Objective:The study was conducted to determine the prevalence of Staphylococcus aureus in cockroaches (Periplaneta americana), and to assess the antimicrobial resistance profiles of the isolated bacteria. Materials and methods: A total of 150 cockroaches (P. americana) were randomly captured from three households and four restaurants in Chittagong City Corporation, Bangladesh during July to December 2014. The cockroaches were transported to the bacteriology laboratory at the Poultry Research and Training Centre (PRTC), Chittagong Veterinary and Animal Sciences University. The isolation and identification of Staphylococcus spp. from the external surface wash and gut homogenates by pooling cockroaches were done by following conventional bacteriological examinations followed by biochemical characterization. The antibiotic susceptibility profiles of the isolates were determined using disc diffusion method. Results: In this study, the overall prevalence of S. aureus was 38% (n=57/150). Higher prevalence of Staphylococcus spp. was observed among the cockroaches from restaurant (49.3%; n=37/75) as compared to those of households (26.7%; n=20/75) having a significant difference (P<0.05). Highest level of resistance by the Staphylococcus spp. was found to Penicillin (68%) followed by Erythromycin (60%), Oxacillin (46%) and Clindamycin (31%). On the other hand, the Staphylococci isolates were highly sensitive to Cephalothin (84%) and Kanamycin (65%). Conclusion:The rational use of antibiotics needs to be adopted in both human and animal medicine practices to prevent the emergence of drug resistant Staphylococcus spp.