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This study examined the molecular properties, biofilm formation, virulence genes, and antimicrobial susceptibility of MRSA isolates from mastitis-infected dairy cow milk in South Korea. Whole-genome sequencing of an ST22-SCC\n                      <jats:italic>mec<\/jats:italic>\n                      IV MRSA strain positive for Panton\u2013Valentine leukocidin (PVL) and toxic shock syndrome toxin-1 (TSST-1) was also performed. Of the 488\n                      <jats:italic>S<\/jats:italic>\n                      .\n                      <jats:italic>aureus<\/jats:italic>\n                      isolates, 30 (6.1%) were identified as MRSA, harboring the\n                      <jats:italic>mecA<\/jats:italic>\n                      gene. MRSA exhibited the highest resistance to kanamycin (66.7%) among non-beta-lactam antibiotics. Multidrug resistance was observed in 83.3% of MRSA isolates. All MRSA strains had the capacity to form biofilm and harbored biofilm-related genes. The primary virulence genes included\n                      <jats:italic>hla<\/jats:italic>\n                      ,\n                      <jats:italic>hlb<\/jats:italic>\n                      ,\n                      <jats:italic>lukED<\/jats:italic>\n                      ,\n                      <jats:italic>seg<\/jats:italic>\n                      ,\n                      <jats:italic>sei<\/jats:italic>\n                      ,\n                      <jats:italic>sem<\/jats:italic>\n                      ,\n                      <jats:italic>sen<\/jats:italic>\n                      , and\n                      <jats:italic>seo<\/jats:italic>\n                      . The ST72-t324-SCC\n                      <jats:italic>mec<\/jats:italic>\n                      IV genotype was the most prevalent. Of note, three ST22-SCC\n                      <jats:italic>mec<\/jats:italic>\n                      IV isolates were positive for PVL and TSST-1. Our findings suggest that the majority of MRSA isolates from milk obtained from dairy cows with mastitis are multidrug-resistant, capable of forming robust biofilms, and harbor multiple virulence genes. The presence of PVL- and TSST-1-positive ST22-SCC\n                      <jats:italic>mec<\/jats:italic>\n                      IV isolates in mastitis-infected bovine milk highlights the need for persistent monitoring to mitigate possible public health risks.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      This study reports on the presence and characteristics of methicillin-resistant\n                      <jats:italic>Staphylococcus aureus<\/jats:italic>\n                      strains in milk from mastitis-infected cows. To our knowledge, this is the first report of a Panton\u2013Valentine leukocidin- and toxic shock syndrome toxin-1-positive ST22-SCC\n                      <jats:italic>mec<\/jats:italic>\n                      IV strain in South Korea.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/spectrum.01197-24","type":"journal-article","created":{"date-parts":[[2024,10,22]],"date-time":"2024-10-22T09:05:53Z","timestamp":1729587953000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":12,"title":["Characteristics of methicillin-resistant\n                    <i>Staphylococcus aureus<\/i>\n                    isolates from bovine mastitis milk in South Korea: molecular characteristics, biofilm, virulence, and antimicrobial resistance"],"prefix":"10.1128","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4478-0147","authenticated-orcid":false,"given":"Hye 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