{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T21:03:20Z","timestamp":1793221400837,"version":"4.5.0"},"reference-count":90,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,7,29]],"date-time":"2025-07-29T00:00:00Z","timestamp":1753747200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100011033","name":"MCIN\/AEI\/10.13039\/501100011033","doi-asserted-by":"publisher","award":["CNS2022-135378"],"award-info":[{"award-number":["CNS2022-135378"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011033","name":"MCIN\/AEI\/10.13039\/501100011033","doi-asserted-by":"publisher","award":["LE044P20"],"award-info":[{"award-number":["LE044P20"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011033","name":"MCIN\/AEI\/10.13039\/501100011033","doi-asserted-by":"publisher","award":["FPU2022-00432"],"award-info":[{"award-number":["FPU2022-00432"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008431","name":"Junta de Castilla y Le\u00f3n (JCyL), Spain","doi-asserted-by":"publisher","award":["CNS2022-135378"],"award-info":[{"award-number":["CNS2022-135378"]}],"id":[{"id":"10.13039\/501100008431","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008431","name":"Junta de Castilla y Le\u00f3n (JCyL), Spain","doi-asserted-by":"publisher","award":["LE044P20"],"award-info":[{"award-number":["LE044P20"]}],"id":[{"id":"10.13039\/501100008431","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008431","name":"Junta de Castilla y Le\u00f3n (JCyL), Spain","doi-asserted-by":"publisher","award":["FPU2022-00432"],"award-info":[{"award-number":["FPU2022-00432"]}],"id":[{"id":"10.13039\/501100008431","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Fundaci\u00f3n General de la Universidad de Le\u00f3n y de la Empresa (FGULEM)","award":["CNS2022-135378"],"award-info":[{"award-number":["CNS2022-135378"]}]},{"name":"Fundaci\u00f3n General de la Universidad de Le\u00f3n y de la Empresa (FGULEM)","award":["LE044P20"],"award-info":[{"award-number":["LE044P20"]}]},{"name":"Fundaci\u00f3n General de la Universidad de Le\u00f3n y de la Empresa (FGULEM)","award":["FPU2022-00432"],"award-info":[{"award-number":["FPU2022-00432"]}]},{"name":"JCyL","award":["CNS2022-135378"],"award-info":[{"award-number":["CNS2022-135378"]}]},{"name":"JCyL","award":["LE044P20"],"award-info":[{"award-number":["LE044P20"]}]},{"name":"JCyL","award":["FPU2022-00432"],"award-info":[{"award-number":["FPU2022-00432"]}]},{"name":"Beca de Colaboraci\u00f3n","award":["CNS2022-135378"],"award-info":[{"award-number":["CNS2022-135378"]}]},{"name":"Beca de Colaboraci\u00f3n","award":["LE044P20"],"award-info":[{"award-number":["LE044P20"]}]},{"name":"Beca de Colaboraci\u00f3n","award":["FPU2022-00432"],"award-info":[{"award-number":["FPU2022-00432"]}]},{"name":"FPU","award":["CNS2022-135378"],"award-info":[{"award-number":["CNS2022-135378"]}]},{"name":"FPU","award":["LE044P20"],"award-info":[{"award-number":["LE044P20"]}]},{"name":"FPU","award":["FPU2022-00432"],"award-info":[{"award-number":["FPU2022-00432"]}]},{"name":"Asociaci\u00f3n Espa\u00f1ola Contra el C\u00e1ncer (AECC)","award":["CNS2022-135378"],"award-info":[{"award-number":["CNS2022-135378"]}]},{"name":"Asociaci\u00f3n Espa\u00f1ola Contra el C\u00e1ncer (AECC)","award":["LE044P20"],"award-info":[{"award-number":["LE044P20"]}]},{"name":"Asociaci\u00f3n Espa\u00f1ola Contra el C\u00e1ncer (AECC)","award":["FPU2022-00432"],"award-info":[{"award-number":["FPU2022-00432"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Background\/Objectives: Listeria monocytogenes is a foodborne pathogen of major public health concern due to its ability to invade host cells and cause severe illness. This study aimed to develop and validate a multi-tiered screening pipeline to identify Bacillus strains with probiotic potential against L. monocytogenes. Methods: A total of 26 Bacillus isolates were screened for antimicrobial activity, gastrointestinal resilience, and host cell adhesion. A fluorescence-based infection assay using mCherry-expressing HCT 116 cells was used to assess cytoprotection against L. monocytogenes NCTC 7973. Eight strains significantly improved host cell viability and were validated by quantification of intracellular CFU. Two top candidates were tested in a murine model of listeriosis. The genome of the lead strain was sequenced to evaluate safety and biosynthetic potential. Results: B. subtilis CECT 8266 completely inhibited intracellular replication of L. monocytogenes in HCT 116 cells, reducing bacterial recovery to undetectable levels. In vivo, it decreased splenic bacterial burden by approximately 6-fold. Genomic analysis revealed eight bacteriocin biosynthetic clusters and silent antibiotic resistance genes within predicted genomic islands, as determined by CARD and Alien Hunter analysis. The strain also demonstrated bile and acid tolerance, as well as strong adhesion to epithelial cells. Conclusions: The proposed pipeline enables efficient identification of probiotic Bacillus strains with intracellular protective activity. B. subtilis CECT 8266 is a promising candidate for translational applications in food safety or health due to its efficacy, resilience, and safety profile.<\/jats:p>","DOI":"10.3390\/antibiotics14080765","type":"journal-article","created":{"date-parts":[[2025,7,29]],"date-time":"2025-07-29T16:16:10Z","timestamp":1753805770000},"page":"765","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Streamlining Bacillus Strain Selection Against Listeria monocytogenes Using a Fluorescence-Based Infection Assay Integrated into a Multi-Tiered Validation Pipeline"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0131-7520","authenticated-orcid":false,"given":"Blanca","family":"Lorente-Torres","sequence":"first","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-4314-4457","authenticated-orcid":false,"given":"Pablo","family":"Casta\u00f1era","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-1582-2779","authenticated-orcid":false,"given":"Helena \u00c1.","family":"Ferrero","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-9642-9850","authenticated-orcid":false,"given":"Sergio","family":"Fern\u00e1ndez-Mart\u00ednez","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3991-0466","authenticated-orcid":false,"given":"Suleiman","family":"Adejoh Ocholi","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Nigeria, University Road 1, Nsukka 410105, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-3238-5565","authenticated-orcid":false,"given":"Jes\u00fas","family":"Llano-Verdeja","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Farzaneh","family":"Javadimarand","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1539-1249","authenticated-orcid":false,"given":"Yaiza","family":"Carnicero-Mayo","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4179-6450","authenticated-orcid":false,"given":"Amanda","family":"Herrero-Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Neural Therapies SL. Edif. Institutos de Investigaci\u00f3n, Planta baja, Local B43. Campus de Vegazana s\/n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alba","family":"Puente-Sanz","sequence":"additional","affiliation":[{"name":"Neural Therapies SL. Edif. Institutos de Investigaci\u00f3n, Planta baja, Local B43. Campus de Vegazana s\/n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene","family":"Sainz Mach\u00edn","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4929-4453","authenticated-orcid":false,"given":"Isabel Karola","family":"Voigt","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Guerrero Villanueva","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1lvaro","family":"L\u00f3pez Garc\u00eda","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Mart\u00edn G\u00f3mez","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James C.","family":"Ogbonna","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Nigeria, University Road 1, Nsukka 410105, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 M.","family":"Gonzalo-Orden","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4242-4701","authenticated-orcid":false,"given":"Jes\u00fas F.","family":"Aparicio","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7763-582X","authenticated-orcid":false,"given":"Luis M.","family":"Mateos","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Instituto de Biolog\u00eda Molecular, Gen\u00f3mica y Prote\u00f3mica (INBIOMIC), Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4926-3891","authenticated-orcid":false,"given":"\u00c1lvaro","family":"Mourenza","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9509-5174","authenticated-orcid":false,"given":"Michal","family":"Letek","sequence":"additional","affiliation":[{"name":"Departamento de Biolog\u00eda Molecular, \u00c1rea de Microbiolog\u00eda, Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Instituto de Desarrollo Ganadero y Sanidad Animal (INDEGSAL), Universidad de Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/S0168-1605(03)00326-X","article-title":"Listeria monocytogenes and Listeriosis: A Review of Hazard Characterisation for Use in Microbiological Risk Assessment of Foods","volume":"92","author":"McLauchlin","year":"2004","journal-title":"Int. J. Food Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2791","DOI":"10.1007\/s00203-020-02004-9","article-title":"Antimicrobial Effect of Spore-Forming Probiotics Bacillus laterosporus and Bacillus megaterium against Listeria monocytogenes","volume":"202","author":"Rajabi","year":"2020","journal-title":"Arch. Microbiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.foodcont.2016.12.016","article-title":"A Review of Listeria monocytogenes: An Update on Outbreaks, Virulence, Dose-Response, Ecology, and Risk Assessments","volume":"75","author":"Buchanan","year":"2017","journal-title":"Food Control"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1089\/fpd.2010.0718","article-title":"Listeria monocytogenes: Antibiotic Resistance in Food Production","volume":"8","author":"Lungu","year":"2011","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.foodcont.2011.06.006","article-title":"Increase over Time in the Prevalence of Multiple Antibiotic Resistance among Isolates of Listeria monocytogenes from Poultry in Spain","volume":"23","author":"Prieto","year":"2012","journal-title":"Food Control"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1007\/s12602-010-9040-9","article-title":"Antilisterial Activity on Poultry Meat of Amylolysin, a Bacteriocin from Bacillus amyloliquefaciens GA1","volume":"2","author":"Halimi","year":"2010","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.1016\/j.micinf.2007.05.011","article-title":"The Epidemiology of Human Listeriosis","volume":"9","author":"Swaminathan","year":"2007","journal-title":"Microbes Infect."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Wilson, A., Gray, J., Scott Chandry, P., and Fox, E.M. (2018). Phenotypic and Genotypic Analysis of Antimicrobial Resistance among Listeria monocytogenes Isolated from Australian Food Production Chains. Genes, 9.","DOI":"10.3390\/genes9020080"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1007\/s12602-010-9042-7","article-title":"Antibacterial Activity of Subtilosin Alone and Combined with Curcumin, Poly-Lysine and Zinc Lactate against Listeria monocytogenes Strains","volume":"2","author":"Amrouche","year":"2010","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1007\/s12602-021-09772-w","article-title":"Bacteriocinogenic Potential of Bacillus amyloliquefaciens Isolated from Kimchi, a Traditional Korean Fermented Cabbage","volume":"13","author":"Jung","year":"2021","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1056\/NEJMoa1907462","article-title":"Outbreak of Listeriosis in South Africa Associated with Processed Meat","volume":"382","author":"Thomas","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"100127","DOI":"10.1016\/j.medmic.2025.100127","article-title":"The Past, Present, Future of Listeria monocytogenes: Understanding the Molecular Pathways, Antibiotic Resistance and Public Health Implications","volume":"25","author":"Anupama","year":"2025","journal-title":"Med. Microecol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"6344","DOI":"10.1038\/s41467-020-20200-5","article-title":"Receptor-Targeted Engineered Probiotics Mitigate Lethal Listeria Infection","volume":"11","author":"Drolia","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1111\/j.1574-6976.2010.00244.x","article-title":"Diversity and Applications of Bacillus Bacteriocins","volume":"35","author":"Abriouel","year":"2011","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.micres.2014.09.004","article-title":"Antibacterial Activity and Genotypic-Phenotypic Characteristics of Bacteriocin-Producing Bacillus subtilis KKU213: Potential as a Probiotic Strain","volume":"170","author":"Khochamit","year":"2015","journal-title":"Microbiol. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.ijfoodmicro.2007.09.001","article-title":"Application of Bacteriocins in Vegetable Food Biopreservation","volume":"121","author":"Settanni","year":"2008","journal-title":"Int. J. Food Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1093\/jac\/dkl495","article-title":"The Spectrum of Antimicrobial Activity of the Bacteriocin Subtilosin A","volume":"59","author":"Shelburne","year":"2007","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1146\/annurev.micro.61.080706.093501","article-title":"Lantibiotics: Peptides of Diverse Structure and Function","volume":"61","author":"Willey","year":"2007","journal-title":"Annu. Rev. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Elshaghabee, F.M.F., Rokana, N., Gulhane, R.D., Sharma, C., and Panwar, H. (2017). Bacillus as Potential Probiotics: Status, Concerns, and Future Perspectives. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.01490"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Choyam, S., Jain, P.M., and Kammara, R. (2021). Characterization of a Potent New-Generation Antimicrobial Peptide of Bacillus. Front. Microbiol., 12.","DOI":"10.3389\/fmicb.2021.710741"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1058","DOI":"10.1016\/j.idairyj.2005.10.026","article-title":"Bacteriocins: Biological Tools for Bio-Preservation and Shelf-Life Extension","volume":"16","author":"Deegan","year":"2006","journal-title":"Int. Dairy J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1007\/s12602-013-9151-1","article-title":"Investigation of the Antimicrobial Activity of Bacillus licheniformis Strains Isolated from Retail Powdered Infant Milk Formulae","volume":"6","author":"Begley","year":"2014","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"5451","DOI":"10.1128\/AEM.00730-09","article-title":"Identification of a Novel Two-Peptide Lantibiotic, Lichenicidin, Following Rational Genome Mining for LanM Proteins","volume":"75","author":"Begley","year":"2009","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Dischinger, J., Josten, M., Szekat, C., Sahl, H.G., and Bierbaum, G. (2009). Production of the Novel Two-Peptide Lantibiotic Lichenicidin by Bacillus licheniformis DSM 13. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0006788"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Saggese, A., De Luca, Y., Baccigalupi, L., and Ricca, E. (2022). An Antimicrobial Peptide Specifically Active against Listeria monocytogenes Is Secreted by Bacillus pumilus SF214. BMC Microbiol., 22.","DOI":"10.1186\/s12866-021-02422-9"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Rani, R.P., Anandharaj, M., Hema, S., Deepika, R., and Ravindran, A.D. (2016). Purification of Antilisterial Peptide (Subtilosin A) from Novel Bacillus tequilensis FR9 and Demonstrate Their Pathogen Invasion Protection Ability Using Human Carcinoma Cell Line. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.01910"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1038\/s41538-022-00130-w","article-title":"African Fermented Foods: Overview, Emerging Benefits, and Novel Approaches to Microbiome Profiling","volume":"6","author":"Obafemi","year":"2022","journal-title":"NPJ Sci. Food"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2570","DOI":"10.1007\/s13197-017-2688-3","article-title":"Importance of the Gastrointestinal Life Cycle of Bacillus for Probiotic Functionality","volume":"54","author":"Bernardeau","year":"2017","journal-title":"J. Food Sci. Technol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"9397","DOI":"10.1039\/D0FO01981J","article-title":"Simulating Human Digestion: Developing Our Knowledge to Create Healthier and More Sustainable Foods","volume":"11","author":"Mackie","year":"2020","journal-title":"Food Funct."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ghasemi, M., Turnbull, T., Sebastian, S., and Kempson, I. (2021). The MTT Assay: Utility, Limitations, Pitfalls, and Interpretation in Bulk and Single-Cell Analysis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222312827"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Van den Bossche, S., Vandeplassche, E., Ostyn, L., Coenye, T., and Crabb\u00e9, A. (2020). Bacterial Interference with Lactate Dehydrogenase Assay Leads to an Underestimation of Cytotoxicity. Front. Cell Infect. Microbiol., 10.","DOI":"10.3389\/fcimb.2020.00494"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"110993","DOI":"10.1016\/j.lwt.2021.110993","article-title":"A Novel Subtilin-like Lantibiotics Subtilin JS-4 Produced by Bacillus subtilis JS-4, and Its Antibacterial Mechanism against Listeria monocytogenes","volume":"142","author":"Wei","year":"2021","journal-title":"LWT"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"107696","DOI":"10.1016\/j.foodcont.2020.107696","article-title":"Identification and Characterization of a Novel Circular Bacteriocin, Bacicyclicin XIN-1, from Bacillus Sp. Xin1","volume":"121","author":"Xin","year":"2021","journal-title":"Food Control"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e00185-21","DOI":"10.1128\/AEM.00185-21","article-title":"Toyoncin, a Novel Leaderless Bacteriocin That Is Produced by Bacillus toyonensis XIN-YC13 and Specifically Targets B. cereus and Listeria monocytogenes","volume":"87","author":"Wang","year":"2021","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Lin, L.Z., Zheng, Q.W., Wei, T., Zhang, Z.Q., Zhao, C.F., Zhong, H., Xu, Q.Y., Lin, J.F., and Guo, L.Q. (2020). Isolation and Characterization of Fengycins Produced by Bacillus amyloliquefaciens JFL21 and Its Broad-Spectrum Antimicrobial Potential Against Multidrug-Resistant Foodborne Pathogens. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.579621"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/j.micres.2012.11.004","article-title":"Inhibitory Activity of Surfactin, Produced by Different Bacillus subtilis subsp. subtilis Strains, against Listeria monocytogenes Sensitive and Bacteriocin-Resistant Strains","volume":"168","author":"Audisio","year":"2013","journal-title":"Microbiol. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"9990","DOI":"10.1021\/acs.jafc.2c02890","article-title":"Thuricins: Novel Leaderless Bacteriocins with Potent Antimicrobial Activity Against Gram-Positive Foodborne Pathogens","volume":"70","author":"Deng","year":"2022","journal-title":"J. Agric. Food Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1462","DOI":"10.4014\/jmb.2208.08006","article-title":"Identification and Characterization of a Bacteriocin from the Newly Isolated Bacillus subtilis HD15 with Inhibitory Effects against Bacillus cereus","volume":"32","author":"Hong","year":"2022","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Wang, Y., Gu, Z., Zhang, S., and Li, P. (2023). Complete Genome Sequencing Revealed the Potential Application of a Novel Weizmannia coagulans PL-W Production with Promising Bacteriocins in Food Preservative. Foods, 12.","DOI":"10.3390\/foods12010216"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1186\/s12934-022-01912-3","article-title":"Three Novel Leaderless Bacteriocins Have Antimicrobial Activity against Gram-Positive Bacteria to Serve as Promising Food Biopreservative","volume":"21","author":"Zhang","year":"2022","journal-title":"Microb. Cell Fact."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"3683","DOI":"10.1007\/s13197-018-3298-4","article-title":"Purification and Characterization of a Broad Spectrum Bacteriocin Produced by a Selected Lactococcus lactis Strain 63 Isolated from Indian Dairy Products","volume":"55","author":"Goyal","year":"2018","journal-title":"J. Food Sci. Technol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Zhang, J., Yang, Y., Yang, H., Bu, Y., Yi, H., Zhang, L., Han, X., and Ai, L. (2018). Purification and Partial Characterization of Bacteriocin Lac-B23, a Novel Bacteriocin Production by Lactobacillus plantarum J23, Isolated from Chinese Traditional Fermented Milk. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.02165"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1007\/s00284-012-0115-6","article-title":"Extracellular Proteins from Lactobacillus plantarum BMCM12 Prevent Adhesion of Enteropathogens to Mucin","volume":"64","author":"Urdaci","year":"2012","journal-title":"Curr. Microbiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1007\/s002530051570","article-title":"Production, Recovery and Purification of Bacteriocins from Lactic Acid Bacteria","volume":"52","author":"Parente","year":"1999","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Sharma, V., Aseri, G., Bhagwat, P., Jain, N., and Ranveer, R.C. (2021). Production, Purification And Characterization Of A Novel Bacteriocin Produced By Bacillus subtilis Vs Isolated From Mango (Mangifera Indica L.). Braz. Arch. Biol. Technol., 64.","DOI":"10.1590\/1678-4324-2021190749"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/j.foodres.2010.09.002","article-title":"Rhamnolipid and Surfactin Inhibit Listeria monocytogenes Adhesion","volume":"44","author":"Abreu","year":"2011","journal-title":"Food Res. Int."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.tim.2007.12.009","article-title":"Bacillus Lipopeptides: Versatile Weapons for Plant Disease Biocontrol","volume":"16","author":"Ongena","year":"2008","journal-title":"Trends Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Nalli, Y., Singh, S., Gajjar, A., Mahizhaveni, B., Dusthackeer, V.N.A., and Shinde, P.B. (2023). Bacillibactin Class Siderophores Produced by the Endophyte Bacillus subtilis NPROOT3 as Antimycobacterial Agents. Lett. Appl. Microbiol., 76.","DOI":"10.1093\/lambio\/ovac026"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"e30974","DOI":"10.1002\/jcp.30974","article-title":"Bacillaene, Sharp Objects Consist in the Arsenal of Antibiotics Produced by Bacillus","volume":"239","author":"Miao","year":"2023","journal-title":"J. Cell Physiol."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Yuan, S., Yong, X., Zhao, T., Li, Y., and Liu, J. (2020). Research Progress of the Biosynthesis of Natural Bio-Antibacterial Agent Pulcherriminic Acid in Bacillus. Molecules, 25.","DOI":"10.3390\/molecules25235611"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Islam, T., Rabbee, M.F., Choi, J., and Baek, K.H. (2022). Biosynthesis, Molecular Regulation, and Application of Bacilysin Produced by Bacillus Species. Metabolites, 12.","DOI":"10.3390\/metabo12050397"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Grahovac, J., Paj\u010din, I., and Vlajkov, V. (2023). Bacillus VOCs in the Context of Biological Control. Antibiotics, 12.","DOI":"10.3390\/antibiotics12030581"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"European Food Safety Authority (2012). Guidance on the Assessment of Bacterial Susceptibility to Antimicrobials of Human and Veterinary Importance. EFSA J., 10, 2740.","DOI":"10.2903\/j.efsa.2012.2740"},{"key":"ref_54","first-page":"e01978","article-title":"A National Survey of the Antibiotic Use, Self-Medication Practices, and Knowledge of Antibiotic Resistance among Graduates of Tertiary Institutions in Nigeria","volume":"23","author":"Popoola","year":"2024","journal-title":"Sci. Afr."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1038\/nrmicro2259","article-title":"Bacterial Competition: Surviving and Thriving in the Microbial Jungle","volume":"8","author":"Hibbing","year":"2010","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Gray, J., Chandry, P.S., Kaur, M., Kocharunchitt, C., Fanning, S., Bowman, J.P., and Fox, E.M. (2021). Colonisation Dynamics of Listeria monocytogenes Strains Isolated from Food Production Environments. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-91503-w"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"786","DOI":"10.3390\/applmicrobiol3030055","article-title":"Perspectives on Using a Competitive Exclusion Approach to Control Listeria monocytogenes in Biological Soil Amendments of Animal Origin (BSAAO): A Review","volume":"3","author":"Wang","year":"2023","journal-title":"Appl. Microbiol."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Akinsemolu, A.A., Onyeaka, H., Odion, S., and Adebanjo, I. (2024). Exploring Bacillus subtilis: Ecology, Biotechnological Applications, and Future Prospects. J. Basic Microbiol., 64.","DOI":"10.1002\/jobm.202300614"},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Vivant, A.L., Garmyn, D., and Piveteau, P. (2013). Listeria monocytogenes, a down-to-Earth Pathogen. Front. Cell Infect. Microbiol., 4.","DOI":"10.3389\/fcimb.2014.00160"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Caulier, S., Nannan, C., Gillis, A., Licciardi, F., Bragard, C., and Mahillon, J. (2019). Overview of the Antimicrobial Compounds Produced by Members of the Bacillus subtilis Group. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.00302"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"101007","DOI":"10.1016\/j.cofs.2023.101007","article-title":"Bacillus species in Food Fermentations: An Underappreciated Group of Organisms for Safe Use in Food Fermentations","volume":"50","author":"Li","year":"2023","journal-title":"Curr. Opin. Food Sci."},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Meade, E., Slattery, M.A., and Garvey, M. (2020). Bacteriocins, Potent Antimicrobial Peptides and the Fight against Multi Drug Resistant Species: Resistance Is Futile?. Antibiotics, 9.","DOI":"10.3390\/antibiotics9010032"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.tifs.2017.08.005","article-title":"Correlation between in Vitro and in Vivo Assays in Selection of Probiotics from Traditional Species of Bacteria","volume":"68","author":"Vinderola","year":"2017","journal-title":"Trends Food Sci. Technol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0168-1605(01)00560-8","article-title":"Bacteriocins: Safe, Natural Antimicrobials for Food Preservation","volume":"71","author":"Cleveland","year":"2001","journal-title":"Int. J. Food Microbiol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1574-695X.2008.00454.x","article-title":"Exported Proteins in Probiotic Bacteria: Adhesion to Intestinal Surfaces, Host Immunomodulation and Molecular Cross-Talking with the Host","volume":"54","author":"Bressollier","year":"2008","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1042\/ETLS20190061","article-title":"Animal Experimentation: Implementation and Application of the 3Rs","volume":"3","author":"Lewis","year":"2019","journal-title":"Emerg. Top. Life Sci."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1186\/1297-9716-43-18","article-title":"Animal Models of Listeriosis: A Comparative Review of the Current State of the Art and Lessons Learned","volume":"43","author":"Hoelzer","year":"2012","journal-title":"Vet. Res."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"2902","DOI":"10.1111\/jam.15738","article-title":"\u2018To Be, or Not to Be\u2019\u2014The Dilemma of \u2018Silent\u2019 Antimicrobial Resistance Genes in Bacteria","volume":"133","author":"Deekshit","year":"2022","journal-title":"J. Appl. Microbiol."},{"key":"ref_69","first-page":"509","article-title":"The Promises and Risks of Probiotic Bacillus Species","volume":"65","author":"Banaszczyk","year":"2018","journal-title":"Acta Biochim. Pol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"13","DOI":"10.2147\/tcrm.2007.3.1.13","article-title":"Efficacy of Bacillus clausii Spores in the Prevention of Recurrent Respiratory Infections in Children: A Pilot Study","volume":"3","author":"Marseglia","year":"2007","journal-title":"Ther. Clin. Risk Manag."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/s12979-015-0051-y","article-title":"Probiotic Strain Bacillus subtilis CU1 Stimulates Immune System of Elderly during Common Infectious Disease Period: A Randomized, Double-Blind Placebo-Controlled Study","volume":"12","author":"Lefevre","year":"2015","journal-title":"Immun. Ageing"},{"key":"ref_72","doi-asserted-by":"crossref","unstructured":"Bravo-Santano, N., Boll, E.J., Capern, L.C., Cieplak, T.M., Keleszade, E., Letek, M., and Costabile, A. (2020). Comparative Evaluation of the Antimicrobial and Mucus Induction Properties of Selected Bacillus Strains against Enterotoxigenic Escherichia coli. Antibiotics, 9.","DOI":"10.3390\/antibiotics9120849"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"e01934-17","DOI":"10.1128\/JCM.01934-17","article-title":"CLSI Methods Development and Standardization Working Group Best Practices for Evaluation of Antimicrobial Susceptibility Tests","volume":"56","author":"Humphries","year":"2018","journal-title":"J. Clin. Microbiol."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"e00374-18","DOI":"10.1128\/mSphere.00374-18","article-title":"Intracellular Staphylococcus aureus Modulates Host Central Carbon Metabolism to Activate Autophagy","volume":"3","author":"Ellis","year":"2018","journal-title":"mSphere"},{"key":"ref_75","doi-asserted-by":"crossref","unstructured":"Bravo-Santano, N., St\u00f6lting, H., Cooper, F., Bileckaja, N., Majstorovic, A., Ihle, N., Mateos, L.M., Calle, Y., Behrends, V., and Letek, M. (2019). Host-Directed Kinase Inhibitors Act as Novel Therapies against Intracellular Staphylococcus aureus. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-41260-8"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1093\/bioinformatics\/btu170","article-title":"Trimmomatic: A Flexible Trimmer for Illumina Sequence Data","volume":"30","author":"Bolger","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Wick, R.R., Judd, L.M., Gorrie, C.L., and Holt, K.E. (2017). Unicycler: Resolving Bacterial Genome Assemblies from Short and Long Sequencing Reads. PLoS Comput. Biol., 13.","DOI":"10.1371\/journal.pcbi.1005595"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"2068","DOI":"10.1093\/bioinformatics\/btu153","article-title":"Prokka: Rapid Prokaryotic Genome Annotation","volume":"30","author":"Seemann","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1236","DOI":"10.1093\/bioinformatics\/btu031","article-title":"InterProScan 5: Genome-Scale Protein Function Classification","volume":"30","author":"Jones","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"D587","DOI":"10.1093\/nar\/gkac963","article-title":"KEGG for Taxonomy-Based Analysis of Pathways and Genomes","volume":"51","author":"Kanehisa","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/s41587-019-0036-z","article-title":"SignalP 5.0 Improves Signal Peptide Predictions Using Deep Neural Networks","volume":"37","author":"Tsirigos","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"1853","DOI":"10.1093\/bioinformatics\/bti303","article-title":"An Improved Hidden Markov Model for Transmembrane Protein Detection and Topology Prediction and Its Applications to Complete Genomes","volume":"21","author":"Kahsay","year":"2005","journal-title":"Bioinformatics"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"W29","DOI":"10.1093\/nar\/gkab335","article-title":"AntiSMASH 6.0: Improving Cluster Detection and Comparison Capabilities","volume":"49","author":"Blin","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_84","first-page":"D517","article-title":"CARD 2020: Antibiotic Resistome Surveillance with the Comprehensive Antibiotic Resistance Database","volume":"48","author":"Alcock","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"ref_85","doi-asserted-by":"crossref","unstructured":"da Silva Filho, A.C., Raittz, R.T., Guizelini, D., De Pierri, C.R., Augusto, D.W., dos Santos-Weiss, I.C.R., and Marchaukoski, J.N. (2018). Comparative Analysis of Genomic Island Prediction Tools. Front. Genet., 9.","DOI":"10.3389\/fgene.2018.00619"},{"key":"ref_86","doi-asserted-by":"crossref","unstructured":"Hart, T., and Moffat, J. (2016). BAGEL: A Computational Framework for Identifying Essential Genes from Pooled Library Screens. BMC Bioinform., 17.","DOI":"10.1186\/s12859-016-1015-8"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1111\/j.1574-6968.2006.00492.x","article-title":"Identification of the Emerging Skin Pathogen Corynebacterium amycolatum Using PCR-Amplification of the Essential divIVA Gene as a Target","volume":"265","author":"Letek","year":"2006","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1038\/s43705-023-00330-9","article-title":"Enhanced Specificity of Bacillus Metataxonomics Using a tuf-Targeted Amplicon Sequencing Approach","volume":"3","author":"Xu","year":"2023","journal-title":"ISME Commun."},{"key":"ref_89","first-page":"W5","article-title":"MAFFT-DASH: Integrated Protein Sequence and Structural Alignment","volume":"47","author":"Rozewicki","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"W293","DOI":"10.1093\/nar\/gkab301","article-title":"Interactive Tree of Life (ITOL) v5: An Online Tool for Phylogenetic Tree Display and Annotation","volume":"49","author":"Letunic","year":"2021","journal-title":"Nucleic Acids Res."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/8\/765\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:18:20Z","timestamp":1760033900000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/14\/8\/765"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,29]]},"references-count":90,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["antibiotics14080765"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics14080765","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,29]]}}}