{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T13:42:24Z","timestamp":1784641344042,"version":"3.55.0"},"reference-count":43,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,30]],"date-time":"2026-01-30T00:00:00Z","timestamp":1769731200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Probiotics from Dog and Cat milk Research Project","award":["K24LD115"],"award-info":[{"award-number":["K24LD115"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Escherichia coli O157:H7 is a pathogenic bacterium that causes severe intestinal infections characterized by inflammation and disruption of the intestinal barrier. Probiotic lactic acid bacteria (LAB) from milk can support intestinal health and combat enteric pathogens; however, the potential of feline milk-derived LAB against E. coli O157:H7 infection remains unclear. In this study, Pediococcus acidilactici (P. acidilactici) M22, isolated from feline milk, was evaluated for probiotic activity in vitro and in vivo in a C57BL\/6 mouse model of Escherichia coli O157:H7 infection. In vitro assays demonstrated that M22 significantly inhibited the adhesion of Escherichia coli O157:H7 to intestinal epithelial cells. For in vivo assessment, C57BL\/6 mice were orally administered M22 prior to infection with E. coli O157:H7. Protective effects were evaluated by monitoring body weight loss, colon length, disease activity index (DAI), myeloperoxidase (MPO) activity, cytokine levels, tight junction protein expression, oxidative stress markers, and gut microbiota composition. M22-treated mice exhibited significantly less body weight loss and lower DAI scores than infected controls. M22 also prevented colon shortening, indicating reduced colonic damage. Probiotic treatment attenuated neutrophil infiltration and mucosal inflammation, as evidenced by decreased colonic MPO activity, reduced levels of pro-inflammatory cytokines, and elevated anti-inflammatory IL-10. Additionally, M22 preserved intestinal barrier function by upregulating tight junction proteins and mitigating infection-induced histopathological changes. M22 supplementation enhanced antioxidant defenses in colonic tissue (lower malondialdehyde, higher superoxide dismutase and glutathione), indicating reduced oxidative stress. Furthermore, gut microbiota analysis (16S rRNA sequencing) revealed that M22 counteracted infection-induced dysbiosis, restoring microbial diversity and a healthy composition (enrichment of beneficial commensals and suppression of harmful bacteria). By safeguarding intestinal integrity and homeostasis, M22 emerges as a promising next-generation probiotic for improving intestinal health in companion animals.<\/jats:p>","DOI":"10.3390\/microorganisms14020332","type":"journal-article","created":{"date-parts":[[2026,1,30]],"date-time":"2026-01-30T14:35:46Z","timestamp":1769783746000},"page":"332","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A Feline Milk-Drived Pediococcus acidilactici M22 Alleviates Escherichia coli O157:H7 Infection Through Anti-Adhesion, Anti-Inflammation, and Microbiota Modulation"],"prefix":"10.3390","volume":"14","author":[{"given":"Xinyu","family":"Gong","sequence":"first","affiliation":[{"name":"Shandong Key Laboratory of Applied Technology for Protein and Peptide Drugs, Institute of Biopharmaceutical Research, Liaocheng University, Liaocheng 252000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue","family":"Wang","sequence":"additional","affiliation":[{"name":"Shandong Key Laboratory of Applied Technology for Protein and Peptide Drugs, Institute of Biopharmaceutical Research, Liaocheng University, Liaocheng 252000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5006-813X","authenticated-orcid":false,"given":"Huiming","family":"Huang","sequence":"additional","affiliation":[{"name":"Shandong Key Laboratory of Applied Technology for Protein and Peptide Drugs, School of Pharmaceutica Sciences and Food Engineering, Liaocheng University, 1 Hunan Street, Liaocheng 252059, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Han","sequence":"additional","affiliation":[{"name":"Shandong Key Laboratory of Applied Technology for Protein and Peptide Drugs, Institute of Biopharmaceutical Research, Liaocheng University, Liaocheng 252000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengping","family":"Wang","sequence":"additional","affiliation":[{"name":"Shandong Key Laboratory of Applied Technology for Protein and Peptide Drugs, Institute of Biopharmaceutical Research, Liaocheng University, Liaocheng 252000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8897-1239","authenticated-orcid":false,"given":"Min","family":"Wen","sequence":"additional","affiliation":[{"name":"Shandong Key Laboratory of Applied Technology for Protein and Peptide Drugs, Institute of Biopharmaceutical Research, Liaocheng University, Liaocheng 252000, China"},{"name":"Pet Nutrition Research and Development Center, Gambol Pet Group Co., Ltd., Liaocheng 252000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"14717","DOI":"10.1038\/s41598-023-41422-9","article-title":"Composition and evolution of the gut microbiota of growing puppies is impacted by their birth weight","volume":"13","author":"Garrigues","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Sperandio, V., and Nguyen, Y. (2012). Enterohemorrhagic E. coli (EHEC) pathogenesis. Front. Cell. Infect. Microbiol., 2.","DOI":"10.3389\/fcimb.2012.00090"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1128\/CMR.00022-13","article-title":"Recent Advances in Understanding Enteric Pathogenic Escherichia coli","volume":"26","author":"Law","year":"2013","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"20180054","DOI":"10.1098\/rstb.2018.0054","article-title":"Behavioural ecology and infectious disease: Implications for conservation of biodiversity","volume":"374","author":"Herrera","year":"2019","journal-title":"Philos. Trans. R. Soc. B Biol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Westr\u00f6m, B., Sureda, E.A., Pierzynowska, K., Pierzynowski, S.G., and P\u00e9rez-Cano, F.-J. (2020). The Immature Gut Barrier and Its Importance in Establishing Immunity in Newborn Mammals. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.01153"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ratsika, A., Codagnone, M.C., O\u2019Mahony, S., Stanton, C., and Cryan, J.F. (2021). Priming for Life: Early Life Nutrition and the Microbiota-Gut-Brain Axis. Nutrients, 13.","DOI":"10.3390\/nu13020423"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5","DOI":"10.4014\/jmb.0908.08007","article-title":"A Brief Overview of Escherichia coli O157:H7 and Its Plasmid O157","volume":"20","author":"Lim","year":"2010","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Gopalakrishna, K.P., and Hand, T.W. (2020). Influence of Maternal Milk on the Neonatal Intestinal Microbiome. Nutrients, 12.","DOI":"10.3390\/nu12030823"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Liu, D., Li, C., Cao, T., Lv, X., Yue, Y., Li, S., Cheng, Y., Liu, F., Huo, G., and Li, B. (2024). Bifidobacterium longum K5 Prevents Enterohaemorrhagic Escherichia coli O157:H7 Infection in Mice through the Modulation of the Gut Microbiota. Nutrients, 16.","DOI":"10.3390\/nu16081164"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Li, Y., Ali, I., Lei, Z., Li, Y., Yang, M., Yang, C., and Li, L. (2023). Effect of a Multistrain Probiotic on Feline Gut Health through the Fecal Microbiota and Its Metabolite SCFAs. Metabolites, 13.","DOI":"10.3390\/metabo13020228"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"10362","DOI":"10.1039\/D0FO02670K","article-title":"Lactobacillus plantarum relieves diarrhea caused by enterotoxin-producing Escherichia coli through inflammation modulation and gut microbiota regulation","volume":"11","author":"Yue","year":"2020","journal-title":"Food Funct."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Gong, X., Wang, X., Chen, L., Wang, Z., Han, J., and Wen, M. (2025). Isolation, probiotic properties, and whole-genome analysis of P. acidilactici M22 from feline milk: A promising candidate for simulated pet milk formulations. Front. Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1664636"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"723","DOI":"10.1016\/j.vaccine.2017.12.026","article-title":"Protective immunity of a Multivalent Vaccine Candidate against piglet diarrhea caused by enterotoxigenic Escherichia coli (ETEC) in a pig model","volume":"36","author":"Zhang","year":"2018","journal-title":"Vaccine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1007\/s12602-017-9312-8","article-title":"In Vitro Evaluation of the Probiotic and Safety Properties of Bacteriocinogenic and Non-Bacteriocinogenic Lactic Acid Bacteria from the Intestines of Nile Tilapia and Common Carp for Their Use as Probiotics in Aquaculture","volume":"10","author":"Kaktcham","year":"2018","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"105507","DOI":"10.1016\/j.jff.2023.105507","article-title":"Preventive of Lacticaseibacillus casei WLCA02 against Salmonella typhimurium infection via strengthening the intestinal barrier and activating the macrophages","volume":"104","author":"Tian","year":"2023","journal-title":"J. Funct. Foods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"101589","DOI":"10.1016\/j.fbio.2022.101589","article-title":"Lactobacillus plantarum KLDS1.0386 with antioxidant capacity ameliorates the lipopolysaccharide-induced acute liver injury in mice by NF-\u03baB and Nrf2 pathway","volume":"47","author":"Dong","year":"2022","journal-title":"Food Biosci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5283","DOI":"10.1038\/s41598-021-84761-1","article-title":"DSS-induced colitis is associated with adipose tissue dysfunction and disrupted hepatic lipid metabolism leading to hepatosteatosis and dyslipidemia in mice","volume":"11","author":"Kwon","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Janicka, P., Stygar, D., Che\u0142mecka, E., Kuropka, P., Mi\u0105\u017cek, A., Studzi\u0144ska, A., Pogorzelska, A., Pala, K., and Ba\u017can\u00f3w, B. (2024). Oxidative Stress Markers and Histopathological Changes in Selected Organs of Mice Infected with Murine Norovirus 1 (MNV-1). Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25073614"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5261","DOI":"10.1128\/AEM.00062-07","article-title":"Na\u00efve Bayesian Classifier for Rapid Assignment of rRNA Sequences into the New Bacterial Taxonomy","volume":"73","author":"Wang","year":"2007","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1421","DOI":"10.1007\/s00253-015-7039-6","article-title":"Denitrifying sulfide removal process on high-salinity wastewaters in the presence of Halomonas sp","volume":"100","author":"Liu","year":"2016","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"i884","DOI":"10.1093\/bioinformatics\/bty560","article-title":"fastp: An ultra-fast all-in-one FASTQ preprocessor","volume":"34","author":"Chen","year":"2018","journal-title":"Bioinformatics"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Hua, Y., Wu, J., Fu, M., Liu, J., Li, X., Zhang, B., Zhao, W., and Wan, C. (2020). Enterohemorrhagic Escherichia coli Effector Protein EspF Interacts with Host Protein ANXA6 and Triggers Myosin Light Chain Kinase (MLCK)-Dependent Tight Junction Dysregulation. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.613061"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Ugalde-Silva, P., Gonzalez-Lugo, O., and Navarro-Garcia, F. (2016). Tight Junction Disruption Induced by Type 3 Secretion System Effectors Injected by Enteropathogenic and Enterohemorrhagic Escherichia coli. Front. Cell. Infect. Microbiol., 6.","DOI":"10.3389\/fcimb.2016.00087"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Vinayamohan, P., Joseph, D., Viju, L.S., Baskaran, S.A., and Venkitanarayanan, K. (2024). Efficacy of Probiotics in Reducing Pathogenic Potential of Infectious Agents. Fermentation, 10.","DOI":"10.3390\/fermentation10120599"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Wang, J., Yang, X., Peng, Y., Zhang, J., Huang, Y., Zhong, Z., Liu, H., Fu, H., Zhou, Z., and Peng, G. (2024). Isolation and in vitro investigation on lactic acid bacteria for potential probiotic properties from cat feces. Front. Vet. Sci., 11.","DOI":"10.3389\/fvets.2024.1495745"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Shah, H., Trivedi, M., Gurjar, T., Sahoo, D.K., Jergens, A.E., Yadav, V.K., Patel, A., and Pandya, P. (2024). Decoding the Gut Microbiome in Companion Animals: Impacts and Innovations. Microorganisms, 12.","DOI":"10.3390\/microorganisms12091831"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Poimenidou, S.V., Skarveli, A., Saxami, G., Mitsou, E.K., Kotsou, M., and Kyriacou, A. (2023). Inhibition of Listeria monocytogenes Growth, Adherence and Invasion in Caco-2 Cells by Potential Probiotic Lactic Acid Bacteria Isolated from Fecal Samples of Healthy Neonates. Microorganisms, 11.","DOI":"10.3390\/microorganisms11020363"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wang, Z., Yang, L., Tang, H., Zhang, K., Chen, Q., Liu, C., Guo, Y., Li, M., Guo, Z., and Li, B. (2022). In vivo evidence of the prevents DSS-induced colitis of Lactiplantibacillus plantarum L15. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.1028919"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Mariani, F., and Roncucci, L. (2017). Role of the Vanins\u2013Myeloperoxidase Axis in Colorectal Carcinogenesis. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18050918"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"e4758","DOI":"10.21769\/BioProtoc.4758","article-title":"Measuring Myeloperoxidase Activity as a Marker of Inflammation in Gut Tissue Samples of Mice and Rat","volume":"13","author":"Hanning","year":"2023","journal-title":"Bio-Protoc."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"790","DOI":"10.1007\/s12031-020-01491-1","article-title":"Cytokines, Masticatory Muscle Inflammation, and Pain: An Update","volume":"70","author":"Ayoub","year":"2020","journal-title":"J. Mol. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Duque, G.A., and Descoteaux, A. (2014). Macrophage Cytokines: Involvement in Immunity and Infectious Diseases. Front. Immunol., 5.","DOI":"10.3389\/fimmu.2014.00491"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5843","DOI":"10.1039\/C9FO01390C","article-title":"Lactobacillus casei LC2W can inhibit the colonization of Escherichia coli O157:H7 in vivo and reduce the severity of colitis","volume":"10","author":"Wang","year":"2019","journal-title":"Food Funct."},{"key":"ref_34","first-page":"1","article-title":"Differential Immune Response of Lactobacillus plantarum 286 Against Salmonella Typhimurium Infection in Conventional and Germ-Free Mice","volume":"Volume 15","author":"Donelli","year":"2021","journal-title":"Advances in Microbiology, Infectious Diseases and Public Health"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"106409","DOI":"10.1016\/j.jff.2024.106409","article-title":"Lactiplantibacillus plantarum ZJ316 alleviates the oxidative stress and cellular apoptosis via modulating Nrf2\/HO-1 signaling pathway","volume":"121","author":"Wu","year":"2024","journal-title":"J. Funct. Foods"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Liu, D., Yue, Y., Ping, L., Sun, C., Zheng, T., Cheng, Y., Huo, G., and Li, B. (2024). Lactobacillus delbrueckii subsp. bulgaricus 1.0207 Exopolysaccharides Attenuate Hydrogen Peroxide-Induced Oxidative Stress Damage in IPEC-J2 Cells through the Keap1\/Nrf2 Pathway. Antioxidants, 13.","DOI":"10.3390\/antiox13091150"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Gong, X., Wang, X., Chen, L., Huang, H., Zhang, N., Han, J., Wang, Z., and Wen, M. (2025). The Prophylactic Protection of P. acidilactici M22 from Feline Milk on S. typhimurium Infection in Mice. Microorganisms, 13.","DOI":"10.3390\/microorganisms13102353"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"103717","DOI":"10.1016\/j.autrev.2024.103717","article-title":"Intestinal mucus barrier: A potential therapeutic target for IBD","volume":"24","author":"Qiao","year":"2025","journal-title":"Autoimmun. Rev."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Yoon, K.-N., Yang, J., Yeom, S.-J., Kim, S.-S., Park, J.-H., Song, B.-S., Eun, J.-B., Park, S.-H., Lee, J.H., and Kim, H.B. (2024). Lactiplantibacillus argentoratensis AGMB00912 protects weaning mice from ETEC infection and enhances gut health. Front. Microbiol., 15.","DOI":"10.3389\/fmicb.2024.1440134"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1016\/j.suronc.2017.07.011","article-title":"The role of intestinal bacteria in the development and progression of gastrointestinal tract neoplasms","volume":"26","author":"Mima","year":"2017","journal-title":"Surg. Oncol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1355","DOI":"10.1007\/s12602-022-09987-5","article-title":"Lactiplantibacillus plantarum Regulated Intestinal Microbial Community and Cytokines to Inhibit Salmonella typhimurium Infection","volume":"15","author":"Liu","year":"2023","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e00304-20","DOI":"10.1128\/AEM.00304-20","article-title":"Chicken Intestinal Mycobiome: Initial Characterization and Its Response to Bacitracin Methylene Disalicylate","volume":"86","author":"Robinson","year":"2020","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1038\/s41587-020-0548-6","article-title":"PICRUSt2 for prediction of metagenome functions","volume":"38","author":"Douglas","year":"2020","journal-title":"Nat. Biotechnol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/2\/332\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T11:34:44Z","timestamp":1770377684000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/2\/332"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,30]]},"references-count":43,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["microorganisms14020332"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14020332","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,30]]}}}