| Summary: | The World Health Organization has identified antibiotic resistance as one of the largest threats to human health and food security. In this study, we compared antibiotic resistance patterns between ESBL-producing <i>Escherichia coli</i> from human clinical diseases and cefotaxime-resistant environmental strains, as well as their potential to be pathogenic. Antibiotic susceptibility was tested amongst clinical isolates (<i>n</i> = 11), hospital wastewater (<i>n</i> = 22), and urban wastewater (<i>n</i> = 36, both influent and treated effluents). Multi-drug resistance predominated (>70%) among hospitalwastewater and urban wastewater influent isolates. Interestingly, isolates from clinical and urban treated effluents showed similar multi-drug resistance rates (~50%). Most hospital wastewater isolates were Phylogroup A, while clinical isolates were predominately B2, with a more diverse phylogroup population in urban wastewater. ESBL characterization of cefotaxime-resistant populations identified <i>bla</i><sub>CTX-M-1</sub> subgroup as the most common, whereby <i>bla</i><sub>KPC</sub> was more associated with ceftazidime and ertapenem resistance. Whole-genome sequencing of a carbapenemase-producing hospital wastewater <i>E. coli</i> strain revealed plasmid-mediated <i>bla</i><sub>KPC-2.</sub> Among cefotaxime-resistant populations, over 60% of clinical and 30% of treated effluent <i>E. coli</i> encoded three or more virulence genes exhibiting a pathogenic potential. Together, the similarity among treated effluent <i>E. coli</i> populations and clinical strains suggest effluents could serve as a reservoir for future multi-drug resistant <i>E. coli</i> clinical infections.
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