{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T07:56:33Z","timestamp":1792223793493,"version":"4.4.0"},"reference-count":105,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2025,6,17]],"date-time":"2025-06-17T00:00:00Z","timestamp":1750118400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100015747","name":"University Grants Commission of Bangladesh","doi-asserted-by":"publisher","award":["UGC-2021-22"],"award-info":[{"award-number":["UGC-2021-22"]}],"id":[{"id":"10.13039\/100015747","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Subclinical mastitis (SCM) poses a significant threat to the global dairy industry, particularly in Bangladesh, where it remains a major constraint in buffalo dairy farming. The rising prevalence of antimicrobial resistant pathogens complicates disease management, resulting reduced milk yield, increased veterinary expenses, compromised animal welfare and potential risk to public health. This study investigated the prevalence and resistance profiles of multidrug-resistant (MDR) and extensively drug-resistant (XDR) <jats:italic>Staphylococcus<\/jats:italic> and <jats:italic>Streptococcus<\/jats:italic> species in raw buffalo milk from SCM cases in Bangladesh. A total of 1,540 quarter milk samples from 385 buffaloes were analyzed, revealing SCM prevalence rates of 67.9% (1046\/1540; 95% CI: 65.6\u201370.3) at the quarter level and 80.8% (311\/385; 95% CI: 76.5\u201384.6) at the animal level. Notable regional variations were observed, with Gowainghat showing the highest prevalence (88.1%; 141\/160). This study did not identify any biologically plausible risk factors for the occurrence of SCM in buffalo. The Modified Whiteside Test and California Mastitis Test confirmed SCM, with culture and biochemical tests identifying 789 (51.2%) <jats:italic>Staphylococcus<\/jats:italic> spp. and 424 (27.5%) <jats:italic>Streptococcus<\/jats:italic> spp. isolates. Polymerase Chain Reaction (PCR) analysis indicated that 72.7% (456\/627) of <jats:italic>Staphylococcus<\/jats:italic> isolates were <jats:italic>Staphylococcus aureus<\/jats:italic>, while the predominant <jats:italic>Streptococcus<\/jats:italic> species included <jats:italic>Streptococcus uberis<\/jats:italic> (32.3%) and <jats:italic>Streptococcus dysgalactiae<\/jats:italic> (14.9%). Resistance gene detection revealed a high prevalence of antimicrobial resistant genes (ARGs), particularly <jats:italic>aac-3(iv)<\/jats:italic> and <jats:italic>tetA,<\/jats:italic> across different buffalo quarters and habitats. Antibiogram profiling demonstrated high susceptibility to tetracycline (80.9; 83.1)\u00a0and trimethoprim-sulfamethoxazole (87.4; 81.9), while significant resistance was noted against ampicillin (88.8; 87.1) and nalidixic acid (68.1; 62.1). MDR was observed in 76.4% (479\/627) of <jats:italic>Staphylococcus<\/jats:italic> spp. and 67.3% (167\/248) of <jats:italic>Streptococcus<\/jats:italic> spp. isolates, with 10.37% (65\/627) and 10.48% (26\/248) classified as possible XDR, respectively. These findings explored high antimicrobial resistance level among <jats:italic>Staphylococcus<\/jats:italic> and <jats:italic>Streptococcus<\/jats:italic> species in subclinical mastitis, highlighting the need for improved management practices and surveillance to mitigate public health risks posed by contaminated milk.<\/jats:p>","DOI":"10.1371\/journal.pone.0324920","type":"journal-article","created":{"date-parts":[[2025,6,17]],"date-time":"2025-06-17T17:41:02Z","timestamp":1750182062000},"page":"e0324920","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":7,"title":["Detection and prevalence of antimicrobial resistance genes in multidrug-resistant and extensively drug-resistant Staphylococcus and Streptococcus species isolated from raw buffalo milk in subclinical mastitis"],"prefix":"10.1371","volume":"20","author":[{"given":"Md. Shahidur Rahman","family":"Chowdhury","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hemayet","family":"Hossain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md. Bashir","family":"Uddin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md. Matiur","family":"Rahman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ferdaus Mohd","family":"Altaf Hossain","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md. Rafiqul","family":"Islam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4861-0030","authenticated-orcid":true,"given":"M. Nazmul","family":"Hoque","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"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2062-8471","authenticated-orcid":true,"given":"Md. Mahfujur","family":"Rahman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,6,17]]},"reference":[{"issue":"2","key":"pone.0324920.ref001","doi-asserted-by":"crossref","first-page":"63","DOI":"10.3329\/sja.v14i2.31246","article-title":"Scopes and opportunities of buffalo farming in Bangladesh: A review","volume":"14","author":"M Hamid","year":"2017","journal-title":"SAARC J Agric"},{"key":"pone.0324920.ref002","article-title":"Subclinical Mastitis of Buffaloes in Asia: Prevalence, Pathogenesis, Risk Factors, Antimicro-bial resistance, and Current Treatment Strategies","author":"MdS Chowdhury","year":"2024","journal-title":"J Anim Sci Technol"},{"issue":"3","key":"pone.0324920.ref003","doi-asserted-by":"crossref","first-page":"305","DOI":"10.3329\/ajmbr.v3i3.34518","article-title":"Dairy buffalo production scenario in Bangladesh: a review","volume":"3","author":"MR Habib","year":"2017","journal-title":"Asian J Med Biol Res"},{"issue":"1","key":"pone.0324920.ref004","doi-asserted-by":"crossref","first-page":"109","DOI":"10.3329\/ralf.v2i1.23042","article-title":"Identification of types of buffaloes available in Kanihari buffalo pocket of Mymensingh district","volume":"2","author":"MMH Sohel","year":"2015","journal-title":"Res Agric Livest Fish"},{"issue":"2","key":"pone.0324920.ref005","doi-asserted-by":"crossref","first-page":"158","DOI":"10.3329\/pa.v29i2.38302","article-title":"Socio-economic status of buffalo farmers and the management practices of buffaloes in plain land of subornachar upazila in Bangladesh","volume":"29","author":"M Rahim","year":"2018","journal-title":"Progress Agric"},{"issue":"1","key":"pone.0324920.ref006","doi-asserted-by":"crossref","first-page":"11704","DOI":"10.1038\/s41598-025-95914-x","article-title":"Emergence of highly virulent multidrug and extensively drug resistant Escherichia coli and Klebsiella pneumoniae in buffalo subclinical mastitis cases","volume":"15","author":"MSR Chowdhury","year":"2025","journal-title":"Sci Rep"},{"key":"pone.0324920.ref007","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1016\/j.rvsc.2021.04.021","article-title":"Global and countrywide prevalence of subclinical and clinical mastitis in dairy cattle and buffaloes by systematic review and meta-analysis","volume":"136","author":"P Krishnamoorthy","year":"2021","journal-title":"Res Vet Sci"},{"issue":"1","key":"pone.0324920.ref008","first-page":"51","article-title":"Prevalence and antibiogram profile of Staphylococcus aureus isolated from milk samples of lactating cows with subclinical mastitis in Gazipur, Bangladesh","volume":"22","author":"S Mostafizur","year":"2018","journal-title":"Ann Bangladesh Agric"},{"issue":"3","key":"pone.0324920.ref009","doi-asserted-by":"crossref","first-page":"1363","DOI":"10.3168\/jds.2011-4503","article-title":"Short communication: Prevalence and risk factors of subclinical mastitis as determined by the California Mastitis Test in water buffaloes (Bubalis bubalis) in Nueva Ecija, Philippines","volume":"95","author":"RT Salvador","year":"2012","journal-title":"J Dairy Sci"},{"issue":"12","key":"pone.0324920.ref010","first-page":"1452","article-title":"Prevalence, economic impact and risk factors associated with mastitis in dairy animals of Punjab","volume":"87","author":"MH Malik","year":"2017","journal-title":"Indian J Anim Sci"},{"issue":"3","key":"pone.0324920.ref011","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1017\/S0022029921000698","article-title":"Occurrence and aetiology of subclinical mastitis in water buffalo in Bangladesh","volume":"88","author":"S Singha","year":"2021","journal-title":"J Dairy Res"},{"key":"pone.0324920.ref012","article-title":"A comprehensive review on subclinical mastitis in dairy animals: Pathogenesis, factors associated, prevalence, economic losses and management strategies","author":"AK Singh","year":"2022","journal-title":"CABI Reviews"},{"issue":"1","key":"pone.0324920.ref013","doi-asserted-by":"crossref","DOI":"10.4142\/jvs.23147","article-title":"Basic concepts, recent advances, and future perspectives in the diagnosis of bovine mastitis","volume":"25","author":"SA Algharib","year":"2024","journal-title":"J Vet Sci"},{"issue":"8","key":"pone.0324920.ref014","doi-asserted-by":"crossref","first-page":"1077","DOI":"10.3390\/antibiotics11081077","article-title":"Environmental Bovine Mastitis Pathogens: Prevalence, Antimicrobial Susceptibility, and Sensitivity to Thymus vulgaris L., Thymus serpyllum L., and Origanum vulgare L. 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