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Whole-genome sequencing (WGS) and bioinformatic analysis were conducted to identify the genomic characteristics. Fourteen\n E. coli<\/jats:italic>\n strains belonging to the same sequence type (ST) 4014, comprising eleven strains retrieved from public databases and three strains from this study, were integrated into a phylogenetic analysis. Conjugation experiments were conducted to evaluate the transferability of the resistance plasmids. Three\n E. coli<\/jats:italic>\n strains exhibited resistance to both tetracyclines and carbapenems, consistent with the presence of\n tet<\/jats:italic>\n (X4) and\n bla<\/jats:italic>\n NDM-1<\/jats:sub>\n genes in their genomes. All strains belonged to the rare ST 4014 and were identified in healthy individuals within a 1-week period. WGS revealed that tet(X4) and\n bla<\/jats:italic>\n NDM-1<\/jats:sub>\n genes were each located on separate plasmids, both exhibiting 100% sequence identity to others previously reported in various bacterial species. Conjugation experiments confirmed the transferability of both\n tet<\/jats:italic>\n (X4) and\n bla<\/jats:italic>\n NDM-1<\/jats:sub>\n -carrying plasmids. Phylogenetic analysis based on cgSNPs revealed limited genetic diversity among the three strains (2\u20136 SNPs), but substantial differences compared to 11 publicly available ST4014 strains (116\u2013172 SNPs). This study reports\n E. coli<\/jats:italic>\n ST4014 strains from healthy individuals harboring conjugative plasmids carrying\n tet<\/jats:italic>\n (X4) and\n bla<\/jats:italic>\n NDM-1<\/jats:sub>\n genes, conferring resistance to tigecycline and carbapenems, respectively. These findings highlight the silent spread of multidrug-resistant strains in community populations and emphasize the need for enhanced surveillance of antimicrobial resistance beyond clinical settings.\n <\/jats:p>\n <\/jats:sec>\n \n IMPORTANCE<\/jats:title>\n \n The emergence of\n Escherichia coli<\/jats:italic>\n strains co-harboring\n tet<\/jats:italic>\n (X4) and\n bla<\/jats:italic>\n NDM-1<\/jats:sub>\n genes in healthy individuals represents a critical public health concern. These genes mediate resistance to tigecycline and carbapenems, two of the few remaining options for treating infections caused by multidrug-resistant gram-negative bacteria. The detection of clonally related ST4014 strains carrying conjugative plasmids encoding both resistance determinants highlights the potential for horizontal gene transfer and silent dissemination of dual-resistance plasmids in community settings. Such colonization among healthy individuals suggests that antimicrobial resistance may be spreading unnoticed beyond hospitals, driven by environmental or foodborne transmission routes. These findings emphasize the urgent need for integrated genomic surveillance and One Health-based interventions encompassing human, animal, and environmental reservoirs to prevent the expansion of high-risk resistance genes and safeguard the clinical efficacy of last-line antibiotics.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1128\/spectrum.03336-25","type":"journal-article","created":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T14:01:27Z","timestamp":1769004087000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":3,"title":["Human intestinal colonization by\n Escherichia coli<\/i>\n ST4014 co-harboring\n tet<\/i>\n (X4) and\n bla<\/i>\n NDM-1<\/sub>\n gene: a potential reservoir for antimicrobial resistance dissemination"],"prefix":"10.1128","volume":"14","author":[{"given":"Huiqiong","family":"Jia","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05m1p5x56","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Laboratory Medicine, The First Affiliated Hospital, Zhejiang University School of Medicine","place":["Hangzhou, China"]},{"name":"Zhejiang Key Laboratory of Clinical In Vitro Diagnostic Techniques","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaocong","family":"Lu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017z00e58","id-type":"ROR","asserted-by":"publisher"}],"name":"The First Clinical College, Chongqing Medical University","place":["Chongqing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanyuan","family":"Jia","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05kvm7n82","id-type":"ROR","asserted-by":"publisher"}],"name":"Suzhou Medical College of Soochow University","place":["Suzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yawen","family":"Yu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00ka6rp58","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuye","family":"Wu","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00ka6rp58","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3186-1968","authenticated-orcid":true,"given":"Danni","family":"Bao","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory, Sanmen People\u2019s Hospital","place":["Taizhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingying","family":"Zhang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00ka6rp58","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3328-5084","authenticated-orcid":true,"given":"Jiehong","family":"Fang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05v1y0t93","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Specialty Agri-products Quality and Hazard Controlling Technology of Zhejiang Province, College of Life Sciences, China Jiliang University","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Butaye","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02zhqgq86","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infectious Diseases and Public Health, Jockey Club College of Veterinary, City University of Hong Kong","place":["Hong Kong, China"]},{"id":[{"id":"https:\/\/ror.org\/00cv9y106","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Pathobiology, Pharmacology and Zoological Medicine, Faculty of Veterinary Medicine, Ghent University","place":["Merelbeke, Belgium"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2516-1129","authenticated-orcid":true,"given":"Jo\u00e3o Pedro 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China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_5_2_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13073-024-01310-x"},{"key":"e_1_3_5_3_2","unstructured":"America IDSo. 2024. IDSA 2024 guidance on the treatment of antimicrobial resistant Gram-negative infections. Available from: https:\/\/www.idsociety.org\/practice-guideline\/amr-guidance\/. Retrieved 8 Dec 2025."},{"key":"e_1_3_5_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.scitotenv.2022.154010"},{"key":"e_1_3_5_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.micres.2024.127730"},{"key":"e_1_3_5_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/spectrum.00196-22"},{"key":"e_1_3_5_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/aac.01945-24"},{"key":"e_1_3_5_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.drup.2024.101123"},{"key":"e_1_3_5_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.drup.2024.101194"}],"container-title":["Microbiology Spectrum"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.03336-25","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/spectrum.03336-25","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T14:06:05Z","timestamp":1772546765000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/spectrum.03336-25"}},"subtitle":[],"editor":[{"given":"Nilton","family":"Lincopan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,3,3]]},"references-count":8,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2026,3,3]]}},"alternative-id":["10.1128\/spectrum.03336-25"],"URL":"https:\/\/doi.org\/10.1128\/spectrum.03336-25","relation":{},"ISSN":["2165-0497"],"issn-type":[{"value":"2165-0497","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,3]]},"assertion":[{"value":"2025-10-16","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-16","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-01-21","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e03336-25"}}