{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:49:35Z","timestamp":1787028575233,"version":"build-2736575974"},"reference-count":29,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T00:00:00Z","timestamp":1761868800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Research, Development, and Innovation Authority (RDIA)-Kingdom of Saudi Arabia","award":["12899-jazzan-2023-JZU-R-2-1-HW"],"award-info":[{"award-number":["12899-jazzan-2023-JZU-R-2-1-HW"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The global rise of antimicrobial resistance represents a critical challenge to public health, with Escherichia coli emerging as one of the most significant contributors due to its high adaptability and prevalence of extended-spectrum \u03b2-lactamase (ESBL) production. Outer membrane vesicles (OMVs), nanoscale structures released by Gram-negative bacteria, have recently been implicated in the dissemination of resistance determinants and direct antibiotic inactivation. This study investigated the role of OMVs derived from ESBL-producing E. coli in mediating resistance to ampicillin. Clinical strains harboring CTX-M-15 resistance genes were cultured under selective pressure, and OMVs were purified via size-exclusion chromatography. Characterization using tunable resistive pulse sensing (TRPS) and cryo-transmission electron microscopy confirmed vesicle integrity, with sizes ranging from 80 to 150 nm. DNA quantification and PCR analysis revealed the presence of CTX-M-15 genes within vesicles, which remained protected from DNase digestion, confirming encapsulation. Functional assays demonstrated \u03b2-lactamase activity within OMVs, with proteinase K treatment indicating localization primarily within vesicles rather than on their surface. Importantly, OMVs inactivated ampicillin in a dose-dependent manner, significantly reducing its efficacy against susceptible E. coli. Disc diffusion and microtiter plate assays confirmed that \u03b2-lactamase-positive OMVs protected susceptible strains from antibiotic killing, promoting bacterial survival and growth. This study uniquely demonstrates that OMVs from CTX-M-15\u2013producing Escherichia coli carry both resistance genes and active \u03b2-lactamase enzymes, thereby facilitating both genetic dissemination and direct antibiotic inactivation. Targeting OMV biogenesis may represent a novel strategy to combat antimicrobial resistance.<\/jats:p>","DOI":"10.3390\/ijms262110601","type":"journal-article","created":{"date-parts":[[2025,10,31]],"date-time":"2025-10-31T05:28:43Z","timestamp":1761888523000},"page":"10601","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Vesicle-Mediated Transfer of CTX-M \u03b2-Lactamase Genes and Proteins Confers Ampicillin Resistance in Escherichia coli"],"prefix":"10.3390","volume":"26","author":[{"given":"Nader","family":"Kameli","sequence":"first","affiliation":[{"name":"Department of Medical Laboratory Technology, College of Nursing and Health Sciences, Jazan University, Jazan 6809, Saudi Arabia"},{"name":"Health Research Center, Jazan University, Jazan 6809, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/S0140-6736(21)02724-0","article-title":"Global burden of bacterial antimicrobial resistance in 2019: A systematic analysis","volume":"399","year":"2022","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1007\/s10096-008-0545-z","article-title":"Epidemiology of bacteremia episodes in a single center: Increase in Gram-negative isolates, antibiotics resistance, and patient age","volume":"27","author":"Marchaim","year":"2008","journal-title":"Eur. 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