary supplementation on growth performance and intestinal barrier function of broiler chickens infected with necrotic enteritis. Anim Feed Sci Technol. 2017;232.(2017)6–15.: 6.
- 40.Quast C, Pruesse E, Yilmaz P, Gerken J, Schweer T, Yarza P, et al. The SILVA ribosomal RNA gene database project.(2012)improved data processing and web-based tools.Nucleic Acids Res.
- 41.Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP. DADA2.(2016)High-resolution sample inference from Illumina amplicon data.Nat Methods.: 581.
- 42.Bolyen E, Rideout JR, Dillon MR, Bokulich NA, Abnet CC, Al-Ghalith GA, et al. Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nat Biotechnol. 2019;37(8).(2019)852–7.: 852.
- 43.Son SH, Kang MG, Kang A, Kang Y, Kim K, Kwak MJ, et al. Gut microbiome and metabolome library construction based on age group using short-read and long-read sequencing techniques in Korean traditional canine species Sapsaree. Front Microbiol. 2024;15.(2024)1486566. https://doi. org/10.3389/fmicb.: 1486566.
- 44.Pang Z, Lu Y, Zhou G, Hui F, Xu L, Viau C, et al. Metaboanalyst 6.0.(2024)towards a unified platform for metabolomics data processing, analysis and interpretation.Nucleic Acids Res.
- 45.Mon KK, Zhu Y, Chanthavixay G, Kern C, Zhou H. Integrative analysis of gut microbiome and metabolites revealed novel mechanisms of intestinalSalmonellacarriage in chicken. Sci Rep. 2020;10.(2020)4809.: 4809.
- 46.Team RC. R.(2016)A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www. r-project.org.
- 47.Gu Z, Eils R, Schlesner M. Complex heatmaps reveal patterns and correlations in multidimensional genomic data. Bioinformatics. 2016;32(18).(2016)org/10.1093/bioinformatics/btw313.: 2847.
- 48.Swift ML. GraphPad prism, data analysis, and scientific graphing. J Chem Inf Comput Sci. 1997;37(2).(1997)411–2.: 411.
- 49.Additives EPo, Feed PoSuiA. Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J. 2012;10(6).(2012)2740.: 2740.
- 50.Rahn K, De Grandis S, Clarke R, McEwen S, Galan J, Ginocchio C, et al. Amplification of aninvAgene sequence ofSalmonellaTyphimurium by polymerase chain reaction as a specific method of detection ofSalmonella. Mol Cell Probes. 1992;6(4).(1992)271–9.: 271.
- 51.Darwin KH, Miller VL. Molecular basis of the interaction ofSalmonellawith the intestinal mucosa. Clin Microbiol Rev. 1999;12(3).(1999)3.405.: 405.
- 52.Parsons BN, Humphrey S, Salisbury AM, Mikoleit J, Hinton JC, Gordon MA, et al. Invasive non-typhoidalSalmonellaTyphimurium ST313 are not host-restricted and have an invasive phenotype in experimentally infected chickens. PLoS Negl Trop Dis. 2013;7(10).(2013)e2487.
- 53.Chang SC, Shen MH, Liu CY, Pu CM, Hu JM, Huang CJ. A gut butyrate-producing bacteriumButyricicoccus pullicaecorumregulates short-chain fatty acid transporter and receptor to reduce the progression of 1,2-dimethylhydrazine-associated colorectal cancer. Oncol Lett. 2020;20(6).(2020)327. https://doi. org/10.3892/ol.: 327.
- 54.Liu X, Mao B, Gu J, Wu J, Cui S, Wang G, et al.Blautia-a new functional genus with potential probiotic properties? Gut Microbes. 2021;13(1).(2021)1–21. https://doi. org/10.1080/19490976.: 1.
- 55.Burrows PB, Godoy-Santos F, Lawther K, Richmond A, Corcionivoschi N, Huws SA. Decoding the chicken gastrointestinal microbiome. BMC Microbiol. 2025;25.(2025)org/10.1186/s12866-024-03690-x.: 35.
- 56.Yuan X, Chen B, Duan Z, Xia Z, Ding Y, Chen T, et al. Depression and anxiety in patients with active ulcerative colitis.(2021)crosstalk of gut microbiota, metabolomics and proteomics.Gut Microbes.: 1987779.
- 57.Leibovitzh H, Lee S-H, Xue M, RaygozaGaray JA, Hernandez-Rocha C, Madsen KL, et al. Altered gut microbiome composition and function are associated with gut barrier dysfunction in healthy relatives of patients with Crohn’s disease. Gastroenterology. 2022;163(5).(2022)1364-76. e10. https://doi. org/10. 1053/j.gastro.: 1364-76.
- 58.Song D, Lee J, Yoo Y, Oh H, Chang S, An J, et al. Effects of probiotics on growth performance, intestinal morphology, intestinal microbiota weaning pig challenged withEscherichia coliandSalmonella enterica. J Anim Sci Technol. 2025;67(1).(2025)2023.e119.: 106.
- 59.Jeong JY, Kim J, Kim M, Kim Y-B, Park C, Song J, et al. Effect of probiotics on growth performance, cytokine levels, and gut microbiome composition of broiler chickens for 7 and 35 days. J Anim Sci Technol. 2025.https.(2025)//doi. org/10.5187/jast.
- 60.Kyoung H, Kang Y, Kim Y, Kim Y, Ahn J, Nam J, et al. ProbioticLactiplantibacillus plantarumimproved growth performance of weaned pigs by enhancing intestinal health and modulating immune responses. J Anim Sci Technol. 2025.https.(2025)//doi. org/10.5187/jast.
- 61.Eum B-G, Elnar AG, Jang Y, Kim G-B. Complete genome sequence ofLigilactobacillus agilisLDTM47, bacteriocin-producing lactic acid bacteria isolated from broiler gastrointestinal tract. J Anim Sci Technol. 2025;67(2).(2025)2024.e29.: 489.
- 62.Parada Venegas D, De la Fuente MK, Landskron G, González MJ, Quera R, Dijkstra G, et al. Short chain fatty acids (SCFAs)-mediated gut epithelial and immune regulation and its relevance for inflammatory bowel diseases. Front Immunol. 2019;10.(2019)277. https://doi. org/10.3389/fimmu.: 277.
- 63.Jyoti, Dey P. Mechanisms and implications of the gut microbial modulation of intestinal metabolic processes. NPJ Metab Health Dis. 2025;3(1).(2025)org/10.1038/s44324-025-00066-1.: 24.
- 64.Comeau AM, Tétart F, Trojet SN, Prère M-F, Krisch H. Phage-antibiotic synergy (PAS).(2007)β-lactam and quinolone antibiotics stimulate virulent phage growth.PLoS ONE.
- 65.Altamirano FLG, Kostoulias X, Subedi D, Korneev D, Peleg AY, Barr JJ. Phage-antibiotic combination is a superior treatment againstAcinetobacter baumanniiin a preclinical study. EBioMedicine. 2022.https.(2022)//doi. org/10. 1016/j.ebiom.
- 66.Unverdi A, Erol HB, Kaskatepe B, Babacan O. Characterization ofSalmonellaphages isolated from poultry coops and its effect with nisin on food bio-control. Food Sci Nutr. 2024;12(4).(2024)1002/fsn3.3956.: 2760.
- 67.Toro H, Price S, McKee S, Hoerr F, Krehling J, Perdue M, et al. Use of bacteriophages in combination with competitive exclusion to reduceSalmonellafrom infected chickens. Avian Dis. 2005;49(1).(2005)118–24.: 118.
- 68.Kumar T, Rajora V, Arora N, Mittal D, Prasad A. In vivo therapeutic evaluation of phage cocktail and probiotic in reducingSalmonellainfection in broilers. Indian J Anim Sci. 2023;93(12).(2023)1150–4.: 1150.
- 69.Elsharkawy MS, Abdelbaki MM, Madkour M, EL Shanawany EE, Aboelenin MM, Wang Q, et al. Synbiotic modulate the host immune response toSalmonellaTyphimurium infection in commercial and indigenous chicken. Ital J Anim Sci. 2024;23(1).(2024)1279–89.: 1279.
- 70.Wishna-Kadawarage RN, Połtowicz K, Hickey RM, Siwek M. Modulation of gene expression in immune-related organs by in ovo stimulation with probiotics and prophybiotics in broiler chickens. J Appl Genet. 2025;66(1).(2025)org/10.1007/s13353-024-00891-y.: 195.
- 71.Zhao H, Comer L, Akram MZ, Corion M, Li Y, Everaert N. Recent advances in the application of microbiota transplantation in chickens. J Anim Sci Biotechnol. 2025;16.(2025)org/10.1186/s40104-025-01233-6.: 91.
- 72.Kogut MH, Fernandez Miyakawa ME. Phenotype alterations in the cecal ecosystem involved in the asymptomatic intestinal persistence of paratyphoidSalmonellain chickens. Animals. 2023;13(18).(2023)2824.: 2824.
- 73.Lacroix-Lamandé S, Bernardi O, Pezier T, Barilleau E, Burlaud-Gaillard J, Gagneux A, et al. DifferentialSalmonellaTyphimurium intracellular replication and host cell responses in caecal and ileal organoids derived from chicken. Vet Res. 2023;54.(2023)org/10.1186/s13567-023-01189-3.: 63.
- 74.Feng Y, Zhang M, Liu Y, Yang X, Wei F, Jin X, et al. Quantitative microbiome profiling reveals the developmental trajectory of the chicken gut microbiota and its connection to host metabolism. iMeta. 2023;2(2).(2023)1002/imt2.105.
- 75.Liu L, Li Q, Yang Y, Guo A. Biological function of short-chain fatty acids and its regulation on intestinal health of poultry. Front Vet Sci. 2021.https.(2021)//doi. org/10.3389/fvets.
- 76.Ducatelle R, Goossens E, Eeckhaut V, Van Immerseel F. Poultry gut health and beyond. Anim Nutr. 2023;13.(2023)240–8. https://doi. org/10. 1016/j.aninu.: 240.
- 77.Lee IK, Gu MJ, Ko KH, Bae S, Kim G, Jin G-D, et al. Regulation of CD4+CD8−CD25+and CD4+CD8+CD25+T cells by gut microbiota in chicken. Sci Rep. 2018;8.(2018)org/10.1038/s41598-018-26763-0.: 8627.
- 78.Saint-Martin V, Guillory V, Chollot M, Fleurot I, Kut E, Roesch F, et al. The gut microbiota and its metabolite butyrate shape metabolism and antiviral immunity along the gut-lung axis in the chicken. Commun Biol. 2024;7(1).(2024)org/10.1038/s42003-024-06815-0.: 1185.
- 79.Shen J, Yang L, You K, Chen T, Su Z, Cui Z, et al. Indole-3-acetic acid alters intestinal microbiota and alleviates ankylosing spondylitis in mice. Front Immunol. 2022.https.(2022)//doi. org/10.3389/fimmu.
- 80.Liu J, Gu H, Jia R, Li S, Chen Z, Zheng A, et al. Effects ofLactobacillus acidophiluson production performance and immunity of broiler chickens and their mechanism. Front Vet Sci. 2025.https.(2025)//doi. org/10.3389/fvets.
- 81.Lyte JM, Jia X, Caputi V, Zhang D, Daniels KM, Phillips GJ, et al. Heat stress in chickens induces temporal changes in the cecal microbiome concomitant with host enteric serotonin responses. Poult Sci. 2025;104(3).(2025)104886. https://doi. org/10. 1016/j.psj.: 104886.
- 82.Pangga GM, Star-Shirko B, Psifidi A, Xia D, Corcionivoschi N, Kelly C, et al. Impact of commercial gut health interventions on caecal metagenome and broiler performance. Microbiome. 2025;13.(2025)org/10.1186/s40168-024-02012-7.: 30.
- 83.Mak PHW, Yin X, Clairmont L, Bean-Hodgins L, Kiarie EG, Tang J, et al. Cecal microbiome in broiler chicken related to antimicrobial feeding and bird’s sex. Can J Microbiol. 2025;71.(2025)org/10.1139/cjm-2024-0190.: 1.
- 84.Hsu BB, Gibson TE, Yeliseyev V, Liu Q, Lyon L, Bry L, et al. Dynamic modulation of the gut microbiota and metabolome by bacteriophages in a mouse model. Cell Host Microbe. 2019;25(6).(2019)803-14. e5. https://doi. org/10. 1016/j.chom.: 803-14.
- 85.Mahmud MR, Tamanna SK, Akter S, Mazumder L, Akter S, Hasan MR, et al. Role of bacteriophages in shaping gut microbial community. Gut Microbes. 2024;16(1).(2024)2390720.: 2390720.
Acknowledgements
We gratefully acknowledged Korea Veterinary Culture Collection (KVCC) for providing KVCC-BA0000422 strain in this research.
Funding
This work was supported by the National Research Foundation of Korea (NRF) grant funded by the National Research Foundation of Korea Grant, funded by the Korean government (MEST) (NRF-2021R1A2C3011051) and by the Korea government (MSIT) (No. RS-2023-00218476).
Ethics Declaration
Ethics approval and consent to participate
All animal procedures followed institutional guidelines and were approved by the Jeonbuk National University (JBNU) IACUC (Approval No. JBNU 2022–091).
Consent for publication
Not applicable.
Competing interests
There are no competing interests in this study.
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