Wastewater is a reservoir for clinically relevant carbapenemase- and 16s rRNA methylase-producing Enterobacteriaceae

The aim of this study was to evaluate wastewater for carbapenemase-producing Enterobacteriaceae (CPE) and 16S rRNA methylase-producing Gram-negative bacteria (MPB) and to assess their occurrence following wastewater treatment. Wastewater samples were collected between June 2015 and March 2016 in the sewage network of the city of Basel (Switzerland) from sites located before and after influx of wastewater from the hospital into the sewage network. Samples were also obtained from the influent and effluent of the receiving wastewater treatment plant. Samples were screened for CPE and MPB using selective media. Escherichia coli and Klebsiella pneumoniae were typed by multilocus sequence typing (MLST). Carbapenemase and 16S rRNA methylase genes were identified by PCR and sequencing. Resistance profiles were determined by the disk diffusion test and Etest. The occurrence of CPE and MPB was increased downstream of hospital wastewater influx. Of 49 CPE isolates, 9 belonged to OXA-48-producing E. coli clone D:ST38, 7 were OXA-48-producing Citrobacter freundii, and 6 were KPC-2- or OXA-48-producing K. pneumoniae belonging to clonal complex 258. NDM (NDM-1, -5 and -9) and VIM (VIM-1) producers were detected sporadically. MPB included ArmA- and RmtB-producing E. coli and Citrobacter spp. Isolates corresponding to strains from wastewater were detected in the effluent of the treatment plant. Conclusively, CPE and MPB, predominantly OXA-48-producing Enterobacteriaceae, are readily detected in wastewater, survive wastewater treatment and are released into the aquatic environment. OXA-48-producers may represent an emerging threat to public health and environmental integrity.