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Wildlife can act as carriers of AMR bacteria and spread them to other wildlife, domestic animals, and humans; thus, they have public health implications. A total of 128 Escherichia coli isolates were obtained from 66 of 217 fecal samples obtained from different wild animals using media without antibiotic supplementation. Antibiograms were performed for 17 antibiotics to determine the phenotypic resistance profile in these isolates. Extended-spectrum \u03b2-lactamase (ESBL) production was tested using the double-disc synergy test, and 29 E. coli strains were selected for whole genome sequencing. In total, 22.1% of the wild animals tested carried multidrug-resistant E. coli isolates, and 0.93% (2\/217) of these wild animals carried E. coli isolates with ESBL-encoding genes (blaCTX-M-65, blaCTX-M-55, and blaEC-1982). The E. coli isolates showed the highest resistance rates to ampicillin and were fully susceptible to amikacin, meropenem, ertapenem, and imipenem. Multiple resistance and virulence genes were detected, as well as different plasmids. The relatively high frequency of multidrug-resistant E. coli isolates in wildlife, with some of them being ESBL producers, raises some concern regarding the potential transmission of antibiotic-resistant bacteria among these animals. Gaining insights into antibiotic resistance patterns in wildlife can be vital in shaping conservation initiatives and developing effective strategies for responsible antibiotic use.<\/jats:p>","DOI":"10.3390\/vetsci11100469","type":"journal-article","created":{"date-parts":[[2024,10,1]],"date-time":"2024-10-01T11:08:47Z","timestamp":1727780927000},"page":"469","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Molecular Characterization of Multidrug-Resistant Escherichia coli from Fecal Samples of Wild Animals"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7317-9123","authenticated-orcid":false,"given":"Carolina","family":"Saben\u00e7a","sequence":"first","affiliation":[{"name":"MicroART-Antibiotic Resistance Team, Department of Veterinary Sciences, University of Tr\u00e1s-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal"},{"name":"Department of Genetics and Biotechnology, University of Tr\u00e1s-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal"},{"name":"Functional Genomics and Proteomics Unit, University of Tr\u00e1s-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal"},{"name":"Associated Laboratory for Green Chemistry, University NOVA of Lisbon, 1099-085 Caparica, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8517-0150","authenticated-orcid":false,"given":"Mario","family":"Romero-Rivera","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University Hospital Ram\u00f3n y Cajal, Instituto Ram\u00f3n y Cajal de Investigaci\u00f3n Sanitaria (IRYCIS), 28034 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raquel","family":"Barbero-Herranz","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University Hospital Ram\u00f3n y Cajal, Instituto Ram\u00f3n y Cajal de Investigaci\u00f3n Sanitaria (IRYCIS), 28034 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3191-7853","authenticated-orcid":false,"given":"Roberto","family":"Sargo","sequence":"additional","affiliation":[{"name":"CRAS\u2014Center for the Recovery of Wild Animals, Veterinary Hospital, University of Tr\u00e1s-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu\u00eds","family":"Sousa","sequence":"additional","affiliation":[{"name":"CRAS\u2014Center for the Recovery of Wild Animals, Veterinary Hospital, University of Tr\u00e1s-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6602-0926","authenticated-orcid":false,"given":"Filipe","family":"Silva","sequence":"additional","affiliation":[{"name":"CRAS\u2014Center for the Recovery of Wild Animals, Veterinary Hospital, University of 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