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Broth microdilution MICs revealed that nearly all (&gt;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\u2013multilocus sequence typing (PFGE-MLST) analysis revealed that nearly all of the isolates belonged to previously described high-risk clones. Sequence type 175 (ST175) was detected in all 9 participating hospitals and accounted for 68% (\n                    <jats:italic>n<\/jats:italic>\n                    = 101) of the XDR isolates, distantly followed by ST244 (\n                    <jats:italic>n<\/jats:italic>\n                    = 16), ST253 (\n                    <jats:italic>n<\/jats:italic>\n                    = 12), ST235 (\n                    <jats:italic>n<\/jats:italic>\n                    = 8), and ST111 (\n                    <jats:italic>n<\/jats:italic>\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                    <jats:italic>n<\/jats:italic>\n                    = 44) were fully sequenced on an Illumina MiSeq. Classical mutational mechanisms, such as those leading to the overexpression of the \u03b2-lactamase AmpC or efflux pumps, OprD inactivation, and\/or quinolone resistance-determining regions (QRDR) mutations, were confirmed in most isolates and correlated well with the resistance phenotypes in the absence of horizontally acquired determinants. Ceftolozane-tazobactam resistance was not detected in carbapenemase-negative isolates, in agreement with sequencing data showing the absence of\n                    <jats:italic>ampC<\/jats:italic>\n                    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 in Spanish hospitals. Finally, other potentially relevant mutations were evidenced, including those in penicillin-binding protein 3 (PBP3), which is involved in \u03b2-lactam (including ceftolozane-tazobactam) resistance, and FusA1, which is linked to aminoglycoside resistance.\n                  <\/jats:p>","DOI":"10.1128\/aac.01589-17","type":"journal-article","created":{"date-parts":[[2017,9,5]],"date-time":"2017-09-05T20:30:20Z","timestamp":1504643420000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":141,"title":["Genomics and Susceptibility Profiles of Extensively Drug-Resistant Pseudomonas aeruginosa Isolates from Spain"],"prefix":"10.1128","volume":"61","author":[{"given":"Ester","family":"del Barrio-Tofi\u00f1o","sequence":"first","affiliation":[{"name":"Department of Microbiology, Intensive Care Unit and Unidad de Investigaci\u00f3n, Hospital Universitari Son Espases, Instituto de Investigaci\u00f3n Illes Balears (IdISBa), 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