Elsevier

Microbial Pathogenesis

Volume 128, March 2019, Pages 1-6
Microbial Pathogenesis

Co-carrying of KPC-2, NDM-5, CTX-M-3 and CTX-M-65 in three plasmids with serotype O89: H10 Escherichia coli strain belonging to the ST2 clone in China

https://doi.org/10.1016/j.micpath.2018.12.033Get rights and content

Highlights

  • O89: H10 E.coli ZSH6 strain co-carried a large number resistance genes.

  • E.coli ZSH6 strain co-carried three plasmids.

  • Three plasmids were pZSH6-blaKPC-2, pZSH6-blaNDM-5and pZSH6-blaCTX-M.

Abstract

Carbapenem-resistant Enterobacteriaceae strains as a new serious threat for the public health have been increasingly reported worldwide. In this study, one multi-resistant Escherichia coli strain ZSH6 which co-carried blaKPC-2, blaNDM-5 and blaCTX-M, was isolated from human blood sample. By using plasmid conjugation experiments, ZSH6 was found to harbor three plasmids carrying the blaNDM-5 gene, the blaKPC-2 and blaCTX-M gene, respectively. Whole-genome sequencing of ZSH6 yielded 122 scaffolds of chromosomal DNA and three circular plasmids including pZSH6-blaKPC-2 (46,319 bp), pZSH6-blaNDM-5 (46,161bp) and pZSH6-blaCTX-M (184,723). The isolate was classified to Sequence Type 2 and to the O89: H10 serotype. The results of genome analyses revealed that ZSH6 carried three virulence factors (capU, gad and iss) and twenty resistance genes [blaKPC-2 blaNDM-5, blaCTX-M-3, blaCTX-M-65, blaTEM-1, floR, tet(A), tet(B), dfrA17, aadA5, sul1, mdf(A), mph(A), erm(B), aph(3′)-Ia, aph(3′)-Ib, aph(4)-Ia, aph(6)-Id, aac(3)-Iva, aac(3)-IId]. Therefore, the co-existence of such a large number of resistance genes in multiple plasmids making ZSH6 highly resistant to almost all kinds of commonly used antibiotics, and brings a serious challenge for resistance control and clinical treatment of infections caused by this bacterium.

Introduction

It is noteworthy that co-carrying of a large number of antibiotics resistance genes by a single pathogenic bacterial isolate has become a major threat to public health [1,2]. Particularly, coproduction of multiple carbapenemases genes (blaNDM and blaKPC) brings a significant challenge for clinical treatment [[3], [4], [5]]. Even more worryingly, most of those resistance genes are found to be carried by a variety of mobile genetic elements (transposons, integrons, integrative and conjugative elements, genomic islands and plasmids), which will profoundly facilitate the inter- and intra-species dissemination of resistance [6,7].

Escherichia coli, a kind of enterobacteriaceae, common opportunistic pathogen normally causes a wide range of infection diseases such as urinary tract infections and other community-acquired infections [[8], [9], [10], [11]]. Especially, the carbapenemases producing E. coli pose a serious threat to public health. Strikingly, the emergence of carbapenem-resistant enterobacteriaceae has been identified as an urgent threat to human health according to the US Centers for Disease Control and Prevention [12]. In addition to the carbapenem-resistant enterobacteriaceae, the carbapenem resistance genes (blaKPCs, blaNDMs, blaIMPs blaVIMs and blaOXA48 like) and other genes (resistance to cephalosporins, chloramphenicol, chromate, rifampicin and sulphonamides) can also be carried by other bacterial species and making them highly resistant to almost all kinds of antibiotics [[13], [14], [15], [16]]. The plasmids carrying the blaKPC-2 or blaNDM gene have been widely detected in enterobacteriaceae species. In this study, we present the genetic characteristics of one E. coli strain ZSH6 (serotype O89: H10) which harbors three different plasmids co-carrying a large number of resistance genes including blaKPC-2, blaNDM and blaCTX-M.

Section snippets

Bacterial strain, antimicrobial susceptibility testing, resistance genes detection

Escherichia coli isolate ZSH06 was obtained from a blood sample of a 82-year-old man,a long-stay patient, admitted to the intensive care unit of Henan People's Hospital (Henan, China) pulmonary tumor with severe pulmonary infection in April 2018. Routine species identification and antimicrobial susceptibility testing were carried out by the Vitek2 system (BioMérieux, France). The minimal inhibitory concentrations (MICs) of ten antimicrobial agents including (imipenem, meropenem, cefepime,

Results and discussion

E. coli strain ZSH6 isolated from the blood sample showed high resistance to all the tested antimicrobial agents (imipenem, meropenem, cefepime, ceftazidime, cefotaxime, ceftriaxone, piperacillin-tazobactam, ampicillin, gentamicin, amikacin and aztreonam) in this study (Table .2). By using resistance genes detection, E. coli strain ZSH6 co-carried the blaKPC-2, blaNDM-5 and blaCTX-M resistance gene. By using the conjugation experiments, three different kinds of transconjugants, one carried the

Conclusions

We identified a ST2, serotype O89: H10 carbapenem-resistant Escherichia coli co-carrying of the blaKPC-2, blaNDM-5 and other resistance genes including blaCTX-M-65, blaCTX-M-3 in three different plasmid. This type of isolates may pose huge threat to public health and is a serious challenge for the resistance control and treating the infections caused by these bacteria.

Conflicts of interest

The authors declare that they have no competing interests.

Nucleotide sequence accession number

This Whole Genome Shotgun project has been deposited at DDBJ/ENA/GenBank under the accession QVOP00000000.

Funding

This research was funded by the National Natural Science Foundationof China (31500114, 31700111) and by a grant from the Sichuan Province Science and Technology project (2016JY0223), and Southwest Medical University Natural Science Foundation [2018LZXNYD-ZK51] and Health and Family Planning Commission of Sichuan Province grant (16PJ538).

References (35)

  • K. Roohollah et al.

    Antimicrobial resistance and integron gene cassette arrays in commensalEscherichia colifrom human and animal sources in IRI

    Gut Pathog.

    (2016)
  • T.J. Johnson et al.

    Pathogenomics of the virulence plasmids of Escherichia coli

    Microbiology & Molecular Biology Reviews Mmbr

    (2009)
  • J. Sarantuya et al.

    Typical enteroaggregative Escherichia coli is the most prevalent pathotype among E. coli strains causing diarrhea in Mongolian children

    J. Clin. Microbiol.

    (2004)
  • K.E. Holt et al.

    Genomic analysis of diversity, population structure, virulence, and antimicrobial resistance in Klebsiella pneumoniae, an urgent threat to public health

    Proc. Natl. Acad. Sci. U. S. A

    (2015)
  • I. Freire Martin et al.

    Sequence analysis of a CTX-M-1 IncI1 plasmid found in Salmonella 4,5,12:i:-, Escherichia coli and Klebsiella pneumoniae on a UK pig farm

    J. Antimicrob. Chemother.

    (2014)
  • G. Peirano et al.

    Characteristics of NDM-1-producing Escherichia coli isolates that belong to the successful and virulent clone ST131

    Antimicrob. Agents Chemother.

    (2011)
  • Y.S. Pan et al.

    Complete sequence of pEC012, a multidrug-resistant IncI1 ST71 plasmid carrying blaCTX-M-65, rmtB, fosA3, floR, and oqxAB in an avian Escherichia coli ST117 strain

    Front. Microbiol.

    (2016)
  • Cited by (13)

    • Detection of multi-drug resistance and AmpC β-lactamase/extended-spectrum β-lactamase genes in bacterial isolates of loggerhead sea turtles (Caretta caretta) from the Mediterranean Sea

      2021, Marine Pollution Bulletin
      Citation Excerpt :

      Bacteria producing these enzymes appear to have a great ability to cause outbreaks (Dhillon and Clark, 2012). The blaCTX−M-3 variant is an ESBL gene first found in the Citrobacter freundii genome and detected in Europe both in hospital and community settings (Gniadkowski et al., 1998; Dutour et al., 2002; Ibrahimagić et al., 2015) and this variant is capable of causing outbreaks worldwide (Kim et al., 2016) especially when associated with other ESBLs variants (Kim et al., 2005; Fu et al., 2019). Livestock as well as wild species seem to be reservoir for this gene (Tacăo et al., 2012; Jiang et al., 2017; Tark et al., 2017).

    • Nosocomial infection by Klebsiella pneumoniae among neonates: a molecular epidemiological study

      2021, Journal of Hospital Infection
      Citation Excerpt :

      The other is genes encoding carbapenemase (e.g. KPC, NDM, and IMP). In addition, reports of CRKP infections have gradually increased in recent years [5,6]. The first case of hypervirulent Klebsiella pneumoniae (hvKp) infection was reported in Taiwan, China, in the 1980s.

    • High incidence and characteristic of PRRSV and resistant bacterial Co-Infection in pig farms

      2020, Microbial Pathogenesis
      Citation Excerpt :

      Sequence alignment showed that plasmid pScEc7-NDM-5 was 46,161 bp in length and belonged to the IncX3 incompatibility plasmid type. Sequence alignment showed that pScEc7-NDM-5 was similar to the plasmid pEc1929 (Accession no: KT824791), pSCM96-2 (Accession no: CP028718) and pZSH6-blaNDM-5 (Accession no: CM010659), which were harbored by the Enterobacteriaceae bacteria [18,19]. Complete sequence of the pScEc7-CTX was circularized by PCR mapping using plasmid pDA33137-178 (Accession no: CP029580.1), pBTR-CTXM (Accession no: MF156697.2) as the templates.

    View all citing articles on Scopus
    1

    Li Fu, Shanmei Wang and Zhikun Zhang and contributed equally to this work.

    View full text