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

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 bla(SPM), bla(VIM_1), bla(NDM_1) and bla(IMP)). 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 bla(IMP). There were no heteroresistant subpopulations in the carbapenem-susceptible group, but three isolates presented heterogeneous subpopulations. The PAP frequency ranged from 2.1 x 10(-4) to 6.9 x 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 x 10(-7) to 2.6 x 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.