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Enterobacter cloacae complex (ECC) is among the most common carbapenem-resistant Enterobacteriaceae (CRE) in China. The emergence of mcr has rendered CRE strains resistant to the last-line antibiotic colistin. This study investigated the prevalence of mcr-9 and mcr-10 in carbapenem-resistant ECC (CRECC) and carbapenem-susceptible ECC (CSECC) in China.The CRECC and CSECC strains were collected from different regions of China. Antimicrobial susceptibility tests, conjugation experiments, whole genome sequencing, bioinformatic analysis, and quantitative RT-PCR were performed to understand the mechanisms of resistance and transmission of mcr in ECC.A total of 534 ECC were collected, among which 57 (10.7%) and 23 (4.3%) were positive for mcr-9 and mcr-10, respectively. The prevalence of mcr-9 in CRECC was significantly higher than that in CSECC (31.8% vs. 3.7%; P0.001), while the prevalence of mcr-10 in CRECC was significantly lower (0.8% vs. 5.5%; P0.05). Most mcr-9-positive strains (n = 45, 78.9%) exhibited multidrug-resistant phenotype, and four (17.4%) of the mcr-10-positive strains exhibited multi-drug resistance. Coexistence of mcr and carbapenemase genes was commonly observed, including 41 (71.9%) mcr-9-positive strains and one (4.3%) mcr-10-positive strain, and the possibility of co-transfer was confirmed by conjugation experiments. The mcr-positive ECC were highly diverse, while most mcr genes were plasmid-encoded, indicating the important role of plasmids in the transmission of mcr in ECC. Furthermore, the expression of mcr-9 was increased after induction by colistin.The widespread mcr genes and co-transfer with carbapenemase genes among ECC strains pose an urgent threat to public health.

Azithromycin resistance in bacterial pathogens has increased worldwide, and Klebsiella pneumoniae (K. pneumoniae) carries a variety of azithromycin resistance encoding genes.Genomic DNA of K. pneumoniae strain 16HN-12 was subjected to whole-plasmid sequencing using both the 150-bp paired-end Illumina NextSeq 500 platform and the long-read Oxford Nanopore Technologies MinION platform. Transferability of the azithromycin-resistance plasmid and the virulence plasmid was assessed by performing the conjugation assay.This study identified an IncB/O/K/Z conjugative plasmid that harboured erm(B) and mph(A) genes from a clinical K. pneumoniae strain. The plasmid was readily able to conjugate to Escherichia coli (E. coli) strain J53 and Salmonella enterica subsp. enterica serovar Typhimurium strain PY1 and promoted phenotypic resistance to azithromycin. Furthermore, the virulence plasmid harboured by this K. pneumoniae strain could be conjugated to E. coli strain EC600 and K. pneumoniae strain WZ1-2 via the help of this resistance plasmid through formation of a fusion plasmid. The fusion process was generated by homologous recombination through a homologous region located in both the virulence plasmid and resistance plasmid.Generation of this kind of conjugative plasmid simultaneously carrying virulence and resistance determinants could accelerate dissemination of these determinants and generate bacterial pathogens encoding these phenotypes. These data provide more information about transmission of azithromycin resistance and virulence determinants and call for action to further investigate and prevent such an evolutionary trend.

The continuous emergence of carbapenem-resistant and hypervirulent Klebsiella pneumoniae (CR-hvKP) poses a great challenge to human health owing to the associated extremely high morbidity and mortality. Klebsiella quasipneumoniae is a newly described bacterial species that is often misidentified as K. pneumoniae. Clinical K. quasipneumoniae strains have been reported worldwide, among which multidrug-resistant lineages have become a severe health problem, while less has been understood on this important pathogen. In this study, we characterised three clinical carbapenem-resistant K. quasipneumoniae subsp. similipneumoniae isolates belonging to sequence type 367 (ST367) and capsular type K1 and containing several virulence genes, including salmochelin (iroBCDN), aerobactin (iucABCDiutA) and regulator of mucoid phenotype (rmpA/A2), as well as some resistance genes, including bla

The TEM-1 β-lactamase can only cleave penicillin and the first-generation cephalosporins but it has evolved to become active against second-, third- and fourth-generation drugs. Through sequence analysis of natural TEM variants and those created by mutagenesis experiments, we described two distinct evolution routes of TEM-1 that has generated over 220 enzyme variants. One began with the Gly

This study aimed to characterise the molecular events underlying formation and evolution of a cointegrate plasmid harbouring bla