Diarrheagenic Escherichia coli (DEC) is an important cause of gastrointestinal infections in both humans and animals, and represents a serious public health concern. Understanding the impact of antibiotic resistance profiles of DEC pathotypes on both populations is critical to inform antibacterial strategies and zoonotic prevention strategies. This study was conducted in Muthanna Governorate, Iraq and. aims to make a comprehensive analysis of antibiotic resistance patterns between DEC isolates from human and animal sources.  A total of 475 clinically confirmed diarrheal samples (125 from children and 350 from lambs) were investigated for E. coli isolation, and the recovered isolates were subjected to molecular detection of five major DEC pathotypes: Enteropathogenic E. coli (EPEC), Enterotoxigenic E. coli (ETEC), Enteroaggregative E. coli (EAEC), Enteroinvasive E. coli (EIEC), and Shiga toxin-producing E. coli (STEC). Antimicrobial susceptibility testing was performed on all confirmed DEC isolates using the Vitek 2 system. The results revealed significant differences in antibiotic resistance patterns between human and animal DEC isolates., A striking contrast with ETEC was displayed - while human ETEC isolates were non-MDRs, all animals ETEC isolates were classified as MDR (P = 0.0003). Additionally, aminoglycoside resistance in ETEC was more in animals (100%) than humans (55.6%), although not significant (P = 0.49). EAEC showed comparable MDR circulation between humans (36.4%) and animals (50%), while STEC remained equally non-MDR in both groups. High sensitivity for carbapenems, polymyxins and quinolones seen in both humans and animals isolates was encouraging, although some animal isolates with the emergence of intermediate resistance in isolates. These findings highlight the need for targeted surveillance and antimicrobial stewardship programs that account for the distinct resistance dynamics between human and animal DEC populations to mitigate the spread of antimicrobial resistance.