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It is becoming increasingly recognized that\n                    <jats:italic>Enterococcus<\/jats:italic>\n                    species cause mastitis in dairy cows. Accurately characterizing the regional epidemiology of enterococcal mastitis, determining its correlations with management variables, and comprehending its effects on udder health all depend on accurate species information. This study investigated the occurrence, antibiotic resistance and virulence factors of\n                    <jats:italic>Enterococcus faecalis<\/jats:italic>\n                    and\n                    <jats:italic>Enterococcus faecium<\/jats:italic>\n                    in cow dung and milk samples from cows with subclinical mastitis. Subclinical mastitis was identified in 39.0% (68\/174) of cows and 27.8% (194\/696) of quarters, based on results from the California Mastitis Test (CMT) and somatic cell counts (SCC), respectively. Matrix-Assisted Laser Desorption\/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) and Polymerase Chain Reaction (PCR) targeting the\n                    <jats:italic>ddl<\/jats:italic>\n                    gene confirmed the predominance of\n                    <jats:italic>E. faecalis<\/jats:italic>\n                    (93%) and\n                    <jats:italic>E. faecium<\/jats:italic>\n                    (6.4%) in milk samples, while cow dung samples yielded only\n                    <jats:italic>E. faecalis<\/jats:italic>\n                    (100%). Notably, among the\n                    <jats:italic>E. faecalis<\/jats:italic>\n                    isolates from milk samples, 17.2% exhibited vancomycin resistance, whereas streptomycin resistance was found in a smaller proportion of isolates (6.8%). All (100%)\n                    <jats:italic>E. faecium<\/jats:italic>\n                    isolates from the same milk samples showed resistance to vancomycin. The findings also revealed that 11 (32.3%) of\n                    <jats:italic>E. faecium<\/jats:italic>\n                    isolates from cow dung were resistant to vancomycin. Multidrug resistance (MDR) was observed in 20.6% of milk and 6.8% of cow dung isolates. The\n                    <jats:italic>vanA<\/jats:italic>\n                    gene was the most prevalent antibiotic resistance gene (ARG), detected in 96% of\n                    <jats:italic>E. faecalis<\/jats:italic>\n                    isolates. Virulence profiling of\n                    <jats:italic>Enterococcus<\/jats:italic>\n                    spp. isolates showed varying gene prevalence in milk (\n                    <jats:italic>asa1<\/jats:italic>\n                    : 33.3%,\n                    <jats:italic>ace<\/jats:italic>\n                    : 12.7%,\n                    <jats:italic>esp<\/jats:italic>\n                    : 10%) and cow dung samples (\n                    <jats:italic>gelE<\/jats:italic>\n                    : 53.2%,\n                    <jats:italic>hyl<\/jats:italic>\n                    : 38.2%). This study has indicated a significant occurrence of antimicrobial-resistant\n                    <jats:italic>E. faecalis<\/jats:italic>\n                    and\n                    <jats:italic>E. faecium<\/jats:italic>\n                    strains obtained from subclinical cattle mastitis. These findings emphasize the role of\n                    <jats:italic>Enterococcus<\/jats:italic>\n                    spp., especially vancomycin-resistant strains, as emerging threats in bovine subclinical mastitis, with possible implications for zoonotic transmission and antimicrobial stewardship in dairy systems.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-32569-8","type":"journal-article","created":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T19:03:56Z","timestamp":1766084636000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Antimicrobial resistance and virulence gene profiles of Enterococcus faecalis and Enterococcus faecium isolated from subclinical bovine mastitis milk and cow dung"],"prefix":"10.1038","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6238-1695","authenticated-orcid":false,"given":"Ntelekwane George","family":"Khasapane","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kgaugelo","family":"Lekota","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mofokeng","family":"Lehlohonolo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jane","family":"Nkhebenyane","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oriel","family":"Thekisoe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsepo","family":"Ramatla","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,18]]},"reference":[{"issue":"3","key":"32569_CR1","doi-asserted-by":"publisher","first-page":"266","DOI":"10.22034\/ijab.v12i3.2304","volume":"12","author":"SM Al-Haider","year":"2024","unstructured":"Al-Haider, S. M., Al-Jubouri, M. O., Imran, S. M. & Marza, A. D. Novel isolation of Enterococcus spp. From common carp, Cyprinus Carpio in iraq: antibiotic resistance and susceptibility to syzygium aromaticum. Int. J. Aquat. Biol. 12 (3), 266\u2013274. https:\/\/doi.org\/10.22034\/ijab.v12i3.2304 (2024).","journal-title":"Int. J. Aquat. Biol."},{"issue":"1","key":"32569_CR2","doi-asserted-by":"publisher","first-page":"3","DOI":"10.3390\/antibiotics12010003","volume":"12","author":"Z Kova\u010devi\u0107","year":"2022","unstructured":"Kova\u010devi\u0107, Z. et al. Is there a relationship between antimicrobial use and antibiotic resistance of the most common mastitis pathogens in dairy cows? Antibiotics 12 (1), 3. https:\/\/doi.org\/10.3390\/antibiotics12010003 (2022).","journal-title":"Antibiotics"},{"issue":"5","key":"32569_CR3","doi-asserted-by":"publisher","first-page":"333","DOI":"10.4236\/aim.2022.125024","volume":"12","author":"HM Abdi","year":"2022","unstructured":"Abdi, H. M., Hassan, M. 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