Carbapenem-resistance mechanisms of multidrug-resistant Pseudomonas aeruginosa

J Med Microbiol. 2013 Sep;62(Pt 9):1317-1325. doi: 10.1099/jmm.0.058354-0. Epub 2013 May 30.

Abstract

Clonal dissemination of multidrug-resistant Pseudomonas aeruginosa (MDRPA) is a major concern worldwide. The aim of this study was to explore the mechanisms leading to the carbapenem resistance of an MDRPA clone. Isolates were obtained from a surgical wound, sputum, urine and a blood culture. Pulsed-field gel electrophoresis (PFGE) showed high genomic homogeneity of these isolates and confirmed the circulation of an endemic clone belonging to serotype O4. Outer membrane protein (OMP) bands were visualized by SDS-PAGE, meropenem accumulation was measured in a bioassay and integrons were detected by PCR. Efflux pumps were studied for several antimicrobial agents and synergic combinations thereof in the presence or absence of both carbonyl cyanide m-chlorophenylhydrazone (CCCP) and Phe-Arg-β-naphthylamide (PAβN) at final concentrations of 10 and 40 mg l(-1), respectively. On OMP electrophoretic profiles, MDRPA showed a reduction of outer membrane porin D (OprD) and PCR demonstrated the presence of a class 1 integron with a cassette encoding aminoglycoside adenyltransferase B (aadB). Meropenem accumulation was slightly higher in bacilli than in the filamentous cells that formed in the presence of antibiotics. Overexpression of the efflux pump MexAB-OprM and a functional MexXY-OprM were detected in all isolates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / metabolism
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Base Sequence
  • Carbapenems / pharmacology*
  • Carbonyl Cyanide m-Chlorophenyl Hydrazone / pharmacology
  • Dipeptides / pharmacology
  • Drug Resistance, Multiple, Bacterial*
  • Drug Synergism
  • Membrane Transport Proteins / genetics
  • Membrane Transport Proteins / metabolism
  • Meropenem
  • Microbial Sensitivity Tests
  • Molecular Sequence Data
  • Porins / genetics
  • Porins / metabolism
  • Pseudomonas aeruginosa / drug effects*
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / metabolism
  • Thienamycins / pharmacology
  • Tobramycin / pharmacology

Substances

  • Anti-Bacterial Agents
  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • Carbapenems
  • Dipeptides
  • Membrane Transport Proteins
  • MexXY protein, Pseudomonas aeruginosa
  • OprM protein, Pseudomonas aeruginosa
  • Porins
  • Thienamycins
  • phenylalanylarginine-naphthylamide
  • OprD protein, Pseudomonas aeruginosa
  • Carbonyl Cyanide m-Chlorophenyl Hydrazone
  • Meropenem
  • Tobramycin

Associated data

  • GENBANK/JF714996
  • GENBANK/JF718319
  • GENBANK/JF727876
  • GENBANK/JF742758