Emergence of two novel tmexCD-toprJ subtypes mediating tigecycline resistance in the megaplasmids from Pseudomonas putida.

The widespread antimicrobial resistance (AMR) problem poses a serious health threat, leaving few drug choices, including tigecycline, to treat multidrug resistance pathogens. However, a plasmid-borne tigecycline resistance gene cluster, tmexCD1-toprJ1, emerged and conferred tigecycline resistance. In this study, we identified two novel subtypes, tmexCD2.3-toprJ2.3 and tmexCD2.4-toprJ1b, obtained from three chicken-origin Pseudomonas putida isolates. Two types of megaplasmids were found as the vital vehicle of these tmexCD-toprJ variants. Phylogenetic and genomic analysis indicated the two variants were mainly distributed in Pseudomonas and acted as an evolved intermediated state precursor of tmexCD2-toprJ2. Further gene cloning assay revealed both the expression of tmexCD2.3-toprJ2.3 and tmexCD2.4-toprJ1b could confer multiple antimicrobial resistance, mediating 8- to 16-fold increase of tigecycline MIC. Importantly, two key nucleotide differences in promoter region influence the promoter activity between PtmexC2.3 and PtmexC2.4, while the downregulation effect of TNfxB on the transcriptional expression level of tmexCD2.3-toprJ2.3 and tmexCD2.4-toprJ1b were observed. The emergency of two novel tmexCD-toprJ variants necessitates preventive measures to curb their spread and highlights concerns about more emerging tmexCD-toprJ variants.