Evidence-based identification and characterization of methicillin-resistant Staphylococcus aureus isolated from subclinical mastitis in dairy buffaloes of Pakistan.

Background: Methicillin-resistant Staphylococcus aureus (MRSA), affecting livestock and human beings, has become a global public health hazard with economic consequences. Aims: The current study was designed to investigate the prevailing MRSAassociated subclinical mastitis and associated risk factors in dairy buffaloes. The study also highlighted the genetic variations and in silico-based proteomic differences among MRSA isolates. Methods: Out of 516 milk samples, 45.93% (237/516) were found positive for subclinical mastitis, while the prevalence of S. aureus was recorded 56.12%. The methicillin resistance in S. aureus isolates was evaluated by oxacillin disc diffusion test and molecular identification of the mecA gene. Results: The results revealed a phenotypic and molecular prevalence of MRSA at 45.11% and 18.79%, respectively. The risk factor analysis revealed that among various assumed risk factors, parity, milking hygiene, milker care during milking, milk yield, housing system, and floor type were significantly associated with subclinical mastitis in buffaloes. The sequencing and phylogenetic analysis showed no significant genetic variations among study isolates and depicted a high similarity with isolates from Africa, USA, India, Italy, Turkey, and Iran. The in-silico protein analysis showed that all sequences had the same protein motifs resembling penicillin protein 2a except Buff-13, whose protein structure resembles alpha-catenin-like protein hmp-1. Conclusion: The current study was the first report of the genotypic characterization and in silico protein analysis of MRSA from dairy buffaloes in Pakistan. The result highlighted the importance of antimicrobial resistance (AMR) and development of control strategies against MRSA infections. [ABSTRACT FROM AUTHOR]