To investigate the capacities of persistence and dissemination of bla_NDM-5 within Escherichia coli and in aquatic environment, we characterized E. coli (sequence type 636) strains B26 and B28 isolated one month apart from the same urban river in Montpellier, France. The two isolates carried a pTsB26 plasmid, which sized 45,495 Kb, harbored bla_NDM-5 gene and belonged to IncX-3 incompatibility group. pTsB26 was conjugative in vitro at high frequency, it was highly stable after 400 generations and it exerted no fitness cost on its host. bla_NDM-5harboring plasmids are widely dispersed in E. coli all around the world, with no lineage specialization. The genomic comparison between B26 and B28 stated that the two isolates probably originated from the same clone, suggesting the persistence of pTsB26 in an E. coli host in aquatic environment.

Carbapenems are β-lactams antimicrobials presenting a broad activity spectrum and are considered as last-resort antibiotic. Since the 2000s, carbapenemase producing Enterobacterales (CPE) have emerged and are been quickly globally spreading. The global dissemination of carbapenemase encoding genes (CEG) within clinical relevant bacteria is attributed in part to its location onto mobile genetic elements. During the last decade, carbapenemase producing bacteria have been isolated from non-human sources including the aquatic environment. Aquatic ecosystems are particularly impacted by anthropic activities, which conduce to a bidirectional exchange between aquatic environments and human beings and therefore the aquatic environment may constitute a hub for CPE and CEG. More recently, the isolation of autochtonous aquatic bacteria carrying acquired CEG have been reported and suggest that CEG exchange by horizontal gene transfer occurred between allochtonous and autochtonous bacteria. Hence, aquatic environment plays a central role in persistence, dissemination and emergence of CEG both within environmental ecosystem and human beings, and deserves to be studied with particular attention.

Aquatic ecosystems subjected to anthropic pressures are likely hotspots for emergence or dissemination of antimicrobial resistant bacteria. The city of Montpellier is located on a Mediterranean climate watershed that undergoes strong demographic pressures. The aim of the study is to explore antimicrobial resistance, particularly those of clinical concern, in urban rivers flowing in this urban area. The method developed herein to explore antimicrobial resistance is based on cultural and molecular approaches completed by hydrological, hydrogeological, climatic, and physico-chemical data. Hospital vicinity and urbanization density significantly increase cultivable bacterial community, fecal bacteria from human origin, and prevalence of β-lactamases and extended-spectrum β-lactamases encoding-genes without an increase in 16S rDNA gene abundance. A total of 22 multidrug Enterobacterales have been isolated. All Escherichia coli (n = 10) and Klebsiella pneumoniae (n = 6) isolated on a made-house media carried β-lactamases genes, bla_CTX-M being the most prevalent (87%), followed by bla_TEM (56%) and bla_SHV (37%), 56% of these strains carrying two or three of these genes. In urban settings, water quality and infectious risk are generally linked to wastewater treatment plants effluents. This study shows that running waters in urbanized area contribute to the dissemination of antimicrobial resistance, making these environments a reservoir for resistant bacteria with important consideration.