{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T05:20:23Z","timestamp":1791523223317,"version":"4.3.4"},"reference-count":54,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2023,5,16]],"date-time":"2023-05-16T00:00:00Z","timestamp":1684195200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001505","name":"Health Research Council of New Zealand","doi-asserted-by":"publisher","award":["17\/372"],"award-info":[{"award-number":["17\/372"]}],"id":[{"id":"10.13039\/501100001505","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>\n                    <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                    causes a wide range of severe infections. Ceftazidime, a cephalosporin, is a key antibiotic for treating infections but a significant proportion of isolates are ceftazidime-resistant. The aim of this research was to identify mutations that contribute to resistance, and to quantify the impacts of individual mutations and mutation combinations. Thirty-five mutants with reduced susceptibility to ceftazidime were evolved from two antibiotic-sensitive\n                    <jats:italic>P<\/jats:italic>\n                    .\n                    <jats:italic>aeruginosa<\/jats:italic>\n                    reference strains PAO1 and PA14. Mutations were identified by whole genome sequencing. The evolved mutants tolerated ceftazidime at concentrations between 4 and 1000 times that of the parental bacteria, with most mutants being ceftazidime resistant (minimum inhibitory concentration [MIC] \u2265 32 mg\/L). Many mutants were also resistant to meropenem, a carbapenem antibiotic. Twenty-eight genes were mutated in multiple mutants, with\n                    <jats:italic>dacB<\/jats:italic>\n                    and\n                    <jats:italic>mpl<\/jats:italic>\n                    being the most frequently mutated. Mutations in six key genes were engineered into the genome of strain PAO1 individually and in combinations. A\n                    <jats:italic>dacB<\/jats:italic>\n                    mutation by itself increased the ceftazidime MIC by 16-fold although the mutant bacteria remained ceftazidime sensitive (MIC &lt; 32 mg\/L). Mutations in\n                    <jats:italic>ampC<\/jats:italic>\n                    ,\n                    <jats:italic>mexR<\/jats:italic>\n                    ,\n                    <jats:italic>nalC<\/jats:italic>\n                    or\n                    <jats:italic>nalD<\/jats:italic>\n                    increased the MIC by 2- to 4-fold. The MIC of a\n                    <jats:italic>dacB<\/jats:italic>\n                    mutant was increased when combined with a mutation in\n                    <jats:italic>ampC<\/jats:italic>\n                    , rendering the bacteria resistant, whereas other mutation combinations did not increase the MIC above those of single mutants. To determine the clinical relevance of mutations identified through experimental evolution, 173 ceftazidime-resistant and 166 sensitive clinical isolates were analysed for the presence of sequence variants that likely alter function of resistance-associated genes.\n                    <jats:italic>dacB<\/jats:italic>\n                    and\n                    <jats:italic>ampC<\/jats:italic>\n                    sequence variants occur most frequently in both resistant and sensitive clinical isolates. Our findings quantify the individual and combinatorial effects of mutations in different genes on ceftazidime susceptibility and demonstrate that the genetic basis of ceftazidime resistance is complex and multifactorial.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0285856","type":"journal-article","created":{"date-parts":[[2023,5,16]],"date-time":"2023-05-16T14:15:14Z","timestamp":1684246514000},"page":"e0285856","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":24,"title":["Ceftazidime resistance in Pseudomonas aeruginosa is multigenic and complex"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6682-9351","authenticated-orcid":true,"given":"Kay 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M.","family":"Patrick","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Craig","family":"Winstanley","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iain L.","family":"Lamont","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,5,16]]},"reference":[{"issue":"3","key":"pone.0285856.ref001","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1111\/apm.13010","article-title":"The environmental occurrence of Pseudomonas aeruginosa","volume":"128","author":"S Crone","year":"2020","journal-title":"APMIS"},{"issue":"3","key":"pone.0285856.ref002","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1586\/eri.13.12","article-title":"Clinical relevance of the ESKAPE pathogens.","volume":"11","author":"JN Pendleton","year":"2013","journal-title":"Expert Rev Anti Infect 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