Evaluation of heteroresistance to polymyxin B among carbapenem-susceptible and -resistant Pseudomonas aeruginosa

J Med Microbiol. 2013 Aug;62(Pt 8):1184-1189. doi: 10.1099/jmm.0.059220-0. Epub 2013 May 22.


One hundred and twenty-four Pseudomonas aeruginosa isolates were selected for antimicrobial susceptibility testing with anti-pseudomonal agents, MIC determination for polymyxin B and metallo-beta-lactamase detection (genes blaSPM, blaVIM-1, blaNDM-1 and blaIMP). According to the imipenem and/or meropenem susceptibility profile, a set of randomly selected isolates (12 isolates carbapenem-susceptible and 12 isolates carbapenem-resistant) were evaluated for heteroresistance to polymyxin B. Heteroresistance testing was performed by plating the isolates onto increasing concentrations of polymyxin B (from 0 to 8.0 mg l(-1)). The population analysis profile (PAP) was defined as the ratio of the number of colony-forming units on the plate with the highest concentration of polymyxin B at which bacterial growth occurred against the number of colony-forming units on the plate without antibiotic. Isolates presenting subpopulations that exhibited growth at polymyxin B concentrations ≥2 mg l(-1) were considered heteroresistant. Isolates containing subpopulations that grew at polymyxin B concentrations at least twice as high as the original MIC but <2 mg l(-1) were considered heterogeneous. Antimicrobial susceptibility testing results indicated a variable degree of susceptibility: high levels of resistance to gentamicin (30.6 %) and imipenem (29.0 %); low levels of resistance to aztreonam (1.6 %) and ciprofloxacin (4.8 %). All isolates were susceptible to polymyxin B: MIC50 and MIC90 were 1 mg l(-1) and 2 mg l(-1), respectively. Thirty-seven isolates (30 %) were carbapenem-resistant. Four isolates resistant to carbapenems were positive for blaIMP. There were no heteroresistant subpopulations in the carbapenem-susceptible group, but three isolates presented heterogeneous subpopulations. The PAP frequency ranged from 2.1×10(-4) to 6.9×10(-8). In the carbapenem-resistant group, one isolate was heteroresistant. Six isolates in this group presented heterogeneous subpopulations. In the resistant population, the PAP frequency ranged from 2.1×10(-7) to 2.6×10(-4). In this study, polymyxin B heteroresistance in P. aeruginosa was uncommon and occurred in only one carbapenem-resistant isolate, despite the fact that several isolates presented heterogeneous subpopulations with increased polymyxin B MICs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Ascitic Fluid / microbiology
  • Bacteremia
  • Brazil
  • Carbapenems / pharmacology
  • Cystic Fibrosis / microbiology
  • Demography
  • Drug Resistance, Bacterial*
  • Genotype
  • Humans
  • Microbial Sensitivity Tests
  • Phenotype
  • Polymyxin B / pharmacology*
  • Pseudomonas Infections / drug therapy
  • Pseudomonas Infections / microbiology*
  • Pseudomonas aeruginosa / drug effects*
  • Pseudomonas aeruginosa / enzymology
  • Pseudomonas aeruginosa / growth & development
  • Pseudomonas aeruginosa / isolation & purification
  • Sputum / microbiology
  • beta-Lactamases / metabolism


  • Anti-Bacterial Agents
  • Carbapenems
  • beta-Lactamases
  • Polymyxin B