Staphylococcus aureus is one of the leading causes of foodborne intoxication. The aim of this study was to isolate and identify S. aureus, determine its microbial load in fish and antimicrobial resistance pattern as well as assessing the knowledge, attitude and practices of food handlers along the fish value chain in Bahir Dar City. A total of 165 samples were collected from fish. S. aureus was isolated and identified by culturing on mannitol salt agar and it was subjected for further biochemical tests. From a total of 165 fish samples examined, 55(33.33%) fish samples were found positive for S. aureus in the three species of fish (Oreochromis niloticus, cypernidie and Clarias gariepinus). S. aureus were found to be positive in cooked, under-cooked and uncooked fish samples 14.28%, 18.51% and 41.8% respectively. Relatively higher mean bacterial count was observed in raw fish samples (3.7×106cfu/g) than cooked fish samples (1.6×104cfu/g). Among165 interviewed fish handlers, only 18.8% of the respondents had knowledge about the existence of foodborne pathogens on the skin, mouth and nose of healthy fish handlers and 42.42% respondents does not have the habit of washing hands before and after handling of fish. Of the eight antimicrobials used against S. aureus by disc diffusion method, susceptibility to ciprofloxacin (100%) and gentamycin (94.5%) was recorded, however, the highest resistance was observed in methicillin (87.27%) and streptomycin (72.7%). Taken together, this study revealed the possibility of public health risks posed by S. aureus in Bahir Dar city. Therefore, Fish handlers should wash hands before and after handling of fish and awareness should be created among fish handlers regarding to good hygienic practices before serving