A cross-sectional study was conducted from November, 2019 to June, 2020 to assess the occurrence and antimicrobial susceptibility patterns of E. coli O157 and Salmonella in raw cow milk collected from smallholder milk producers, dairy farms, milk collection centers and cafeterias in and around Bahir Dar city, Ethiopia. Samples were collected using a simple random sampling technique and analyzed using the recommended standard procedures to isolate and identify the pathogen. Additionally, a structured questionnaire survey was conducted to assess the status of hygienic practices in the smallholder milk producers, dairy farms, milk collection centers and cafeterias. The data was processed and analyzed by using SPSS version 20.0 software. Descriptive statistical analysis such as percentage, chi-square and fishery exact test of various risk factor and dependent variables. Out of 150 raw milk samples, 70 smallholder milk producers, 29 dairy farms, 35 milk collection centers and 16 cafeterias examined.The occurrence rate E. coli O157 and Salmonella was 9 (6%) and 7(4.7%), respectively. The occurrence of E. coli O157 was highest in smallholder milk producers (7.15). While, lowest in dairy farms (3.4). Similarly, the occurrence of Salmonella was highest in cafeteria (6.3%). While, lowest in dairy farms (3.4%). The antimicrobial susceptibility profile showed that all isolates were 100% susceptible to Gentamicin and Ciprofloxacin, and resistant to Ampicillin. While,11% of E. coli O157 and 42.9 % of Salmonella isolates were found to be multidrug resistance. Physicians within the area should consider Ciprofloxacin and Gentamicin as first choice drugs within the treatment of clinical diseases associated with Escherichia coli O157 and Salmonella. It is recommended training should be provided on hygienic practices for the stakeholders involved in the milk value chain, and therefore the proper pasteurization of milk to maintain the safety and quality of milk for consumers in the area. Keywords: Antimicrobial resistance, Milk, Occurrence, Escherichia coli O157, Salmonella, H ygienic practice