{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T14:43:05Z","timestamp":1768833785527,"version":"3.49.0"},"reference-count":54,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,11]],"date-time":"2021-03-11T00:00:00Z","timestamp":1615420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100013288","name":"LIFE programme","doi-asserted-by":"publisher","award":["European Union LIFE Programme under Grant Agree- 597 ment LIFE14 ENV\/PT\/000739 - LIFE Impetus (https:\/\/life-impetus.eu\/)"],"award-info":[{"award-number":["European Union LIFE Programme under Grant Agree- 597 ment LIFE14 ENV\/PT\/000739 - LIFE Impetus (https:\/\/life-impetus.eu\/)"]}],"id":[{"id":"10.13039\/100013288","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Wastewater treatment plants (WWTPs) are significant reservoirs of bacterial resistance. This work aims to identify the determinants of resistance produced by Gram-negative bacteria in the influent and effluent of two WWTPs in Portugal. A total of 96 wastewater samples were obtained between 2016 and 2019. The numbers of total aerobic and fecal contamination bacteria were evaluated, and genomic features were searched by polymerase chain reaction (PCR) and Next-Generation Sequencing (NGS). Enterobacteriaceae corresponded to 78.6% (n = 161) of the 205 isolates identified by 16sRNA. The most frequent isolates were Escherichia spp. (57.1%, n = 117), followed by Aeromonas spp. (16.1%, n = 33) and Klebsiella spp. (12.7%, n = 26). The remaining 29 isolates (14.1%) were distributed across 10 different genera. Among the 183 resistant genes detected, 54 isolates produced extended spectrum \u03b2-lactamases (ESBL), of which blaCTX-M-15 was predominant (37 isolates; 68.5%). A KPC-3 carbapenemase-producing K. oxytoca was identified (n = 1), with blaKPC-3 included in a transposon Tn4401 isoform b. A higher number of virulence genes (VG) (19 genes) was found in the E. coli 5301 (O25b-ST131-B2) isolate compared with a commensal E. coli 5281 (O25b-ST410-A) (six genes). Both shared five VG [Enterobactin; Aerobactin, CFA\/1 (clade \u03b1); Type1 (clade \u03b31); Type IV]. In conclusion, this work highlights the role of relevant clinical bacteria in WWTPs, such as KPC-3-producing K. oxytoca, and, for the first time, a CTX-M-15-producing Ochromobactrum intermedium, a human opportunistic pathogen, and a SED-1-producing Citrobacter farmeri, an uncommon CTX-M-type extended-spectrum beta-lactamase.<\/jats:p>","DOI":"10.3390\/microorganisms9030576","type":"journal-article","created":{"date-parts":[[2021,3,11]],"date-time":"2021-03-11T20:17:40Z","timestamp":1615493860000},"page":"576","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["An Update on Wastewater Multi-Resistant Bacteria: Identification of Clinical Pathogens Such as Escherichia coli O25b:H4-B2-ST131-Producing CTX-M-15 ESBL and KPC-3 Carbapenemase-Producing Klebsiella oxytoca"],"prefix":"10.3390","volume":"9","author":[{"given":"Elsa","family":"Mesquita","sequence":"first","affiliation":[{"name":"Water Quality and Treatment Laboratory, Urban Water Unit, Hydraulics and Environment Department, The Laborat\u00f3rio Nacional de Engenharia Civil (LNEC)\u2014National Civil Engineering Laboratory, 1700-066 Lisboa, Portugal"}]},{"given":"Rita","family":"Ribeiro","sequence":"additional","affiliation":[{"name":"Water Quality and Treatment Laboratory, Urban Water Unit, Hydraulics and Environment Department, The Laborat\u00f3rio Nacional de Engenharia Civil (LNEC)\u2014National Civil Engineering Laboratory, 1700-066 Lisboa, Portugal"}]},{"given":"Carla J. C.","family":"Silva","sequence":"additional","affiliation":[{"name":"Dire\u00e7\u00e3o de Opera\u00e7\u00f5es de Saneamento, ETAR de Beirolas, Rua Chen He, 1990-513 Sacav\u00e9m, Portugal"}]},{"given":"Rita","family":"Alves","sequence":"additional","affiliation":[{"name":"Dire\u00e7\u00e3o de Opera\u00e7\u00f5es de Saneamento, ETAR de Beirolas, Rua Chen He, 1990-513 Sacav\u00e9m, Portugal"}]},{"given":"Rita","family":"Baptista","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio do Sotavento da \u00c1guas do Algarve, Rua do Repouso, n\u00b0 10, 8000-302 Faro, Portugal"}]},{"given":"S\u00edlvia","family":"Condinho","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio do Sotavento da \u00c1guas do Algarve, Rua do Repouso, n\u00b0 10, 8000-302 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5705-366X","authenticated-orcid":false,"given":"Maria Jo\u00e3o","family":"Rosa","sequence":"additional","affiliation":[{"name":"Water Quality and Treatment Laboratory, Urban Water Unit, Hydraulics and Environment Department, The Laborat\u00f3rio Nacional de Engenharia Civil (LNEC)\u2014National Civil Engineering Laboratory, 1700-066 Lisboa, Portugal"}]},{"given":"Jo\u00e3o","family":"Perdig\u00e3o","sequence":"additional","affiliation":[{"name":"Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade Lisboa, Av. Gama Pinto, 1649-003 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3735-8554","authenticated-orcid":false,"given":"C\u00e1tia","family":"Caneiras","sequence":"additional","affiliation":[{"name":"Microbiology Research Laboratory of Environmental Health (EnviHealthMicro Lab), Institute of Environmental Health (ISAMB), Faculty of Medicine, Universidade de Lisboa, 1649-028 Lisboa, Portugal"},{"name":"Institute of Preventive Medicine and Public Health (IMP&amp;SP), Faculty of Medicine, Universidade de Lisboa, 1649-028 Lisboa, Portugal"},{"name":"Department of Microbiology and Immunology, Faculty of Pharmacy, Av. Gama Pinto, 1649-003 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0616-4650","authenticated-orcid":false,"given":"Aida","family":"Duarte","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Pharmacy, Av. Gama Pinto, 1649-003 Lisboa, Portugal"},{"name":"Centro de Investiga\u00e7\u00e3o Interdisciplinar Egas Moniz, Instituto Universit\u00e1rio Egas Moniz, 2829-511 Monte da Caparica, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1139\/er-2015-0069","article-title":"Antibiotic resistance genes as an emerging environmental contaminant","volume":"24","author":"Sanderson","year":"2016","journal-title":"Environ. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/j.eimc.2018.11.001","article-title":"Virulence and resistance determinants of Klebsiella pneumoniae isolated from a Portuguese tertiary university hospital centre over a 31-year period","volume":"37","author":"Caneiras","year":"2019","journal-title":"Enferm. Infecc. Microbiol. 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