Genomics and Susceptibility Profiles of Extensively Drug-Resistant Pseudomonas aeruginosa Isolates from Spain (vol 61, e01589-17, 2017)

Abstract

This study assessed the molecular epidemiology, resistance mechanisms, and susceptibility profiles of a collection of 150 extensively drug-resistant (XDR) Pseudomonas aeruginosa clinical isolates obtained from a 2015 Spanish multicenter study, with a particular focus on resistome analysis in relation to ceftolozane-tazobactam sus ceptibility. Broth microdilution MICs revealed that nearly all ( 95%) of the isolates were nonsusceptible to piperacillin-tazobactam, ceftazidime, cefepime, aztreonam, imipenem, meropenem, and ciprofloxacin. Most of them were also resistant to tobramycin (77%), whereas nonsusceptibility rates were lower for ceftolozane-tazobactam (31%), amikacin (7%), and colistin (2%). Pulsed-field gel electrophoresis–multilocus sequence typing (PFGE-MLST) analysis revealed that nearly all of the isolates belonged to previously de scribed high-risk clones. Sequence type 175 (ST175) was detected in all 9 participating hospitals and accounted for 68% (n 101) of the XDR isolates, distantly followed by ST244 (n 16), ST253 (n 12), ST235 (n 8), and ST111 (n 2), which were detected only in 1 to 2 hospitals. Through phenotypic and molecular methods, the presence of horizontally acquired carbapenemases was detected in 21% of the isolates, mostly VIM (17%) and GES enzymes (4%). At least two representative isolates from each clone and hospital (n- 44) were fully sequenced on an Illumina MiSeq. Classical mutational mech anisms, such as those leading to the overexpression of the b-lactamase AmpC or efflux pumps, OprD inactivation, and/or quinolone resistance-determining regions (QRDR) mu tations, were confirmed in most isolates and correlated well with the resistance phenotypes in the absence of horizontally acquired determinants. Ceftolozane-tazobactam re sistance was not detected in carbapenemase-negative isolates, in agreement with sequencing data showing the absence of ampC mutations. The unique set of mutations responsible for the XDR phenotype of ST175 clone documented 7 years earlier were found to be conserved, denoting the long-term persistence of this specific XDR lineage n Spanish hospitals. Finally, other potentially relevant mutations were evidenced, includ ing those in penicillin-binding protein 3 (PBP3), which is involved in -lactam (including ceftolozane-tazobactam) resistance, and FusA1, which is linked to aminoglycoside resis tance.