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Abstract Class I integrons have garnered significant attention due to pivotal roles in the dissemination of antimicrobial resistance genes (ARGs), which impose risks to public health and food safety. Here, the prevalence and characteristics of class I integrons in Escherichia coli isolates derived from food-producing animals and human patients were assessed. Of 721 E. coli isolates collected from human patients (113), pigs (298), and poultry (310), 93 (12.90%) carried the class I integrase gene (intI1). Multilocus sequence typing identified 39 sequence types from 93 intI1-postive isolates, including three novel types. Sequence analysis revealed that 59 classical class I integrons encompassed six distinct gene cassettes arrangements [dfrA17-aadA5, dfrA12-aadA2, dfrA1-aadA1, dfrA7, aac(6’)-Ib, and aadA1-aac(3)-VIa]. Six insertion sequences (IS1, IS6, IS21, IS91, IS110, and IS256) and one transposon (Tn3) were harbored in proximity to the integrons. A comparison with sequences retrieved from the National Center for Biotechnology Information database demonstrated that E. coli isolates with integron sequences were detected in various food-producing animals and human hosts in environmental niches across Asia, Europe, and North America. These findings indicate the potential risk of ARG transmission between food-producing animals and humans by bacteria populations and provide useful baseline data for monitoring of ARGs.