First Report of NDM and VIM Coproducing Klebsiella pneumoniae in Tunisia and Emergence of Novel Clones.

Metallo-β-lactamase (MBL) producing bacteria constitute nowadays a serious global concern worldwide. The purpose of our present study was to characterize molecular features of MBL producing bacteria and to identify the existing clones in our area. Thirteen MBL-producing-Klebsiella pneumoniae were detected in clinical samples from patients hospitalized in the Military hospital of Tunisia during 2017. The molecular research by polymerase chain reaction and sequencing of gene encoding MBL enzymes showed that only two types were identified in our study: blaNDM-1 and blaVIM-1 genes detected, respectively, in eight and six isolates. An association between these two MBL genes (blaNDM-1+blaVIM-1) has been observed in one of our isolates. Other β-lactamase types (CTXM-15/4 isolates; SHV/2 isolates; OXA-48/3 isolates) were detected in association with New Delhi metallo-beta-lactamase (NDM) and/or Verona Integron-Mediated Metallo-β-lactamase (VIM) enzymes. Furthermore, these isolates were resistant to other antimicrobial agents, including gentamicin [aac(3)-II/11 isolates], tetracycline (tetB or tetA/2 isolates), chloramphenicol (cmlA and/or floR/3 isolates), streptomycin (aadA/5 isolates), and sulfonamides (sul1 or sul2 or sul3/4isolates). The Multilocus Sequence Typing revealed 10 different Sequence types (ST) of which 7 novel ST: ST147 (3 isolates), ST101 (1 isolate), ST630 (1 isolate), ST3485 (1 isolate), ST3486 (1 isolate), ST3487 (1 isolate), ST3488 (1 isolate), ST3489 (1 isolate), ST3490 (1 isolate), ST3491 (2 isolates). Our study provides new data about MBL producing K. pneumoniae in Tunisia. Thus, we report for the first time the coexpression of blaNDM-1 and blaVIM-1 in our country and also we describe seven novel ST of MBL producing K. pneumoniae in the world.

[1]  R. Kaneva,et al.  Multicentre investigation of carbapenemase-producing Klebsiella pneumoniae and Escherichia coli in Bulgarian hospitals - Interregional spread of ST11 NDM-1-producing K. pneumoniae. , 2019, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[2]  C. Torres,et al.  Metallo-β-lactamases and class D carbapenemases in south-east Tunisia: Implication of mobile genetic elements in their dissemination. , 2018, International journal of antimicrobial agents.

[3]  V. Cheng,et al.  Occurrence of Highly Conjugative IncX3 Epidemic Plasmid Carrying blaNDM in Enterobacteriaceae Isolates in Geographically Widespread Areas , 2018, Front. Microbiol..

[4]  A. Tsakris,et al.  Hospital outbreak due to a Klebsiella pneumoniae ST147 clonal strain co-producing KPC-2 and VIM-1 carbapenemases in a tertiary teaching hospital in Northern Greece. , 2018, International journal of antimicrobial agents.

[5]  R. Bonomo,et al.  Carbapenemase-Producing Organisms : A Global Scourge ! , 2018 .

[6]  N. Klibi,et al.  Occurrence and Characterization of Carbapenemase-Producing Enterobacteriaceae in a Tunisian Hospital. , 2018, Microbial drug resistance.

[7]  A. Slim,et al.  An Outbreak of NDM-1-Producing Klebsiella pneumoniae, Associated with OmpK35 and OmpK36 Porin Loss in Tunisia. , 2018, Microbial drug resistance.

[8]  E. Petinaki,et al.  Emergence of sequence type 11 Klebsiella pneumoniae coproducing NDM-1 and VIM-1 metallo-β-lactamases in a Greek hospital. , 2017, Diagnostic microbiology and infectious disease.

[9]  P. François,et al.  Emergence of blaNDM-7–Producing Enterobacteriaceae in Gabon, 2016 , 2017, Emerging infectious diseases.

[10]  Dayang Zou,et al.  Diversity of New Delhi metallo-beta-lactamase-producing bacteria in China. , 2017, International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases.

[11]  T. W. Boo,et al.  An Irish outbreak of New Delhi metallo-β-lactamase (NDM)-1 carbapenemase-producing Enterobacteriaceae: increasing but unrecognized prevalence. , 2016, The Journal of hospital infection.

[12]  G. Zarfel,et al.  Molecular characterization of class b carbapenemases in advanced stage of dissemination and emergence of class d carbapenemases in Enterobacteriaceae from Croatia. , 2016, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[13]  Robert A Bonomo,et al.  B1-Metallo-β-Lactamases: Where Do We Stand? , 2016, Current drug targets.

[14]  E. Roilides,et al.  Molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae in Greece. , 2016, Future microbiology.

[15]  P. Nordmann,et al.  The difficult-to-control spread of carbapenemase producers among Enterobacteriaceae worldwide. , 2014, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[16]  M. Timinouni,et al.  First report of a Klebsiella pneumoniae strain coproducing NDM‐1, VIM‐1 and OXA‐48 carbapenemases isolated in Morocco , 2013, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[17]  T. Palzkill.  Metallo‐β‐lactamase structure and function , 2013, Annals of the New York Academy of Sciences.

[18]  S. Hong,et al.  Multiplex PCR for Rapid Detection of Genes Encoding Class A Carbapenemases , 2012, Annals of laboratory medicine.

[19]  T. Walsh,et al.  VIM and IMP metallo‐β‐lactamases and other extended‐spectrum β‐lactamases in Escherichia coli and Klebsiella pneumoniae from environmental samples in a Tunisian hospital , 2011, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[20]  N. Woodford,et al.  Multiplex PCR for rapid detection of genes encoding acquired metallo-beta-lactamases. , 2006, The Journal of antimicrobial chemotherapy.

[21]  L. Dijkshoorn,et al.  Distribution and in-vitro transfer of tetracycline resistance determinants in clinical and aquatic Acinetobacter strains. , 2000, Journal of medical microbiology.

[22]  G. Paul,et al.  Nucleotide sequences of the genes coding for the TEM-like beta-lactamases IRT-1 and IRT-2 (formerly called TRI-1 and TRI-2). , 1994, FEMS microbiology letters.

[23]  W. Naija,et al.  ST147 NDM-1-producing Klebsiella pneumoniae spread in two Tunisian hospitals , 2017, The Journal of antimicrobial chemotherapy.

[24]  T. Walsh.  The emergence and implications of metallo-β-lactamases in Gram-negative bacteria , 2005 .