{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T10:01:46Z","timestamp":1753869706674,"version":"3.41.2"},"reference-count":37,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,5,15]],"date-time":"2025-05-15T00:00:00Z","timestamp":1747267200000},"content-version":"vor","delay-in-days":44,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["MicrobiologyOpen"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Field collections of marine invertebrates are often accompanied by delays in preservation, which may impact microbiome composition. Here, we tested the effects of delayed preservation and relaxation methods on microbiome diversity and composition in the colonial ascidian <jats:italic>Trididemnum solidum<\/jats:italic> using 16S rRNA amplicon sequencing. Replicate samples collected from Belizean reefs were either (1) immediately preserved in ethanol (\u201ccontrol\u201d), (2) held in ambient seawater for 3\u2009h before preservation (\u201cSW\u201d), or (3) held in ambient seawater with menthol (a common pre\u2010preservation relaxation technique for ascidian identification) for 3\u2009h before preservation (\u201cSW\u2009+\u2009M\u201d). All <jats:italic>T. solidum<\/jats:italic> microbiomes were different from ambient seawater bacterioplankton and dominated by the same microbial taxa, including the genera <jats:italic>Thalassobaculum<\/jats:italic>, <jats:italic>Tistrella<\/jats:italic>, and <jats:italic>Synechocystis<\/jats:italic>. However, the 3\u2010h delay in sample preservation (SW) significantly reduced microbiome richness compared to controls (<jats:italic>p<\/jats:italic>\u2009=\u20090.028), while menthol treatment (SW\u2009+\u2009M) mitigated this diversity loss (<jats:italic>p<\/jats:italic>\u2009=\u20090.208). Microbial composition at the community level did not differ significantly for either delayed preservation method compared to controls (SW <jats:italic>p<\/jats:italic>\u2009=\u20090.054, SW\u2009+\u2009M <jats:italic>p<\/jats:italic>\u2009=\u20090.052). Taxon\u2010level shifts were rare but did occur, most notably a bloom of the facultatively anaerobic gammaproteobacterium <jats:italic>Catenococcus<\/jats:italic> that was 37x (SW) and 197x (SW\u2009+\u2009M) more abundant in delayed preservations. After a 3\u2010h preservation delay (SW), only 122 microbial taxa (1.85% of total) exhibited significantly differential abundances with controls, with menthol treatment (SW\u2009+\u2009M) reducing taxon\u2010level shifts to 65 taxa (0.98%). Our results showed that brief delays in preservation did not significantly alter community\u2010level microbiome composition and dominant taxa, with menthol exposure counteracting minor microbiome shifts associated with preservation delays.<\/jats:p>","DOI":"10.1002\/mbo3.70019","type":"journal-article","created":{"date-parts":[[2025,5,16]],"date-time":"2025-05-16T04:50:59Z","timestamp":1747371059000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Microbial Distortion? Impacts of Delayed Preservation on Microbiome Diversity and Composition in a Marine Invertebrate"],"prefix":"10.1002","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2376-230X","authenticated-orcid":false,"given":"Brenna","family":"Hutchings","sequence":"first","affiliation":[{"name":"Department of Biology &amp; Marine Biology, Center for Marine Science University of North Carolina Wilmington Wilmington North Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8737-6729","authenticated-orcid":false,"given":"Susanna","family":"L\u00f3pez\u2010Legentil","sequence":"additional","affiliation":[{"name":"Department of Biology &amp; Marine Biology, Center for Marine Science University of North Carolina Wilmington Wilmington North Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lauren","family":"Stefaniak","sequence":"additional","affiliation":[{"name":"Department of Marine Science Coastal Carolina University Conway South Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marie","family":"Nydam","sequence":"additional","affiliation":[{"name":"Life Sciences Concentration Soka University of America Aliso Viejo California USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9276-0020","authenticated-orcid":false,"given":"Patrick M.","family":"Erwin","sequence":"additional","affiliation":[{"name":"Department of Biology &amp; Marine Biology, Center for Marine Science University of North Carolina Wilmington Wilmington North Carolina USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,5,15]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2001.tb10745.x"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1000080107"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41596-019-0264-1"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0093386"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/ismej.2013.188"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-07425-z"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejar.2021.11.007"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/aem.47.1.49-55.1984"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmars.2021.805202"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.16537"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/s40168-020-00835-8"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1744-7410.2008.00153.x"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1017\/S0025315406013002"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00227-006-0378-1"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/jf011476k"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11356-022-22652-2"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0023938"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2015.01022"},{"key":"e_1_2_10_20_1","first-page":"247","volume-title":"Coral Reef Ascidians of New Caledonia","author":"Monniot C.","year":"1991"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/mbo3.1405"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1462-2920.2006.01209.x"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.2307\/1541381"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1099\/ijsem.0.005585"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0080307"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep06986"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.01541-09"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1111\/1462-2920.16249"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jembe.2010.11.004"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00508734"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.3391\/mbi.2016.7.1.13"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0037118"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.49.6.2474-2478.2005"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1021\/np200543z"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1111\/ele.13122"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms10040811"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja301735a"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1515\/biolog-2016-0102"}],"container-title":["MicrobiologyOpen"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/mbo3.70019","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,21]],"date-time":"2025-05-21T11:30:44Z","timestamp":1747827044000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/mbo3.70019"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4]]},"references-count":37,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2025,4]]}},"alternative-id":["10.1002\/mbo3.70019"],"URL":"https:\/\/doi.org\/10.1002\/mbo3.70019","archive":["Portico"],"relation":{},"ISSN":["2045-8827","2045-8827"],"issn-type":[{"type":"print","value":"2045-8827"},{"type":"electronic","value":"2045-8827"}],"subject":[],"published":{"date-parts":[[2025,4]]},"assertion":[{"value":"2024-08-30","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-05-01","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-05-15","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70019"}}