{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T04:41:15Z","timestamp":1788410475125,"version":"build-2803163510"},"reference-count":108,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,11,6]],"date-time":"2023-11-06T00:00:00Z","timestamp":1699228800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>The rise in extended-spectrum beta-lactamase (ESBL)-producing <jats:italic>Enterobacteriaceae<\/jats:italic> in dairy cattle farms poses a risk to human health as they can spread to humans through the food chain, including raw milk. This study was designed to determine the status, antimicrobial resistance, and pathogenic potential of ESBL-producing -<jats:italic>E. coli<\/jats:italic> and -<jats:italic>Klebsiella<\/jats:italic> spp. isolates from bulk tank milk (BTM).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Thirty-three BTM samples were collected from 17 dairy farms and screened for ESBL-<jats:italic>E. coli<\/jats:italic> and -<jats:italic>Klebsiella<\/jats:italic> spp. on CHROMagar ESBL plates. All isolates were confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and subjected to antimicrobial susceptibility testing and whole genome sequencing (WGS).<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Ten presumptive ESBL-producing bacteria, eight <jats:italic>E. coli<\/jats:italic>, and two <jats:italic>K. pneumoniae<\/jats:italic> were isolated. The prevalence of ESBL-<jats:italic>E. coli<\/jats:italic> and -<jats:italic>K. pneumoniae<\/jats:italic> in BTM was 21.2% and 6.1%, respectively. ESBL-<jats:italic>E. coli<\/jats:italic> were detected in 41.2% of the study farms. Seven of the ESBL-<jats:italic>E. coli<\/jats:italic> isolates were multidrug resistant (MDR). The two ESBL-producing <jats:italic>K. pneumoniae<\/jats:italic> isolates were resistant to ceftriaxone. Seven ESBL-<jats:italic>E. coli<\/jats:italic> strains carry the <jats:italic>bla<\/jats:italic><jats:sub>CTX-M<\/jats:sub> gene, and five of them co-harbored <jats:italic>bla<\/jats:italic><jats:sub>TEM-1<\/jats:sub>. ESBL-<jats:italic>E. coli<\/jats:italic> co-harbored <jats:italic>bla<\/jats:italic><jats:sub>CTX-M<\/jats:sub> with other resistance genes, including <jats:italic>qnr<\/jats:italic>B<jats:italic>19<\/jats:italic>, <jats:italic>tet<\/jats:italic>(A), <jats:italic>aad<\/jats:italic>A1, <jats:italic>aph(3\u2019\u2019)<\/jats:italic>-Ib, <jats:italic>aph(6)<\/jats:italic>-Id), <jats:italic>floR<\/jats:italic>, <jats:italic>sul2<\/jats:italic>, and chromosomal mutations (<jats:italic>gyrA, gyrB, parC, parE, and pmrB<\/jats:italic>). Most <jats:italic>E. coli<\/jats:italic> resistance genes were associated with mobile genetic elements, mainly plasmids. Six sequence types (STs) of <jats:italic>E. coli<\/jats:italic> were detected. All ESBL-<jats:italic>E. coli<\/jats:italic> were predicted to be pathogenic to humans. Four STs (three ST10 and ST69) were high-risk clones of <jats:italic>E. coli<\/jats:italic>. Up to 40 virulence markers were detected in all <jats:italic>E. coli<\/jats:italic> isolates. One of the <jats:italic>K. pneumoniae<\/jats:italic> was ST867; the other was novel strain. <jats:italic>K. pneumoniae<\/jats:italic> isolates carried three types of beta-lactamase genes (<jats:italic>bla<\/jats:italic><jats:sub>CTX-M<\/jats:sub>, <jats:italic>bla<\/jats:italic><jats:sub>TEM-1<\/jats:sub> and <jats:italic>bla<\/jats:italic><jats:sub>SHV<\/jats:sub>). The novel <jats:italic>K. pneumoniae<\/jats:italic> ST also carried a novel IncFII(K) plasmid ST.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Detection of high-risk clones of MDR ESBL-<jats:italic>E. coli<\/jats:italic> and ESBL-<jats:italic>K. pneumoniae<\/jats:italic> in BTM indicates that raw milk could be a reservoir of potentially zoonotic ESBL-<jats:italic>E. coli<\/jats:italic> and -<jats:italic>K. pneumoniae<\/jats:italic>.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2023.1283165","type":"journal-article","created":{"date-parts":[[2023,11,6]],"date-time":"2023-11-06T16:10:34Z","timestamp":1699287034000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":21,"title":["Molecular epidemiology and pathogenomics of extended-spectrum beta-lactamase producing- Escherichia coli and - Klebsiella pneumoniae isolates from bulk tank milk in Tennessee, USA"],"prefix":"10.3389","volume":"14","author":[{"given":"Benti D.","family":"Gelalcha","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruwaa I.","family":"Mohammed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aga E.","family":"Gelgie","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oudessa","family":"Kerro Dego","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,11,6]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1221","DOI":"10.2147\/IDR.S241073","article-title":"Prevalence of virulence genes and their association with antimicrobial resistance among Pathogenic E. coli isolated from Egyptian patients with different clinical infections","volume":"13","author":"Abd El-Baky","year":"2020","journal-title":"Infect Drug Resist."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"e02430","DOI":"10.1128\/AEM.02430-17","article-title":"Molecular epidemiology of dairy cattle-associated Escherichia coli carrying blaCTX-M genes in Washington state","volume":"84","author":"Afema","year":"2018","journal-title":"Appl. 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