The most significant morphological, biochemical, and antimicrobial susceptibility traits of typical E. coli bacteria were shown by the study. Swab samples were collected from the colons of 20 fowl from various veterinary clinics in Babylon in December 2023, following the observation of clinical indications, selective growth on EMB and MacConkey agar, and positive biochemical reactions like the indole test and TSI agar fermentation made it easier to identify the bacteria. The investigation gave a thorough characteriszation of Escherichia coli, confirming that isolates demonstrated resistance to widely prescribed antibiotics, emphasising the necessity of judicious antibiotic use and heightened surveillance.