{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T18:41:41Z","timestamp":1788374501711,"version":"build-2803163510"},"reference-count":35,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2023,2,22]],"date-time":"2023-02-22T00:00:00Z","timestamp":1677024000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"DOI":"10.13039\/100000061","name":"Fogarty International Center","doi-asserted-by":"publisher","award":["TW008650- Student Stipend (NIH D43 Forgarty Fellow GOHi-OHEART-OSU)"],"award-info":[{"award-number":["TW008650- Student Stipend (NIH D43 Forgarty Fellow GOHi-OHEART-OSU)"]}],"id":[{"id":"10.13039\/100000061","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>As safe agents of last resort, carbapenems are reserved for the treatment of infections caused by multidrug-resistant organisms. The impact of \u03b2-lactam antibiotics, cefotaxime, and meropenem on the frequency and diversity of carbapenemase-producing organisms recovered from environmental samples has not been fully established. Therefore, this methodological study aimed at determining \u03b2-lactam drugs used in selective enrichment and their impact on the recovery of carbapenemase-producing <jats:italic>Enterobacterales<\/jats:italic> (CPE) from untreated wastewater. We used a longitudinal study design where 1L wastewater samples were collected weekly from wastewater treatment plant (WWTP) influent and quarterly from contributing sanitary sewers in Columbus, Ohio USA with 52 total samples collected. Aliquots of 500 mL were passed through membrane filters of decreasing pore sizes to enable all the water to pass through and capture bacteria. For each sample, the resulting filters were placed into two modified MacConkey (MAC) broths, one supplemented with 0.5 \u03bcg\/mL of meropenem and 70 \u03bcg\/mL of ZnSO<jats:sub>4<\/jats:sub> and the other supplemented with 2 \u03bcg\/mL cefotaxime. The inoculated broth was then incubated at 37\u00b0 C overnight, after which they were streaked onto two types of correspondingly-modified MAC agar plates supplemented with 0.5 \u03bcg\/mL and 1.0 \u03bcg\/mL of meropenem and 70 \u03bcg\/mL of ZnSO<jats:sub>4<\/jats:sub> and incubated at 37\u00b0C overnight. The isolates were identified based on morphological and biochemical characteristics. Then, up to four distinct colonies of each isolate\u2019s pure culture per sample were tested for carbapenemase production using the Carba-NP test. Matrix-assisted laser desorption\/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) MALDI-TOF MS was used to identify carbapenemase-producing organisms. In total 391 Carba-NP positive isolates were recovered from the 52 wastewater samples: 305 (78%) isolates had <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub>, 73 (19%) carried <jats:italic>bla<\/jats:italic><jats:sub>NDM<\/jats:sub>, and 14 (4%) harbored both <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> and <jats:italic>bla<\/jats:italic><jats:sub>NDM<\/jats:sub> resistance genes. CPE genes of both <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> and <jats:italic>bla<\/jats:italic><jats:sub>NDM<\/jats:sub> were recovered in both types of modified MAC broths, with 84 (21%) having a <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> gene<jats:sub>,<\/jats:sub> 22 (6%) carrying <jats:italic>bla<\/jats:italic><jats:sub>NDM<\/jats:sub> and 9 (2%) harbored both a <jats:italic>bla<\/jats:italic><jats:sub>KPC<\/jats:sub> and <jats:italic>bla<\/jats:italic><jats:sub>NDM<\/jats:sub> of isolates recovered from MAC medium incorporated with 0.5ug\/mL meropenem and 70ug\/mL ZnSO<jats:sub>4<\/jats:sub>. The most prevalent isolates were <jats:italic>Klebsiella pneumoniae<\/jats:italic>, <jats:italic>Escherichia coli<\/jats:italic>, and <jats:italic>Citrobacter spp<\/jats:italic>.<\/jats:p>","DOI":"10.1371\/journal.pone.0281909","type":"journal-article","created":{"date-parts":[[2023,2,22]],"date-time":"2023-02-22T18:31:34Z","timestamp":1677090694000},"page":"e0281909","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Frequency and diversity of carbapenemase-producing Enterobacterales recovered from untreated wastewater impacted by selective media containing cefotaxime and meropenem in Ohio, USA"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6100-9090","authenticated-orcid":true,"given":"Rael J.","family":"Too","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George 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E.","family":"Wittum","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,2,22]]},"reference":[{"issue":"12","key":"pone.0281909.ref001","doi-asserted-by":"crossref","first-page":"2241","DOI":"10.1007\/s10096-018-3365-9","article-title":"Related carbapenemase-producing Klebsiella isolates detected in both a hospital and associated aquatic environment in Sweden","volume":"37","author":"FA Khan","year":"2018","journal-title":"Eur J Clin Microbiol Infect Dis"},{"issue":"1","key":"pone.0281909.ref002","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/S1473-3099(18)30708-4","article-title":"Attributable deaths and disability-adjusted life-years caused by infections with antibiotic-resistant bacteria in Switzerland","volume":"19","author":"M Gasser","year":"2019","journal-title":"Lancet Infect 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