Detection of an IncA/C plasmid encoding VIM-4 and CMY-4 β-lactamases in Klebsiella oxytoca and Citrobacter koseri from an inpatient in a cardiac rehabilitation unit.

A 62-year-old patient was transferred to the cardiac rehabilitation unit of the I.R.C.C.S. Fondazione S. Maugeri after undergoing a heart transplantation at the Acute Care Hospital I.R.C.C.S. S. Matteo of Pavia. On 1 August 2013 and during hospitalization in the rehabilitation unit, Klebsiella oxytoca and Citrobacter koseri clinical isolates were simultaneously recovered from the patient's preputial swab. Both the K. oxytoca and C. koseri strains were carbapenem- resistant by MicroScan System (Beckman Coulter). Carbapenem-resistant K. pneumoniae had previously been reported in the same rehabilitation facility. The aim of the study was to identify the carbapenem resistance mechanisms among the enterobacterial species recovered. Phenotypic screening tests useful to detect the β-lactamases/carbapenemases were performed. Carbapenem MICs were obtained by Etest. AmpC and MBL encoding genes were identified by PCR and sequencing. Conjugation assays and plasmid characterization were performed. Both of the K. oxytoca and C. koseri isolates were multi drug resistant, showing resistance to amoxicillin-clavulanic acid, three generation cephalosporins, ertapenem (K. oxytoca MIC, >32 mg/L; C. koseri MIC, 4 mg/L), imipenem (K. oxytoca MIC, 4 mg/L; C. koseri MIC, 12 mg/L), thrimethoprim sulphamethoxazole and gentamicin. Susceptibility was retained to fluoroquinolones, colistin and tigecycline. Molecular characterization confirmed the co-presence of blaCMY-4 and blaVIM-4 determinants in a 150 Kb transferable plasmid of IncA/C group. This case is the first detection in Italy of the K. oxytoca and C. koseri clinical isolates co-producing the CMY-4 and VIM-4 enzymes.

[1]  A. Piazza,et al.  Emergence of a VIM-1 MBL and CTX-M-15 ESbL-producing Klebsiella pneumoniae clone from acute and rehabilitation hospitals in Italy. , 2013, The new microbiologica.

[2]  A. Pantosti,et al.  Epidemic diffusion of KPC carbapenemase-producing Klebsiella pneumoniae in Italy: results of the first countrywide survey, 15 May to 30 June 2011. , 2013, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[3]  S. Bracco,et al.  Multifocal diffusion of a KPC-3 producing ST512 K. pneumoniae clone in Northern Italy. , 2013, The new microbiologica.

[4]  F. Tryposkiadis,et al.  Detection of Citrobacter koseri carrying beta-lactamase KPC-2 in a hospitalised patient, Greece, July 2011. , 2011, Euro surveillance : bulletin Europeen sur les maladies transmissibles = European communicable disease bulletin.

[5]  S. Pournaras,et al.  Dissemination of Clinical Isolates of Klebsiella oxytoca Harboring CMY-31, VIM-1, and a New OXY-2-Type Variant in the Community , 2011, Antimicrobial Agents and Chemotherapy.

[6]  K. Miyamoto,et al.  Laboratory Surveillance for Prospective Plasmid-Mediated AmpC β-Lactamases in the Kinki Region of Japan , 2010, Journal of Clinical Microbiology.

[7]  W. Melchers,et al.  Prevalence, Molecular Characterization, and Phenotypic Confirmation of Extended-Spectrum Beta-Lactamases in Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca at the Radboud University Nijmegen Medical Centre in The Netherlands , 2017 .

[8]  Alessandra Carattoli,et al.  Resistance Plasmid Families in Enterobacteriaceae , 2009, Antimicrobial Agents and Chemotherapy.

[9]  M. Castanheira,et al.  Emergence of a Plasmid Mediated blaVIM-1 in Citrobacter koseri: Report from the SENTRY Antimicrobial Surveillance Program (Italy) , 2009, Journal of chemotherapy.

[10]  G. Rossolini,et al.  First Countrywide Survey of Acquired Metallo-β-Lactamases in Gram-Negative Pathogens in Italy , 2008, Antimicrobial Agents and Chemotherapy.

[11]  A. Gales,et al.  Outbreak of OXY-2-Producing Klebsiella oxytoca in a Renal Transplant Unit , 2008, Journal of Clinical Microbiology.

[12]  J. L. Muñoz Bellido,et al.  Prevalence of clinical isolates of Escherichia coli and Klebsiella spp. producing multiple extended-spectrum beta-lactamases. , 2007, Diagnostic microbiology and infectious disease.

[13]  V. Miriagou,et al.  Replicon Typing of Plasmids Encoding Resistance to Newer β-Lactams , 2006, Emerging infectious diseases.

[14]  Roderick C. Jones,et al.  Outbreak of catheter-associated Klebsiella oxytoca and Enterobacter cloacae bloodstream infections in an oncology chemotherapy center. , 2005, Archives of internal medicine.

[15]  G. Rossolini,et al.  Emergence in Klebsiella pneumoniae and Enterobacter cloacae Clinical Isolates of the VIM-4 Metallo-β-Lactamase Encoded by a Conjugative Plasmid , 2004, Antimicrobial Agents and Chemotherapy.

[16]  N. Hanson,et al.  Detection of Plasmid-Mediated AmpC β-Lactamase Genes in Clinical Isolates by Using Multiplex PCR , 2002, Journal of Clinical Microbiology.

[17]  G. Rossolini,et al.  Emerging Extended-Spectrum β-Lactamases in Proteus mirabilis , 2002, Journal of Clinical Microbiology.

[18]  P. Savelkoul,et al.  Identification of Epidemic Strains ofAcinetobacter baumannii by Integrase Gene PCR , 2001, Journal of Clinical Microbiology.

[19]  T. Doran,et al.  The role of Citrobacter in clinical disease of children: review. , 1999, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[20]  V. Jarlier.  [Beta-lactamase producers and other bacteria: which ones to take into consideration and when? The concept of beta-lactamase inhibitor]. , 1998, Presse medicale.

[21]  G. P. Harding,et al.  A general method for detecting and sizing large plasmids. , 1995, Analytical biochemistry.