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Background  The spread of  Enterobacteriaceae  producing both carbapenemases and Mcr, encoded by plasmid-mediated colistin resistance genes, has become a serious public health problem worldwide. This study describes three clinical isolates of  Enterobacter cloacae  complex co-harboring  bla  IMP-1  and  mcr-9  that were resistant to carbapenem but susceptible to colistin.    Methods  Thirty-two clinical isolates of  E. cloacae  complex non-susceptible to carbapenems were obtained from patients at 14 hospitals in Japan. Their minimum inhibitory concentrations (MICs) were determined by broth microdilution methods and E-tests. Their entire genomes were sequenced by MiSeq and MinION methods. Multilocus sequence types were determined and a phylogenetic tree constructed by single nucleotide polymorphism (SNP) alignment of whole genome sequencing data.    Results  All 32 isolates showed MICs of ≥2 μg/ml for imipenem and/or meropenem. Whole-genome analysis revealed that all these isolates harbored  bla  IMP-1 , with three also harboring  mcr-9 . These three isolates showed low MICs of 0.125 μg/ml for colistin. In two of these isolates,  bla  IMP-1  and  mcr-9  were present on two separate plasmids, of sizes 62 kb and 280/290 kb, respectively. These two isolates did not possess a  qseBC  gene encoding a two-component system, which is thought to regulate the expression of  mcr-9 . In the third isolate, however, both  bla  IMP-1  and  mcr-9  were present on the chromosome.    Conclusion  The  mcr-9  is silently distributed among carbapenem-resistant  E. cloacae  complex isolates, of which are emerging in hospitals in Japan. To our knowledge, this is the first report of isolates of  E. cloacae  complex harboring both  bla  IMP-1  and  mcr-9  in Japan.

Emergence of carbapenem-resistant and colistin-susceptible Enterobacter cloacae complex co-harboring blaIMP-1 and mcr-9 in Japan