Background: Foodborne illness and death due to foodborne diseases caused by highlyrndangerous pathogens is common in the world; especially it is widespread in developing nations.rnIn Ethiopia, the consumption of meat as a raw is well practiced among the people, which is arnpotential cause of foodborne illness. Cattle are considered as reservoir of pathogens like thernhighly virulent Escherichia coli O157:H7 (E. coli O157:H7). Low standard of meat handling andrnsafety practices, lack of protecting personal hygiene among workers of meat supply chain andrnlack of awareness about E. coli O157:H7-caused infection are major problems in Ethiopia.rnFurthermore, there is paucity of information regarding the epidemiology of the pathogen alongrnthe beef carcass supply chain at Addis Ababa Abattoir Enterprise (AAAE). Therefore, thernpurpose of this study was to isolate E. coli O157: H7 and determine the site of contamination ofrnraw beef meat at AAAE and its carcass supply chain of butcher shops in Yeka sub-city, AddisrnAbaba, Ethiopia rnMethodology: A total of 210 samples from carcass swab, raw minced beef meat and pooledrnenvironmental sources were collected along the supply chain. Collected samples were cultured tornisolate E. coli O157: H7 and identified using biochemical tests and Biolog bacterial identificationrnsystem. Antimicrobial susceptibility tests were performed on E. coli O157: H7 isolates using thernKirby-Bauer disk diffusion method on Muller-Hinton agar. Observation was used to assess thernoverall hygiene and sanitation of the beef-selling environment, and a questionnaire was used tornassess the workers' knowledge, attitude, and practice. Following data entry into an Excelrnspreadsheet, the data was exported to and analyzed using statistical software (SPSS versionrn26.0). Descriptive statistics and the Chi-square (Xrn2rn) test were used, and a p-value (p< 0.05) wasrnconsidered statistically significant. rnResult: The overall prevalence of E. coli O157:H7 was 2/210 (0.95%) and the two isolates wererndetected from knife swab sample at slaughter house and sample from minced raw beef meat atrnbutcher shop. In the case of the antimicrobial susceptibility test, both isolates were susceptible tornciprofloxacin, gentamicin, meropenem, chloramphenicol, amoxicillin clavulanate andrncefotaxime, but resistant to sulfamethoxazole + trimethoprim, amoxicillin, streptomycin andrnampicillin. rnConclusion: Overall, although the prevalence of E. coli O157:H7 was low, the results indicatedrnthat the contamination detected from raw beef meat from butcher shop and the utensil in thernabattoir is a matter of concern that calls for an effort to improve the hygienic status. Hence, tornensure the hygienic quality of meat, everyone involved in carcass slaughter, distribution andrnserving to the customers in the chain should be trained for sanitary and hygienic practices ofrnthese meat supply services including providing basic trainings to the personnel involved in thernindustry.