| Summary: | <i>S. aureus</i>, particularly methicillin-resistant <i>S. aureus</i>, has been recognized as a main cause of bovine mastitis and food poisoning. This study investigated the prevalence, antibiotic resistance, and phage-based biocontrol of <i>S. aureus</i> and methicillin-resistant <i>S. aureus</i> isolated from raw milk of cows with subclinical mastitis. The results showed that the prevalence of <i>S. aureus</i> and methicillin-resistant <i>S. aureus</i> was 12% (48/400) and 1.5% (6/400), respectively. The <i>S. aureus</i> isolates were highly resistant to penicillin (72.92%), erythromycin (43.75%), and tetracycline (39.58%). Out of 48 <i>S. aureus</i> isolates, 6 were identified as methicillin-resistant strains. Among them, one isolate was found to harbor the <i>sea</i> gene. A total of 5 phages were recovered from 50 pork and 50 chicken meat samples, 1 from pork and 4 from chicken meat samples. Phage PSA2 capable of lysing all 6 methicillin-resistant isolates was selected for characterization. The use of phage PSA2 completely inactivated methicillin-resistant <i>S. aureus</i> SA33 in raw milk at both 24 °C and 4 °C, indicating its potential as a promising antibacterial agent in controlling methicillin-resistant <i>S. aureus</i> in raw milk and treating bovine mastitis.
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