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The planktonic dinoflagellate genus Centrodinium has been understudied, with the type species C. elongatum remaining undocumented since the original description. Here, we report C. elongatum isolated from Mazatlán, Mexican Pacific. In the chains, the posterior daughter cell with an incomplete apical horn shows the morphology of C. elongatum, while the anterior daughter cell with complete epitheca corresponds to C. pulchrum. For the first time, a species of Centrodinium sensu stricto (highly laterally flattened species with horns) was cultured. An unarmored life stage, known as Murrayella ovalis, derived from the spheroplast after ecdysis. In the rDNA molecular phylogenies, C. elongatum (=C. pulchrum) nested as basal to morphologically similar species (C. eminens and C. intermedium) and as a sister group of a former Murrayella species, C. punctatum. C. elongatum differs from C. eminens and C. intermedium in the chain formation, second apical (2′) plate not being divided, horns with coarse poroid ornamentation, and missing prominent distal spinules. The taxonomy of Centrodinium sensu stricto is revised, with a discussion in the identities of C. complanatum, C. eminens, and C. maximum. The name C. deflexum is restored as a senior synonym of C. intermedium and C. ovale. [ABSTRACT FROM AUTHOR]

The Haruku Strait in the Central Moluccas is a potential marine resource located eastern Indonesia. Small pelagic fishes have been noted in abundance in the waters of this strait. Their abundance is mainly associated with the ample amount of plankton organisms serving as food for the former. To address the abundance of phytoplankton in Haruhu Strait, a study was carried out in September 2022. Sampling was achieved using a sample bottle from a depth of 1 meter. After that, the sample was filtered with a Kitahara net. Nineteen genera of phytoplankton were found in Haruku Strait during the study period. The phytoplankton abundance varied between 4,5 x 102 – 1,65 x 103 cells/l. Phytoplankton consists of Bacillariophyceae (Diatom) with 63% of the total genera, Dinoflagellata with 32 % of the total genera, and Cyanobacteria with 5%, while Chaetoceros, Rhizosolenia, Gonyaulax, Coscinodiscus, and Ceratium were the predominant phytoplankton genera in this area. The distribution pattern forms 8 groups. [ABSTRACT FROM AUTHOR]

Due to the increase in nanoplastics (NPs) abundance in aquatic environments, their effects on phytoplankton have aroused large research attention. In this study, 100 nm sized polystyrene NPs were chosen to investigate their effecting performance and mechanisms on a typical dinoflagellates Alexandrium tamarense. The results indicated the population growth and photosynthetic efficiencies of A. tamarense were significantly inhibited by NPs exposure, as well as the increase in cellular total carotenoids and paralytic shellfish toxins (PSTs). Meanwhile, the cellar ROS levels increased, corresponding to the increased activities or contents of multiple antioxidant components, including SOD, CAT, GPX, GR, GSH and GSSG. The transcriptional results support the physiological–biochemical results and further revealed the down-regulation of genes encoding the light reaction centers (PSI and PSII) and up-regulation of genes encoding the antioxidant components. Up-regulation of genes encoding key enzymes of the Calvin cycle and glycolytic pathway together with the TCA cycle could accelerate organic carbon and ATP production for A. tamarense cells resistant to NPs stress. Finally, more Glu and acetyl-CoA produced by the enhanced GSH cycle and the glycolytic pathway, respectively, accompanied by the up-regulation of Glu and Arg biosynthesis genes supported the increase in the PST contents under NPs exposure. This study established a data set involving physiological–biochemical changes and gene information about marine dinoflagellates responding to NPs, providing a data basis for further evaluating the ecological risk of NPs in marine environments. [ABSTRACT FROM AUTHOR]

The dinoflagellate Alexandrium catenella is a well-known paralytic shellfish toxin producer that forms harmful algal blooms, repeatedly causing damage to Chilean coastal waters. The causes and behavior of algal blooms are complex and vary across different regions. As bacterial interactions with algal species are increasingly recognized as a key factor driving algal blooms, the present study identifies several bacterial candidates potentially associated with Chilean Alexandrium catenella. This research narrowed down the selection of bacteria from the Chilean A. catenella culture using antibiotic treatment and 16S rRNA metabarcoding analysis. Subsequently, seawater from two Chilean coastal stations, Isla Julia and Isla San Pedro, was monitored for two years to detect Alexandrium species and the selected bacteria, utilizing 16S and 18S rRNA gene metabarcoding analyses. The results suggested a potential association between Alexandrium species and Spongiibacteraceae at both stations. The proposed candidate bacteria within the Spongiibacteraceae family, potentially engaging in mutualistic relationships with Alexandrium species, included the genus of BD1-7 clade, Spongiibbacter, and Zhongshania. [ABSTRACT FROM AUTHOR]

Many dinoflagellate species are bioluminescent, which is one of the anti-predation mechanisms in these species. In addition, dinoflagellate species experience a wide range of salinities in the ocean. However, the effects of salinity on their bioluminescence intensity has only been investigated for one species. Here, we explored the effect of salinity on the bioluminescence intensity of the heterotrophic dinoflagellate Noctiluca scintillans NSDJ2010 feeding on the chlorophyte Dunaliella salina, the heterotrophic dinoflagellate Polykrikos kofoidii PKJH1607 feeding on the dinoflagellate Alexadrium minutum, and the autotrophic dinoflagellate Alexandrium mediterraneum AMYS1807. Moreover, to determine the cell volume and growth effects on bioluminescence intensity, the cell volume and growth rate of three bioluminescent dinoflagellates were simultaneously investigated. The mean 200-s-integrated bioluminescence intensity (BL) per cell, equivalent to the total bioluminescence, of N. scintillans, P. kofoidii, and A. mediterraneum was significantly affected by salinity and increased with increasing salinity from 10 to 40. The results of the present study suggest that the total bioluminescence of N. scintillans, P. kofoidii, and A. mediterraneum in offshore and oceanic waters is greater than that in estuarine waters. [ABSTRACT FROM AUTHOR]

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]

A comparison of three analytical methods (the indirect GC‐FPD and MIMS, and direct LC‐MS/MS) for dimethylsulfoniopropionate (DMSP) measurements was conducted to assess their accuracy and reliability. The three methods showed a linear response but are distinguished by their linearity range, the largest being for MIMS. All three methods showed good precision on Alexandrium minutum samples (2–12%). The variability between the three methods when comparing analyses of A. minutum replicates was 11%, with the DMSP measurements by LC‐MS/MS being the highest. This result also confirms that indirect DMSP measurement after hydrolysis for GC or MIMS methods does not lead to an overestimation of DMSP values in A. minutum. A special focus was made on the more recent LC‐MS/MS method including further assays in sample preparation and storage from cultures of the dinoflagellate A. minutum. Dinoflagellate cells should be harvested by gentle filtration (< 5 cm Hg) or slow centrifugation (500 × g) to retrieve the largest DMSP pool. For the LC‐MS/MS method, MeOH used for cell extraction should be added prior to freezing (to prevent DMSP degradation). Samples will then be stable in frozen storage for at least 2 months. Finally, direct and indirect methods are complementary for identifying the exact DMSP fraction among dimethylsulfide‐producing compounds that compose total and particulate DMSP pools issued from newly screened organisms or environmental samples. [ABSTRACT FROM AUTHOR]

The cyst-defined extant Spiniferites elongatus Reid 1974 and Spiniferites membranaceus (Rossignol 1964) Sarjeant 1970 are environmentally significant fossil-species of the Quaternary, the former often dominating polar and subpolar assemblages. Following cyst incubation experiments and the establishment of cultures, these species were emended to incorporate information on their motile stages, and transferred to the non-fossil genus Gonyaulax Diesing 1866, as Gonyaulax elongata (Reid 1974) Ellegaard et al. 2003 and Gonyaulax membranacea (Rossignol 1964) Ellegaard et al. 2003. This unified approach to dinoflagellate nomenclature severs an important link with the fossil-genus Spiniferites Mantell 1850. We have applied dual nomenclature, as sanctioned by the International Code of Nomenclature for algae, fungi and plants, in returning these species to their previous assignments as Spiniferites elongatus and Spiniferites membranaceus, and we propose Gonyaulax ovum (Gaarder 1954) comb. nov., emend. and Gonyaulax lewisiae sp. nov., respectively, as their equivalent non-fossil species. A distinctive morphotype initially described as Rottnestia amphicavata var. amphicavata Dobell & Norris in Harland et al. 1980 is proposed as Spiniferites elongatus forma amphicavata stat. nov. [ABSTRACT FROM AUTHOR]

Dual nomenclature in dinoflagellates is supported under the current nomenclatural code for algae, fungi and plants and allows a fossil-defined (usually cyst) species to bear a name other than that of its equivalent non-fossil species, as established for example by incubation experiments. Two names can then apply to the same cyst morphotype, reflecting the separate but overlapping concepts and criteria used for fossil- and non-fossil taxa. Fossil-species are normally and logically assigned to fossil-genera and non-fossil species to non-fossil genera, a practice that facilitates dual nomenclature. Inconsistencies and ambiguities arise when binomials combine the names of fossil- with non-fossil taxa. Examples of this hybridised nomenclature and its consequences are examined, with problems identified and potential solutions discussed. Accordingly, a new non-fossil genus Lingulaulax is proposed with Lingulaulax polyedra (von Stein 1883) comb. nov. as its type and equivalent to the fossil-species Lingulodinium machaerophorum (Deflandre & Cookson 1955) Wall 1967, along with the new combination Lingulaulax milneri (Murray & Whitting 1899); the genus Lingulodinium Wall 1967 is retained in its exclusively fossil status. The non-fossil name Gonyaulax ellegaardiae Mertens et al. 2015 is validly published herein. [ABSTRACT FROM AUTHOR]

The marine dinoflagellate Alexandrium is known to form harmful algal blooms (HABs) and produces saxitoxin (STX) and its derivatives (STXs) that cause paralytic shellfish poisoning (PSP) in humans. Cell growth and cellular metabolism are affected by environmental conditions, including nutrients, temperature, light, and the salinity of aquatic systems. Abiotic factors not only engage in photosynthesis, but also modulate the production of toxic secondary metabolites, such as STXs, in dinoflagellates. STXs production is influenced by a variety of abiotic factors; however, the relationship between the regulation of these abiotic variables and STXs accumulation seems not to be consistent, and sometimes it is controversial. Few studies have suggested that abiotic factors may influence toxicity and STXs-biosynthesis gene (sxt) regulation in toxic Alexandrium, particularly in A. catenella, A. minutum, and A. pacificum. Hence, in this review, we focused on STXs production in toxic Alexandrium with respect to the major abiotic factors, such as temperature, salinity, nutrients, and light intensity. This review informs future research on more sxt genes involved in STXs production in relation to the abiotic factors in toxic dinoflagellates. [ABSTRACT FROM AUTHOR]

Many dinoflagellates produce bioluminescence. To estimate the intensity of bioluminescence produced by populations of the heterotrophic dinoflagellates Noctiluca scintillans and Polykrikos kofoidii and autotrophic dinoflagellate Alexandrium mediterraneum in Korean waters, we measured cellular bioluminescence intensity as a function of water temperature and calculated population bioluminescence intensity with cell abundances and water temperature. The mean 200-second-integrated bioluminescence intensity per cell (BLcell) of N. scintillans satiated with the chlorophyte Dunaliella salina decreased continuously with increasing water temperature from 5 to 25°C. However, the BLcell of P. kofoidii satiated with the mixotrophic dinoflagellate Alexandrium minutum continuously increased from 5 to 15°C but decreased at temperatures exceeding this (to 30°C). Similarly, the BLcell of A. mediterraneum continuously increased from 10 to 20°C but decreased between 20 and 30°C. The difference between highest and lowest BLcell of N. scintillans, P. kofoidii, and A. mediterraneum at the tested water temperatures was 3.5, 11.8, and 21.0 times, respectively, indicating that water temperature clearly affected BLcell. The highest estimated population bioluminescence intensity (BLpopul) of N. scintillans in Korean waters in 1998-2022 was 4.22 × 1013 relative light unit per liter (RLU L-1), which was 1,850 and 554,000 times greater than that of P. kofoidii and A. mediterraneum, respectively. This indicates that N. scintillans populations produced much brighter bioluminescence in Korean waters than the populations of P. kofoidii or A. mediterraneum. [ABSTRACT FROM AUTHOR]

This study investigated recent seasonal changes in the phytoplankton community in the southern coast of Korea, analyzing monthly samples collected at four stations (St.1-4) in Jinhae Bay from 2017 to 2022. Environmental factors except nutrients were similar each station. Water temperature ranged from 6.0 to 29.0°C and DO ranged from 2.7 to 11.6 mg L-1 with salinity ranging from 25.9 to 33.8 psu. There was no significant differences in environmental factors between stations except for nutrients. Phytoplankton cell concentrations ranged from 5.0×10³ cells L-1 to 7×105 cells L-1, with patterns similar to chlorophyll-a at each site. Phytoplankton taxa identified here were 42 genera and 77 species, including 49 diatoms, 27 dinoflagellates, and one Dictyochophyceae species. Diatoms dominated throughout all monitoring stations and seasons; however, dinoflagellates exhibited sporadic patterns during spring seasons. Nine harmful phytoplankton were identified, including one diatome Psuedo-nitzschia and eight dinoflagellate Alexandrium spp., Akashiwo sanguinea, Margalefidinium polykrikoides, Dinophysis spp., Gymnodinium spp., Pheopolykrikos hartmannii, Polykrikos kofoidii and Prorocentrum spp. The appearance pattern of the harmful phytoplankton showed a high correlation with seasonal factors. This study provides fundamental data on the composition of phytoplankton and their relationship with environmental factors in Jinhae Bay. In addition, they may be useful for understanding long-term changes in harmful phytoplankton in the southern coasts of Korea. [ABSTRACT FROM AUTHOR]

Extant species of the dinoflagellate genus Gonyaulax are capable of producing resting cysts morphologically similar to different cyst-based genera, and their cyst-theca relationships are far from resolved. Here we have carried out germination experiments on several living cysts that resemble Spiniferites delicatus from the subtropical regions of China and Mexico. Both cyst and theca morphology were examined by light and scanning electron microscopy. A new species, Gonyaulax pospelovana, is described, characterized by a cingulum displacement and an overhang of twice its width, and two short antapical spines. The cysts of G. pospelovana had a granular surface and gonal processes with petaloid tips. Maximum likelihood and Bayesian inference analyses based on LSU and SSU rRNA gene sequences revealed that strains identified as G. pospelovana were monophyletic, forming a sister clade to Gonyaulax ellegaardiae and several presumable strains of Gonyaulax spinifera. One Chinese strain and two Mexican strains of G. pospelovana were examined for yessotoxin production using LC-MS/MS, but were not found to produce a detectable amount of toxins. Metabarcoding targeting the 18S V4 rRNA gene was performed on monthly collected samples in Xiamen Bay, China. A ZOTU (zero-radius operational taxonomic units) was detected that was consistent with G. pospelovana. Its maximum abundance was recorded in summer. Additionally, an OTU was identified as G. pospelovana from the Tara Oceans metabarcoding data, which occurred in the Indian and Pacific oceans at temperatures ranging from 28°C to 31°C, suggesting that it is a warm water species. [ABSTRACT FROM AUTHOR]

In recent years, environmental DNA (eDNA) has emerged as a promising tool for detecting microorganisms in the natural environment. One specific application of eDNA technology is detecting and monitoring Harmful Algal Blooms (HABs), which can cause significant ecological and economic damage. In addition, quantitative analysis using eDNA is limited by the lack of optimized and validated Standard Curves. In this study, we aimed to optimize and validate a concentration-based Standard Curve for the quantitative analysis of HABs using eDNA. Instead of using DNA dilutions, which are commonly used in quantitative PCR (qPCR), the toxic red tide Alexandrium catenella was cultured and counted under a microscope, and Standard Curves were prepared by DNA extraction after dilution. By adjusting experimental conditions such as concentration, PCR amplification cycle, and specific primer design, we obtained an optimal Standard Curve with an R2 value close to 1. This Standard Curve showed similar results to those obtained using DNA dilution-based methods. Simultaneous quantification of eDNA from clonal Alexandrium catenella cultured in the laboratory and Alexandrium species from field samples showed a higher correlation with cell dilution than with DNA dilution. These results provide evidence that each Alexandrium spp. cells have a different amount of DNA. Consequently, when using a cell dilution-based Standard Curve to quantify Alexandrium spp. the Standard Curve is better determined by a dilution method based on cell counting. Overall, this study provides a useful method to optimize and validate concentration-based Standard Curves for eDNA-based quantitative analysis of HABs. [ABSTRACT FROM AUTHOR]

Species of the dinoflagellate genus Alexandrium are marine and planktonic forms, widely distributed, and some are recognized to form harmful algal blooms and to produce saxitoxins causing Paralytic Shellfish Poisoning in humans, and other toxins. We studied the species composition of Alexandrium in tropical and subtropical coastal areas of the Mexican Pacific: from the southern Gulf of California to the Gulf of Tehuantepec. Eleven Alexandrium species were identified, described, and illustrated using light microscopy, and occasionally scanning electron microscopy for certain species. Additionally, the genetic characterization of seven strains and four species, was performed, using sequences of the D1/D2 LSU rDNA and ITS regions. We identified the species (morphospecies): Alexandrium affine, A. gaarderae, A. globosum, A. leei, A. margalefii, A. minutum, A. monilatum, A. pseudogonyaulax, A. tamarense, A. tamiyavanichii, and A. tropicale. Of these, A. affine, A. leei, A. minutum, A. monilatum, A. pseudogonyaulax, A. tamarense and A. tamiyavanichii have been widely recognized as harmful algae. Alexandrium gaarderae, A. globosum and A. tropicale are new records for the Mexican Pacific. This is the first morphological documentation of A. pseudogonyaulax. Future studies of the genus might increase its species richness if more cultures are established and metabarcoding approach is used. [ABSTRACT FROM AUTHOR]

In red-tide dynamics, one species often predominates microalgal communities after severe competition among several species. In Korean waters, Prorocentrum donghaiense, Tripos fusus, Alexandrium fraterculus, and Margalefidinium polykrikoides form red tides, which sometimes cause massive fish kills and economic losses. When two of these species were cultivated together, M. polykrikoides inhibited the growth of A. fraterculus, P. donghaiense, and T. fusus at a M. polykrikoides abundance of 610, 2230, and 410 cells mL−1, respectively. Alexandrium fraterculus inhibited the growth of P. donghaiense and T. fusus at a A. fraterculus abundance of 840 and 10 cells mL−1, respectively. Prorocentrum donghaiense inhibited the growth of T. fusus at a P. donghaiense abundance of 2020 cells mL−1. Our field data on the abundance of these species in 2014 show that the abundances of A. fraterculus, P. donghaiense, and T. fusus were less than 10 cells mL−1 when that of M. polykrikoides exceeded 500 cells mL−1. Therefore, the red-tide formation of one of these four species can be affected by the high abundance of the others, which is an example of density-dependent inhibition. [ABSTRACT FROM AUTHOR]

The dinoflagellate genus Alexandrium hosts many toxigenic species with remarkable negative influence on ecosystems and human health by developing harmful algal blooms (HABs). Despite 34 Alexandrium species having been characterized with unique taxonomical features, relatively few Alexandrium species have been identified and traced in ecological studies, largely due to the limited resolution of morphology-based methodologies and demanding workload associated with ecological studies. Thus, both diversity and spatial–temporal dynamics of Alexandrium species in marine ecosystems remain poorly resolved and understood, leading to severe under appreciation of Alexandrium species in ecosystems. In this project, metabarcoding of the 18S rDNA V4 region was applied for the first time to quantitatively ascertain biodiversity and spatial–temporal dynamics of Alexandrium species in Jiaozhou Bay, a semi-enclosed model ecosystem in Qingdao, China. Eight known Alexandrium species (A. affine, A. andersonii, A. catenella, A. leei, A. minutum, A. ostenfeldii, A. pacificum, and A. pohangense), one unidentified Alexandrium species, and possibly three additional Alexandrium species (A. hiranoi, A. pseudogonyaulax, A. insuetum) were successfully identified in this study, compared to only four previously reported Alexandrium species in Jiaozhou Bay, demonstrating the competitive strength of metabarcoding technology in ecological research of Alexandrium species. Quantitative analysis showed that most Alexandrium species displayed strong temporal dynamics with high abundance in months with relative high temperature. In particular, Alexandrium affine displayed a strong preference from June to December, peaking in September, suggesting that temperature is an important driving factor for the temporal dynamics of Alexandrium species. However, Alexandrium catenella displayed preference from January to May. Metabarcoding analysis could be widely applied for detection and study of spatial–temporal dynamics of Alexandrium species. [ABSTRACT FROM AUTHOR]

The marine dinoflagellate Alexandrium is known to form harmful algal blooms (HABs) and produces saxitoxin (STX) and its derivatives (STXs) that cause paralytic shellfish poisoning (PSP) in humans. Cell growth and cellular metabolism are affected by environmental conditions, including nutrients, temperature, light, and the salinity of aquatic systems. Abiotic factors not only engage in photosynthesis, but also modulate the production of toxic secondary metabolites, such as STXs, in dinoflagellates. STXs production is influenced by a variety of abiotic factors; however, the relationship between the regulation of these abiotic variables and STXs accumulation seems not to be consistent, and sometimes it is controversial. Few studies have suggested that abiotic factors may influence toxicity and STXs-biosynthesis gene (sxt) regulation in toxic Alexandrium, particularly in A. catenella, A. minutum, and A. pacificum. Hence, in this review, we focused on STXs production in toxic Alexandrium with respect to the major abiotic factors, such as temperature, salinity, nutrients, and light intensity. This review informs future research on more sxt genes involved in STXs production in relation to the abiotic factors in toxic dinoflagellates.

In the framework of a monitoring program of harmful microalgae from the marine coastal waters of Buenos Aires Province to mitigate the impacts of harmful algal blooms on the aquaculture and marine life and to protect human health, a strain of Alexandrium affine (LPCc012) was isolated. Alexandrium affine is a worldwide distributed species reported as harmful algal bloom-forming, with toxin- and non-toxin-producing strains. Field and cultured materials from Buenos Aires Province were examined using light and scanning electron microscopy, morphologically and genetically (LSU rDNA partial sequencing) characterized, and compared with material from different regions worldwide. Toxin analyses using ultra-performance liquid chromatography with fluorescence detection and liquid chromatography-mass spectrometry methods failed to detect any paralytic or lipophilic shellfish toxins. This is the first record of A. affine from Argentina. [ABSTRACT FROM AUTHOR]

The Galician Rías (NW Iberian Peninsula) are an important shellfish aquaculture area periodically affected by toxic episodes often caused by dinoflagellates such as Dinophysis acuminata and Alexandrium minutum, among others. In turn, water discolorations are mostly associated with non-toxic organisms such as the heterotrophic dinoflagellate Noctiluca scintillans, a voracious non-selective predator. The objective of this work was to study the biological interactions among these dinoflagellates and their outcome in terms of survival, growth and toxins content. To that aim, short experiments (4 days) were carried out on mixed cultures with N. scintillans (20 cells mL−1) and (i) one strain of D. acuminata (50, 100 and 500 cells mL−1) and (ii) two strains of A. minutum (100, 500 and 1000 cells mL−1). Cultures of N. scintillans with two A. minutum collapsed by the end of the assays. Both D. acuminata and A. minutum exposed to N. scintillans arrested its growth, though feeding vacuoles in the latter rarely contained any prey. Toxin analyses at the end of the experiment showed an increase in intracellular OA levels in D. acuminata and a significant reduction in PSTs in both A. minutum strains. Neither OA nor PSTs were detected in N. scintillans. Overall, the present study indicated that the interactions among them were ruled by negative allelopathic effects. [ABSTRACT FROM AUTHOR]

The armoured dinoflagellate Alexandrium can be found throughout many of the world's temperate and tropical marine environments. The genus has been studied extensively since approximately half of its members produce a family of potent neurotoxins, collectively called saxitoxin. These compounds represent a significant threat to animal and environmental health. Moreover, the consumption of bivalve molluscs contaminated with saxitoxin poses a threat to human health. The identification of Alexandrium cells collected from sea water samples using light microscopy can provide early warnings of a toxic event, giving harvesters and competent authorities time to implement measures that safeguard consumers. However, this method cannot reliably resolve Alexandrium to a species level and, therefore, is unable to differentiate between toxic and non-toxic variants. The assay outlined in this study uses a quick recombinase polymerase amplification and nanopore sequencing method to first target and amplify a 500 bp fragment of the ribosomal RNA large subunit and then sequence the amplicon so that individual species from the Alexandrium genus can be resolved. The analytical sensitivity and specificity of the assay was assessed using seawater samples spiked with different Alexandrium species. When using a 0.22 µm membrane to capture and resuspend cells, the assay was consistently able to identify a single cell of A. minutum in 50 mL of seawater. Phylogenetic analysis showed the assay could identify the A. catenella, A. minutum, A. tamutum, A. tamarense, A. pacificum, and A. ostenfeldii species from environmental samples, with just the alignment of the reads being sufficient to provide accurate, real-time species identification. By using sequencing data to qualify when the toxic A. catenella species was present, it was possible to improve the correlation between cell counts and shellfish toxicity from r = 0.386 to r = 0.769 (p ≤ 0.05). Furthermore, a McNemar's paired test performed on qualitative data highlighted no statistical differences between samples confirmed positive or negative for toxic species of Alexandrium by both phylogenetic analysis and real time alignment with the presence or absence of toxins in shellfish. The assay was designed to be deployed in the field for the purposes of in situ testing, which required the development of custom tools and state-of-the-art automation. The assay is rapid and resilient to matrix inhibition, making it suitable as a potential alternative detection method or a complementary one, especially when applying regulatory controls. [ABSTRACT FROM AUTHOR]

Gonyaulax polygramma, a bloom-forming dinoflagellate, has been repeatedly observed along the southeastern Arabian Sea in recent years. During our study in October 2021, a patch of reddish-brown water was observed in the nearshore waters off Kannur (southwest coast of India) and later identified as Gonyaulax polygramma using scanning electron microscopy (SEM) and HPLC-based phytoplankton marker pigments. Gonyaulax polygramma accounted for 99.4% of the phytoplankton abundance at the bloom location, with high concentrations of peridinin and chlorophyll-a at the study site. High concentration of SiO42− was observed at the bloom site, while other nutrients were lower than the previously reported values. The bloom of Gonyaulax polygramma also resulted in high concentrations of dimethylsulfide, an anti-greenhouse gas, at the bloom site. In addition to onsite observation, Sentinel-3 satellite data was also used in the detection and validation of the observed bloom using the NDCI index. From the satellite image, it was evident that the bloom persisted at the mouth of the rivers during the study period. Since the red tide of Gonyaulax polygramma has been observed recurrently in the southeastern Arabian Sea, it is proposed to use satellites to detect and monitor the bloom on a routine basis. [ABSTRACT FROM AUTHOR]

The emerging threat of ciguatera poisoning (CP) in Europe has been associated with fish captured in the Canary Islands (Spain) and Selvagens Islands (Portugal). The first are heavily populated islands where numerous scientific studies have been carried out. Conversely, the Selvagens Islands as a nature reserve with low human pressure have been rarely surveyed in terms of the marine benthic microalgae, including the epiphytic ciguatera-causing dinoflagellate species. To investigate the harmful microalgal diversity of the Selvagens Islands, a scientific cruise to these remote islands took place in September, 2018. The Gambierdiscus species composition and distribution, and the associated epiphytic dinoflagellate community, were assessed using artificial substrate devices. Gambierdiscus cells were found in all samples, reaching concentrations of up to 725 cells 100 cm–2. G. australes was the only species identified after morphological and molecular analysis of the retrieved cultures. Species identification was confirmed by molecular characterization based on the LSU D8–D10 region. Nevertheless, phylogenetic studies indicated that some strains diverged from the G. australes clade suggesting genetic differentiation. Toxicity was estimated by neuro-2a cell-based assay in four strains, ranging from 2.46–83 fg of CTX1B eq. cell–1. The epiphytic dinoflagellate community that co-occurred with Gambierdiscus comprised other toxic or potentially toxic dinoflagellates, such as Ostreopsis, Prorocentrum, Amphidinium and Coolia species. Oceanographic and meteorological data were also obtained to characterize the occurrence of Gambierdiscus. This study is the first stage in understanding the role of the Selvagens Islands in the incubation and proliferation of the ciguatera-causing dinoflagellates Gambierdiscus in the NE Atlantic. The Selvagens Islands are a ciguatera hotspot in Europe. Gambierdiscus australes was the only species observed in the Selvagens Islands. Strains diverging from the G. australes clade suggest genetic differentiation. [ABSTRACT FROM AUTHOR]

Phytoplankton‐bacteria interactions represent the evolution of complex cross‐kingdom networks requiring niche specialization of diverse microbes. Unraveling this co‐evolutionary process has proven challenging because microbial partnerships are complex, and their assembly can be dynamic as well as scale‐ and taxon‐dependent. Here, we monitored long‐term experimental evolution of phytoplankton‐bacteria interactions by reintroducing the intact microbiome into an axenized dinoflagellate Alexandrium tamarense to better understand microbiome assembly dynamics and how microbiome composition could shift and stabilize over 15 months. We examined host functioning by growth rate, photosynthetic capability, cell size, and other physiological signatures and compared it to associated microbial communities determined by 16S rRNA gene sequences. Our results showed that microbiome reconstitution did not restore the intact microbiome, instead a distinct microbial community shift to Roseobacter clade was observed in the re‐established cultures. In‐depth comparisons of microbial interactions revealed no apparent coupling between host physiology and specific bacterial taxa, indicating that highly represented, abundant taxa might not be essential for host functioning. The emergence of highly divergent Roseobacter clade sequences suggests fine‐scale microbial dynamics driven by microdiversity could be potentially linked to host functioning. Collectively, our results indicate that functionally comparable microbiomes can be assembled from markedly different, highly diverse bacterial taxa in changing environments. [ABSTRACT FROM AUTHOR]

HABs pose a threat to coastal ecosystems, the economic sector and human health, and are expanding globally. However, their influence on copepods, a major connector between primary producers and upper trophic levels, remains essentially unknown. Microalgal toxins can eventually control copepod survival and reproduction by deterring grazing and hence reducing food availability. We present several 24-h experiments in which the globally distributed marine copepod, Acartia tonsa, was exposed to different concentrations of the toxic dinoflagellate, Alexandrium minutum, grown under three N:P ratios (4:1, 16:1 and 80:1), with the simultaneous presence of non-toxic food (the dinoflagellate Prorocentrum micans). The different N:P ratios did not affect the toxicity of A. minutum, probably due to the low toxicity of the tested strain. Production of eggs and pellets as well as ingested carbon appeared to be affected by food toxicity. Toxicity levels in A. minutum also had an effect on hatching success and on the toxin excreted in pellets. Overall, A. minutum toxicity affected the reproduction, toxin excretion and, to an extent, the feeding behavior of A. tonsa. This work indicates that even short-term exposure to toxic A. minutum can impact the vital functions of A. tonsa and might ultimately pose serious threats to copepod recruitment and survival. Still, further investigation is required for identifying and understanding, in particular, the long-term effects of harmful microalgae on marine copepods. [ABSTRACT FROM AUTHOR]

The more frequent occurrence of marine harmful algal blooms (HABs) and recent problems with newly-described toxins in Puget Sound have increased the risk for illness and have negatively impacted sustainable access to shellfish in Washington State. Marine toxins that affect safe shellfish harvest because of their impact on human health are the saxitoxins that cause paralytic shellfish poisoning (PSP), domoic acid that causes amnesic shellfish poisoning (ASP), diarrhetic shellfish toxins that cause diarrhetic shellfish poisoning (DSP) and the recent measurement of azaspiracids, known to cause azaspiracid poisoning (AZP), at low concentrations in Puget Sound shellfish. The flagellate, Heterosigma akashiwo, impacts the health and harvestability of aquacultured and wild salmon in Puget Sound. The more recently described flagellates that cause the illness or death of cultivated and wild shellfish, include Protoceratium reticulatum, known to produce yessotoxins, Akashiwo sanguinea and Phaeocystis globosa. This increased incidence of HABs, especially dinoflagellate HABs that are expected in increase with enhanced stratification linked to climate change, has necessitated the partnership of state regulatory programs with SoundToxins, the research, monitoring and early warning program for HABs in Puget Sound, that allows shellfish growers, Native tribes, environmental learning centers and citizens, to be the "eyes on the coast". This partnership enables safe harvest of wholesome seafood for consumption in the region and helps to describe unusual events that impact the health of oceans, wildlife and humans. [ABSTRACT FROM AUTHOR]

Alien invasive algae along with ship ballast water have posed a serious threat to China's marine ecological security. A clear understanding of the geographical distribution of invasive species and their response to climate change can provide a scientific basis for their prevention and control. In this study, combined with environmental variables and distribution data, MaxEnt was used to predict the potential geographical distribution and change trend of Alexandrium in the four major sea areas of China currently, 2040s (2040-2050) and 2090s (2090-2100), reveal the dominant environmental factors affecting the geographical distribution, analyze the migration trends of centroids the highly suitable area, and clarify the key control sea areas. The results showed that: 1) The AUC values of all models were significantly higher than random models, and the Kappa statistics of all models were higher than "general", demonstrated that the prediction results were available. 2) The most important environmental variable affecting the geographical distribution of Alexandrium was the Temperature. Range (bio24), and its suitable range was 23.43-31.52°C. 2) At present, in the corresponding sea areas of Nantong, Yancheng, Ningbo, Jiaxing, Dongying, Binzhou, Cangzhou, Tianjin, Tangshan, Qinhuangdao, Shenzhen, Dongguan and Zhongshan, special attention should be paid to the control of Alexandrium. 3) Under the future climate change scenarios, excepted in the South China Sea, the total and highly suitable area of Alexandrium showed an increasing trend. 4) Under climate change scenarios, in the Yellow Sea, the centroids of the highly suitable area would move to the southeast. In the Bohai Sea and the East China Sea, the centroid showed a trend of moving to different latitudes and directions. In the South China Sea, the centroids of of the highly suitable area and total suitable area of Alexandrium would both move to the southwest. Our results can provide a theoretical basis for the prevention and control of Alexandrium. [ABSTRACT FROM AUTHOR]

(1) Background: Harmful algal blooms (HABs) can negatively impact marine ecosystems, but few studies have evaluated the microbial diversity associated with HABs and its potential role in the fates of these proliferations. (2) Methods: Marine prokaryotic diversity was investigated using high-throughput sequencing of the 16S rRNA gene during the bloom declines of two dinoflagellates detected in the summer of 2019 along the northern and southern Tunisian coasts (South Mediterranean Sea). The species Gymnodinium impudicum (Carthage, Tunis Gulf) and Alexandrium minutum (Sfax, Gabes Gulf) were identified using microscopy and molecular methods and were related to physicochemical factors and prokaryotic compositions. (3) Results: The abundance of G. impudicum decreased over time with decreasing phosphate concentrations. During the G. impudicum bloom decay, prokaryotes were predominated by the archaeal MGII group (Thalassarchaeaceae), Pelagibacterales (SAR11), Rhodobacterales, and Flavobacteriales. At Sfax, the abundance of A. minutum declined with decreasing phosphate concentrations and increasing pH. At the A. minutum peak, prokaryotic communities were largely dominated by anoxygenic phototrophic sulfur-oxidizing Chromatiaceae (Gammaproteobacteria) before decreasing at the end of the survey. Both the ubiquitous archaeal MGII group and Pelagibacterales were found in low proportions during the A. minutum decay. Contrary to the photosynthetic Cyanobacteria, the photo-autotrophic and -heterotrophic Rhodobacterales and Flavobacteriales contents remained stable during the dinoflagellate bloom decays. (4) Conclusions: These results indicated changes in prokaryotic community diversity during dinoflagellate bloom decays, suggesting different bacterial adaptations to environmental conditions, with stable core populations that were potentially able to degrade HABs. [ABSTRACT FROM AUTHOR]

Based on biological traits, the ecological quality status under the pressure of two harmful algal bloom (HAB) species was evaluated using functional units (FUs) of periphytic protozoan communities. Five treatments with different concentrations of Alexandrium tamarense and Gymnodinium catenatum, i.e., 100, 102, 103, 104, and 105 cells ml−1, were used. A total of 20 FUs were identified from 25 test protozoan species. Among these FUs, vagile algivores with large sizes showed a decreasing trend (i.e., in diversity and abundance) with increasing concentrations of algae, while vagile bacterivores and non-selectives with small sizes dominated at concentrations of 104 cells ml−1 of both algal species. Ellipse tests on pair-wise functional distinctness indices revealed a significant departure of test protozoan communities from an expected functional distinctness breadth when algal concentrations exceeded 104 cells ml−1. Based on these findings, it was concluded that FUs of periphytic protozoa may be a useful tool for evaluating the effects of HABs on ecological quality status in marine ecosystems. [ABSTRACT FROM AUTHOR]

To explore the effects of eutrophic deep waters offshore on red-tide outbreaks, waters were collected from 0, 20, and 40 m depths at an offshore station in the South Sea of Korea, and the growth rate of each major red-tide dinoflagellate Prorocentrum donghaiense, Tripos furca, Alexandrium fraterculus, and Margalefidinium polykrikoides in the waters was measured. No species grew at dissolved inorganic nitrogen concentrations (DIN) of < 7.0–8.4 µM, but the growth rates of all four species rapidly increased and became saturated at 12–15 µM DIN. On July 31, 2020, the DIN was 4.1–4.6 µM in 0–10 m depth waters but 8.9–18.4 µM in 20–50 m depth waters. Under these circumstances, considering that calculated reachable depths of P. donghaiense, T. furca, A. fraterculus, and M. polykrikoides were 10, 15, 24, and 52 m, respectively, A. fraterculus and M. polykrikoides can reach the deep waters and grow, but P. donghaiense and T. furca cannot. However, if the deep waters intrude and ascend by 10–20 m relative to that on July 31, four species can reach deep waters and grow, whereas if the deep waters retreat and descend by 10–20 m, only M. polykrikoides can reach the deep waters and grow. Red-tide outbreaks by M. polykrikoides in the South Sea in 2014 occurred after the deep water retreat. Thus, the position of the eutrophic deep waters, affected by their intrusion and retreat and solar insolation, can affect outbreaks of harmful M. polykrikoides red tides. [ABSTRACT FROM AUTHOR]

Outbreaks of paralytic shellfish poisoning caused by the toxic dinoflagellate Alexandrium catenella (Dinophyceae) are currently a serious global problem both from economic and food hygiene perspectives. In Chile, A. catenella was first recorded in 1972 and is currently the main harmful algae species in the country, in terms of seafood security. As no relevant microsatellite markers for Chilean A. catenella populations were available, we isolated 23 new polymorphic microsatellite loci of A. catenella from Chile, of which 15 are applicable to both Chilean and Japanese populations. We found a strong genetic break between Chilean and Japanese A. catenella populations (FST=0.298, P<0.001), consistent with the morphological differences between them. In contrast to the Japanese population, for which genetic diversity ranged from 0.268 to 0.937, the Chilean population had a lower genetic diversity, ranging between 0.065 and 0.512. This study successfully developed A. catenella microsatellite markers that can be used to investigate its genetic spatial and temporal bloom diversity and assess further genetic connectivity between the population from southern and northern Chilean areas. [ABSTRACT FROM AUTHOR]

Six strains of three different Gonyaulax species were established by isolating cells from the Korean coastal area and the East China Sea, and their morphologies and molecular phylogenies based on SSU and LSU rRNA gene sequences were examined. In addition, the growth responses of the Gonyaulax species to changes in temperature and salinity were investigated. Based on morphological features and phylogenetic positions, Gonyaulax whaseongensis and G. polygramma were identified, and G. geomunensis sp. nov. is proposed in this study. These species displayed the plate formula typical for Gonyaulax, but G. polygramma and G. geomunensis sometimes showed a small intercalary plate (1a) surrounded by plates 2', *3' and 3". G. geomunensis was morphologically characterized by an S-type ventral organization, descending with a displacement of one cingulum width and bearing one, two or three prominent antapical spines of similar size. The cell surface, which was distinct from other Gonyaulax species, was thick and heavily reticulated into numerous polygonal areas. The reticulation was deeply excavated. The phylogenetics revealed that G. geomunensis and G. whaseongensis belong to different clades, and that there are two ribotypes of G. polygramma, which were morphologically indistinguishable. These species had a close phylogenetic relationship to G. hyalina, and all of them were characterized by dextral torsion. G. whaseongensis, G. polygramma and G. geomunensis had different growth responses to changes of temperature and salinity, respectively, indicating that morphological and phylogenetic identification of Gonyaulax species can be supported by ecological niches. [ABSTRACT FROM AUTHOR]

Bivalve mollusk production represents the principal aquaculture activity in Sardinia (40°03′ N, 9°05′ E). In 2021, 859 water samples and 1270 mollusk samples were analyzed. The species Alexandrium minutum caused the accumulation of Paralytic Shellfish Toxins (PST) in three samples of bivalve mollusks. Dinophysis acuminata complex caused the accumulation of lipophilic toxins (LTs) belonging to the okadaic acid group (OAs) in 18 samples of bivalve mollusks. The research of paralytic shellfish toxins (PSTs) in shellfish samples has been carried out with LC-FLD, as mentioned in the AOAC 2005 Official Method 2005.06. The determination of LTs was carried out by LC-MS/MS analysis. DTX2, belonging to the group of OA toxins, was detected for the first time in Sardinia, in mussels sampled in Tortolì. The presence of Dinophysis and Prorocentrum species was correlated with the accumulation of the OA toxin group in bivalve mollusks, showing a certain repeatability at certain times of the year in the areas included in the study. The results of the present study can help to plan and organize more effective bio-monitoring sampling strategies. [ABSTRACT FROM AUTHOR]

Despite theoretical expectations, marine microeukaryote population are often highly structured and the mechanisms behind such patterns remain to be elucidated. These organisms display huge census population sizes, yet genotyping usually requires clonal strains originating from single cells, hindering proper population sampling. Estimating allelic frequency directly from population wide samples, without any isolation step, offers an interesting alternative. Here, we validate the use of meta‐transcriptome environmental samples to determine the population genetic structure of the dinoflagellate Alexandrium minutum. Strain and meta‐transcriptome based results both indicated a strong genetic structure for A. minutum in Western Europe, to the level expected between cryptic species. The presence of numerous private alleles, and even fixed polymorphism, would indicate ancient divergence and absence of gene flow between populations. Single nucleotide polymorphisms (SNPs) displaying strong allele frequency differences were distributed throughout the genome, which might indicate pervasive selection from standing genetic variation (soft selective sweeps). However, a few genomic regions displayed extremely low diversity that could result from the fixation of adaptive de novo mutations (hard selective sweeps) within the populations. [ABSTRACT FROM AUTHOR]

The interactions of parasitic fungi with their phytoplankton hosts in the marine environment are mostly unknown. In this study, we evaluated the diversity of Chytridiomycota in phytoplankton communities dominated by dinoflagellates at several coastal locations in the NW Mediterranean Sea and demonstrated the most prominent interactions of these parasites with their hosts. The protist community in seawater differed from that in sediment, with the latter characterized by a greater heterogeneity of putative hosts, such as dinoflagellates and diatoms, as well as a chytrid community more diverse in its composition and with a higher relative abundance. Chytrids accounted for 77 amplicon sequence variants, of which 70 were found exclusively among different blooming host species. The relative abundance of chytrids was highest in samples dominated by the dinoflagellate genera Ostreopsis and Alexandrium, clearly indicating the presence of specific chytrid communities. The establishment of parasitoid‐host co‐cultures of chytrids and dinoflagellates allowed the morphological identification and molecular characterization of three species of Chytridiomycota, including Dinomyces arenysensis, as one of the most abundant environmental sequences, and the discovery of two other species not yet described. [ABSTRACT FROM AUTHOR]

Alexandrium blooms in the northwest area of the Bohai Sea (Qinhuangdao coastal area), China, produce large amounts of toxins that could be enriched in shellfish and consequently harm human bodies. To understand the succession of the phytoplankton community structure during Alexandrium bloom events in the northwest area of the Bohai Sea off Qinhuangdao from April 2 to May 7, 2019, microscopy observations and high-performance chromatography (HPLC)-pigment analysis were performed. Sixty species of phytoplankton were identified, mainly diatoms and dinoflagellates. The abundance of Alexandrium reached the maximum on April 16 (3.3×103 cells/L). HPLC-pigment CHEMTAX analysis showed that the phytoplankton community was composed mainly of diatoms, dinoflagellates, prasinophytes, and cryptophytes. Diatoms were the main contributor to the total Chl-a pool. There was a downward trend for the proportion of diatom biomass to the total Chl-a pool, followed by an upward trend. The proportion of dinoflagellate biomass showed the opposite trend, whereas that of the prasinophyte biomass presented an obvious increasing trend. Temperature, nutrients, and nutrient structures were the main factors on the distribution of different phytoplankton groups in the study area as shown in the redundancy analysis. This work illustrates the succession of phytoplankton community structures during Alexandrium blooms and provided a theoretical basis for studies on the mechanism underlying the outbreak of harmful algal blooms in sea areas. [ABSTRACT FROM AUTHOR]

Blooms of marine planktonic dinoflagellates are usual along the Pacific coast of Mexico, sometimes producing toxic red tides. The thecate dinoflagellate Gonyaulax areolata was found to produce a red tide event in the tropical Mexican Pacific. The species was originally described in 1911 by Kofoid and Michener without an illustration. The morphology of G. areolata was studied here by LM and SEM. Photosynthetic, solitary cells were fairly small, polyhedral, with a short apical protrusion and 'shoulders' on the epitheca, and two short, asymmetrical antapical spines. The cingulum was approximately median, cavozone, with the ends fairly displaced but with no, or only slight, overhang. The thecae were strongly ornamented with striae, areolae and pores. The plate formula was Po, 4ʹ, Q, 6ʹʹ, 6c, 4s, 6ʹʹʹ, 1p, 1ʹʹʹʹ, as recently attributed to some species of Gonyaulax. Details of important plates are given. Gonyaulax areolata seems to be morphologically closely related to other small species with a slight cingulum displacement and overhang, such as G. minima and G. striata, and more distantly related to G. spinifera. The maximum concentration of G. areolata was 1.74 × 106 cells l–1, in association with lower numbers of other dinoflagellates and diatoms. Yessotoxins were not detected. Oceanographic conditions during the bloom indicated moderate to weak upwelling, confirmed by both relatively low surface temperature and high Chl a concentration (14 µg l–1). [ABSTRACT FROM AUTHOR]

• Lethal and sublethal effects of A. pseudogonyaulax on algae, zooplankton and fish gill cells. • First report of fish-killing potency of the harmful dinoflagellate A. pseudogonyaulax. • Adverse effects of A. pseudogonyaulax were mainly driven by unknown bioactive extracellular compounds and not by goniodomins. • Intrastrain variabilities highlight the limited understanding of factors modulating the lytic activity of Alexandrium spp. The dinoflagellate Alexandrium pseudogonyaulax , a harmful algal bloom species, is currently appearing in increasing frequency and abundance across Northern European waters, displacing other Alexandrium species. This mixotrophic alga produces goniodomins (GDs) and bioactive extracellular substances (BECs) that may pose a threat to coastal ecosystems and other marine resources. This study demonstrated the adverse effects of A. pseudogonyaulax on four marine trophic levels, including microalgae (Rhodomonas salina), microzooplankton (Polykrikos kofoidii) and mesozooplankton (Acartia tonsa), as well as fish gill cells (RTgill-W1, Oncorhynchus mykiss), ultimately leading to enhanced mortality and cell lysis. Furthermore, cell-free supernatants collected from A. pseudogonyaulax cultures caused complete loss of metabolic activity in the RTgill-W1 cell line, indicating ichthyotoxic properties, while all tested GDs were much less toxic. In addition, cell-free supernatants of A. pseudogonyaulax led to cell lysis of R. salina , while all tested GDs were non-lytic. Finally, reduced egg hatching rates of A. tonsa eggs exposed to cell-free supernatants of A. pseudogonyaulax and impaired mobility of P. kofoidii and A. tonsa exposed to A. pseudogonyaulax were also observed. Altogether, bioassay results suggest that the toxicity of A. pseudogonyaulax is mainly driven by BECs and not by GDs, although further research into factors modulating the lytic activity of Alexandrium spp. are needed. [Display omitted] [ABSTRACT FROM AUTHOR]

• A lexandrium catenella produced paralytic shellfish toxins and allelochemicals. • Allelochemicals caused negative effects on protists and metazoan microzooplankton. • co-occurring competitors alleviated negative effects on microzooplankton. • mathematical model predicted experimental outcome of complex food web dynamics 5. The production of allelochemicals by the toxigenic dinoflagellate Alexandrium catenella is one of the suggested mechanisms to facilitate its bloom formation and persistence by outcompeting other phototrophic protists and reducing grazing pressure. In Southern California, toxic events caused by A. catenella and paralytic shellfish toxins (PSTs) regularly impact coastal ecosystems; however, the trophic interactions and mechanisms promoting this species in a food web context are still not fully understood. In the present study, we combined a dynamical mathematical model with laboratory experiments to investigate potential toxic and allelochemical effects of an A. catenella strain isolated off the coast of Los Angeles, Southern California, on competitors and a common zooplankton consumer. Experiments were conducted using three toxigenic strains of A. catenella , comparing the new Californian isolate (Alex Cal) to two strains previously described from the North Sea, a lytic (Alex2) and non-lytic (Alex5) strain, testing for donor density-dependent effects on two phytoplankton species (Rhodomonas salina, Tetraselmis sp.) and on the rotifer Brachionus plicatilis. Bioassays revealed a steep decline in competitor and consumer populations with increasing Alex Cal concentrations, indicating an intermediate lytic activity compared to the North Sea strains (lytic Alex2 and non-lytic Alex5). The rotifer fed and grew well on the PST- toxic, but non-lytic Alex5 strain, while its survival significantly decreased with increasing concentrations of the two lytic strains Alex Cal and Alex 2, indicating that negative effects on the rotifer were mediated by allelochemicals rather than PST-toxins. Mixed culture experiments including both competitors and consumers demonstrated that the intensity of allelochemical effects not only depended on the A. catenella density but also on the target density. Negative effects on grazers were alleviated by co-occurring competitors with a lower sensitivity to allelochemicals, thus reducing harmful compounds and allowing grazing control on the dinoflagellate to come into effect again. Results from mixed culture experiments were supported by the mathematical approach used in this study which was calibrated with data from simple monoculture growth, pairwise competition and predator-prey experiments, demonstrating the applicability of this model approach to predict the outcome of more complex food web dynamics at the community level. [Display omitted] [ABSTRACT FROM AUTHOR]

• We performed a systematic quantitative review of Alexandrium – Acartia interactions. • Alexandrium and its toxins do not have effects on food ingestion and egg production. • Alexandrium toxins have negative effects on hatching (bi-phasic) and mortality. • Our results support that chronically exposed copepod populations have improved physiological performance relative to naïve populations, in a wide geographical scale. • Literature data showed very large dispersion for the variables ingestion rate and egg production. The study of interactions between copepods of the genus Acartia and toxic dinoflagellates of the genus Alexandrium has been an important topic during the last four decades. Feeding behavior and physiological responses of copepods have been studied in laboratory and field experiments, sometimes with contradictory results. More recently, an evolutionary adaptive mechanism leading to enhanced tolerance of Alexandrium toxins in a population of Acartia experiencing chronic exposure to these dinoflagellates has been reported. In the present work, we collected data from the existing studies on the effects of Alexandrium on feeding, reproduction and mortality of Acartia. With these data, we performed a systematic review consisting of a secondary analysis employing general or generalized linear models, weighting data from different studies by the reciprocal of their standard deviation. Our first aim was to overcome shortcomings of individual studies: limited ranges of the variables and overlooked variables (experiment length, population adaptation). These shortcomings could have led to inconsistent conclusions by missing heterogeneous patterns in copepod responses and in the interactions between variables. Our second aim was to test the enhanced physiological performance of chronically exposed relative to naïve copepod populations over a wide geographic range. We found that the feeding rate is enhanced by increased food biomass, irrespective of the food type. Toxins do not have a clear effect on egg production and have a bi-phasic effect on egg hatching success, which was negative above a specific threshold. Toxins also increased mortality. Experiment length had a positive effect on egg production and negative on egg hatching. Naïve copepod populations showed consistently lower ingestion of Alexandrium and egg hatching rates, thereby supporting the spread of the aforementioned mechanism across populations over a wide geographic range. [ABSTRACT FROM AUTHOR]

Filter-feeding bivalves can accumulate paralytic shellfish toxins (PST) produced by toxic microalgae, which may induce oxidative stress and lipid peroxidation. Peroxisomal acyl-coenzyme A oxidases (ACOXs) are key enzymes functioning in maintaining redox and lipid homeostasis, but their roles in PST response in bivalves are less understood. Herein, a total of six and six ACOXs were identified in the Chlamys farreri and Patinopecten yessoensis genome, respectively, and the expansion of ACOX1s was observed. Gene expression analysis revealed an organ/tissue-specific expression pattern in both scallops, with all ACOXs being predominantly expressed in the two most toxic organs, digestive glands and kidneys. The regulation patterns of scallop ACOXs after exposure to different PST-producing algaes Alexandrium catenella (ACDH) and A. minutum (AM-1) were revealed. After ACDH exposure, more differentially expressed genes (DEGs) were identified in C. farreri digestive glands (three) and kidneys (five) than that in P. yessoensis (two), but the up-regulated DEGs showed similar expression patterns in both species. In C. farreri, three DEGs were found in both digestive glands and kidneys after AM-1 exposure, with two same CfACOX1s being acutely and chronically induced, respectively. Notably, these two CfACOX1s also showed different expression patterns in kidneys between ACDH (acute response) and AM-1 (chronic response) exposure. Moreover, inductive expression of CfACOXs after AM-1 exposure was observed in gills and mantles, and all DEGs in both tissues were up-regulated and their common DEGs exhibited both acute and chronic induction. These results indicate the involvement of scallop ACOXs in PST response, and their plasticity expression patterns between scallop species, among tissues, and between the exposure of different PST analogs. [ABSTRACT FROM AUTHOR]

The armoured dinoflagellate Alexandrium can be found throughout many of the world’s temperate and tropical marine environments. The genus has been studied extensively since approximately half of its members produce a family of potent neurotoxins, collectively called saxitoxin. These compounds represent a significant threat to animal and environmental health. Moreover, the consumption of bivalve molluscs contaminated with saxitoxin poses a threat to human health. The identification of Alexandrium cells collected from sea water samples using light microscopy can provide early warnings of a toxic event, giving harvesters and competent authorities time to implement measures that safeguard consumers. However, this method cannot reliably resolve Alexandrium to a species level and, therefore, is unable to differentiate between toxic and non-toxic variants. The assay outlined in this study uses a quick recombinase polymerase amplification and nanopore sequencing method to first target and amplify a 500 bp fragment of the ribosomal RNA large subunit and then sequence the amplicon so that individual species from the Alexandrium genus can be resolved. The analytical sensitivity and specificity of the assay was assessed using seawater samples spiked with different Alexandrium species. When using a 0.22 µm membrane to capture and resuspend cells, the assay was consistently able to identify a single cell of A. minutum in 50 mL of seawater. Phylogenetic analysis showed the assay could identify the A. catenella, A. minutum, A. tamutum, A. tamarense, A. pacificum, and A. ostenfeldii species from environmental samples, with just the alignment of the reads being sufficient to provide accurate, real-time species identification. By using sequencing data to qualify when the toxic A. catenella species was present, it was possible to improve the correlation between cell counts and shellfish toxicity from r = 0.386 to r = 0.769 (p ≤ 0.05). Furthermore, a McNemar’s paired test performed on qualitative data highlighted no statistical differences between samples confirmed positive or negative for toxic species of Alexandrium by both phylogenetic analysis and real time alignment with the presence or absence of toxins in shellfish. The assay was designed to be deployed in the field for the purposes of in situ testing, which required the development of custom tools and state-of-the-art automation. The assay is rapid and resilient to matrix inhibition, making it suitable as a potential alternative detection method or a complementary one, especially when applying regulatory controls.

Bivalve molluscs are filter-feeding organisms that can accumulate paralytic shellfish toxins (PST) through ingesting toxic marine dinoflagellates. While the effects of PST accumulation upon the physiology of bivalves have been documented, the underlying molecular mechanism remains poorly understood. In this study, transcriptomic analysis was performed in the gills of Zhikong scallop (Chlamys farreri) after 1, 3, 5, 10, and 15 day(s) exposure of PST-producing dinoflagellate Alexandrium minutum. Higher numbers of differentially expressed genes (DEGs) were detected at day 1 (1538) and day 15 (989) than that at day 3 (77), day 5 (82), and day 10 (80) after exposure, and most of the DEGs were only regulated at day 1 or day 15, highlighting different response mechanisms of scallop to PST-producing dinoflagellate at different stages of exposure. Functional enrichment results suggested that PST exposure induced the alterations of nervous system development processes and the activation of xenobiotic metabolism and substance transport processes at the acute and chronic stages of exposure, respectively, while the immune functions were inhibited by PST and might ultimately cause the activation of apoptosis. Furthermore, a weighted gene co-expression network was constructed, and ten responsive modules for toxic algae exposure were identified, among which the yellow module was found to be significantly correlated with PST content. Most of the hub genes in the yellow module were annotated as solute carriers (SLCs) with eight being OCTN1s, implying their dominant roles in regulating PST accumulation in scallop gills. Overall, our results reveal the gene set responding to and involved in PST accumulation in scallop gills, which will deepen our understanding of the molecular mechanism of bivalve resistance to PST. [ABSTRACT FROM AUTHOR]

The uncontrolled growth of harmful algal blooms (HABs) can negatively impact the environment and pose threats to human health and aquatic ecosystems. Titanium dioxide (TiO2) is known to be effective in killing harmful algae through flocculation and sedimentation. However, TiO2 in a dispersed form can harm other non-target marine organisms, which has raised concerns by environmentalists and scientists. This research seeks to explore the utility of immobilized titanium oxide as a photocatalyst for mitigation of HABs, where the Alexandrium minutum bloom was used as a model system herein. Chitosan was modified with 0.2 wt.% TiO2 (Chi/TiO2 (x mL; x = 1, 3 and 5 mL) and the corresponding films were prepared via solvent casting method. Scanning electron microscope (SEM) images of the films reveal a highly uneven surface. X-ray diffraction (XRD) analysis indicates the reduction in chitosan crystallinity, where the presence of TiO2 was negligible, in accordance with its dispersion within the chitosan matrix. The photocatalytic mitigation of A.minutum was carried out via a physical approach in a laboratory-scale setting. The negative surface charge of the films was observed to repel the negatively charged A.minutum causing fluctuation in the removal efficiency (RE). The highest RE (76.1 ± 13.8%) was obtained when Chi/TiO2 (1 mL) was used at 72 h, where the hydroxyl radicals generated were inferred to contribute to the deactivation of the algae cells by causing oxidative stress. An outcome of this study indicates that such hybrid films have the potential to replace the non-immobilized (dispersed) TiO2 for HAB mitigation. However, further investigation is required to deploy these films for field applications at a larger scale. [ABSTRACT FROM AUTHOR]

Growing interest and affordability of environmental DNA and RNA (eDNA and eRNA) approaches for biodiversity assessments and monitoring of complex ecosystems have led to the emergence of manifold protocols for nucleic acids (NAs) isolation and processing. Although there is no consensus on a standardized workflow, the common practice for water samples is to concentrate NAs via filtration using varying pore size membranes. Using the smallest pore is assumed to be most efficient for NAs capture from a wide range of material (including sub‐cellular particles); however, a trade‐off must occur between detection of a meaningful molecular signal and cost/time effort when processing samples using fine pore membranes.Comparative studies involving formal efficiency assessments are lacking, which restricts informed decision‐making around an optimized sampling approach for applications such as biosurveillance (i.e. detection and monitoring of target taxa—nuisance organisms, endangered and indicator taxa or other species of economic or cultural importance). Here, we present an experimental study using an easily cultured microalgal species (Alexandrium pacificum) to test different filter membranes for capturing NAs in the context of cost/time effort and cell fractions encountered in nature (whole cells, partially lysed and naked NAs).The results showed no statistically significant difference between membrane types for capturing target eDNA signal from intact and partially lysed cell treatments. In terms of time effort and volume processed, higher efficiency ratings were obtained with the larger pore size (5 μm) cellulose membranes. Positively charged nylon demonstrated enhanced capture of naked NAs, and especially eRNA signal, across treatments.Our findings support using coarse pore size filters for adequate capture of target NA signal (from both eDNA and eRNA) with less processing time. The framework presented here can provide a quick and robust feasibility check and comparative assessment of new and existing NA processing technologies, and allows sufficient control over multiple parameters, including physical–chemical water properties, temporal scales, and concentration and type of input material. [ABSTRACT FROM AUTHOR]

Blooms of many dinoflagellates, including several harmful algal bloom (HAB) species, are seeded and revived through the germination of benthic resting cysts. Temperature is a key determinant of cysts' germination rate, and temperature-germination rate relationships are therefore fundamental to understanding species' germling cell production, cyst bed persistence, and resilience to climate warming. This study measured germination by cysts of the HAB dinoflagellate Alexandrium catenella using a growing degree-day (DD) approach that accounts for the time and intensity of warming above a critical temperature. Time courses of germination at different temperatures were fit to lognormal cumulative distribution functions for the estimation of the median days to germination. As temperature increased, germination times decreased hyperbolically. DD scaling collapsed variability in germination times between temperatures after cysts were oxygenated. A parallel experiment demonstrated stable temperature-rate relationships in cysts collected during different phases of seasonal temperature cycles in situ over three years. DD scaling of the results from prior A. catenella germination studies showed consistent differences between populations across a wide range of temperatures and suggests selective pressure for different germination rates. The DD model provides an elegant approach to quantify and compare the temperature dependency of germination among populations, between species, and in response to changing environmental conditions. [ABSTRACT FROM AUTHOR]

The taxonomy of the extant dinoflagellate genus Gonyaulax is challenging since its thecate morphology is rather conservative. In contrast, cysts of Gonyaulax are varied in morphology and have been related with the fossil‐based genera Spiniferites and Impagidinium. To better understand the systematics of Gonyaulax species, we performed germination experiments on cysts that can be identified as S. ristingensis, an unidentified Spiniferites with petaloid processes here described as Spiniferites pseudodelicatus sp. nov. and Impagidinium variaseptum from Chinese and Portuguese waters. Despite marked differences in cyst morphology, motile cells of S. pseudodelicatus and I. variaseptum are indistinguishable from Gonyaulax baltica. Motile cells hatched from S. ristingensis are morphologically similar to G. baltica as well but differ in the presence of one pronounced antapical spine. Three new species, Gonyaulax amoyensis (cyst equivalent S. pseudodelicatus), Gonyaulax bohaiensis (cyst equivalent I. variaseptum), and Gonyaulax portimonensis (cyst equivalent S. ristingensis), were erected. In addition, a new ribotype (B) of G. baltica was reported from South Korea and a bloom of G. baltica ribotype B is reported from New Zealand. Molecular phylogeny based on LSU and SSU rRNA gene sequences revealed that Gonyaulax species with minute or short antapical spines formed a well‐resolved clade, whereas species with two pronounced antapical spines or lack of antapical spines formed the sister clade. Six strains of four above species were examined for yessotoxin production by liquid chromatography coupled with tandem mass spectrometry, and very low concentrations of yessotoxin were detected for one G. bohaiensis strain. [ABSTRACT FROM AUTHOR]

Harmful algal bloom (HAB) species Alexandrium catenella and Dinophysis acuminata are associated with paralytic shellfish poisoning (PSP) and diarrhetic shellfish poisoning (DSP) in humans, respectively. While PSP and DSP have been studied extensively, less is known about the effects of these HAB species or their associated toxins on shellfish. This study investigated A. catenella and D. acuminata toxicity in a larval oyster (Crassostrea virginica) bioassay. Larval activity and mortality were examined through 96-h laboratory exposures to live HAB cells (10–1000 cells/mL), cell lysates (1000 cells/mL equivalents), and purified toxins (10,000 cells/mL equivalents). Exposure to 1000 cells/mL live or lysed D. acuminata caused larval mortality (21.9 ± 7.0%, 10.2 ± 4.0%, respectively) while exposure to any tested cell concentration of live A. catenella, but not lysate, caused swimming arrest and/or mortality in >50% of larvae. Exposure to high concentrations of saxitoxin (STX) or okadaic acid (OA), toxins traditionally associated with PSP and DSP, respectively, had no effect on larval activity or mortality. In contrast, pectenotoxin-2 (PTX2) caused rapid larval mortality (49.6 ± 5.8% by 48 h) and completely immobilized larval oysters. The results indicate that the toxic effects of A. catenella and D. acuminata on shellfish are not linked to the primary toxins associated with PSP and DSP in humans, and that PTX2 is acutely toxic to larval oysters. [ABSTRACT FROM AUTHOR]

The toxigenic Alexandrium species can secrete paralytic shellfish toxins (PSTs) into seawater during their growth, potentially threatening marine organisms and human health. The release of intracellular toxins during the growth cycle of Alexandrium spp. and the uptake of dissolved PSTs by shellfish are still not clear. Three strains of A. minutum GY-H46, A. minutum ATDH and A. pacificum ATHK were used to explore the dynamic variations in profile of intra- and extracellular toxins during their growth cycles. And a comparative study was carried out on the accumulation and tissue distribution of PSTs in mussels Mytilus unguiculatus exposed to the dissolved and particulate toxins during a 5-days period. Results showed that the toxin cell quotas increased rapidly during the early exponential growth period, then decreased gradually and plateaued at the stationary growth period for ATHK and ATDH, while gradually increased during the stationary growth period for GY-H46. The toxin cell quotas for GY-H46, ATDH and ATHK ranged from 8.3 to 21.5, 5.3 to 11.7 and 7.4 to 9.3 fmol cell−1, respectively, during the whole growth cycle. The contents of extracellular toxins increased gradually throughout the growth of GY-H46, ATDH and ATHK, and the molar mass percentage of extracellular toxins ranged from 53% to 72%, 20% to 46% and 36% to 68%, respectively. Mussels can actively uptake the dissolved PSTs from seawater medium, and mainly accumulate toxins in the digestive glands. The accumulation level of PSTs from the dissolved phase by mussels was about 1% of that from feeding on microalgal cells. The results of this study will contribute to a comprehensive understanding of the dynamic release of intracellular PSTs during microalgal growth and the accumulation of dissolved toxins by shellfish. [Display omitted] • The toxin cell quotas for the three Alexandrium spp. were markedly different. • The three Alexandrium spp. secreted toxins during the whole growth cycle. • The extracellular toxins underwent chemical transformations. • Mussels can actively uptake the dissolved PSTs from seawater. • The accumulation level of PSTs was low in mussels exposed to the dissolved toxins. [ABSTRACT FROM AUTHOR]