Genomic characterization of carbapenemase-producing Enterobacterales from wastewater reveals the convergence of KPC-2 and GES-16 in Brazil

Abstract

Carbapenemase-producing Enterobacterales (CPE) pose a significant public health concern due to the limited therapeutic options and increasing dissemination outside clinical settings. Wastewater treatment plants (WWTPs) have been proposed as relevant environmental reservoirs for antimicrobial-resistant bacteria and mobile genetic elements. In this study, we performed genomic characterization of CPE strains recovered from raw wastewater samples from the influents of different WWTPs. Antimicrobial susceptibility testing revealed multidrug resistance, including coresistance to carbapenems and polymyxins, among Klebsiella pneumoniae , Enterobacter asburiae , and Enterobacter kobei strains. Whole-genome sequencing identified the convergence of the bla KPC-2 and bla GES-16 genes, as well as the presence of bla GES-5 in E. kobei sequence type (ST) 540 strains. Moreover, the bla KPC-2 gene was detected in K. pneumoniae strains belonging to high-risk clones ST11 (capsular types KL64 and KL15) and ST307 (capsular type KL102), and E. asburiae ST384. The bla GES-5 and bla GES-16 genes were associated with class 1 integrons, while the bla KPC-2 gene was embedded within transposons (Tn 4401a , Tn 4401i , and Tn 3 -like) and insertion sequences (IS Kpn27 and IS Kpn6 ). Notably, genomic analyses and literature review demonstrated that the bla GES-16 gene remains unique to Brazil. The putative pathogenic potential of carbapenem-resistant K. pneumoniae ST11 was also assessed. These findings support the environmental circulation of clinically relevant Enterobacterales genotypes and emphasize the potential role of WWTPs as conduits for CPE dissemination, if not adequately operated. Therefore, genomic surveillance in extra-hospital settings may contribute to a better understanding of antimicrobial resistance ecology and inform One Health mitigation strategies.

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