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One of these mechanisms is the production of outer membrane vesicles (OMVs), nanoscale structures that bud off from bacterial cells into the extracellular space. OMVs can deliver bioactive cargoes, including enzymes, RNA and DNA, enabling functions such as cell-to-cell communication, nutrient acquisition and immunomodulation. However, the role of OMVs in beneficial host-associated microbiomes remains unclear. Here, we investigated OMV production in the gut bacteria of the western honeybee (Apis mellifera), which forms a highly conserved and stable microbial community. Using electron microscopy, fluorescence labelling and nanoparticle tracking analysis, we detected OMV production in every Gram-negative species of the bee gut microbiota that we investigated. Vesicles were observed in gut contents of wild and laboratory-inoculated bees, but absent in bees lacking a microbiota. OMVs contained nucleic acids, with more RNA than DNA. Bacterial strains varied in OMV properties, including abundance, size and zeta potential. These findings indicate that OMVs are likely significant mediators of interbacterial and host\u2013microbe interactions in the bee gut.<\/jats:p>","DOI":"10.1098\/rsbl.2026.0327","type":"journal-article","created":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:08:32Z","timestamp":1791936512000},"source":"Crossref","is-referenced-by-count":0,"title":["Discovery and characterization of outer membrane vesicles produced by the bee gut microbiota"],"prefix":"10.1098","volume":"22","author":[{"given":"Rodney P.","family":"Eyles","sequence":"first","affiliation":[{"name":"Gulbenkian Institute for Molecular Medicine 1 , Lisbon, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7999-3217","authenticated-orcid":false,"given":"Waldan K.","family":"Kwong","sequence":"additional","affiliation":[{"name":"Gulbenkian Institute for Molecular Medicine 1 , Lisbon, Portugal"},{"name":"Faculty of Medicine, University of Lisbon 2 , Lisbon, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"175","published-online":{"date-parts":[[2026,10,14]]},"reference":[{"key":"2026102010515939200_RSBL20260327C1","doi-asserted-by":"crossref","first-page":"e12507","DOI":"10.1002\/jev2.12507","article-title":"The activity of the quorum sensing regulator HapR is modulated by the bacterial extracellular vesicle (BEV)-associated protein ObfA of Vibrio cholerae","volume":"13","author":"Ebenberger","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"2026102010515939200_RSBL20260327C2","doi-asserted-by":"crossref","first-page":"R167","DOI":"10.1530\/EJE-14-0874","article-title":"Mechanisms in endocrinology: gut microbiota in patients with type 2 diabetes mellitus","volume":"172","author":"Allin","year":"2015","journal-title":"Eur. J. Endocrinol."},{"key":"2026102010515939200_RSBL20260327C3","doi-asserted-by":"crossref","first-page":"101806","DOI":"10.1016\/j.pneurobio.2020.101806","article-title":"Microbiota-gut-brain axis in health and disease: is NLRP3 inflammasome at the crossroads of microbiota-gut-brain communications?","volume":"191","author":"Pellegrini","year":"2020","journal-title":"Prog. Neurobiol."},{"key":"2026102010515939200_RSBL20260327C4","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1186\/s41232-022-00224-8","article-title":"Helicobacter pylori infection in the stomach induces neuroinflammation: the potential roles of bacterial outer membrane vesicles in an animal model of Alzheimer's disease","volume":"42","author":"Park","year":"2022","journal-title":"Inflamm. Regen."},{"key":"2026102010515939200_RSBL20260327C5","doi-asserted-by":"crossref","first-page":"14888","DOI":"10.1038\/ncomms14888","article-title":"A Pseudomonas T6SS effector recruits PQS-containing outer membrane vesicles for iron acquisition","volume":"8","author":"Lin","year":"2017","journal-title":"Nat. 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Microbes Infect."},{"key":"2026102010515939200_RSBL20260327C8","doi-asserted-by":"crossref","first-page":"e66988","DOI":"10.7554\/eLife.66988","article-title":"A novel decoy strategy for polymyxin resistance in Acinetobacter baumannii","volume":"10","author":"Park","year":"2021","journal-title":"eLife"},{"key":"2026102010515939200_RSBL20260327C9","doi-asserted-by":"crossref","first-page":"e00792-17","DOI":"10.1128\/JB.00792-17","article-title":"Vibrio cholerae outer membrane vesicles inhibit bacteriophage infection","volume":"200","author":"Reyes-Robles","year":"2018","journal-title":"J. Bacteriol."},{"key":"2026102010515939200_RSBL20260327C10","doi-asserted-by":"crossref","first-page":"e1005672","DOI":"10.1371\/journal.ppat.1005672","article-title":"A novel mechanism of host-pathogen interaction through sRNA in bacterial outer membrane vesicles","volume":"12","author":"Koeppen","year":"2016","journal-title":"PLoS Pathog."},{"key":"2026102010515939200_RSBL20260327C11","doi-asserted-by":"crossref","first-page":"1478","DOI":"10.1128\/IAI.01365-15","article-title":"Cytotoxic and inflammatory responses induced by outer membrane vesicle-associated biologically active proteases from Vibrio cholerae","volume":"84","author":"Mondal","year":"2016","journal-title":"Infect. Immun."},{"key":"2026102010515939200_RSBL20260327C12","first-page":"1","article-title":"Outer membrane vesicle-associated lipase FtlA enhances cellular invasion and virulence in Francisella tularensis LVS","volume":"6","author":"Chen","year":"2017","journal-title":"Emerg. 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