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Identification of Outer Membrane Vesicles as a New Vehicle Mediating Antibiotic Resistance Gene Transfer in Campylobacter

Mengyu Zhao,M. He,Xiaolong Lin,Ke Wu,Fang Yang,Xinggui Chen,Haozheng Li,Hongning Wang,Yizhi Tang

Published 2025 in Journal of Extracellular Vesicles

ABSTRACT

ABSTRACT The emergence and worldwide dissemination of antibiotic resistance genes (ARGs) compromise antibiotic therapy and are a major public health crisis. Horizontal gene transfer (HGT) plays a major role in the spread of ARGs among bacterial pathogens. Outer membrane vesicles (OMVs), which are membrane‐bound particles and naturally released by Gram‐negative bacteria, have been reported to carry a variety of cargos such as DNA, proteins and lipids. However, it remains unknown whether OMVs mediate transfer of ARGs in Campylobacter, an important foodborne pathogen whose resistance to antibiotics poses a serious threat to public health. To close this knowledge gap, we determined the role of OMVs in ARG transfer. Using a non‐conjugative plasmid (pRY112), we demonstrated that OMVs successfully transferred the plasmid from Campylobacter coli to Campylobacter jejuni. Additionally, OMVs transferred chromosomally encoded florfenicol resistance from a clinical C. coli isolate (SH89) to C. jejuni. The OMV‐mediated transfer is independent of natural transformation as both DNase I treatment (for digestion of external‐free DNA) and use of a strain deficient of natural transformation as the recipient strain did not affect OMV‐mediated ARG transfer. Transmission electron microscopy revealed direct fusion between OMVs and recipient bacterial membranes, suggesting membrane fusion as the mechanism for OMV‐mediated DNA transfer. Furthermore, we showed that OMVs derived from strains expressing a functionally‐enhanced CmeB (FE‐CmeB) transiently protect florfenicol‐susceptible C. jejuni against selection by the antibiotic. Together, these findings indicate that OMVs mediate the transfer of both plasmid‐ and chromosome‐encoded ARGs in Campylobacter and define OMVs as a novel pathway for Campylobacter to acquire antibiotic resistance via HGT.

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