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The present study was conducted to study the antibiogram pattern with special reference to multi drug resistant S. aureus in buffaloes of eastern plain zone of Uttar Pradesh, India. A total of 200 milk samples were collected from the buffaloes of Sultanpur and Faizabad district of U.P. and processed for isolation of Staphylococcus aureus. S. aureus was successfully isolated from 140 milk samples using Mannitol Salt Agar. Out of 140 isolates none of the S. aureus isolates from mastitic milk were 100% sensitive to any antibiotic. Multi Drug Resistance was observed in 100% buffaloes of eastern plain zone for atleast 2 drugs. Highest 133 isolates (95%) were resistant to Cefotaxime, 120 isolates (85.72%) were resistant to Ampicillin, 113 isolates (80.72%) were resistant to Ceftazidime-Tazobactam, 27 isolates (18.58%) showed resistance against Gentamicin and Methicillin, 33 isolates (23.58%) showed resistance against Cloxacillin followed by Tetracycline where 20 isolates (14.28%) showed only resistance. Least resistance (33 isolates, 9.29%) was exhibited for cloxacillin, that proved to be the most effective antibiotic. Among the 140 S. aureus isolated from mastitic milk, 27 isolates (19.28%) exhibited resistance against 2 antibiotics, 20 (14.28%) isolates were resistance to 3 antibiotics, 33 isolates (23.58%) were resistant to 4 antibiotics. 40 (28.57%) isolates were resistant to 5 antibiotics. Seven (5%) isolates each were resistant to 6 and 7 antibiotics. 6 isolates (4.28%) were resistant to maximum 8 antibiotics On antibiogram mapping it was found that, the most effective antibiotic against S. aureus in this region in subclinical mastitis was Ofloxacin followed by Tetracycline, Methicillin and Gentamicin.