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Mastitis is a major concern for lactating animals causing mild to severe mammary gland infection. A questionnaire-based cross-sectional survey was conducted in 30 commercial and subsistent farms in Pothohar region of Rawalpindi district, Punjab Pakistan. Where randomly 300 lactating bovine milk samples were tested to California Mastitis Test. Three hundred environmental samples were also collected from the immediate premises surrounding of the udder. Data entry and validation was performed through Epi-Data. Data analysis was performed through SPSS. Chi-square and regression analysis were conducted. For molecular epidemiology sequence analysis was conducted to genetically characterize the isolated bacterial strains. An overall prevalence of subclinical mastitis was 68.6% (cattle and buffalo) on California Mastitis Test (CMT). Several risk factors were identified as associated with the occurrence of subclinical mastitis. Penicillin group was found highly resistant (92.5%) while Quinolone (Moxifloxacin 95%) group was recorded to be the most sensitive (91.7%). On sequence analysis, our isolates (mecA/MRSA/Env/2018/Pak; mecA/MRSA/Bovines/2018/Pak) were found mostly like MRSA strains isolated from Pork samples in China and Humans in India. Our isolates showed unique substitutions in comparison to previously reported MRSA strains from different regions of the world. It can be concluded that the higher prevalence and emerging pattern of mecA gene carrying MRSA strains in farm animals is an alarming problem not only to animals but also for public health. Antibiotic failure in the veterinary field in the treatment of mastitis could be mostly contributed by the higher prevalence of MRSA strains in the field. These results highlight immediate attention to be focused on the control strategies to be implemented to control the spread of these pathogens amongst the animal population and also to the higher risk human population.
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