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The study was carried out in 1299 lactating buffaloes to determine prevalence and their antimicrobial sensitivity of causative organisms of mastitis in Haryana state, India. Out of 4452 quarter milk samples tested, 1503 (33.76%) and 809 (18.17%) cases were found positive for subclinical mastitis and clinical mastitis, respectively. Majority of clinical cases of mastitis were of chronic nature, occurring during first two parities and first five months of lactation. Staphylococci (51.16%) were the most prevalent organism followed by Streptococci (37.94%), E. coli (8.41%) and Corynebacterium pyogenes (1.62%). All the organisms except E. coli revealed high sensitivity against Ceftriaxone and Cefaperazone. E. coli isolates showed multiple resistance patterns. Regular monitoring of udder health and determination of antimicrobial sensitivity of causative agents prior to initiation of mastitis treatment is suggested.