| Summary: | Transmission of pathogenic microorganisms in the last decades has been considered a significant health hazard and pathogenic <i>E. coli</i>, particularly antibiotic-resistant strains, have long been identified as a zoonotic problem. This study aimed to investigate multidrug resistant pathogenic <i>E. coli</i> isolates from wild birds, chickens, and environment in selected Orang Asli and Malay villages in Peninsular Malaysia. The bacteriological culture-based technique, disc diffusion method, and multiplex Polymerase Chain Reaction (mPCR) assay was used to determine the occurrence of pathogenic <i>E. coli</i> strains in the several samples in the study. <i>E. coli</i> isolates showed a variety of multi-drug resistant (MDR) antibiotypes and Enteropathogenic <i>E. coli</i> (EPEC) and Enteroinvasive <i>E. coli</i> (EIEC) were the most predominantly identified pathogenic <i>E. coli</i> strains. The findings of this study demonstrated the significance of animal reservoirs and the environment as sources of pathogenic <i>E. coli,</i> resistant bacteria, and resistance genes. Hence, there is a need for adoption of a practical surveillance approach on MDR pathogens to control foodborne contamination.
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