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Background  Carbapenemases produced by Enterobacterales are often encoded by genes on transferable plasmids and represent a major healthcare problem, especially if the plasmids contain additional antibiotic resistance genes. As part of Dutch national surveillance, 50 medical microbiological laboratories submit their Enterobacterales isolates suspected of carbapenemase production to the National Institute for Public Health and the Environment for characterization. All isolates for which carbapenemase production is confirmed are subjected to next-generation sequencing.    Objectives  To study the molecular characteristics of a genetic cluster of  Enterobacter cloacae  complex isolates collected in Dutch national surveillance in the period 2015–20 in the Netherlands.    Methods  Short- and long-read genome sequencing was used in combination with MLST and pan-genome MLST (pgMLST) analyses. Automated antimicrobial susceptibility testing (AST), the Etest for meropenem and the broth microdilution test for colistin were performed. The carbapenem inactivation method was used to assess carbapenemase production.    Results  pgMLST revealed that nine  E. cloacae  complex isolates from three different hospitals in the Netherlands differed by <20 alleles and grouped in a genetic cluster termed EclCluster-013. Seven isolates were submitted by one hospital in 2016–20. EclCluster-013 isolates produced carbapenemase and were from ST78, a globally disseminated lineage. EclCluster-013 isolates harboured a 316 078 bp IncH12 plasmid carrying the  bla  VIM-1  carbapenemase and the novel  mcr-9  colistin resistance gene along with genes encoding resistance to different antibiotic classes. AST showed that EclCluster-013 isolates were MDR, but susceptible to meropenem (<2 mg/L) and colistin (<2 mg/L).    Conclusions  The EclCluster-013 reported here represents an MDR  E. cloacae  complex ST78 strain containing an IncH12 plasmid carrying both the  bla  VIM-1  carbapenemase and the  mcr-9  colistin resistance gene.

A genetic cluster of MDR Enterobacter cloacae complex ST78 harbouring a plasmid containing blaVIM-1 and mcr-9 in the Netherlands