Background: In general, dealing with healthy food reduces the risk of opportunistic pathogens, and due to the increasing growth in the restaurant industry in Iraq, with the scarcity of previous evaluations in this subject, this study comes at the right time. Objective: This study aimed to assessment the efficacy of training on good hygiene practices (GHP) to reduce potential restaurants contamination by E. coli O157:H7. Methodology: A total of 160 samples were collected from ten different restaurants in Al-Kharkh area, Baghdad, in which 80 samples were taken before implementing GHP and 80 samples after applying GHP. These samples consisted of the following categories: beef burger products before and after grilling, knives, refrigerators, food cutting boards, tables, and workers (hands, nails, and clothes). Results: There was E. coli O157:H7 in 40% (4/10) of restaurants before GHP was implemented. Cultural, biochemical, and molecular-based methods confirmed the presence of E. coli O157:H7 in five of eighty samples (6.25%), including raw beef burger (3/10, 30%), knives (1/10, 10%), and cutting boards (1/10, 10%). There was no evidence of E. coli O157:H7 in 80 samples after GHP training, demonstrating its effectiveness. Conclusion: Poor restaurant hygiene aids spreading E. coli O157:H7. GHP, in addition to personal hygiene, are crucial; otherwise, E. coli O157:H7 strain could contaminate equipment and food.