There is little information about carbapenemase-producing (CP) Klebsiella oxytoca. We characterized CP K. oxytoca isolates collected in some Spanish hospitals from January 2016 to October 2017. During the study period, 139 unduplicated CP K. oxytoca isolates were identified; of them, 80 were studied in detail. Carbapenemases and ESBLs were identified by PCR and sequencing. The genetic relationship was studied by PFGE. WGS was carried out in 12 representative isolates, and WGS data were used to construct a phylogenetic tree, to identify the resistome, and to reconstruct plasmids carrying carbapenemases genes.Forty-eight (60%) isolates produced VIM-1, 30 (37.5%) OXA-48, 3 (3.7%) KPC-2, 2 (2.5%) KPC-3 and one (1.2%) NDM-1; four isolates co-produced two carbapenemases each. By PFGE, 69 patterns were obtained from the 80 CP K. oxytoca; four well-defined clusters were detected: Cluster 1 consisted of 11 OXA-48-producing isolates, the other three clusters consisted of five, three and three VIM-1-producing isolates. In the 12 sequenced isolates, the average number of acquired resistance genes was higher in VIM-1-producing isolates (10.8) than in OXA-48-producing isolates (2.3). All 12 isolates had genes of the blaOXY-2 genotype and by MLST most belonged to the ST2. The plasmid types carrying carbapenemases genes were IncL, IncHI2, IncFII, IncN, IncA/C2 and IncP6.The emergence of CP K. oxytoca in Spain was principally due to the clonal and polyclonal spread of VIM-1 and OXA-48-producing isolates carried in IncL, IncHI2, IncFIB and IncN plasmids. CP K. oxytoca mainly belonged to the ST2 and to the genotype blaOXY-2.
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