1. Phytoplankton diversity in the Northern Adriatic Sea : insights and inconsistencies from microscopy and metabarcodingGiorgia Montali, Francesca Neri, Elisa Banchi, Federica Cerino, Timotej Turk Dermastia, Janja Francé, Patricija Mozetič, 2026, original scientific article Abstract: Phytoplankton plays a fundamental role in marine ecosystems and is widely used to assess environmental change. In this study, light microscopy and DNA metabarcoding approaches were used to investigate phytoplankton communities at three long-term monitoring sites in the northern Adriatic Sea. By combining the two approaches, more than 500 species were recorded. Metabarcoding detected substantially higher diversity, particularly among small and fragile organisms that are difficult to identify under a microscope, whereas microscopy was better for identifying larger species with distinctive morphology. Because species contain different amounts of genetic material, DNA data can overestimate or underestimate their true contribution to the community. Indeed, after applying correction factors that account for these differences, the DNA-based estimates became more consistent with the microscopy results for several phytoplankton groups. The differences among the samples were influenced more by the approach used than by the sampling location. Overall, combining these two approaches provides an accurate and robust picture of phytoplankton communities. In this way, it is possible to improve the reliability of marine biodiversity assessments and to support better environmental monitoring and management in coastal seas. Keywords: phytoplankton, diversity, environmental DNA, Long-Term Ecological Research, amplicon sequencing, North Adriatic Sea Published in DiRROS: 25.03.2026; Views: 289; Downloads: 220
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2. Microalgal communities in mucilage aggregates (Northern Adriatic Sea, summer 2024) based on microscopy and metabarcodingMarika Ubaldi, Francesca Neri, Giorgia Montali, Tiziana Romagnoli, Aurora Tomasini, Federica Cerino, Timotej Turk Dermastia, Patricija Mozetič, Janja Francé, Camilla Spoto, Stefano Accoroni, Cecilia Totti, 2026, original scientific article Abstract: The mucilage phenomenon consists of the appearance of large gelatinous aggregates floating in the water column. In summer 2024, this event has reappeared in the Northern Adriatic Sea (NAS) on a large scale. This study provides an integrated characterization of the microalgal community within mucilage aggregates and surrounding waters in two NAS areas (Gulf of Trieste and Conero Riviera) using light microscopy (LM), metabarcoding (MB) based on ribosomal 18S V4 and V9 markers, and scanning electron microscopy (SEM) for selected taxa identification. Mucilage aggregates acted as dynamic microbial hotspots, hosting a rich diatom community, with abundances 1–2 orders of magnitude higher than in the surrounding water. Dominant diatom species were Cylindrotheca closterium, Nitzschia spp., Nitzschia gobbii, and Thalassionema nitzschioides. Some phytoflagellates (e.g., Tetraselmis spp.) and dinoflagellates (e.g., Karlodinium veneficum, Pseliodinium fusus, and Wangodinium sinense) were detected exclusively by MB, while LM and SEM revealed species missed by other approaches. Gonyaulax fragilis, one of the species indicated as an important mucus producer, was present at the onset and throughout the phenomenon, as detected by LM and MB. The integrated approach improves knowledge of microalgal communities in NAS mucilage. Keywords: mucilage, Northern Adriatic Sea, phytoplankton, microalgae, metabarcoding, microscopy, Gonyaulax fragilis, Harmful Algal Blooms Published in DiRROS: 27.01.2026; Views: 444; Downloads: 356
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3. Comprehensive insights into Pseudo-nitzschia research in the Adriatic Sea : diverse perspectives and emerging discoveriesTimotej Turk Dermastia, Janja Francé, Jasna Arapov, Francesca Neri, Stefano Accoroni, Cecilia Totti, Federica Cerino, Maria Immacolata Ferrante, Fabrizio Bernardi Aubry, Stefania Finotto, Jelena Godrijan, Dragana Drakulović, Patricija Mozetič, 2025, review article Abstract: Pseudo-nitzschia, a globally distributed diatom genus, dominates phytoplankton communities in biomass and abundance, with some species producing domoic acid (DA), the neurotoxin responsible for Amnesic Shellfish Poisoning (ASP). The Adriatic Sea has become a hub of Pseudo-nitzschia research over the past 15 years. This review synthesizes multidisciplinary research across Adriatic sub-basins, integrating morphological, genetic, and ecological data. While ASP has never been documented in the Adriatic, DA was occasionally detected in shellfish. The production was linked to toxic species such as P. multistriata, P. calliantha, and P. galaxiae, which were found to produce DA in cultures from Adriatic isolates. Despite progress, LM-based monitoring remains prevalent, underestimating species complexity and toxigenic potential. Electron microscopy and genetic barcoding have nonetheless revealed new insights into the species composition of this genus in Adriatic. A total of 14 species have been reported so far with prominent regional disparity: the northern and middle Adriatic host higher diversity, contrasting with the southern Adriatic, where only five species were confirmed, and research remains sparse. Seasonal dynamics show summer-autumn peaks in abundance, though interannual and regional variability is notable. We present recent advances from the Adriatic including the discovery of Pseudo-nitzschia-infecting viruses, population genomic insights in P. multistriata, and metabarcoding-driven revelations of cryptic diversity. Finally, we highlight critical knowledge gaps, including the ecological drivers of toxicity and the impacts of oligotrophication on community shifts. Collaborative, high-resolution methodologies (e.g., omics, automated imaging) are urged to address emerging challenges under climate change and anthropogenic pressures. Keywords: Adriatic, Pseudo-nitzschia, diversity, ecology Published in DiRROS: 14.04.2025; Views: 1144; Downloads: 583
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4. Productivity, pressure, and new perspectives : impacts of the COVID-19 pandemic on marine early-career researchersAmanda Schadeberg, Eleanor Ford, Alina M. Wieczorek, Louse C. Gammage, María López-Acosta, Ivana Buselic, Timotej Turk Dermastia, Marcos Fontela, Cristina Galobart, Natalia Llopis Monferrer, Marek Lubośny, Stefania Piarulli, Giuseppe Suaria, 2022, original scientific article Abstract: The worldwide disruption caused by the beginning of the COVID-19 pandemic has dramatically impacted the activities of marine scientists working towards the goals of the UN Ocean Decade. As in other disciplines, marine early-career researchers (ECRs) are essential contributors to the development of novel and innovative science. Based on a survey of 322 of our peers, we show that the pandemic negatively impacted marine ECRs in ways that further exacerbate existing structural challenges such as social isolation, job insecurity, and short-term contracts, competitive funding, and work pressure. Furthermore, we find that the success and wellbeing of marine ECRs depends heavily on networking opportunities, gaining practical experience, collecting data, and producing publications, all of which were disrupted by the pandemic. Our analysis shows that those in the earliest stages of their careers feel most vulnerable to long-term career disadvantage as a result of the pandemic. This paper contributes to the empirical body of work about the impacts of the pandemic on marine science and offers recommendations on how marine ECRs should be supported to achieve the UN Ocean Decade’s goal of producing “the science we need for the ocean we want”.
Keywords: early career scientists, intersectional, institutional responses, marine science, reflexive science, SARS-CoV-2, UN Ocean Decade Published in DiRROS: 12.03.2025; Views: 989; Downloads: 949
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5. Discovery of novel and known viruses associated with toxigenic and non-toxigenic bloom forming diatoms from the Northern Adriatic SeaTimotej Turk Dermastia, Denis Kutnjak, Ion Gutiérrez-Aguirre, Corina P. D. Brussaard, Katarina Bačnik, 2024, original scientific article Abstract: Algal blooms impact trophic interactions, community structure and element fluxes. Despite playing an important role in the demise of phytoplankton blooms, only few viruses infecting diatoms have been cultured. Pseudo-nitzschia is a widespread diatom genus that commonly blooms in coastal waters and contains toxin-producing species. This study describes the characterization of a novel virus infecting the toxigenic species Pseudo-nitzschia galaxiae isolated from the northern Adriatic Sea. The ssRNA virus PnGalRNAV has 29.5 nm ± 1.2 nm icosahedral virions and a genome size of 8.8 kb. It belongs to the picorna-like Marnaviridae family and shows high specificity for P. galaxiae infecting two genetically and morphologically distinct strains. We found two genetically distinct types of this virus and screening of the global virome database revealed matching sequences from the Mediterranean region and China, suggesting its global distribution. Another virus of similar shape and size infecting Pseudo-nitzschia calliantha was found, but its genome could not be determined. In addition, we have obtained and characterized a new virus that infects Chaetoceros tenuissimus. The replicase protein of this virus is very similar to the previously described ChTenDNAV type-II virus, but it has a unique genome and infection pattern. Our study is an important contribution to the collective diatom virus culture collection and will allow further investigation into how these viruses control diatom bloom termination, carbon export and toxin release in the case of Pseudo-nitzschia. Keywords: algal blooms, diatoms, virus, PnGalRNAV, genome, Adriatic Sea, marine virology, phytoplankton ecology, marine biology Published in DiRROS: 23.10.2024; Views: 1462; Downloads: 1498
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6. Toxicity of the Diatom Genus Pseudo-nitzschia (Bacillariophyceae) : insights from toxicity tests and genetic screening in the Northern Adriatic SeaTimotej Turk Dermastia, Sonia Dall’Ara, Jožica Dolenc, Patricija Mozetič, 2022, original scientific article Abstract: Diatoms of the genus Pseudo-nitzschia H.Peragallo are known to produce domoic acid (DA), a toxin involved in amnesic shellfish poisoning (ASP). Strains of the same species are often classified as both toxic and nontoxic, and it is largely unknown whether this difference is also genetic. In the Northern Adriatic Sea, there are virtually no cases of ASP, but DA occasionally occurs in shellfish samples. So far, three species—P. delicatissima (Cleve) Heiden, P. multistriata (H. Takano) H. Takano, and P. calliantha Lundholm, Moestrup, & Hasle—have been identified as producers of DA in the Adriatic Sea. By means of enzme-linked immunosorbent assay (ELISA), high-performance liquid chromatography with UV and visible spectrum detection (HPLC-UV/VIS), and liquid chromatography with tandem mass spectrometry (LC-MS/MS), we reconfirmed the presence of DA in P. multistriata and P. delicatissima and detect for the first time in the Adriatic Sea DA in P. galaxiae Lundholm, & Moestrup. Furthermore, we attempted to answer the question of the distribution of DA production among Pseudo-nitzschia species and strains by sequencing the internal transcribed spacer (ITS) phylogenetic marker and the dabA DA biosynthesis gene and coupling this with toxicity data. Results show that all subclades of the Pseudo-nitzschia genus contain toxic species and that toxicity appears to be strain dependent, often with geographic partitioning. Amplification of dabA was successful only in toxic strains of P. multistriata and the presence of the genetic architecture for DA production in non-toxic strains was thus not confirmed. Keywords: Adriatic, dabA, domoic acid, Pseudo-nitzschia galaxiae Published in DiRROS: 16.07.2024; Views: 1216; Downloads: 710
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7. Inter-comparison of marine microbiome sampling protocolsFrancisco Pascoal, Maria Paola Tomasino, Roberta Piredda, Grazia Marina Quero, Luís Torgo, Julie Poulain, Tinkara Tinta, Timotej Turk Dermastia, 2023, original scientific article Abstract: Research on marine microbial communities is growing, but studies are hard to compare because of variation in seawater sampling protocols. To help researchers in the inter-comparison of studies that use different seawater sampling methodologies, as well as to help them design future sampling campaigns, we developed the EuroMarine Open Science Exploration initiative (EMOSE). Within the EMOSE framework, we sampled thousands of liters of seawater from a single station in the NW Mediterranean Sea (Service d'Observation du Laboratoire Arago [SOLA], Banyuls-sur-Mer), during one single day. The resulting dataset includes multiple seawater processing approaches, encompassing different material-type kinds of filters (cartridge membrane and flat membrane), three different size fractionations (>0.22 µm, 0.22–3 µm, 3–20 µm and >20 µm), and a number of different seawater volumes ranging from 1 L up to 1000 L. We show that the volume of seawater that is filtered does not have a significant effect on prokaryotic and protist diversity, independently of the sequencing strategy. However, there was a clear difference in alpha and beta diversity between size fractions and between these and “whole water” (with no pre-fractionation). Overall, we recommend care when merging data from datasets that use filters of different pore size, but we consider that the type of filter and volume should not act as confounding variables for the tested sequencing strategies. To the best of our knowledge, this is the first time a publicly available dataset effectively allows for the clarification of the impact of marine microbiome methodological options across a wide range of protocols, including large-scale variations in sampled volume. Keywords: marine microbiome, standardized sampling, inter-comparison, amplicon sequencing, microbial diversity, seawater sampling Published in DiRROS: 12.07.2024; Views: 1466; Downloads: 1019
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8. Evaluation of the rbcL marker for metabarcoding of marine diatoms and inference of population structure of selected generaTimotej Turk Dermastia, Ivano Vascotto, Janja Francé, David Stanković, Patricija Mozetič, 2023, original scientific article Abstract: Diatoms are one of the most important phytoplankton groups in the world’s oceans. There are responsible for up to 40% of the photosynthetic activity in the Ocean, and they play an important role in the silicon and carbon cycles by decoupling carbon from atmospheric interactions through sinking and export. These processes are strongly influenced by the taxonomic composition of diatom assemblages. Traditionally, these have been assessed using microscopy, which in some cases is not reliable or reproducible. Next-generation sequencing enabled us to study diversity in a high-throughput manner and uncover new distribution patterns and diversity. However, phylogenetic markers used for this purpose, such as various 18S rDNA regions, are often insufficient because they cannot distinguish between some taxa. In this work, we demonstrate the performance of the chloroplast-encoded rbcL marker for metabarcoding marine diatoms compared to microscopy and 18S-V9 metabarcoding using a series of monthly samples from the Gulf of Trieste (GoT), northern Adriatic Sea. We demonstrate that rbcL is able to detect more taxa compared to 18S-V9 metabarcoding or microscopy, while the overall structure of the diatom assemblage was comparable to the other two methods with some variations, that were taxon dependent. In total, 6 new genera and 22 new diatom species for the study region were identified. We were able to spot misidentification of genera obtained with microscopy such as Pseudo-nitzschia galaxiae, which was mistaken for Cylindrotheca closterium, as well as genera that were completely overlooked, such as Minidiscus and several genera from the Cymatosiraceae family. Furthermore, on the example of two well-studied genera in the region, namely Chaetoceros and particularly Pseudo-nitzschia, we show how the rbcL method can be used to infer even deeper phylogenetic and ecologically significant differences at the species population level. Despite a very thorough community analysis obtained by rbcL the incompleteness of reference databases was still evident, and we shed light on possible improvements. Our work has further implications for studies dealing with taxa distribution and population structure, as well as carbon and silica flux models and networks. Keywords: rbcL, metabarcoding, monitoring, diatoms, population genetics, Pseudo-nitzschia, Adriatic Published in DiRROS: 12.07.2024; Views: 1455; Downloads: 823
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9. Ecological time series and integrative taxonomy unveil seasonality and diversity of the toxic diatom Pseudo-nitzschia H. Peragallo in the northern Adriatic SeaTimotej Turk Dermastia, Federica Cerino, David Stanković, Janja Francé, Andreja Ramšak, Magda Tušek-Žnidarič, Alfred Beran, Vanessa Natali, Marina Cabrini, Patricija Mozetič, 2020, original scientific article Abstract: Pseudo-nitzschia H. Peragallo (1900) is a globally distributed genus of pennate diatoms that are important components of phytoplankton communities worldwide. Some members of the genus produce the neurotoxin domoic acid, so regular monitoring is in place. However, the identification of toxic members in routine samplings remains problematic. In this study, the diversity and seasonal occurrence of Pseudo-nitzschia species were investigated in the Gulf of Trieste, a shallow gulf in the northern Adriatic Sea. We used time series data from 2005 to 2018 to describe the seasonal and inter-annual occurrence of the genus in the area and its contribution to the phytoplankton community. On average, the genus accounted for about 15 % of total diatom abundance and peaked in spring and autumn, with occasional outbreaks during summer and large inter-annual fluctuations. Increased water temperature and decreased salinity positively affected the presence of some members of the genus, while strong effects could be masked by an unsuitable definition of the species complexes used for monitoring purposes. Therefore, combining morphological (TEM) and molecular analyses by sequencing the ITS, 28S and rbcL markers, eight species were identified from 83 isolated monoclonal strains: P. calliantha, P. fraudulenta, P. delicatissima, P. galaxiae, P. mannii, P. multistriata, P. pungens and P. subfraudulenta. A genetic comparison between the isolated strains and other strains in the Mediterranean was carried out and rbcL was inspected as a potential barcode marker in respect to our results. This is the first study in the Gulf of Trieste on Pseudo-nitzschia time series from a long-term ecological research (LTER) site coupled with molecular data. We show that meaningful ecological conclusions can be drawn by applying integrative methodology, as opposed to the approach that only considers species complexes. The results of this work will provide guidance for further monitoring efforts as well as research activities, including population genetics and genomics, associated with seasonal distribution and toxicity profiles. Keywords: Pseudo-nitzschia, morphology, phylogeny, seasonality, time series, Adriatic Sea Published in DiRROS: 18.03.2020; Views: 4938; Downloads: 1732
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