IncN1 ST7 Epidemic Plasmid Carrying blaIMP-4 in One ST85-Type Klebsiella oxytoca Clinical Isolate with Porin Deficiency.

PURPOSE: Klebsiella oxytoca is an opportunistic pathogen causing nosocomial infections. This study was designed to characterize the genomic features of a carbapenem-resistant K. oxytoca strain and analyze its molecular characteristics. MATERIALS AND METHODS: The strain wzx-IMP was isolated from the blood of a 2-year-old girl diagnosed with acute myeloid leukemia-M7. Species identification was performed, and the minimal inhibitory concentration of the strain was measured. Multilocus sequence typing was performed to identify the subtypes of K. oxytoca. The transfer capacity of the blaIMP-4-harboring plasmid was investigated by conjugation experiments, and the genome characteristics of the strain were examined using whole-genome sequencing. RESULTS: wzx-IMP belongs to the ST85 type and is resistant to imipenem and meropenem, which harbored the blaIMP-4 gene. The blaIMP-4 gene was located in an IS26-associated class 1 integron of pwzx_IMP, which contains conserved IncN1-type backbone regions with a replication gene and its accessory structure for plasmid replication. The blaIMP-4-carrying plasmid in wzx-IMP was successfully transferred to Escherichia coli EC600 by conjugation. Whole-genome sequencing showed that the wzx-IMP isolate included the blaOXY-1-1 gene, accompanied by OmpK36 absence. CONCLUSION: We report an ST85-type carbapenem-resistant K. oxytoca strain, which produces blaIMP-4 located in an IncN1-type plasmid and accompanied by OmpK36 porin deficiency.

The Formation of Two Hybrid Plasmids Mediated by IS26 and Tn6952 in Salmonella enterica Serotype Enteritidis.

To characterize the formation mechanism and characteristics of two cointegrate plasmids in Salmonella enterica serotype Enteritidis strain S13, plasmids from strain S13 and three corresponding transconjugants were subjected to whole genome sequencing and analyzed using bioinformatics tools. The traits of two fusion plasmids in transconjugants were characterized by stability and conjugation experiments. Sequence analysis indicated that strain S13 contained four plasmids, including mcr-1-bearing pS13-1, bla CTX-M-55-carrying pS13-2, tet(M)-bearing pS13-3, and floR-carrying pS13-4. IncN1-F33:A-:B- plasmid pS13-2, respectively, fused with IncFI:A-:B- plasmid pS13-3 and IncX1 plasmid pS13-4, which generated two cointegrate plasmids, designated pS13D and pS13F, which involved in two intermolecular replicative mechanisms mediated by IS26 and the novel transposon Tn6952 (ΔTnAS3-IS26-ΔISEcp1-ramA-ΔIS26-ΔTnAS1), respectively. This is the first report of the fusion of the IncN1-F33:A-:B- plasmid and IncFI:A-:B- plasmid mediated by IS26, and with IncX1 plasmid mediated by Tn6952. The formation and evolution of cointegrate plasmids could expand the resistance and host spectrum of fusion plasmids.

Identification of blaVIM-1 Gene in ST307 and ST661 Klebsiella pneumoniae Clones in Italy: Old Acquaintances for New Combinations.

In the past years, the Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae sequence type 258 (ST258) became an important worldwide spread nosocomial pathogen. Recent evidence shows that the global epidemiology is changing, with the rise of new lineages. In this study we report the microbiological and genomic features of two VIM-1-producing K. pneumoniae isolates belonging to the emerging ST307. Two extensively drug-resistant K. pneumoniae strains, collected between May and June 2017, were confirmed as blaVIM positive by GeneXpert system. The whole-genome sequencing revealed that both KpV_S_1 and KpV_S_2 isolates harbored blaVIM-1 and blaCTX-M-15 genes, besides qnrS1 and qnrB1, strB, mphA, tetR, and tetA determinants. KpV_S_1 and KpV_S_2 isolates belonged to ST661 and ST307, respectively. Both STs have been recently reported as responsible of outbreaks in several European countries. The detection of blaVIM-1 gene in nonpredominant K. pneumoniae clones in a hospital setting should alert on the changing of the epidemiological situation in Italy, usually endemic reservoir of KPC enzyme.

Structural genomics of pNDM-BTR harboring In191 and Tn6360, and other bla NDM-carrying IncN1 plasmids.

AIM: To characterize a conjugative bla NDM-1-carrying plasmid pNDM-BTR from a clinical Escherichia coli isolate. MATERIALS & METHODS: The complete nucleotide sequence of pNDM-BTR was determined using next-generation sequencing technology. Comparative genomic analysis of bla NDM-carrying IncN1 plasmids, including pNDM-BTR, was performed, and the antimicrobial resistance phenotypes were determined. RESULTS: pNDM-BTR contained three accessory modules, namely IS26, a novel Tn3-family transposon Tn6360 and the dfrA14 region composed of In191, ecoRII-ecoRIImet and ΔIS1X2. The relatively small IncN1 backbones could integrate massive accessory modules, most of which were integrated at two 'hotspots'. These IncN1 plasmids contained distinct profiles of accessory modules, which included those carrying various resistance genes. CONCLUSION: This study provides a deeper insight into horizontal transfer of resistance genes among IncN1 plasmids.