The agar disc diffusion test was performed to determine the activity of selected antibiotics against Staphylococcus aureus strains obtained from various sources in Dhaka, Bangladesh. The 80 S. aureus strains used were isolated from 4 categories of sources: milk and food, normal skin microflora of humans, patient samples, and hospital environment swabs; 20 samples were obtained from each category. Ten antibiotics commonly used for various purposes were chosen: penicillin-G, ampicillin, streptomycin, tetracycline, amoxicillin, neomycin, ciprofloxacin, vancomycin, cefaclor, and bacitracin. It was seen that 1.25% (vancomycin) to 82.5% (penicillin) of the S. aureus strains are resistant to the antibiotics used against them. Patient and hospital environment samples exhibited a higher resistance rate than food and normal microflora isolates. Multi-drug resistance, defined as resistance to penicillin and 4 other groups of antibiotics, was found in 35% of the isolates. Of these, three of the hospital environment samples were resistant to at least 8 antibiotics, including one isolate which was vancomycin-resistant. Over 40% of the strains are resistant to 5 or more of the antibiotics tested. 80% of the isolates tested were susceptible to ciprofloxacin, vancomycin, cefaclor and bacitracin.