A cross sectional study was conducted between February, 2014 and April, 2014 to isolate and identify Staphylococcus from dairy cattle farms and municipal abattoir; and to evaluate antimicrobial sensitivity for isolates in and around Asella, Ethiopia. An over all of 181 samples were collected and processed from nine dairy cattle farms (87) and seven municipal abattoir visits (94). Accordingly, 42 (23.2%) udder milk, 9 (5.0%) tank milk, 9 (5.0%) polled bucket swab, 9 (5.0%) tank swab, 9 (5.0%) polled hand swab, 9 (5.0%) polled nasal swab, from dairy cattle farms; and 66 (36.5%) meat swab, 7 (3.9%) polled knife swab, 7 (3.9%) polled slaughter line swab, 7 (3.9%) polled hand swab and 7 (3.9%) polled nasal swab from municipal abattoir visits were collected. The result showed the overall proportion of Staphylococcus was 89 (49.2%). Staphylococcal species were more predominant in abattoir 50/94 (53.2%) than farms 39/87 (44.8%), but there was no significant difference between them because p>0.05 at 95% confidence interval. Also high proportion of Staphylococcus was isolated from polled farm nasal swab 8/9 (88.9%), but this difference between sample type and the presence of Staphylococcus is not significant, because p-value (0.303) is greater than 0.05 at 0.05 level. Up on isolation and identification 35 (19.3%), 6 (3.3%), 24 (13.3%), 24 (13.3%) were S. aureus, S. intermedius, S. hyicus and Coagulase Negative Staphylococci (CNS), respectively. From total positive samples, 55 isolates were tested for antimicrobial susceptibility to different 15 antimicrobial discs. The comparative efficacies of antimicrobials used indicates Gentamycin, Kanamycin, Chloramphenicol, Ciprofloxacillin, and Sulphamethoxazole trimethoprim, were the most effective antibiotics where by 94.5%, 89.1%, 81.8%, 81.8%, and 81.8% respectively. Good hygienic practices should be followed both in dairy cattle farms and municipal abattoir including working personnel and equipment's used; and antimicrobials susceptibility test should be carried out at regular intervals to find out the development of resistance against the most commonly applied antibiotics.
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