Resistance mechanisms of multiresistant Pseudomonas aeruginosa strains from Germany and correlation with hypermutation

Abstract

In this study, we analyzed the mechanisms of multiresistance for 22 clinical multiresistant and clonally different Pseudomonas aeruginosa strains from Germany. Twelve and 10 strains originated from cystic fibrosis (CF) and non-CF patients, respectively. Overproduction of the efflux systems MexAB-OprM, MexCD-OprJ, MexEF-OprN, and MexXY-OprM was studied. Furthermore, loss of OprD, alterations in type II topoisomerases, AmpC overproduction, and the presence of 25 acquired resistance determinants were investigated. The presence of a hypermutation phenotype was also taken into account. Besides modifications in GyrA (91%), the most frequent mechanisms of resistance were MexXY-OprM overproduction (82%), OprD loss (82%), and AmpC overproduction (73%). Clear differences between strains from CF and non-CF patients were found: numerous genes coding for aminoglycoside-modifying enzymes and located, partially in combination with beta-lactamase genes, in class 1 integrons were found only in strains from non-CF patients. Furthermore, multiple modifications in type II topoisomerases conferring high quinolone resistance levels and overexpression of MexAB-OprM were exclusively detected in multiresistant strains from non-CF patients. Correlations of the detected phenotypes and resistance mechanisms revealed a great impact of efflux pump overproduction on multiresistance in P. aeruginosa. Confirming previous studies, we found that additional, unknown chromosomally mediated resistance mechanisms remain to be determined. In our study, 11 out of 12 strains and 3 out of 10 strains from CF patients and non-CF patients, respectively, were hypermutable. This extremely high proportion of mutator strains should be taken into consideration for the treatment of multiresistant P. aeruginosa.

FIG. 1.

Structures of class 1 integrons detected exclusively in multiresistant P. aeruginosa clones from non-CF patients. Sequence analysis (with the BLASTX program) revealed 79% amino acid sequence homology with QecE (NP_044260) (*), 45% amino acid sequence homology with hypothetical protein MED121_04115 of Marinomonas sp. strain MED121 (ZP_01077982) (**), and 57% amino acid sequence homology with the RecA/RadA recombinase of Yersinia bercovieri (ZP_00823172) (***). 5′-CS, 5′ conserved segment; 3′-CS, 3′ conserved segment.