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Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The co-existence phenomenon of antibiotic resistance genes (ARGs), particularly of last-resort antibiotics in multi-drug resistant (MDR) bacteria, is of particular concern in the least studied bacterial species.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In 2023, strain M2 was isolated from the sludge sample at a commercial bovine farm in Hebei province, China, using a MacConkey plate containing meropenem. PCR amplification and Sanger sequencing verified it co-carrying <jats:italic>bla<\/jats:italic><jats:sub>NDM<\/jats:sub> and <jats:italic>tet<\/jats:italic>(X) genes. It was classified within the Acinetobacter genus by MALDI-TOF-MS and 16S rDNA analyses. Whole-genome sequencing (WGS) was performed on the Oxford Nanopore platform, with species-level identification via ANI and dDDH. Antimicrobial susceptibility testing was performed against 20 antibiotics. Conjugation assays employed the filter-mating method using <jats:italic>E. coli<\/jats:italic> J53 and <jats:italic>Salmonella<\/jats:italic> LGJ2 as recipients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>This strain was confirmed as a novel species of <jats:italic>Acinetobacter<\/jats:italic> genus, showing resistance to meropenem, ampicillin, ceftazidime, cefepime, gentamicin, kanamycin, fosfomycin, imipenem, ertapenem, and tetracycline. Despite carrying <jats:italic>tet<\/jats:italic>(X3), it remained susceptible to tigecycline, omadacycline, and doxycycline. The genome carried 11 ARG types, multiple metal resistance genes (MRGs), and virulence factor (VF) genes. The <jats:italic>bla<\/jats:italic><jats:sub>NDM-1<\/jats:sub> was located in a skeleton, IS<jats:italic>Aba125-bla<\/jats:italic><jats:sub>NDM-1<\/jats:sub>-<jats:italic>ble<\/jats:italic><jats:sub>MBL<\/jats:sub>-<jats:italic>trpF<\/jats:italic>, which was carried by an IS<jats:italic>Aba<\/jats:italic>14-mediated rolling-circle-like structure in pM2-2-NDM-1 (rep_cluster_481). Integrative and conjugative element (ICE) and multiple pd<jats:italic>if<\/jats:italic> modules (driven by the XerCD site-specific recombination (XerCD SSR) system), which were associated with the mobilization of resistance determinants, were identified in this plasmid. Chromosomal <jats:italic>tet<\/jats:italic>(X3) was mediated by IS<jats:italic>Vsa3<\/jats:italic>, forming a skeleton, IS<jats:italic>Vsa3-XerD-tet<\/jats:italic> (X3)<jats:italic>-res-ISVsa3<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>The co-occurrence of <jats:italic>bla<\/jats:italic><jats:sub>NDM<\/jats:sub> and <jats:italic>tet<\/jats:italic>(X) in a novel species of the <jats:italic>Acinetobacter<\/jats:italic> genus hints that substantial undiscovered bacteria co-carrying high-risk ARGs are concealing in the agroecological system, which should cause particular concern.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2025.1594982","type":"journal-article","created":{"date-parts":[[2025,7,2]],"date-time":"2025-07-02T05:53:00Z","timestamp":1751435580000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["A strain defined as a novel species in the Acinetobacter genus co-harboring chromosomal associated tet(X3) and plasmid associated blaNDM-1 from a beef cattle farm in Hebei, China"],"prefix":"10.3389","volume":"15","author":[{"given":"Qing","family":"Wang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanming","family":"Wei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad","family":"Shoaib","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanhua","family":"Qiu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guonian","family":"Dai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Honglin","family":"Lin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weiwei","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiyu","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,7,2]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1131","DOI":"10.1038\/s41467-022-28819-2","article-title":"Role of mobile genetic elements in the global dissemination of the carbapenem resistance gene bla NDM","volume":"13","author":"Acman","year":"2022","journal-title":"Nat. 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