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Lipids are essential molecular components of EVs but at the moment the knowledge about the lipid composition and the function of lipids in EVs is limited and as for now none lipidomic studies in <jats:italic>Giardia<\/jats:italic> EVs was described. Therefore, the focus of the current study was to conduct, for the first time, the characterization of the polar lipidome, namely phospholipid and sphingolipid profiles of <jats:italic>G<\/jats:italic>. <jats:italic>lamblia<\/jats:italic> trophozoites, microvesicles (MVs) and exosomes, using C18\u2014Liquid Chromatography\u2014Mass Spectrometry (C18-LC-MS) and Tandem Mass Spectrometry (MS\/MS). A total of 162 lipid species were identified and semi-quantified, in the trophozoites, or in the MVs and exosomes belonging to 8 lipid classes, including the phospholipid classes phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), cardiolipins (CL), the sphingolipid classes sphingomyelin (SM) and ceramides (Cer), and cholesterol (ST), and 3 lipid subclasses that include lyso PC (LPC), lyso PE (LPE) and lyso PG (LPG), but showing different abundances. This work also identified, for the first time, in <jats:italic>G<\/jats:italic>. <jats:italic>lamblia<\/jats:italic> trophozoites, the lipid classes CL, Cer and ST and subclasses of LPC, LPE and LPG. Univariate and multivariate analysis showed clear discrimination of lipid profiles between trophozoite, exosomes and MVs. The principal component analysis (PCA) plot of the lipidomics dataset showed clear discrimination between the three groups. Future studies focused on the composition and functional properties of <jats:italic>Giardia<\/jats:italic> EVs may prove crucial to understand the role of lipids in host-parasite communication, and to identify new targets that could be exploited to develop novel classes of drugs to treat giardiasis.<\/jats:p>","DOI":"10.1371\/journal.pone.0291292","type":"journal-article","created":{"date-parts":[[2023,9,8]],"date-time":"2023-09-08T17:55:04Z","timestamp":1694195704000},"page":"e0291292","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":7,"title":["Lipidome of extracellular vesicles from Giardia lamblia"],"prefix":"10.1371","volume":"18","author":[{"given":"Clarissa Perez","family":"Faria","sequence":"first","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9727-9906","authenticated-orcid":true,"given":"Barbara","family":"Ferreira","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4642-8657","authenticated-orcid":true,"given":"\u00c1gata","family":"Louren\u00e7o","sequence":"additional","affiliation":[]},{"given":"In\u00eas","family":"Guerra","sequence":"additional","affiliation":[]},{"given":"T\u00e2nia","family":"Melo","sequence":"additional","affiliation":[]},{"given":"Pedro","family":"Domingues","sequence":"additional","affiliation":[]},{"given":"Maria do Ros\u00e1rio Marques","family":"Domingues","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9846-6754","authenticated-orcid":true,"given":"Maria Teresa","family":"Cruz","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4957-7831","authenticated-orcid":true,"given":"Maria do C\u00e9u","family":"Sousa","sequence":"additional","affiliation":[]}],"member":"340","published-online":{"date-parts":[[2023,9,8]]},"reference":[{"key":"pone.0291292.ref001","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1128\/CMR.00067-12","article-title":"Transmission and epidemiology of zoonotic protozoal diseases of companion animals","volume":"26","author":"KJ Esch","year":"2013","journal-title":"Clin Microbiol Rev"},{"key":"pone.0291292.ref002","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.vetpar.2004.09.008","article-title":"The zoonotic significance and molecular epidemiology of Giardia and giardiasis","volume":"126","author":"RC Thompson","year":"2004","journal-title":"Vet. 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