{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T00:05:11Z","timestamp":1761609911618,"version":"build-2065373602"},"reference-count":64,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2015,6,30]],"date-time":"2015-06-30T00:00:00Z","timestamp":1435622400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Available genomic data for the toxic, bloom-forming, benthic Ostreopsis spp. are traditionally obtained from isolates rather than from individuals originally present in environmental samples. Samples from the final phase of the first reported Ostreopsis bloom in European North Atlantic waters (Algarve, south coast of Portugal) were studied and characterized, using a culture-independent approach. In the first instance, a  microscopy-based analysis revealed the intricate complexity of the samples. Then, we evaluated the adequacy of commonly used molecular tools (i.e., primers and nuclear ribosomal markers) for the study of Ostreopsis diversity in natural samples. A PCR-based methodology previously developed to identify\/detect common Ostreopsis species was tested, including one new combination of existing PCR primers. Two sets of environmental rRNA sequences were obtained, one of them (1052 bp) with the newly tested primer set. These latter sequences encompass both the ITS1-5.8S-ITS2 region and the D1\/D2 domain of the LSU rRNA gene, leading us to an accurate identification of ITS2. In turn, this allowed us to predict and show for the first time the ITS2 secondary structure of Ostreopsis. With  92 bp in length and a two-helix structure, the ITS2 of this genus revealed to be unique among the dinoflagellates. Both the PCR approach as the phylogenetic analyses allowed to place the Ostreopsis cells observed in the samples within the O. cf. ovata phylospecies\u2019 complex, discarding the presence of O. cf. siamensis. The (phylo)genetic results point out a certain level of nucleotide sequence divergence, but were inconclusive in relation to a possible geographic origin of the O. cf. ovata population from the Algarve\u2019s bloom.<\/jats:p>","DOI":"10.3390\/toxins7072514","type":"journal-article","created":{"date-parts":[[2015,6,30]],"date-time":"2015-06-30T11:40:13Z","timestamp":1435664413000},"page":"2514-2533","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Culture-Independent Study of the Late-Stage of a Bloom of the Toxic Dinoflagellate Ostreopsis cf. ovata: Preliminary Findings Suggest Genetic Differences at the Sub-Species Level and  Allow ITS2 Structure Characterization"],"prefix":"10.3390","volume":"7","author":[{"given":"Vitor","family":"Ramos","sequence":"first","affiliation":[{"name":"CIIMAR\/CIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research,  University of Porto, Rua dos Bragas 289, 4050-123 Porto, Portugal"}]},{"given":"Daniele","family":"Salvi","sequence":"additional","affiliation":[{"name":"CIBIO, Centro de Investiga\u00e7\u00e3o em Biodiversidade e Recursos Gen\u00e9ticos, InBIO,  Universidade do Porto, Campus Agr\u00e1rio de Vair\u00e3o, 4485-661 Vair\u00e3o, Portugal"}]},{"given":"Jo\u00e3o","family":"Machado","sequence":"additional","affiliation":[{"name":"CIIMAR\/CIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research,  University of Porto, Rua dos Bragas 289, 4050-123 Porto, Portugal"}]},{"given":"Micaela","family":"Vale","sequence":"additional","affiliation":[{"name":"CIIMAR\/CIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research,  University of Porto, Rua dos Bragas 289, 4050-123 Porto, Portugal"}]},{"given":"Joana","family":"Azevedo","sequence":"additional","affiliation":[{"name":"CIIMAR\/CIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research,  University of Porto, Rua dos Bragas 289, 4050-123 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3585-2417","authenticated-orcid":false,"given":"Vitor","family":"Vasconcelos","sequence":"additional","affiliation":[{"name":"CIIMAR\/CIMAR\u2014Interdisciplinary Centre of Marine and Environmental Research,  University of Porto, Rua dos Bragas 289, 4050-123 Porto, Portugal"},{"name":"Departamento de Biologia, Faculdade de Ci\u00eancias, Universidade do Porto, Rua do Campo Alegre, 4169-007 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2015,6,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1016\/j.toxicon.2010.05.010","article-title":"World-wide occurrence of the toxic dinoflagellate genus Ostreopsis Schmidt","volume":"57","author":"Rhodes","year":"2011","journal-title":"Toxicon"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"408","DOI":"10.1016\/j.toxicon.2010.11.019","article-title":"Trends in Ostreopsis proliferation along the Northern Mediterranean coasts","volume":"57","author":"Mangialajo","year":"2011","journal-title":"Toxicon"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/j.hal.2006.02.005","article-title":"The presence of the potentially toxic genera Ostreopsis and Coolia (Dinophyceae) in the north Aegean sea, Greece","volume":"5","author":"Aligizaki","year":"2006","journal-title":"Harmful Algae"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.hal.2012.05.003","article-title":"Morphometric analysis of Ostreopsis cf. ovata cells in relation to environmental conditions and bloom phases","volume":"19","author":"Accoroni","year":"2012","journal-title":"Harmful Algae"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e57627","DOI":"10.1371\/journal.pone.0057627","article-title":"Quantitative PCR method for enumeration of cells of cryptic species of the toxic marine dinoflagellate Ostreopsis spp. in coastal waters of Japan","volume":"8","author":"Hariganeya","year":"2013","journal-title":"PLoS ONE"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1111\/j.0022-3646.1996.01053.x","article-title":"Further SEM study of marine dinoflagellates: The genus Ostreopsis (Dinophyceae)","volume":"32","author":"Faust","year":"1996","journal-title":"J. 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