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In\n                    <jats:italic>Giardia duodenalis<\/jats:italic>\n                    , EVs contribute to parasite survival, host\u2013pathogen interactions, and immune modulation. In this study, EVs were isolated from trophozoites grown in two serum\u2010free media of distinct nutritional complexity (TYI\u201033 and DMEM) and characterized to define their molecular composition, pathogenic potential, and immunogenic properties. Nanoparticle tracking analysis showed vesicles predominantly 51\u2013150\u00a0nm in diameter. Proteomic profiling identified 227 proteins in TYI\u201033 EVs and 97 in DMEM EVs, including molecules involved in metabolism, antioxidant defence, and virulence. Among these, giardipains 1\u20133, enolase, fructose\u2010bisphosphate aldolase, ornithine carbamoyltransferase, and arginine deiminase were detected. TYI\u201033 EVs induced disruption of epithelial junctions and apoptotic\u2010like changes in IEC\u20106 monolayers, consistent with contact\u2010independent cytopathic activity. To assess their immunoprotective capacity, gerbils were intranasally immunized with TYI\u201033 EVs and challenged with\n                    <jats:italic>G. duodenalis<\/jats:italic>\n                    . Immunized animals showed significant reductions in intestinal trophozoite burden and faecal cyst shedding at 7, 14, and 21 days post\u2010infection, accompanied by increased serum IL\u201017F and IL\u201022. Collectively, these findings indicate that\n                    <jats:italic>Giardia<\/jats:italic>\n                    EVs, while carrying virulence\u2010associated factors, also possess strong immunogenic properties, supporting their potential as a novel mucosal vaccine platform against giardiasis.\n                  <\/jats:p>","DOI":"10.1002\/jex2.70155","type":"journal-article","created":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T17:54:13Z","timestamp":1781200453000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Extracellular Vesicles of\n                    <i>Giardia duodenalis<\/i>\n                    : Unravelling Their Virulence Factors and Potential to Induce Protection Against Experimental 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