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However, the composition and stability of long RNA species, particularly messenger RNA (mRNAs) and long non\u2010coding RNAs (lncRNAs), remain largely unexplored in teleosts. Pre\u2010analytical factors such as input\u00a0plasma volume and storage conditions may influence RNA recovery, especially in low\u2010input samples like fish plasma. Here, we evaluated the effects of input sample volume and freezing storage on EV\u2010RNA yield and integrity in gilthead sea bream (\n                    <jats:italic>Sparus aurata)<\/jats:italic>\n                    plasma using the exoRNeasy commercial kit. Both Maxi\u00a0(\u223c2\u00a0mL) and Midi (\u223c1\u00a0mL) kit\u00a0formats produced comparable RNA recovery, indicating flexibility in plasma volume requirements. Freezing did not alter the overall RNA biotype composition or the identity of the dominant transcripts but resulted in a higher proportion of shorter and incomplete\u00a0isoforms, suggesting selective susceptibility to degradation of long RNAs during freezing storage.\n                  <\/jats:p>\n                  <jats:p>Using Oxford Nanopore Technologies long\u2010read sequencing, we generated the first long RNA\u2010resolved EV transcriptome for this species. EV cargo was dominated by ribosomal and metabolic mRNAs, consistent with conserved vertebrate EV signatures, and 302 distinct lncRNAs were identified, two of which accounted for nearly 90% of lncRNA\u00a0reads. Predicted interactions between abundant lncRNAs, EV mRNAs, and their\u00a0encoded ribonucleoproteins suggest coordinated packaging of RNA\u2013protein modules.<\/jats:p>\n                  <jats:p>Overall, this study validates exoRNeasy for fish plasma and provides the first comprehensive long\u2010RNA EV landscape in a teleost, providing a valuable framework for future molecular and functional studies in non\u2010model fish species.<\/jats:p>","DOI":"10.1002\/jex2.70175","type":"journal-article","created":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T10:56:32Z","timestamp":1792061792000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Evaluating Plasma\u2010Handling Effects on Extracellular Vesicle RNA Yield and Cargo Profiles in Fish: A Methodological Insight"],"prefix":"10.1002","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0572-9549","authenticated-orcid":false,"given":"Antonio","family":"Espuela\u2010Ortiz","sequence":"first","affiliation":[{"name":"Department of Animal Biology, Soil Science and Geology Universidad de La Laguna  San Crist\u00f3bal de La Laguna Tenerife Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Estrella","family":"Ant\u00f3n\u2010Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Department of Animal Biology, Soil Science and Geology Universidad de La Laguna  San Crist\u00f3bal de La Laguna Tenerife Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Louis","family":"Bonnefous","sequence":"additional","affiliation":[{"name":"Department of Animal Biology, Soil Science and Geology Universidad de La Laguna  San Crist\u00f3bal de La Laguna Tenerife Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. Virginia","family":"Mart\u00edn","sequence":"additional","affiliation":[{"name":"Canary Islands Oceanographic Center (COC) IEO\u2010CSIC  Santa Cruz de Tenerife Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Salvador","family":"Jer\u00e9z","sequence":"additional","affiliation":[{"name":"Canary Islands Oceanographic Center (COC) IEO\u2010CSIC  Santa Cruz de Tenerife Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deiene","family":"Rodr\u00edguez\u2010Barreto","sequence":"additional","affiliation":[{"name":"Department of Animal Biology, Soil Science and Geology Universidad de La Laguna  San Crist\u00f3bal de La Laguna Tenerife Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,15]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.70113"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13287\u2010024\u201004005\u20107"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12864\u2010018\u20104982\u20105"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jproteome.8b00479"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0301\u2010472X(02)00791\u20109"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.imlet.2007.07.014"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1261\/rna.052886.115"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12158"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.15670"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms22126417"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.3402\/jev.v3.23430"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598\u2010020\u201057497\u20107"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1080\/20013078.2018.1481321"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1177\/0300985813505114"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41573\u2010022\u201000521\u20104"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12144"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1093\/gigascience\/giab008"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1201\/9780429246593"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0136133"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12043"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.3390\/ncrna3010010"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nbt.2022.03.001"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1039\/D3MH00717K"},{"key":"e_1_2_11_25_1","doi-asserted-by":"crossref","unstructured":"Fox J. 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