Abstract
The purpose of this work was to study extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (ESBL-EC) in freshwaters, hospital effluents, and wastewaters during two sampling campaigns in 2021. Water sampling was performed at 24 stations in the Ourthe watershed in Belgium. A total of 644 ESBL (n=642) and AmpC (n=2) E. coli strains were isolated. Disk-diffusion assays were performed following the EUCAST's recommendations. All strains were tested for the presence of blaCTX-M-1, blaCTX-M-2, and blaCTX-M-9 gene groups by PCR. Genes belonging to blaCTX-M-1 and blaCTX-M-9 groups were detected, respectively, in 73.6% and 14.9% of the strains. No blaCTX-M-2 group's gene was found. A subset of strains (n=40) was selected for whole genome sequencing. Escherichia coli serotype O18: H7 ST 1463 was predominant (n=14) in the sequenced strains and showed pathogenicity in the Galleria mellonella larvae model. β-lactamase genes identified were blaCTX-M (n=21), with blaCTX-M-15 mostly represented (n=15), as well as blaTEM (n=11), blaOXA (n=7), blaSHV (n=9), and carbapenemase (CP) genes were observed in several strains-blaKPC-3 (n=19), blaNDM-1 (n=1), blaVIM-1 (n=2), and blaOXA-244 (n=2)-even from freshwaters. ESBL-EC are widely distributed in the aquatic environment in Belgium and contain a variety of ESBL and CP genes.
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