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Several coastal countries including France have experienced serious and increasing problems related to Pseudo‐nitzschia toxic blooms. These toxic blooms occur in estuarine and coastal waters potentially subject to fluctuations in salinity. In this study, we document for the first time the viability, growth, photosynthetic efficiency, and toxin production of two strains of Pseudo‐nitzschia australis grown under conditions with sudden salinity changes. Following salinity variation, the two strains survived over a restricted salinity range of 30–35, with favorable physiological responses, as the growth, effective quantum yield and toxin content were high compared to the other conditions. In addition, high cellular quotas of domoic acid (DA) were observed at a salinity of 40 for the strain IFR‐PAU‐16.1 in comparison with the other strain IFR‐PAU‐16.2 where the cell DA content was directly released into the medium. On the other hand, the osmotic stress imposed at lower salinities, 20 and 10, resulted in cell lysis and a sudden DA leakage in the medium. Intra‐specific variability was observed in growth and toxin production, with the strain IFR‐PAU‐16.1 apparently able to withstand higher salinities than the strain IFR‐PAU‐16.2. On the whole, DA does not appear to act as an osmolyte in response to sudden salinity changes. Since most of the shellfish harvesting areas of bivalve molluscs in France are located in areas where the salinity generally varies between 30 and 35, Pseudo‐nitzschia australis blooms might potentially impact public health and commercial shellfish resources in these places. [ABSTRACT FROM AUTHOR]

The new benthic toxic dinoflagellate, Ostreopsis fattorussoi sp. nov., is described from the Eastern Mediterranean Sea, Lebanon and Cyprus coasts, and is supported by morphological and molecular data. The plate formula, Po, 3′, 7″, 6c, 7s, 5‴, 2′′′′, is typical for the Ostreopsis genus. It differs from all other Ostreopsis species in that (i) the curved suture between plates 1′ and 3′ makes them approximately hexagonal, (ii) the 1′ plate lies in the left half of the epitheca and is obliquely orientated leading to a characteristic shape of plate 6″. The round thecal pores are bigger than the other two Mediterranean species ( O. cf. ovata and O. cf. siamensis). O. fattorussoi is among the smallest species of the genus ( DV: 60.07 ± 5.63 μm, AP: 25.66 ± 2.97 μm, W: 39.81 ± 5.05 μm) along with O. ovata. Phylogenetic analyses based on the LSU and internal transcribed spacer rDNA shows that O. fattorussoi belongs to the Atlantic/Mediterranean Ostreopsis spp. clade separated from the other Ostreopsis species. Ostreopsis fattorussoi produces OVTX-a and structural isomers OVTX-d and -e, O. cf. ovata is the only other species of this genus known to produce these toxins. The Lebanese O. fattorussoi did not produce the new palytoxin-like compounds (ovatoxin-i, ovatoxin-j1, ovatoxin-j2, and ovatoxin-k) that were previously found in O. fattorussoi from Cyprus. The toxin content was in the range of 0.28-0.94 pg · cell−1. On the Lebanon coast, O. fattorussoi was recorded throughout the year 2015 (temperature range 18°C-31.5°C), with peaks in June and August. [ABSTRACT FROM AUTHOR]

The two most toxic Gambierdiscus species identified from the Caribbean are G. excentricus and G. silvae. These species are the primary causes of ciguatera fish poisoning and likely contribute disproportionately to the toxicity of marine food webs. While Gambierdiscus species are difficult to distinguish using light or scanning electron microscopy, reliable species-specific molecular identification methods have been developed and used successfully to identify a number of other Gambierdiscus species. Corresponding speciesspecific assays are not yet available for G. excentricus and G. silvae, which imposes limitations on species identification and related ecological studies. The following note describes species-specific polymerase chain reaction assays for G. excentricus and G. silvae that can be used for these purposes. [ABSTRACT FROM AUTHOR]

Toxin-producing blooms of dinoflagellates in the genus Alexandrium have plagued the inhabitants of the Salish Sea for centuries. Yet the environmental conditions that promote accelerated growth of this organism, a producer of paralytic shellfish toxins, is lacking. This study quantitatively determined the growth response of two Alexandrium isolates to a range of temperatures and salinities, factors that will strongly respond to future climate change scenarios. An empirical equation, derived from observed growth rates describing the temperature and salinity dependence of growth, was used to hindcast bloom risk. Hindcasting was achieved by comparing predicted growth rates, calculated from in situ temperature and salinity data from Quartermaster Harbor, with corresponding Alexandrium cell counts and shellfish toxin data. The greatest bloom risk, defined at μ >0.25 d−1, generally occurred from April through November annually; however, growth rates rarely fell below 0.10 d−1. Except for a few occasions, Alexandrium cells were only observed during the periods of highest bloom risk and paralytic shellfish toxins above the regulatory limit always fell within the periods of predicted bloom occurrence. While acknowledging that Alexandrium growth rates are affected by other abiotic and biotic factors, such as grazing pressure and nutrient availability, the use of this empirical growth function to predict higher risk time frames for blooms and toxic shellfish within the Salish Sea provides the groundwork for a more comprehensive biological model of Alexandrium bloom dynamics in the region and will enhance our ability to forecast blooms in the Salish Sea under future climate change scenarios. [ABSTRACT FROM AUTHOR]

Knowledge concerning the ability of microalgae to produce metabolites of interest such as toxins or high-value secondary metabolites requires exhaustive details to be supplied on how the research was conducted. These should include the microalgal species and strain characterization, the culture conditions, the cell density, and physiological state at the time of harvesting, the harvesting method, the sample pre-treatment protocol, and the subsequent instrumental analytical separation/detection system. In this comment, we discuss issues that affect algal research from an analytical chemistry perspective, particularly (i) the need to specify detection capabilities of the entire method (i.e., limits of detection or threshold detection levels), which we illustrate in relation to classification of a species or strain as being 'toxin producing' or 'non-toxin producing'; and (ii) the requirements that have to be satisfied to confirm a microalgal species (new or not) as a producer of a particular chemical of interest for phycologists, which again we illustrate in relation to toxins. A successful collaboration among phycologists and analytical chemists will only be achieved as a result of a synergistic collaboration between the two disciplines, with a reciprocal understanding at least at a background level. [ABSTRACT FROM AUTHOR]

Species belonging to the potentially harmful diatom genus Pseudo-nitzschia, isolated from 16 localities (31 sampling events) in the coastal waters of south-eastern Australia, were examined. Clonal isolates were characterized by (i) light and transmission electron microscopy; (ii) phylogenies, based on sequencing of nuclear-encoded ribosomal deoxyribonucleic acid ( rDNA) regions and, (iii) domoic acid ( DA) production as measured by liquid chromatography-mass spectrometry ( LC- MS/ MS). Ten taxa were unequivocally confirmed as Pseudo-nitzschia americana, P. arenysensis, P. calliantha, P. cuspidata, P. fraudulenta, P. hasleana, P. micropora, P. multiseries, P. multistriata, and P. pungens. An updated taxonomic key for south-eastern Australian Pseudo-nitzschia is presented. The occurrence of two toxigenic species, P. multistriata (maximum concentration 11 pg DA per cell) and P. cuspidata (25.4 pg DA per cell), was documented for the first time in Australia. The Australian strains of P. multiseries, a consistent producer of DA in strains throughout the world, were nontoxic. Data from 5,888 water samples, collected from 31 oyster-growing estuaries (2,000 km coastline) from 2005 to 2009, revealed 310 regulatory exceedances for 'Total Pseudo-nitzschia,' resulting in six toxic episodes. Further examination of high-risk estuaries revealed that the ' P. seriata group' had highest cell densities in the austral summer, autumn, or spring (species dependent), and lowest cell densities in the austral winter, while the ' P. delicatissima group' had highest in winter and spring. [ABSTRACT FROM AUTHOR]

Periodic and seasonal exposure to high light is a common occurrence for many near-shore and estuarine phytoplankton. Rapid acclimatization to shifts in light may provide an axis by which some species of phytoplankton can outcompete other microalgae. Patterns of photoacclimation and photosynthetic capacity in the raphidophyte Heterosigma akashiwo (Hada) Hada ex Hara et Chihara isolated from the mid-Atlantic of the United States were followed in continuous cultures at low- and high-light intensities, followed by reciprocal shifts to the opposite light level. The maximum quantum yield ( F v /F m) as well as the photosynthetic cross-section (σ PSII) of photosystem II was higher in high-light cultures compared to low-light cultures. Significant diurnal variability in photochemistry and photoprotection was noted at both light levels, and high-light-acclimated cultures displayed greater variability in photoprotective pathways. When shifted from low to high light, there was only a slight and temporary decline in maximum quantum yield, while cell specific growth more than doubled within 24 h. Rapid acclimation to high light was facilitated by short-term photoprotection (nonphotochemical quenching), reduced PSII reaction center connectivity, and electron transport. Short-term increases in de-epoxidated xanthophyll pigments contributed to nonphotochemical protection, but lagged behind initial increases in nonphotochemical quenching and were not the primary pathway of photoprotection in this alga. By 48 h, photochemistry of cultures shifted from low to high light resembled long-term high-light-acclimated cultures. This isolate of H. akashiwo appears well poised to exploit rapid shifts in light by using unique cellular adjustments in light harvesting and photochemistry. [ABSTRACT FROM AUTHOR]

Cell counts are the standard measure to quantify harmful algae and the basis for decisions on measures necessary to protect human health. Molecular detection methods have been developed for a range of algal species and genera, but these methods generally quantify DNA or RNA, and corresponding cell numbers are inferred based on the assumption that the cellular nucleic acid content is constant over time and in different conditions. Here, we tested this assumption for ichthyotoxic flagellates of the genus Pseudochattonella (Dictyochophyceae) under different light, temperature, salinity, and nutrient conditions. Our results show changes in cellular RNA contents of nearly one order of magnitude depending on the condition and also the time of exposure, rendering it difficult to anticipate per-cell RNA yields even if environmental conditions are known. However, cellular RNA content was positively correlated with cell size and growth rate across our experiments, and total RNA was comparable to cell number as a predictor for total biovolume. These results demonstrate the importance of considering the variability of RNA levels for comparisons with cell counts and provide a valuable aid for the interpretation of data from RNA-based detection methods. [ABSTRACT FROM AUTHOR]

Heterokont members of the Pelagophyceae form the massive brown tides that have continually plagued the coastal regions of the eastern U.S. seaboard and the Gulf of Mexico. To gain a better understanding of the photosynthetic competence that may be linked to their success in forming massive blooms, we sequenced the chloroplast genomes of two pelagophytes: Aureococcus anophagefferens Hargraves et Sieburth and Aureoumbra lagunensis D. A. Stockw., DeYoe, Hargraves et P. W. Johnson. The chloroplast genomes of A. anophagefferens (89,599 bp) and Ar. lagunensis (94,346 bp) are significantly smaller than those of six other stramenopiles sequenced to date. The structure (or configuration) is partially due to the absence of the large inverted repeats common in chloroplast genomes. Eight of 10 small and tandem repeats from the A. anophagefferens and Ar. lagunensis genomes are adjacent to genes coding for photosynthetic or energy production functions, implying that these domains may have functional constraints. High genomic synteny, a multigene phylogenetic analysis, and a synapomorphic change in the form of an attenuated psbA gene confirm that A. anophagefferens and Ar. lagunensis are closely related taxa. Finally, the presence of three light-independent chl-biosynthesis genes in the chloroplast of Ar. lagunensis, but absence in the chloroplast and nuclear genomes of A. anophagefferens, suggests the persistence of a more ancient (i.e., dark-adaptive) potential in Ar. lagunensis but not in A. anophagefferens. Whether the presence of both chl-biosynthesis pathways in Ar. lagunensis contributes to the ability of this organism to sustain prolonged bloom (continuously for ∼8 years) under reduced light conditions, but not A. anophagefferens (a few months), remains an open question. [ABSTRACT FROM AUTHOR]

Imaging FlowCytobot (IFCB) combines video and flow cytometric technology to capture images of nano- and microplankton (∼10 to >100 μm) and to measure the chlorophyll fluorescence associated with each image. The images are of sufficient resolution to identify many organisms to genus or even species level. IFCB has provided >200 million images since its installation at the entrance to the Mission-Aransas estuary (Port Aransas, TX, USA) in September 2007. In early February 2008, Dinophysis cells (1–5 · mL−1) were detected by manual inspection of images; by late February, abundance estimates exceeded 200 cells · mL−1. Manual microscopy of water samples from the site confirmed that D. cf. ovum F. Schütt was the dominant species, with cell concentrations similar to those calculated from IFCB data, and toxin analyses showed that okadaic acid was present, which led to closing of shellfish harvesting. Analysis of the time series using automated image classification (extraction of image features and supervised machine learning algorithms) revealed a dynamic phytoplankton community composition. Before the Dinophysis bloom, Myrionecta rubra (a prey item of Dinophysis) was observed, and another potentially toxic dinoflagellate, Prorocentrum, was observed after the bloom. Dinophysis cell-division rates, as estimated from the frequency of dividing cells, were the highest at the beginning of the bloom. Considered on a daily basis, cell concentration increased roughly exponentially up to the bloom peak, but closer inspection revealed that the increases generally occurred when the direction of water flow was into the estuary, suggesting the source of the bloom was offshore. [ABSTRACT FROM AUTHOR]

Since 1998, a heterokont flagellate initially named Chattonella aff . verruculosa has formed recurrent extensive blooms in the North Sea and the Skagerrak, causing fish mortalities. Cells were isolated from the 2001 bloom off the south coast of Norway, and monoalgal cultures were established and compared with the Chattonella verruculosa Y. Hara et Chihara reference strain NIES 670 from Japan. The cells in Norwegian cultured isolates were very variable in size and form, being large oblong (up to 34 μm long) to small rounded (5–9 μm in diameter) with two unequal flagella, numerous chloroplasts, and mucocysts. The SSU and partial LSU rDNA sequences of strains from Norway and Japan were compared and differed by 0.4% (SSU) and 1.3% (LSU), respectively. Five strains from Norway were identical in the LSU rDNA region. Phylogenetic analyses based on heterokont SSU and concatenated SSU + LSU rDNA sequences placed C. aff. verruculosa and the Japanese C. verruculosa within the clade of Dictyochophyceae, with the picoflagellate Florenciella parvula Eikrem as the closest relative. Ultrastructure, morphology, and pigment composition supported this affinity. We propose the name Verrucophora farcimen sp. et gen. nov. for this flagellate and systematically place it within the class Dictyochophyceae. Our studies also show that C. verruculosa from Japan is genetically and morphologically different but closely related to V. farcimen. The species is transferred from the class Raphidophyceae to the class Dictyochophyceae and renamed Verrucophora verruculosa. We propose a new order, Florenciellales, to accommodate V. farcimen, V. verruculosa, and F. parvula. [ABSTRACT FROM AUTHOR]

Harmful algal blooms (HABs) have occurred with increasing frequency in recent years with eutrophication and other anthropogenic alterations of coastal ecosystems. Many of these blooms severely alter or degrade ecosystem function, and are referred to here as ecosystem disruptive algal blooms (EDABs). These blooms are often caused by toxic or unpalatable species that decrease grazing rates by planktonic and benthic herbivores, and thereby disrupt the transfer of nutrients and energy to higher trophic levels, and decrease nutrient recycling. Many factors, such as nutrient availability and herbivore grazing have been proposed to separately influence EDAB dynamics, but interactions among these factors have rarely been considered. Here we discuss positive feedback interactions among nutrient availability, herbivore grazing, and nutrient regeneration, which have the potential to substantially influence the dynamics of EDAB events. The positive feedbacks result from a reduction of grazing rates on EDAB species caused by toxicity or unpalatability of these algae, which promotes the proliferation of the EDAB species. The decreased rates also lower grazer-mediated recycling of nutrients and thereby decrease nutrient availability. Since many EDAB species are well-adapted to nutrient-stressed environments and many exhibit increased toxin production and toxicity under nutrient limitation, positive feedbacks are established which can greatly increase the rate of bloom development and the adverse effects on the ecosystem. An understanding of how these feedbacks interact with other regulating factors, such as benthic/pelagic nutrient coupling, physical forcing, and life cycles of EDAB species provides a substantial future challenge. [ABSTRACT FROM AUTHOR]

Salinity varies widely in coastal areas that often have a high abundance ofPseudo-nitzschiaH. Peragallo.Pseudo-nitzschiais abundant in Louisiana waters, and high cellular domoic acid has been observed in natural samples but no human illness has been reported. To assess the threat of amnesic shellfish poisoning (ASP), we examined the effect of salinity onPseudo-nitzschiaoccurrence in the field and growth in the laboratory with special emphasis on the salinity range where oysters are harvested (10–20 psu). In Louisiana coastal waters,Pseudo-nitzschiaspp. occurred over a salinity range of 1 to>35 psu, but they occurred more frequently at higher rather than lower salinities. Seven species were identified, including toxigenic species occurring at low salinities. In culture studies, seven clones of three species grew over a salinity range of 15 to 40 psu, some grew at salinities down to 6.25 psu, and most grew at salinities up to 45 psu. Tolerance of low salinities decreased fromPseudo-nitzschia delicatissima(Cleve) Heiden toP. multiseries(Hasle) Hasle toP. pseudodelicatissima(Hasle) Hasleemend. Lundholm, Hasle et Moestrup. In conclusion, althoughPseudo-nitzschiawas more prevalent in the field and grew better in the laboratory at higher salinities, it grew and has been observed at low salinities. Therefore, the probability of ASP from consumption of oysters harvested from the low salinity estuaries of the northern Gulf of Mexico is low but not zero; animal mortality events from toxin vectors other than oysters at higher salinity on the shelf are more likely. [ABSTRACT FROM AUTHOR]

Water quality, microbial contamination, prior fish health, and variable results have been major impediments to identifying the cause and mechanism of fish mortality in standard aquarium-format Pfiesteria bioassays. Therefore, we developed a sensitive 96-h larval fish bioassay for assessing Pfiesteria spp. pathogenicity using six-well tissue culture plates and 7-day-old larval cyprinodontid fish. We used the assay to test pathogenicity of several clonal lines of Pfiesteria piscicida Steidinger and Burkholder and P. shumwayae Glasgow and Burkholder that had been cultured with algal prey for 2 to 36 months. The P. shumwayae cultures exhibited 80%–100% cumulative mortality in less than 96 h at initial zoospore densities of approximately 1000 cells·mL-1 . No fish mortalities occurred with P. piscicida at identical densities or in controls. In a dose-response assay, we demonstrated a strong positive correlation between dinospore density and fish mortality in a highly pathogenic culture of P. shumwayae , generating a 96-h LD50 of 108 zoospores·mL-1 . Additionally, we applied the assay to evaluate a 38-L P. shumwayae bioassay that was actively killing fish and compared results with those from exposures of juvenile tilapia (Oreochromis niloticus ) in a 500-mL assay system. Water from the fish-killing 38-L assay was filtered and centrifuged to produce fractions dominated by dinoflagellates, bacteria, or presumed ichthyotoxin (cell-free fraction). After 96 h, the larval fish assay exhibited 50%–100% cumulative mortality only in fractions containing dinoflagellates, with no mortalities occurring in the other fractions. The 500-mL bioassay with tilapia produced inconsistent results and demonstrated no clear correlation between mortality and treatment. The new larval fish bioassay was demonstrated as a highly effective method to verify and evaluate dinoflagellate pathogenicity. [ABSTRACT FROM AUTHOR]

Water quality, microbial contamination, prior fish health, and variable results have been major impediments to identifying the cause and mechanism of fish mortality in standard aquarium-format Pfiesteria bioassays. Therefore, we developed a sensitive 96-h larval fish bioassay for assessing Pfiesteria spp. pathogenicity using six-well tissue culture plates and 7-day-old larval cyprinodontid fish. We used the assay to test pathogenicity of several clonal lines of Pfiesteria piscicida Steidinger and Burkholder and P. shumwayae Glasgow and Burkholder that had been cultured with algal prey for 2 to 36 months. The P. shumwayae cultures exhibited 80%–100% cumulative mortality in less than 96 h at initial zoospore densities of approximately 1000 cells·mL-1 . No fish mortalities occurred with P. piscicida at identical densities or in controls. In a dose-response assay, we demonstrated a strong positive correlation between dinospore density and fish mortality in a highly pathogenic culture of P. shumwayae , generating a 96-h LD50 of 108 zoospores·mL-1 . Additionally, we applied the assay to evaluate a 38-L P. shumwayae bioassay that was actively killing fish and compared results with those from exposures of juvenile tilapia (Oreochromis niloticus ) in a 500-mL assay system. Water from the fish-killing 38-L assay was filtered and centrifuged to produce fractions dominated by dinoflagellates, bacteria, or presumed ichthyotoxin (cell-free fraction). After 96 h, the larval fish assay exhibited 50%–100% cumulative mortality only in fractions containing dinoflagellates, with no mortalities occurring in the other fractions. The 500-mL bioassay with tilapia produced inconsistent results and demonstrated no clear correlation between mortality and treatment. The new larval fish bioassay was demonstrated as a highly effective method to verify and evaluate dinoflagellate pathogenicity. [ABSTRACT FROM AUTHOR]

The two most toxic Gambierdiscus species identified from the Caribbean are G. excentricus and G. silvae. These species are the primary causes of ciguatera fish poisoning and likely contribute disproportionately to the toxicity of marine food webs. While Gambierdiscus species are difficult to distinguish using light or scanning electron microscopy, reliable species-specific molecular identification methods have been developed and used successfully to identify a number of other Gambierdiscus species. Corresponding species-specific assays are not yet available for G. excentricus and G. silvae, which imposes limitations on species identification and related ecological studies. The following note describes species-specific polymerase chain reaction assays for G. excentricus and G. silvae that can be used for these purposes., (© 2019 Phycological Society of America.)

Knowledge concerning the ability of microalgae to produce metabolites of interest such as toxins or high-value secondary metabolites requires exhaustive details to be supplied on how the research was conducted. These should include the microalgal species and strain characterization, the culture conditions, the cell density, and physiological state at the time of harvesting, the harvesting method, the sample pre-treatment protocol, and the subsequent instrumental analytical separation/detection system. In this comment, we discuss issues that affect algal research from an analytical chemistry perspective, particularly (i) the need to specify detection capabilities of the entire method (i.e., limits of detection or threshold detection levels), which we illustrate in relation to classification of a species or strain as being "toxin producing" or "non-toxin producing"; and (ii) the requirements that have to be satisfied to confirm a microalgal species (new or not) as a producer of a particular chemical of interest for phycologists, which again we illustrate in relation to toxins. A successful collaboration among phycologists and analytical chemists will only be achieved as a result of a synergistic collaboration between the two disciplines, with a reciprocal understanding at least at a background level., (© 2013 Phycological Society of America.)