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This study investigated the prevalence, antibiotic resistance profiles, and virulence traits of MRSA from SCM-affected riverine buffaloes in Jamalpur, Bangladesh. A total of 344 milk samples were screened using the California Mastitis Test (CMT) and Modified Whiteside Test (MWST). Among the milk samples, 46.5% were positive for SCM by CMT. Culture, biochemical tests, and PCR confirmed 73 (21.2%)\n                    <jats:italic>Staphylococcus<\/jats:italic>\n                    spp., of which 30 (41.1%) were identified as\n                    <jats:italic>S. aureus<\/jats:italic>\n                    and 43 (58.9%) as non-aureus staphylococci (NAS). Among the 30\n                    <jats:italic>S. aureus<\/jats:italic>\n                    -positive isolates, 10 (33.3%) were identified as methicillin-resistant\n                    <jats:italic>S. aureus<\/jats:italic>\n                    (MRSA), corresponding to a prevalence of 2.9% among the total milk samples. The MRSA isolates exhibited high multidrug resistance, especially to tetracycline (80%) and cefoxitin (80%), and commonly harbored resistance genes such as\n                    <jats:italic>tetA<\/jats:italic>\n                    (80%),\n                    <jats:italic>aac(3)-iv<\/jats:italic>\n                    (70%), and\n                    <jats:italic>sul1<\/jats:italic>\n                    (50%). Virulence genes\n                    <jats:italic>hla<\/jats:italic>\n                    (66.7%) and\n                    <jats:italic>sea<\/jats:italic>\n                    (50%) were frequently detected, while\n                    <jats:italic>icaA<\/jats:italic>\n                    was found in 23.3% of MRSA. Notably, 60% of MRSA isolates were categorized as XDR based on international standard definitions, while 60% were biofilm producers with high MARI values up to 0.92, indicating severe resistance potential. These findings underscore a significant burden of MDR\/XDR MRSA with virulence potential in buffalo SCM, posing serious risks to animal and public health.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-17476-2","type":"journal-article","created":{"date-parts":[[2025,10,2]],"date-time":"2025-10-02T09:53:11Z","timestamp":1759398791000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Biofilm production and virulence traits among extensively drug-resistant and methicillin-resistant Staphylococcus aureus from buffalo subclinical mastitis in Bangladesh"],"prefix":"10.1038","volume":"15","author":[{"given":"Md Shahidur Rahman","family":"Chowdhury","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Mosharof","family":"Hosen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Hemayet","family":"Hossain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Rafiqul","family":"Islam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Bashir","family":"Uddin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Masudur","family":"Rahman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Mukter","family":"Hossain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Mahfujur","family":"Rahman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,10,2]]},"reference":[{"key":"17476_CR1","doi-asserted-by":"publisher","first-page":"63","DOI":"10.3329\/sja.v14i2.31246","volume":"14","author":"M Hamid","year":"2017","unstructured":"Hamid, M., Siddiky, M., Rahman, M. & Hossain, K. 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