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The toxin‐producing dinoflagellate Alexandrium pseudogonyaulax has become increasingly abundant in northern European waters, replacing other Alexandrium species. A. pseudogonyaulax produces goniodomins and lytic substances, which can be cytotoxic toward other organisms, including fish, but we still know little about the environmental conditions influencing its growth and toxicity. Here, we investigated the impacts of different nitrogen sources and light intensities, common bottom‐up drivers of bloom formation, on the growth and toxin content of three A. pseudogonyaulax strains isolated from the Danish Limfjord. While the growth rates were significantly influenced by nitrogen source and light intensity, the intracellular toxin contents only showed strong differences between the exponential and stationary growth phases. Moreover, the photophysiological response of A. pseudogonyaulax showed little variation across varying light intensities, while light‐harvesting pigments were significantly more abundant under low light conditions. This study additionally highlights considerable physiological variability between strains, emphasizing the importance of conducting laboratory experiments with several algal strains. A high physiological plasticity toward changing abiotic parameters points to a long‐term establishment of A. pseudogonyaulax in northern European waters. [ABSTRACT FROM AUTHOR]

Elevated pCO2 and warming may promote algal growth and toxin production, and thereby possibly support the proliferation and toxicity of harmful algal blooms (HABs). Here, we tested whether empirical data support this hypothesis using a meta‐analytic approach and investigated the responses of growth rate and toxin content or toxicity of numerous marine and estuarine HAB species to elevated pCO2 and warming. Most of the available data on HAB responses towards the two tested climate change variables concern dinoflagellates, as many members of this phytoplankton group are known to cause HAB outbreaks. Toxin content and toxicity did not reveal a consistent response towards both tested climate change variables, while growth rate increased consistently with elevated pCO2. Warming also led to higher growth rates, but only for species isolated at higher latitudes. The observed gradient in temperature growth responses shows the potential for enhanced development of HABs at higher latitudes. Increases in growth rates with more CO2 may present an additional competitive advantage for HAB species, particularly as CO2 was not shown to enhance growth rate of other non‐HAB phytoplankton species. However, this may also be related to the difference in representation of dinoflagellate and diatom species in the respective HAB and non‐HAB phytoplankton groups. Since the proliferation of HAB species may strongly depend on their growth rates, our results warn for a greater potential of dinoflagellate HAB development in future coastal waters, particularly in temperate regions. [ABSTRACT FROM AUTHOR]

Phytoplankton and other protists in the 3 µm to 50 µm size fraction are grazed on by zooplankton and form the base of Arctic marine food webs essential for local indigenous communities. Anthropogenic climate change is increasing stratification over much of the Arctic Ocean and surrounding seas, but the influence of stratification on protist communities in more coastal regions along Eastern Hudson Bay, Hudson Strait and Ungava Bay is little known. We used 18S rRNA and rDNA amplicon sequencing during two consecutive summers (2017 and 2018) and detailed water column properties to compare the 3 µm to 50 µm protist communities under contrasting stratification regimes in the Eastern Hudson Bay Complex. We found that the surface mixed layer in Eastern Hudson Bay, which is under the influence of river runoff, was strongly stratified and dominated by mixotrophic and bacterivorous taxa, mostly the dinoflagellates Heterocapsa rotundata and Gymnodiniales spp., and a more diatom-dominated community at the Subsurface Chlorophyll Maximum (SCM), which persisted in deeper colder and more saline water. The massive sequencing effort retrieved seven putative toxic algae from the upper warmer waters of eastern Hudson Bay. These included Pseudo-nitzschia spp. and potentially harmful dinoflagellates, most notably Alexandrium sp. The persistent weaker stratification conditions in Hudson Strait and Ungava Bay in summer favored a different diatom community, dominated by Chaetoceros spp. and Thalassiosira spp., and small photosynthetic flagellates including Phaeocystis pouchetii and Micromonas polaris. As freshwater input increases and stratification intensifies in the Arctic, our findings suggest the summer dinoflagellate-based community seen in coastal Hudson Bay may also be favored in other regions receiving increased river runoff. These conditions could also favor harmful algal events. The Hudson Strait and Ungava Bay protist communities were found nearer the surface and consisted of diverse species able to profit from ongoing nutrient input due to tidal mixing. These results suggest greater resilience in this and other tidally influenced coastal Arctic Bays lacking larger rivers inputs.

Phytoplankton plays an essential role in the biogeochemical cycle because it is at the top of the food chain and is a source of oxygen. Eutrophication causes coastal areas to deteriorate as industrialization accelerates, leading to harmful algal blooms (HABs), severely affecting human and ecological health. The frequency and extent of HAB events potentially may increase due to climate change. HAB outbreaks have led to substantial losses for major coastal economies globally, and therefore have emerged as a critical research focus in environmental sciences. However, the lack of an overview of diverse factors influencing HABs complicates the cause identification and the effective countermeasure development for HAB occurrence, thereby impeding the formulation of targeted strategies for prediction and mitigation. Therefore, this review summarizes the influential factors affecting HABs in coastal areas, including water quality factors (nutrients, salinity, stratification, and biological factors) and climatological factors (temperature, pH and pCO2 , and irradiance and light). Recent work with several harmful algae species suggested that warmer temperatures combined with nutrient variation, stronger stratification, and ocean acidification may increase the growth of some toxic dinoflagellate species. Although the effects of factors vary for different species and locations, the intensification of anthropogenic activities and climate change likely will increase the frequency, outbreak scale, and severity of most coastal HABs. Because predicting HABs is crucial for understanding the factors and synergy affecting their growth and minimizing losses for decision makers and stakeholders, we reviewed models for predicting HABs, including process-based models, traditional statistical-empirical models, and data-driven machine learning models. Predicting HABs becomes more challenging as the spatial distribution of harmful algae is influenced by future climate patterns. This review paper presents a comprehensive overview of the various factors impacting HABs in coastal areas, serving as a valuable resource for decision makers and researchers to design targeted research and mitigation strategies. [ABSTRACT FROM AUTHOR]

Highlights: [•] Planktonic copepods are major grazers of harmful algae (HA) in the sea. [•] Copepods are major entry points for vectorial intoxication of pelagic food webs with HA toxins. [•] This paper reviews copepod:HA grazing studies primarily over the last decade. [•] It examines the extent to which HA toxins are copepod grazing deterrents, selective feeding on HA, impact of grazing on HA bloom development, and other issues. [•] The review also focuses on biogeographic aspects of copepod:HA grazer interactions. [Copyright &y& Elsevier]

Abstract: The Delaware Inland Bays (DIB) have experienced harmful algal blooms of dinoflagellates and raphidophytes in recent years. We used quantitative polymerase chain reaction (QPCR) techniques to investigate the community dynamics of three DIB dinoflagellates (Karlodinium veneficum, Gyrodinium instriatum, and Prorocentrum minimum) and one raphidophyte (Heterosigma akashiwo) at a single site in the DIB (IR-32) in summer 2006 relative to salinity, temperature and nutrient concentrations. We also carried out complementary laboratory culture studies. New primers and probes were developed and validated for the 18S rRNA genes in the three dinoflagellates. K. veneficum, H. akashiwo, and G. instriatum were present in almost all samples throughout the summer of 2006. In contrast, P. minimum was undetectable in late June through September, when temperatures ranged from 20 to 30°C (average 25.7°C). Dissolved nutrients ranged from 0.1 to 2.8μM PO4 3− (median=0.3μM), 0.7–30.2μM NO x (median=12.9μM), and 0–19.4μM NH4 + (median=0.7μM). Dissolved N:P ratios covered a wide range from 2.6 to 177, with a median of 40. There was considerable variability in occurrence of the four species versus nutrients, but in general P. minimum and H. akashiwo were most abundant at higher (>40) N:P ratios and dissolved nitrogen concentrations, while K. veneficum and G. instriatum were most abundant at low dissolved N:P ratios (<20) and dissolved nitrogen concentrations<10μM. The semi-continuous laboratory competition experiment used mixed cultures of K. veneficum, P. minimum, and H. akashiwo grown at dissolved N:P ratios of 5, 16, and 25. At an N:P of 16 and 25 P. minimum was the dominant alga at the end of the experiment, even at a temperature that was much higher than that at which this alga was found to bloom in the field (27°C). P. minimum and H. akashiwo had highest densities in the N:P of 25. K. veneficum grew equally well at all three N:P ratios, and was co-dominant at times at an N:P of 5. H. akashiwo had the lowest densities of the three algae in the laboratory experiment. Laboratory and field results showed both interesting similarities and significant differences in the influences of important environmental factors on competition between these harmful algal species, suggesting the need for more work to fully understand HAB dynamics in the DIB. [Copyright &y& Elsevier]

The spatial distribution of harmful microalgal taxa along the coasts of the Apulia region (Mediterranean Sea) based on results of eight years (2012–2019) of routine monitoring program and a series of sporadic observations is presented. A total number of 69 potentially harmful taxa were found during the study period. Occurrence, abundance and richness of harmful taxa (toxic, potentially toxic and high biomass producers) varied along the Apulian coasts. The occurrence of harmful species was significantly higher where most of anthropogenic pressures overlap than only a few or no pressures existed. The physical alteration of coast is the most important pressure determining this pattern. Despite the variety and the abundances of the harmful microalgae, to our knowledge, no human health problems or risks have been ever recorded, nor were full-blown consequences on marine organisms such as fish kills during algal blooms. However, blooms coupled with water discoloration phenomena could become a big issue to tourism and recreational activities that have locally important socio-economic value. • Harmful microalgae occurrence, abundance and richness varied along the Apulian coasts (Mediterranean Sea). • HABs occurrence along the whole coast increased linearly over time. • HABs occurrence was significantly higher where most of anthropogenic pressures overlap than only a few or no pressures existed. • Physical alteration of coast is the most important pressure determining occurrence pattern. [ABSTRACT FROM AUTHOR]

Harmful events caused by benthic microalgae have recently garnered a high level of attention in temperate areas, upon reports of Ostreopsis species causing fauna damage, food intoxication and respiratory illness. A resurgence of interest has also occurred in ciguatera fish poisoning (CFP), a syndrome associated with the consumption of reef fish contaminated by Ciguatoxins produced by the dinoflagellate genus Gambierdiscus in tropical and subtropical areas. To foster research on events caused by benthic microalgae, the SCOR-IOC Global Ecology and Oceanography on Harmful Algal Blooms (GEOHAB) program started a Core Research Project on Benthic Harmful Algal Blooms (BHABs). During an Open Science Meeting (OSM) in 2010, scientists studying benthic and planktonic microalgae identified the main gaps of knowledge and the research, technology and infrastructure needs in the field of the ecology and oceanography of BHABs. The issues addressed during the OSM covered the state of taxonomy, sampling methods, biogeography, genetic diversity, ecology and ecophysiology of BHABs, along with the identification of research priorities and approaches to be taken in order to improve understanding and prediction of BHABs. [ABSTRACT FROM AUTHOR]

Abstract: This paper describes the results of a harmful algal bloom (HAB) monitoring effort in the Indian River Lagoon. The goal of the study was to describe spatial and temporal variability in the distribution, frequency of occurrence, and composition of HABs, along with an examination of potential driving factors, such as hydrologic conditions and nutrient concentrations. Six sampling sites in the northern lagoon were selected for the study. The composition and abundance of the phytoplankton community was determined microscopically. Water column parameters measured in the study included salinity, water temperature, Secchi depth, total phosphorus, and total nitrogen. Dinoflagellates, diatoms or cyanobacteria dominated the phytoplankton communities in terms of biovolume at all six sampling sites. Five potential toxin producing species were observed at bloom levels during the study period, including the diatom Pseudo-nitzschia calliantha and the dinoflagellates Pyrodinium bahamense var. bahamense, Prorocentrum rathymum, Cochlodinium polykrikoides, and Karlodinium veneficum. The saxitoxin-producing dinoflagellate P. bahamense var. bahamense had the highest biovolume observed over the study period, 33.9×106 μm3 ml−1, and was present in almost half of the samples collected. Three non-toxic HAB species were observed at bloom levels of biovolume, including Akashiwo sanguinea, Peridinium quinquecorne, and Kryptoperidinium foliaceum. As part of this study, a statistical approach to estimating the probability of detecting HAB events was explored, using three common and important HAB species in the IRL, P. bahamense var. bahamense, A. sanguinea and P. calliantha, as exemplars. The potential driving factors for HAB events are discussed within the context of the hydrological, meteorological and watershed characteristics of the lagoon. [Copyright &y& Elsevier]

Harmful algal blooms (HABs) are a naturally occurring global phenomena that have the potential to impact fisheries, leisure and ecosystems, as well as posing a significant hazard to animal and human health. There is significant interest in the development and application of methodologies to study all aspects of the causative organisms and toxins associated with these events. This paper reports the first application of nanopore sequencing technology for the detection of eukaryotic harmful algal bloom organisms. The MinION sequencing platform from Oxford Nanopore technologies provides long read sequencing capabilities in a compact, low cost, and portable format. In this study we used the MinION to sequence long-range PCR amplicons from multiple dinoflagellate species with a focus on the genus Alexandrium. Primers applicable to a wide range of dinoflagellates were selected, meaning that although the study was primarily focused on Alexandrium the applicability to three additional genera of toxic algae, namely; Gonyaulax , Prorocentrum , and Lingulodinium was also demonstrated. The amplicon generated here spanned approximately 3 kb of the rDNA cassette, including most of the 18S, the complete ITS1, 5.8S, ITS2 and regions D1 and D2 of the 28S. The inclusion of barcode genes as well as highly conserved regions resulted in identification of organisms to the species level. The analysis of reference cultures resulted in over 99% of all sequences being attributed to the correct species with an average identity above 95% from a reference list of over 200 species (see Supplementary Material 1). The use of mock community analysis within environmental samples highlighted that complex matrices did not prevent the ability to distinguish between phylogenetically similar species. Successful identification of causative organisms in environmental samples during natural toxic events further highlighted the potential of the assay. This study proves the suitability of nanopore sequencing technology for taxonomic identification of harmful algal bloom organisms and acquisition of data relevant to the World Health Organisations "one health" approach to marine monitoring. [ABSTRACT FROM AUTHOR]

• First dated phylogeny of dinophytes at a global scale. • Molecular phylogenetics based on concatenated rRNA sequences. • Congruence between fossil record and molecular diversification. • Groups including harmful dinophytes predate the Cretaceous–Paleogene boundary. Unicellular dinophytes include a considerable number of harmful algae and potent producers of toxins. The dinophyte fossil record is one of the richest among protists and indicates a geologically old origin of the group dating back to the Triassic. Besides of these records, very few molecular dating studies of dinophytes have been published to date, precluding an understanding of the diversification history of these organisms. In the present study, we used first appearances in the fossil record for the best-represented dinophyte lineages, namely Gonyaulacales and Peridiniales, to calibrate a molecular phylogeny. It is inferred from ribosomal RNA sequence data covering a representative taxon sampling of all currently recognised lineages. Dinophytes may have started diversifying during main tectonic events of the supercontinent Pangaea, witnessing and surviving some of the biggest mass extinction events on Earth. Groups including harmful dinophytes originated at different points in time, but they all predate the Cretaceous–Paleogene boundary. Our chronogram provides a first time frame and may stimulate studies in future bringing molecular phylogenetics of dinophytes and their impressive fossil record together in more detail. [ABSTRACT FROM AUTHOR]

The toxin-producing dinoflagellate Alexandrium minutum is responsible for the outbreaks of harmful algae bloom (HABs). It is a widely distributed species and is responsible for producing paralytic shellfish poisoning toxins. However, the information associated with the environmental adaptation pathway and toxin biosynthesis in this species is still lacking. Therefore, this study focuses on the functional characterization of A. minutum unigenes obtained from transcriptome sequencing using the Illumina Hiseq 4000 sequencing platform. A total of 58,802 (47.05%) unigenes were successfully annotated using public databases such as NCBI-Nr, UniprotKB, EggNOG, KEGG, InterPRO and Gene Ontology (GO). This study has successfully identified key features that enable A. minutum to adapt to the marine environment, including several carbon metabolic pathways, assimilation of various sources of nitrogen and phosphorus. A. minutum was found to encode homologues for several proteins involved in saxitoxin biosynthesis, including the first three proteins in the pathway of saxitoxin biosynthesis, namely sxtA, sxtG and sxtB. The comprehensive transcriptome analysis presented in this study represents a valuable resource for understanding the dinoflagellates molecular metabolic model regarding nutrient acquisition and biosynthesis of saxitoxin.

Pinnatoxin G (PnTx-G) is a marine cyclic imine toxin produced by the dinoflagellate Vulcanodinium rugosum, frequently detected in edible shellfish from Ingril Lagoon (France). As other pinnatoxins, to date, no human poisonings ascribed to consumption of PnTx-G contaminated seafood have been reported, despite its potent antagonism at nicotinic acetylcholine receptors and its high and fast-acting toxicity after intraperitoneal or oral administration in mice. The hazard characterization of PnTx-G by oral exposure is limited to a single acute toxicity study recording lethality and clinical signs in non-fasted mice treated by gavage or through voluntary food ingestion, which showed differences in PnTx-G toxic potency. Thus, an acute toxicity study was carried out using 3 h-fasted CD-1 female mice, administered by gavage with PnTx-G (8−450 µg kg−1). At the dose of 220 µg kg−1 and above, the toxin induced a rapid onset of clinical signs (piloerection, prostration, hypothermia, abdominal breathing, paralysis of the hind limbs, and cyanosis), leading to the death of mice within 30 min. Except for moderate mucosal degeneration in the small intestine recorded at doses of 300 µg kg−1, the toxin did not induce significant morphological changes in the other main organs and tissues, or alterations in blood chemistry parameters. This acute oral toxicity study allowed to calculate an oral LD50 for PnTx-G equal to 208 μg kg−1 (95% confidence limits: 155−281 µg kg−1) and to estimate a provisional NOEL of 120 µg kg−1.

Hatcheries nationwide suffer from unexplained acute production failures, termed crashes. The microbiota of oysters relates to larval health with previous studies showing that some bacterial species have positive and others negative effects on oyster health. To investigate microbial correlates of crashes, we collected samples from every batch of oyster larvae (Crassostrea virginica) produced by the Horn Point Laboratory Oyster Hatchery since 2021 and analyzed the microbiota of 15 of those batches over their duration in the hatchery, from age of 3 to 5 days until either harvest or complete die off of the batch. Across events, die-offs generally became evident at or after six days of age. We found that the microbiota of oyster larvae appears to respond to die-off events with crashed batches having fundamentally different microbiota than good batches at age 7 to 9 and 9 to 12 days. Crashed batches were often taken over by microeukaryotes and bacterial taxa from the Protobacteria and Bacteroidetes phyla. However, this presumably opportunistic community differed between batches. Observed Vibrio species level groups did not appear to be oyster pathogens and appeared to respond to, rather than precede, crashes. The microbiota of 3 to 5 day old larvae were statistically related to whether a die-off occurred later in the larval batches' life, only when the taxa were first agglomerated to family level. The detection of two microbial species not previously known to associate with oysters, along with an increased presence of Dinophyceae, predominantly the toxin-producing Gyrodinium jinhaense, in 3 to 5 day old oyster larvae was statistically linked with subsequent batch crashes. This study suggests that the health of larval oysters shapes their microbiome. Conversely, it provides hints that the microbiome of larvae, and perhaps harmful algae, may drive hatchery crashes. [ABSTRACT FROM AUTHOR]

Dinoflagellates are essential components in marine ecosystems, and they possess two dissimilar flagella to facilitate movement. Dinoflagellates are major components of marine food webs and of extreme importance in balancing the ecosystem energy flux in oceans. They have been reported to be the primary cause of harmful algae bloom (HABs) events around the world, causing seafood poisoning and therefore having a direct impact on human health. Interestingly, dinoflagellates in the genus Symbiodinium are major components of coral reef foundations. Knowledge regarding their genes and genome organization is currently limited due to their large genome size and other genetic and cytological characteristics that hinder whole genome sequencing of dinoflagellates. Transcriptomic approaches and genetic analyses have been employed to unravel the physiological and metabolic characteristics of dinoflagellates and their complexity. In this review, we summarize the current knowledge and findings from transcriptomic studies to understand the cell growth, effects on environmental stress, toxin biosynthesis, dynamic of HABs, phylogeny and endosymbiosis of dinoflagellates. With the advancement of high throughput sequencing technologies and lower cost of sequencing, transcriptomic approaches will likely deepen our understanding in other aspects of dinoflagellates’ molecular biology such as gene functional analysis, systems biology and development of model organisms.

Marine dinoflagellate species in the genus Alexandrium are well known to produce paralytic shellfish poison as well as common coastal species with cosmopolitan distribution. However, few studies on the feeding of copepods on Alexandrium species have been conducted. The toxic dinoflagellate Alexandrium pseudogonyaulax contains goniodomin A and causes red tides in many countries. To investigate the relationship between the toxic dinoflagellate A. pseudogonyaulax and the calanoid copepods Acartia spp., we quantified the ingestion rates of Acartia spp. feeding on A. pseudogonyaulax as a function of prey concentration. Additionally, we estimated grazing coefficients by integrating data from field observations of Acartia spp. and coexisting A. pseudogonyaulax with laboratory measurements of ingestion rates obtained during this investigation. Furthermore, we compared the ingestion rates of Acartia spp. and other predators feeding on Alexandrium species as previously reported. The ingestion rates of Acartia spp. on A. pseudogonyaulax increased continuously with increasing mean prey concentration. The highest values among the ingestion rate of Acartia spp. feeding on A. pseudogonyaulax was 3,407 cells predator-1 d-1 (4,872 ng C predator-1 d-1) at the given prey concentration. The calculated grazing coefficients for Acartia spp. on A. pseudogonyaulax in Shiwha Bay, Korea, were up to 0.073 d-1. The results of this study suggest that A. pseudogonyaulax may decrease or maintain the population of Acartia spp. in marine food webs. [ABSTRACT FROM AUTHOR]

In this paper are given to know toxic species with public health importance, samples were obtained during 2010-2012 in the coast of Chiapas, México, by net (20 μm mesh) in vertical hauls (up to 15 m), fixed with Lugol’s solution and studied by light microscope bright field, Twenty-four species were documented to be present in the study área: Gymnodinium (1); Alexandrium (5); Pyrodinium (1); Phalacroma (2); Dinophysis (4);Prorocentrum (4); Karenia (4); Protoceratium (1); Gonyaulax (1); Lingulodinium (1), It was found corresponding with 64-89% of the species from coastal waters of México.

Simple Summary: Dinoflagellates are important primary producers known to biosynthesize metabolites of interest and toxins and form Harmful Algae Blooms (HABs). Water conditions such as nutrient availability, anthropogenic contaminants or pH impact dinoflagellate toxin productions, and HABs' formation remains unclear. In this review, we present the recent contributions of OMICs approaches to the investigation of dinoflagellate responses to water chemical stressors. Transcriptomic and proteomic studies highlight whole-cell strategies to cope with nutrient deficiencies. Metabolomic studies offer a great view of toxin, lipid and sugar productions under stressors. However, the confrontation of different OMICs studies is tedious, as approaches are conducted in different species. As for other model organisms, it would be interesting to use multi-OMIC approaches to build a complete view of dinoflagellate responses to chemical stressors. Overcoming the complex genome of dinoflagellates and increasing their genomic resources is therefore essential to push further. The combination of OMICs studies will provide a much-needed global view of molecular processes, which is essential to optimize the production of dinoflagellate metabolites of interest and identify markers of HABs' formation and toxin production events. Dinoflagellates are important primary producers known to form Harmful Algae Blooms (HABs). In water, nutrient availability, pH, salinity and anthropogenic contamination constitute chemical stressors for them. The emergence of OMICs approaches propelled our understanding of dinoflagellates' responses to stressors. However, in dinoflagellates, these approaches are still biased, as transcriptomic approaches are largely conducted compared to proteomic and metabolomic approaches. Furthermore, integrated OMICs approaches are just emerging. Here, we report recent contributions of the different OMICs approaches to the investigation of dinoflagellates' responses to chemical stressors and discuss the current challenges we need to face to push studies further despite the lack of genomic resources available for dinoflagellates. [ABSTRACT FROM AUTHOR]

Oyster culture is a sustainable solution to food production. However, this activity can be severely impacted by the presence and proliferation of harmful microalgae such as the benthic dinoflagellates Prorocentrum hoffmannianum and Ostreopsis cf. ovata. This study aimed to evaluate the in vitro effects of P. hoffmannianum and O. cf. ovata on immune system cells (hemocytes) of the native cultured oyster Crassostrea gasar. The direct toxicity of both dinoflagellates was first evaluated assessing hemocyte viability exposed to eight concentrations of each HAB species. No reduction in hemocyte viability was found with the exposure to cell culture or the crude extract of P. hoffmannianum , but O. cf. ovata culture induced hemocyte death in a concentration-dependent manner. Ostreopsis cf. ovata concentration that promoted half of maximal reduction in hemocyte viability (EC 50) was 779 cells mL−1. Posteriorly, hemocytes were exposed to both dinoflagellate cells and crude extracts to investigate their effects on hemocyte functional parameters. Despite no direct toxicity of the dinoflagellate cells, P. hoffmannianum extract caused a threefold increase in ROS production and decreased the phagocytosis rate by less than half. Ostreopsis cf. ovata cells and crude extracts also triggered an increase in ROS production (two-fold), but the phagocytosis rate was reduced (by half) only in response to the two lower cell concentrations. These results indicate a harmful potential of both dinoflagellates through a direct toxicity (only for O. cf. ovata) and functional impairment of hemocytes (both species) which could expose C. gasar oyster to opportunistic infections. • Cultures of Prorocentrum hoffmannianum did not cause any direct toxicity in C. gasar hemocytes. • Intracellular compounds from P. hoffmaninanum can affect C. gasar hemocytes. • Direct toxicity of Ostreopsis cf. ovata was demonstrated on C. gasar hemocytes. • O. cf. ovata caused an increase in mortality and ROS production and a decrease in phagocytosis. [ABSTRACT FROM AUTHOR]

Harmful Algal Blooms (HABs) have been classified depending on the causative organism and its impacts: non-toxic HAB (microalgae capable of affecting tourism and causing oxygen deficiency, which generates mortality of marine organisms), toxic HAB (microalgae capable of transferring toxins to the food chain), and ichthyotoxic HAB (microalgae capable of generating mechanical damage in fish). HABs represent a worldwide problem and have apparently increased in frequency, intensity, and geographic distribution at different latitudes. This review details the occurrence of HAB events in the Southeast Pacific, Chile, over a 65-year period, analysing two of the three types of HAB described: toxic and ichthyotoxic HABs. For this, we conducted a review from many different scientific sources and from the written press and social media, that have mentioned HAB events in the country. In Chile, the microalgae involved in HAB events are dinoflagellate (52%), diatoms (33%) and silicoflagellate (10%), with a total of 41 species and/or genera described in the literature. A total of 501 HAB events were recorded in Chile between 1956 and 2021, where 240 (47.9%), 238 (47.5%), 14 (2.7%), 8 (1.5%) and 1 (0.2%) event were caused by diatoms, dinoflagellate, silicoflagellate, raphidophycean and haptophyte, respectively. An apparent increase in the frequency of HAB events is observed since the first record in 1956, with a maximum of 46 events during the years 2017 and 2019. The highest incidence in fish is caused by the group of silicoflagellate, raphidophycean and haptophyte (23 events), where 10 events caused mortalities in salmon with an incidence rate of 43.4%. Unlike what is observed with diatoms and dinoflagellate, the events associated with these groups are less frequent, but hold a much higher salmon mortality rate. During the last 65 years, HAB's geographic extent shows an apparent trend to increase south-to-north. However, the identification of events is closely linked to the areas where much of the country's aquaculture is located and, therefore, it could be biased. In turn, it is observed that the apparent increase in HAB events could be associated with a greater monitoring effort after major events (e.g., after the 2016 HAB event). On the other hand, it is also recognized a lack of knowledge about harmful algae throughout the Chilean Humboldt Current system, particularly in the northern regions, such as Atacama and Coquimbo. Therefore, the total number of blooms that have occurred in fjords and channels, particularly those that have caused minor economic impacts for artisanal fishermen and the salmon and mussel farming sector, might be underestimated. [ABSTRACT FROM AUTHOR]

Highlights • Temperature and nutrient conditions affected growth and toxin production. • Production of okadaic acid (OA) was induced in all conditions in Prorocentrum hoffmannianum. • Toxin production was enhanced in P-limited conditions at 27 °C. • OA concentrations were 1 order of magnitude higher than in other DSP species. • No OA esters were detected in this strain of Prorocentrum hoffmannianum. Abstract The benthic dinoflagellate Prorocentrum hoffmannianum M.A. Faust is typical of tropical warm waters and produces biotoxins responsible for diarrhetic shellfish poisoning (DSP). In this study, the effect of temperature and nutrient limitation on growth and toxin production of P. hoffmannianum isolated from field samples collected in the Florida Keys was investigated. Batch culture experiments were ran at two temperatures (i.e. 21 ± 0.1 and 27 ± 0.1 °C) and under nitrogen-limited (14.7 μmol L−1 N-NO 3 - and 18.1 μmol L−1 P-PO 4 3-) and phosphorus-limited (441 μmol L−1 N-NO 3 - and 0.6 μmol L−1 P-PO 4 3-) levels with respect to control nutrient conditions (441 μmol L−1 N-NO 3 -and 18.1 μmol L−1 P-PO 4 3-). Both temperature and nutrient conditions significantly affected growth rates and maximum yield of P. hoffmannianum with the maximum values being recorded at the higher temperature and in the replete medium. Production of okadaic acid was induced under all conditions (from 13.5 to 859.8 pg cell−1), with values up to one order of magnitude higher than those observed in other DSP toxin producing species. Toxin production was enhanced under P limitation at 27 °C, corroborating the theory that toxin production is modulated by cell physiological conditions, which are in turn affected by a wide spectrum of factors, including environmental stressors such as nutrient availability. Toxin fraction released in the growth medium was negligible. No okadaic acid esters were detected in this strain of P. hoffmannianum. [ABSTRACT FROM AUTHOR]

A dinoflagellate identified as Prorocentrum dentatum, P. shikokuense or P. donghaiense is responsible of massive harmful events. Blooms of a species identified as P. shikokuense have been recently reported in the Mediterranean Sea, and an exotic origin, tentatively introduced by ballast waters from Asia, has been hypothesized. The molecular data based on the small-, large subunit, and internal transcriber spacers ribosomal RNA gene (SSU-, LSU-, ITS rRNA) sequences confirmed P. shikokuense in the Mediterranean Sea. The Mediterranean ribotype is identical to a subtropical North Atlantic ribotype, and with slight divergence from the numerous sequences from the Pacific Ocean. To revisit the relationship between P. shikokuense (=P. donghaiense) and P. obtusidens, we provide the first micrographs of P. dentatum and P. obtusidens, the latter collected from the type locality. Our observations indicate that P. dentatum, P. obtusidens, and P. shikokuense are three different species. Their diagnostic morphological characters are: Prorocentrum dentatum is 44-60 µm long, leaf-shaped, pointed and central posterior end, conspicuous anterior shoulder; P. obtusidens is 33-41 µm long, irregular parallelepiped, almost parallel valve margins, eccentric and pointed posterior end, moderate anterior shoulder; and P. shikokuense is <25 µm long, sunflower seed-shaped, round and centric posterior end, inconspicuous anterior shoulder. [ABSTRACT FROM AUTHOR]

Harmful algal blooms (HABs) and marine pathogens – like Vibrio spp. – are increasingly common due to climate change. These stressors affect the growth, viability and development of bivalve larvae. Little is known, however, about the potential for interactions between these two concurrent stressors. While some mixed exposures have been performed with adult bivalves, no such work has been done with larvae which are generally more sensitive. This study examines whether dinoflagellates and bacteria may interactively affect the viability and immunological resilience of blue mussel Mytilus edulis larvae. Embryos were exposed to environmentally relevant concentrations (100, 500, 2500 & 12,500 cells ml −1 ) of a dinoflagellate ( Alexandrium minutum , Alexandrium ostenfeldii , Karenia mikimotoi , Protoceratium reticulatum , Prorocentrum cordatum , P. lima or P. micans ), a known pathogen ( Vibrio coralliilyticus/neptunius -like isolate or Vibrio splendidus ; 10 5 CFU ml −1 ), or both. After five days of exposure, significant (p < 0.05) adverse effects on larval viability and larval development were found for all dinoflagellates (except P. cordatum ) and V. splendidus . Yet, despite the individual effect of each stressor, no significant interactions were found between the pathogens and harmful algae. The larval viability and the phenoloxidase innate immune system responded independently to each stressor. This independence may be related to a differential timing of the effects of HABs and pathogens. [ABSTRACT FROM AUTHOR]

Blooms of the harmful dinoflagellate Karenia brevis on the West Florida Shelf (WFS), Gulf of Mexico, are hypothesized to initiate in association with the colonial cyanobacterium Trichodesmium spp. and benefit from dissolved organic nitrogen (DON) release derived from N2-fixation by the cyanobacteria. Previous studies have detected DON release using direct experimental measurements, but there have been few studies that have followed nutrient release by in situ blooms of Trichodesmium and the associated plankton community. It was determined that long-term Trichodesmium spp. and Karenia brevis abundances on the WFS were related, following a 2-month lag. A separate Eulerian study of a Trichodesmium erythraeum bloom event was conducted over 9 days on the Great Barrier Reef. Concentrations of T. erythraeum increased over the course of the study, with coincident increases in dinoflagellate abundance and decreases in diatom abundance. Inside the bloom, concentrations of NH4+, PO43−, and DON increased significantly. The copepod grazer Macrosetella gracilis also increased in abundance as T. erythraeum numbers increased, contributing to nutrient release. Copepod grazing rates were measured, and N release rates estimated. Together, these studies show that Trichodesmium blooms have consequences for dinoflagellate abundance at both seasonal and ephemeral scales via direct and indirect N release. [ABSTRACT FROM AUTHOR]

Exploring mixotrophy of dinoflagellate species is critical to understanding red-tide dynamics and dinoflagellate evolution. Some species in the dinoflagellate genus Karenia have caused harmful algal blooms. Among 10 Karenia species, the mixotrophic ability of only two species, Karenia mikimotoi and Karenia brevis, has been investigated. These species have been revealed to be mixotrophic; however, the mixotrophy of the other species should be explored. Moreover, although K. mikimotoi was previously known to be mixotrophic, only a few potential prey species have been tested. We explored the mixotrophic ability of Karenia bicuneiformis, Karenia papilionacea, and Karenia selliformis and the prey spectrum of K. mikimotoi by incubating them with 16 potential prey species, including a cyanobacterium, diatom, prymnesiophyte, prasinophyte, raphidophyte, cryptophytes, and dinoflagellates. Cells of K. bicuneiformis, K. papilionacea, and K. selliformis did not feed on any tested potential prey species, indicating a lack of mixotrophy. The present study newly discovered that K. mikimotoi was able to feed on the common cryptophyte Teleaulax amphioxeia. The phylogenetic tree based on the large subunit ribosomal DNA showed that the mixotrophic species K. mikimotoi and K. brevis belonged to the same clade, but K. bicuneiformis, K. papilionacea, and K. selliformis were divided into different clades. Therefore, the presence or lack of a mixotrophic ability in this genus may be partially related to genetic characterizations. The results of this study suggest that Karenia species are not all mixotrophic, varying from the results of previous studies. [ABSTRACT FROM AUTHOR]

Abstract The saxitoxin-producing dinoflagellate Alexandrium minutum is a well-known microalga that causes paralytic shellfish poisoning (PSP) in many coastal regions of the world. In this study, we measured the valve movements of cultivated mussels Mytilus galloprovincialis feeding on toxic A. minutum (n = 29 mussels, shell length = 67.1 ± 3.2 mm, x̅ ± SD) or a morphologically-similar, but toxin-free phytoplankton, Heterocapsa triquetra (n = 24 mussels, shell length = 68.3 ± 2.9 mm). Phytoplankton inoculations were conducted in three sequential "pulses" intended to increase microalgal cell concentrations in a stepwise manner up to ~5000 cells l−1 over a 9-h period. M. galloprovincialis was connected to a non-invasive valvometry apparatus that concurrently measured the magnitude of valve openness once every 0.1 s. It was found that M. galloprovincialis tended to keep its valves open over the course of the experiment, regardless of the phytoplankton species present in water. Standard valve opening metrics, such as the opening duration and opening amplitude, were not significantly affected by the species of phytoplankton. However, the frequency of brief and partial valve closures (microclosures) was significantly influenced by phytoplankton species (P <.01). M. galloprovincialis subjected to toxic A. minutum exhibited 20.3 ± 0.4 (x̅ ± SEM) microclosures per 3-h pulse period, whereas those exposed to the control H. triquetra exhibited 7.9 ± 0.4 (x̅ ± SEM) microclosures. This response was detectable over the 3 h following the first inoculation pulse that provided a phytoplankton concentration of 1000 cells l−1. Our findings are consistent with growing evidence that bivalves are sensitive to very low concentrations of harmful microalgae. Deploying in situ valvometry sensors with real-time monitoring capabilities may provide an early warning of harmful algal blooms. Highlights • The behavioral response of mussels to toxic Alexandrium minutum was assessed. • Valve microclosures were more frequent in the presence of these toxic cells. • The response was manifested within 3 h at low cell concentrations. • The response might serve as an early warning of toxic A. minutum proliferations. [ABSTRACT FROM AUTHOR]

Harmful blooms of the toxic dinoflagellate Ostreopsis cf. ovata have been a recurrent phenomenon along the Mediterranean coasts in the last decades. Since first recorded, there has been a widespread belief that the extension of this dinoflagellate, well-known in tropical areas, to higher latitudes was due to global warming and the general rise of seawater temperature. Blooms of O. cf. ovata along the Conero Riviera (northern Adriatic Sea) occur between the end of the summer and the beginning of the autumn since 2006. The Ostreopsis cf. ovata abundances collected from its first record to today were analysed to better define the interannual trend of this phenomenon and its possible linking to certain climate change predictors. A significant increasing trend in the magnitude of Ostreopsis phenomenon was observed up to year 2012, then a stabilization at relatively low values was observed. This trend does not follow the incessant increase in water temperature observed during the last three decades in the Adriatic Sea, but rather recalls patterns seen in invasive species, although the provenience of O. cf. ovata in the Mediterranean Sea is still unresolved. Even if the Ostreopsis bloom in this area seems to slightly lessen in the last decade, Ostreopsis abundances still reach values of up to 103 cells cm−2 which could be harmful to human health. [ABSTRACT FROM AUTHOR]

Prorocentrum shikokuense (formerly P. donghaiense) is a pivotal dinoflagellate species associating with the HABs in the East China Sea. The complexity of its large nuclear genome hindered us from understanding its genomic characteristics. Full-length transcriptome sequencing offers a practical solution to decipher the physiological mechanisms of a species without the reference genome. In this study, we employed single-molecule real-time (SMRT) sequencing technology to sequence the full-length transcriptome of Prorocentrum shikokuense. We successfully generated 41.73 Gb of clean SMRT sequencing reads and isolated 105,249 non-redundant full-length non-chimeric reads. Our trial has led to the identification of 11,917 long non-coding RNA transcripts, 514 alternative splicing events, 437 putative transcription factor genes from 17 TF gene families, and 34,723 simple sequence repeats. Additionally, a total of 78,265 open reading frames were identified, of them 15,501 were the protein coding sequences. This dataset is valuable for annotating P. shikokuense genome, and will contribute significantly to the in-depth studies on the molecular mechanisms underlining the dinoflagellate bloom formation. [ABSTRACT FROM AUTHOR]

Ocean warming and acidification may significantly alter the distribution and intensity of harmful algal blooms as well as their effects on marine food webs. Estimating such effects rely, in part, on understanding the physiological response of individual algal species to controlled laboratory simulations of climate change conditions. Here we report the physiological response of the harmful dinoflagellate Karlodinium veneficum to the combined effects of elevated temperature and CO2 (29°C/1000 ppm CO2). We first examined these effects by comparing ambient control (25°C/441 ppm CO2) and elevated conditions under short-term (~20 generations) growth. Next, we compared the short-term elevated condition to a longer-term (~200 generations) growth scenario under the same elevated temperature and CO2. Under the short-term elevated conditions, K. veneficum growth declined, cell toxicity increased, and saturated and mono-unsaturated fatty acid (FA) composition varied significantly from ambient conditions. Meanwhile, after ~ 200 generations of growth under elevated temperature and CO2, K. veneficum carbon assimilation, growth, and cell toxicity were significantly higher than the short-term elevated treatment. Further, while total saturated FA declined, essential fatty acids increased and likely represented an adaptive temporal response to long-term exposure to high temperature and CO2. Such shifts in FA profiles and cell toxicity may possibly alter K. veneficum nutritional quality as prey and its mixotrophic behavior, thereby affecting the energy and mass transfer through the marine food webs as well as bloom dynamics. [ABSTRACT FROM AUTHOR]

The occurrence of harmful algal blooms (HABs) throughout the world has increased and poses a large threat to human health, fishery resources and tourism industries. The genus Alexandrium includes a number of toxic species associated with HABs. Therefore, it is very important to rapidly detect and monitor the harmful algae, such as Alexandrium genus. In this study, a standard curve of plasmid containing 18S rDNA-28S rDNA region from Alexandrium catenella was constructed and 5.8S rDNA sequence served as the primer of the real-time PCR. Cultured A. catenella, Alexandrium affine, Alexandrium lusitanicum and Alexandrium minutum samples were analyzed by real-time PCR using the same set of primers simultaneously. Using microscopy cells counts, 5.8S rDNA copies per cell and total DNA per cell were estimated. This assay method is promising for rapid detection of large number of Alexandrium samples. [ABSTRACT FROM AUTHOR]

• Benthic Prorocentrum from coastal and oceanic sites of the Southwest Atlantic studied. • Morphology and molecular (LSU rDNA and ITS) data of Prorocentrum species are provided. • Abundance of benthic Prorocentrum species was determined at 14 sites. • Fernando de Noronha is a hotspot of biodiversity, with 10 species registered. • A latitudinal gradient in global benthic Prorocentrum species richness is evident. In the tropical and subtropical South Atlantic Ocean, studies on the taxonomy and abundance of benthic harmful algae are scarce and the region has been largely under investigated. In this study, morphological descriptions, molecular (LSU rDNA and ITS region) and abundance data of benthic Prorocentrum species from the tropical and subtropical Southwest Atlantic and three oceanic islands are presented. Moreover, a review of global benthic Prorocentrum species richness and distribution is presented. Eleven benthic Prorocentrum species were found in Brazil. Morphological and molecular data on P. borbonicum, P. hoffmannianum, P. lima species complex and P. rhathymum were provided. Prorocentrum panamense, P. cf. caipirignum, P. cf. concavum, P. cf. norrisianum, P. emarginatum / fukuyoi / sculptile complex and two not identified species were observed using scanning electron and/or light microscopy, and morphological descriptions are presented. Prorocentrum lima species complex was found at all investigated sites, in abundances up to 2 × 104 cells g−1 FW at the Northeast Brazil, while maximum abundance of all the remaining species did not exceed 1 × 103 cells g−1 FW. The Fernando de Noronha archipelago can be considered a hotspot of benthic Prorocentrum species diversity, with ten species registered. Data compiled in the literature review shows a clear latitudinal gradient with higher species richness in tropical and subtropical regions relative to temperate areas. It is also evident that there is a bias caused by taxonomic impediment and an uneven sampling effort, with many regions still to be investigated using a combined morphological and molecular effort. Therefore, the current knowledge on the global distribution of benthic Prorocentrum species is likely underestimated. [ABSTRACT FROM AUTHOR]