Staphylococcus aureus is associated with poor food handling and sanitation practices responsible for notorious for causing human diseases. Using a cross-sectional study, it was isolated along Asella Municipal Beef Abattoir line and assessed for antimicrobial resistance profiles. A total of 470 samples consisting of 400 beef carcasses swab and 70 environmental (personal hand, apron, knife, splitting axil, hooks and meat wrapping plastic swab samples and cleaning water) samples were collected. Randomly selected isolates from each location were tested for 12 different antimicrobial agent using disk diffusion techniques. An overall 171 (36.4%) samples for which 137 (34.3%) from the carcass swab and 34 (48.6%) from Abattoir facilities were positive for S. aureus. The isolation was ranged from 30% in clean water sample to 60% each from personal hand swab and Hook swab sample. Although resistance was not observed for bacitracin, chloramphenicol, kanamycin and sulfamethozathole, the resistance of 77.5%, 45.0%, 36.3%, 28.8%, 26.3%, 21.3%, 20.0% and 7.5% to amoxicillin, cloxacillin, ampicillin, penicillin-G, vancomycin, erythromycin, tetracycline and doxycycline were observed in descending order. One, two, three, four and five drugs resistant isolates were observed in 14.1%, 33.3%, 29.5%, 19.2% and 3.8% of the resistant isolates. Resistant isolate to amoxicillin, cloxacillin, ampicillin and penicillin-G, were frequently observed along the majorities of sampling location. The findings indicate the risk of public in acquiring drug resistant, staphylococcal infections enter food poisoning. Implementing line based hygienic operation during beef carcass processing and transporting could minimize carcass contamination with S. aureus including the resistant one.