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The study evaluates antimicrobial resistance (AMR) and virulence characteristics of Enterococcus spp. isolated from bovine milk. Bile esculin agar was used to assess 1471 milk samples, followed by colony identification, gram staining, catalase tests, and 45 \u00b0C incubation. PCR analysis targeted E. faecalis and E. faecium in characteristic Enterococcus spp. colonies, with MALDI-TOF used for negative samples. Multiple tests, including disk diffusion, chromogenic VRE agar for vancomycin resistance, Vancomycin Etest\u00ae for MIC determination, and PCR for virulence factors (cylA, esp, efaA, ace, asa1, gelE, and hyl genes), were performed. Out of 100 identified strains, E. durans (30.66%), E. faecium (26.28%), and E. faecalis (18.25%) were predominant. AMR in Enterococcus spp. varied, with the highest rates against rifampicin (27%), tetracycline (20%), and erythromycin (18%). Linezolid (5%), vancomycin, ciprofloxacin, and teicoplanin (3% each) had lower prevalence. E. faecium and E. faecalis showed high AMR to rifampicin, erythromycin, and tetracycline. Thirty-two strains (18.98%) grew on VRE Chromoselect agar, while 4 (2 E. faecalis and 2 E. faecium) showed vancomycin resistance by MIC values. E. faecalis carried gelE (45.5%) and asa1 (36%), and E. gallinarum had 9.1% with the asa1 gene. Detecting resistant Enterococcus in bovine milk supports control strategies for enterococci on dairy farms, highlighting AMR concerns in the food chain.<\/jats:p>","DOI":"10.3390\/antibiotics12081243","type":"journal-article","created":{"date-parts":[[2023,7,28]],"date-time":"2023-07-28T07:35:24Z","timestamp":1690529724000},"page":"1243","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":33,"title":["The Emergence of Antimicrobial Resistance and Virulence Characteristics in Enterococcus Species Isolated from Bovine Milk"],"prefix":"10.3390","volume":"12","author":[{"given":"Beatriz Rizzo","family":"Paschoalini","sequence":"first","affiliation":[{"name":"Department of Food Science and Nutrition, School of Food Engineering, University of Campinas, Campinas 13083-862, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7799-1979","authenticated-orcid":false,"given":"Karen Vanessa Munive","family":"Nu\u00f1ez","sequence":"additional","affiliation":[{"name":"Department of Food Science and Nutrition, School of Food Engineering, University of Campinas, Campinas 13083-862, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juliana Takahashi","family":"Maffei","sequence":"additional","affiliation":[{"name":"Department of Food Science and Nutrition, School of Food Engineering, University of Campinas, Campinas 13083-862, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5127-0762","authenticated-orcid":false,"given":"H\u00e9lio","family":"Langoni","sequence":"additional","affiliation":[{"name":"Department of Animal Production and Preventive Veterinary Medicine, Faculty of Veterinary Medicine and Animal Science, S\u00e3o Paulo State University, Botucatu 18618-681, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3349-7431","authenticated-orcid":false,"given":"Felipe Freitas","family":"Guimar\u00e3es","sequence":"additional","affiliation":[{"name":"Department of Animal Production and Preventive Veterinary Medicine, Faculty of Veterinary Medicine and Animal Science, S\u00e3o Paulo State University, Botucatu 18618-681, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8753-1020","authenticated-orcid":false,"given":"Clarice","family":"Gebara","sequence":"additional","affiliation":[{"name":"Food Research Center, School of Veterinary Medicine and Animal Science, Federal University of Goi\u00e1s, Campus Road, Goi\u00e2nia 74690-900, GO, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0613-6780","authenticated-orcid":false,"given":"Natylane Eufransino","family":"Freitas","sequence":"additional","affiliation":[{"name":"Food Research Center, School of Veterinary Medicine and Animal Science, Federal University of Goi\u00e1s, Campus Road, Goi\u00e2nia 74690-900, GO, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4273-3494","authenticated-orcid":false,"given":"Marcos Veiga","family":"dos Santos","sequence":"additional","affiliation":[{"name":"Department of Animal Nutrition and Production, School of Veterinary Medicine and Animal Sciences, University of S\u00e3o Paulo (USP), Pirassununga 13635-900, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos Eduardo","family":"Fidelis","sequence":"additional","affiliation":[{"name":"Department of Animal Nutrition and Production, School of Veterinary Medicine and Animal Sciences, University of S\u00e3o Paulo (USP), Pirassununga 13635-900, SP, 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