Paralytic Shellfish Poisoning Toxins Research Articles

Determination of hydrophilic marine biotoxins in bivalve molluscs by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS)

This work describes a method for the determination of paralytic shellfish toxins in mussels (M. galloprovincialis), oysters (Crassostrea gigas), and clams (Ruditapes decussatus) samples. The acetic acid extraction method is used for simultaneous analysis of Paralytic Shellfish Poisoning (PSP) and the separation of these compounds was achieved on ethylene bridged hybrid (BEH) Amide column with gradient mobile phase composed of acetonitrile (MeCN) and water (as eluent A and eluent B) both containing 2 mM ammonium formate and 50 mM formic acid. The method specificity, linearity, sensitivity, precision, and accuracy were successfully evaluated towards the detection of 13 hydrophilic marine biotoxins analyzed together (single run) in different sample matrixes such as mussels. oysters. and clams. Calibration curves were found to be linear (R2 ≥ 0.98). average Recovery% of 98.5–99.9%. limits of detection (LODs): 0.16–1.92 µg·kg−1; limits of quantitation (LOQs): 0.54–6.41 µg·kg−1) and the relative standard deviation (RSD%) of the precision was ≤ 2.8%. The method was applied for the determination of PSP in shellfish collected from several Tunisian locations. All LC-MS/MS results of analyzed molluscs collected from different Tunisia locations, reveal no PSP toxin group contamination and that the amount found does not exceeding the threshold of 800 µg STXequ·kg−1.

Growth and paralytic shellfish poisoning toxin production by a Mexican dinoflagellate strain of Alexandrium tamiyavanichii Balech (1994) under different nutrient conditions

Alexandrium tamiyavanichii is a marine dinoflagellate known to produce Paralytic Shellfish Poisoning (PSP) toxin. Thus, a strain was isolated from La Paz Bay, Baja California Sur, Mexico and used to explore whether stress conditions, such as phosphorus limitation (PL) and nitrogen enrichment (NE) modulate population growth and PSP toxin production in the GSe medium. Growth kinetics showed that the PL treatment produced a 3.4-fold increase in cell density versus control at day 30 of the culture cycle. The highest PSP concentration was found in the control culture (309 fmol cell−1) on day 21. Saxitoxin (STX) was the main analog in all the treatments (> 40 % mol). In conclusion, PL and NE treatments promoted growth kinetics in the species studied but did not affect the PSP toxin production. For the first time, the present research describes A. tamiyavanichii high toxicity strain isolated from Mexican coasts relative to the South-Atlantic strains.

Expansion risk of the toxic dinoflagellate Gymnodinium catenatum blooms in Chinese waters under climate change

The paralytic shellfish poison toxin (PST)-producing dinoflagellate, Gymnodinium catenatum, frequently blooms in China, posing a threat to food safety and human health. To understand the drivers of G. catenatum blooms and predict potential habitats for G. catenatum under climate change, samples from occurrence localities and environmental datasets from multiple agencies were aggregated and used to model the habitat suitability of G. catenatum in the China Sea using a maximum entropy model (Maxent). The accumulated variable contributions for the Maxent model were defined to measure the importance of key predictors in the model. The most important environmental variables were distance to the coastline, depth of seawater, and long-term average of the minimum annual temperature. This highlights the main reasons why G. catenatum blooms always occur in coastal waters. Occurrence probabilities higher than 0.66 were defined as habitats with high suitability for shellfish management and aquaculture. Projected habitats with high suitability in Haizhou Bay, coastal waters along the western Taiwan Strait, and Bohai Bay remained stable with increasing temperature by 2100, regardless of the IPCC Representative Concentration Pathways (RCPs). However, those in the China Sea would be reduced overall, leading to a northward movement of the center of integrated habitats. Habitats with a spatial area of >6000 km2 in the Bohai Sea, Yellow Sea, and South China Sea and >23,000 km2 in the East China Sea would be exposed to high risk under low greenhouse gas emission scenarios (RCP2.6).

Definition of a saxitoxin (STX) binding code enables discovery and characterization of the anuran saxiphilin family.

American bullfrog (Rana castesbeiana) saxiphilin (RcSxph) is a high-affinity ‘toxin sponge’ protein thought to prevent intoxication by saxitoxin (STX), a lethal bis-guanidinium neurotoxin that causes paralytic shellfish poisoning (PSP) by blocking voltage-gated sodium channels (NaVs). How specific RcSxph interactions contribute to STX binding has not been defined and whether other organisms have similar proteins is unclear. Here, we use mutagenesis, ligand binding, and structural studies to define the energetic basis of Sxph:STX recognition. The resultant STX ‘recognition code’ enabled engineering of RcSxph to improve its ability to rescue NaVs from STX and facilitated discovery of ten new frog and toad Sxphs. Definition of the STX binding code and Sxph family expansion among diverse Anurans separated by ∼140 million years of evolution provides a molecular basis for understanding the roles of toxin sponge proteins in toxin resistance and for developing novel proteins to sense or neutralize STX and related PSP toxins.

Modelling the Spatial and Temporal Dynamics of Paralytic Shellfish Toxins (PST) at Different Scales: Implications for Research and Management.

Harmful algal blooms, in particular recurrent blooms of the dinoflagellate Alexandrium catenella, associated with paralytic shellfish poisoning (PSP), frequently limit commercial shellfish harvests, resulting in serious socio-economic consequences. Although the PSP-inducing species that threaten the most vulnerable commercial species of shellfish are very patchy and spatially heterogeneous in their distribution, the spatial and temporal scales of their effects have largely been ignored in monitoring programs and by researchers. In this study, we examined the spatial and temporal dynamics of PSP toxicity in the clam (Ameghinomya antiqua) in two fishing grounds in southern Chile (Ovalada Island and Low Bay). During the summer of 2009, both were affected by an intense toxic bloom of A. catenella (up to 1.1 × 106 cells L-1). Generalized linear models were used to assess the potential influence of different environmental variables on the field detoxification rates of PSP toxins over a period of 12 months. This was achieved using a four parameter exponential decay model to fit and compare field detoxification rates per sampling site. The results show differences in the spatial variability and temporal dynamics of PSP toxicity, given that greater toxicities (+10-fold) and faster detoxification (20% faster) are observed at the Ovalada Island site, the less oceanic zone, and where higher amounts of clam are annually produced. Our observations support the relevance of considering different spatial and temporal scales to obtain more accurate assessments of PSP accumulation and detoxification dynamics and to improve the efficacy of fisheries management after toxic events.

Definition of a saxitoxin (STX) binding code enables discovery and characterization of the anuran saxiphilin family

American bullfrog (Rana castesbeiana) saxiphilin (RcSxph) is a high-affinity "toxin sponge" protein thought to prevent intoxication by saxitoxin (STX), a lethal bis-guanidinium neurotoxin that causes paralytic shellfish poisoning (PSP) by blocking voltage-gated sodium channels (NaVs). How specific RcSxph interactions contribute to STX binding has not been defined and whether other organisms have similar proteins is unclear. Here, we use mutagenesis, ligand binding, and structural studies to define the energetic basis of Sxph:STX recognition. The resultant STX "recognition code" enabled engineering of RcSxph to improve its ability to rescue NaVs from STX and facilitated discovery of 10 new frog and toad Sxphs. Definition of the STX binding code and Sxph family expansion among diverse anurans separated by ∼140 My of evolution provides a molecular basis for understanding the roles of toxin sponge proteins in toxin resistance and for developing novel proteins to sense or neutralize STX and related PSP toxins.

Effect of Different N:P Ratios on the Growth, Toxicity, and Toxin Profile of Gymnodinium catenatum (Dinophyceae) Strains from the Gulf of California

The harmful microalgae Gymnodinium catenatum is a unique naked dinoflagellate that produces paralytic shellfish poisoning toxins (PSTs). This species is common along the coasts of the Mexican Pacific and is responsible for paralytic shellfish poisoning, which has resulted in notable financial losses in both fisheries and aquaculture. In the Gulf of California, G. catenatum has been related to mass mortality events in fish, shrimp, seabirds, and marine mammals. In this study, the growth, toxin profiles, and toxin content of four G. catenatum strains isolated from Bahía de La Paz (BAPAZ) and Bahía de Mazatlán (BAMAZ) were evaluated with different N:P ratios, keeping the phosphorus concentration constant. All strains were cultivated in semi-continuous cultures (200 mL, 21.0 °C, 120 µmol photon m−2s−1, and a 12:12 h light-dark cycle) with f/2 + Se medium using N:P ratios of: 4:1, 8:1, 16:1, 32:1, and 64:1. Paralytic toxins were analyzed by HPLC with fluorescence detection. Maximum cellular abundance and growth were obtained at an N:P ratio of 64:1 (3188 cells mL−1 and 0.34 div day−1) with the BAMAZ and BAPAZ strains. A total of ten saxitoxin analogs dominated by N-sulfocarbamoyl (60–90 mol%), decarbamoyl (10–20 mol%), and carbamoyl (5–10 mol%) toxins were detected. The different N:P ratios did not cause significant changes in the PST content or toxin profiles of the strains from both bays, although they did affect cell abundance.

Distribution of phycotoxins in Última Esperanza Province during the PROFAN expedition 2019

Harmful Algae Blooms pose an increasing threat to the public health and economic stability of Southern Chile, particularly to the aquaculture industries. This fieldwork performed during the PROFAN expedition from 12th to 22nd November 2019 extends the knowledge on the distribution of marine toxin-producing species in the difficult to access Última Esperanza Province in the Magallanes Region. Paralytic Shellfish Poisoning toxins with high relative abundances of saxitoxin and lipophilic toxins dominated by yessotoxins, pectenotoxins and domoic acid were detected at nearly each sampling station. The respective toxin-producing organisms are mainly from the genus Alexandrium and Dinophysis. Furthermore, the first detection of pinnatoxin-G (PnTx-G) in Chilean waters strongly indicates the presence of the dinoflagellate Vulcanodinium rugosum.

Spatiotemporal distribution of paralytic shellfish poisoning (PSP) toxins in Beagle Channel (South America) during 2005–2017

Toxigenic algal blooms have been reported since 1886 in the Beagle Channel, at the southern limit of South America, mainly associated with the occurrence of paralytic shellfish poisoning (PSP) toxins produced by dinoflagellates of the genus Alexandrium. Toxic outbreaks have serious impacts on marine wildlife and ecosystem services, particularly on fisheries and mussel aquaculture. Within the framework of the Shellfish Toxicity Monitoring Program carried out close to Almanza in Beagle Channel eastern coastal waters, here we analyzed the spatiotemporal patterns of PSP toxins in the Magellan mussel Aulacomya ater and the blue mussel Mytilus edulis, measured by the mouse bioassay for 12 years, between 2005 and 2017. PSP outbreaks occurred mostly during summer and their duration was significantly longer than for outbreaks detected during spring and winter. A marked interannual variability in their magnitude was also evident. The largest outbreaks were detected in summers 2010–2011, in extreme warm and low-wind conditions, reaching up to 5600 µg saxitoxin equivalent per 100 g of tissue, causing a closure of shellfish fisheries of up to 200 days. Small scale spatial variability in PSP was also found, the highest values being observed in the main mussel culture areas, and the lowest in the natural mussel beds. PSP toxicities for the Magellan mussel were significantly higher than for the blue mussel. Natural detoxification in mussels from Beagle Channel followed an exponential decay, with mean detoxification rates of 3.5% toxin day−1, lower than those from elsewhere in the world for similar PSP toxin concentrations, which means a longer time to attain concentration limits safe for human consumption. Understanding PSP dynamics is important to avoid large economic losses related to halting mussel production.

Progress on the investigation and monitoring of marine phycotoxins in China

Marine phycotoxins associated with paralytic shellfish poisoning (PSP), diarrhetic shellfish poisoning (DSP), amnesic shellfish poisoning (ASP), neurotoxic shellfish poisoning (NSP), ciguatera fish poisoning (CFP), tetrodotoxin (TTX), palytoxin (PLTX) and neurotoxin β-N-methylamino-L-alanine (BMAA) have been investigated and routinely monitored along the coast of China. The mouse bioassay for monitoring of marine toxins has been progressively replaced by the enzyme-linked immunosorbent assay (ELISA) and liquid chromatography tandem mass spectrometry (LC-MS/MS), which led to the discovery of many new hydrophilic and lipophilic marine toxins. PSP toxins have been detected in the whole of coastal waters of China, where they are the most serious marine toxins. PSP events in the Northern Yellow Sea, the Bohai Sea and the East China Sea are a cause of severe public health concern. Okadaic acid (OA) and dinophysistoxin-1 (DTX1), which are major toxin components associated with DSP, were mainly found in coastal waters of Zhejiang and Fujian provinces, and other lipophilic toxins, such as pectenotoxins, yessotoxins, azaspiracids, cyclic imines, and dinophysistoxin-2(DTX2) were detected in bivalves, seawater, sediment, as well as phytoplankton. CFP events mainly occurred in the South China Sea, while TTX events mainly occurred in Jiangsu, Zhejiang and Fujian provinces. Microalgae that produce PLTX and BMAA were found in the phytoplankton community along the coastal waters of China.

The value of toxin profiles in the chemotaxonomic analysis of paralytic shellfish toxins in determining the relationship between British Alexandrium spp. and experimentally contaminated Mytilus sp.

Although phytoplankton is ubiquitous in the world's oceans some species can produce compounds that cause damaging effects in other organisms. These include the toxins responsible for paralytic shellfish poisoning, which, in UK waters, are produced by dinoflagellates from the Alexandrium genus. Within Great Britain (GB) a monitoring programme exists to detect this harmful genus as well as the Paralytic Shellfish Poisoning (PSP) toxins in the flesh of shellfish from classified production areas. The techniques used for toxin analysis allow for detailed analysis of the toxin profiles present in contaminated shellfish. It is possible to compare the toxin profiles of contaminated shellfish with the profiles from toxin producing algae and use this information to infer the causative microalgal species responsible for the contamination. This study sought to evaluate the potential for this process within the GB monitoring framework. Two species of toxic Alexandrium, A. catenella from Scotland and A. minutum from Southern England, were fed to mussels (Mytilus sp.) under controlled conditions. The toxin profile in mussels derived from feeding on each species independently, when mixed and when introduced sequentially was analysed and compared to the source algal cultures using K means cluster analysis. Toxin profiles in contaminated shellfish clustered with those of the causative algae and separately from one another during toxin accumulation and, where A. catenella was the sole toxin source, during depuration. During depuration after feeding with A. minutum and where mixed or sequential feeding was undertaken deviant toxin profiles were observed. Finally, data generated within this experimental study were compared to monitoring data from the GB official control programme. These data indicated that the causative algal species in sole source contaminations could be inferred from toxin profile analysis. This technique will be of benefit within monitoring programmes to enhance the value of data with minimal additional expense, where the toxin profiles of causative microalgae have been well described.

Paralytic Shellfish Poisoning (PSP) in Mussels from the Eastern Cantabrian Sea: Toxicity, Toxin Profile, and Co-Occurrence with Cyclic Imines.

In the late autumn of 2018 and 2019, some samples taken by the official monitoring systems of Cantabria and the Basque Country were found to be paralytic shellfish poisoning (PSP)-positive using a mouse bioassay. To confirm the presence of PSP toxins and to obtain their profile, these samples were analyzed using an optimized version of the Official Method AOAC 2005.06 and using LC–MS/MS (HILIC). The presence of some PSP toxins (PSTs) in that geographical area (~600 km of coast) was confirmed for the first time. The estimated toxicities ranged from 170 to 983 µg STXdiHCl eq.·kg−1 for the AOAC 2005.06 method and from 150 to 1094 µg STXdiHCl eq.·kg−1 for the LC–MS/MS method, with a good correlation between both methods (r2 = 0.94). Most samples contained STX, GTX2,3, and GTX1,4, and some also had NEO and dcGTX2. All of the PSP-positive samples also contained gymnodimine A, with the concentrations of the two groups of toxins being significantly correlated. The PSP toxin profiles suggest that a species of the genus Alexandrium was likely the causative agent. The presence of gymnodimine A suggests that A. ostenfeldii could be involved, but the contribution of a mixture of Alexandrium species cannot be ruled out.

On paralytic shellfish poisoning producing capacity of <italic>Alexandrium tamarense</italic>

In order to explore the capacity of paralytic shellfish poisoning (PSP) production in <italic>Alexandrium tamarense</italic>, the algae strain was cultivated in different space volumes, salinities and nutrient concentrations with controlling other conditions unchanged. After culturing for a period of time, a certain number of algal cells were collected by centrifugation to analyze and calculate the toxin content and profiles by liquid chromatography tandem-mass spectrometry (LC-MS/MS). Results showed that: 1) <italic>A</italic>. <italic>tamarense</italic> could produce at least 13 PSP components, of which the main components were GTX1,GTX4, GTX5, C1 and C2, accounting for 97.2% of the total toxin content, and the content of GTX1 was as high as 40%. At the same time, there was a significant difference in the minimum number of <italic>A</italic>. <italic>tamarense</italic> cells required for extraction and the quantitative detection of each component. For example, when the number of algal cells used to extract and analyze toxin profiles was less than 4×10³, none of the 13 PSP components could be quantitatively detected. When the number of algae cells used to extract and analyze toxin profiles was between 4×10³ and 5.9×10⁶, only part of the toxin components could be quantitatively detected. All these 13 PSP components could be quantified only when the number of algae cells used to extract and analyze toxin profiles was higher than 5.9×10⁶. 2) The PSP producing capacity of <italic>A</italic>.<italic>tamarense</italic> cultured in salinity 25 and 35 was significantly higher than that cultured in salinity 15, the PSP producing capacity of <italic>A</italic>. <italic>tamarense</italic> cultured in the flask of 1 L was significantly higher than that cultured in the flask of 0.1 L, and cultured in the oligotrophic group was significantly higher than that cultured in the eutrophic group. From these results, we could know that <italic>A</italic>. <italic>tamarense</italic> had a rich toxin profile, and contained at least 13 PSP toxin components with known structures. <italic>A</italic>.<italic>tamarense</italic> could be used for basic research of PSP toxin, and it was also the best theoretical algae strain for separating and extracting GTX1. It showed stronger capacity of PSP production when cultivated in the flask of 1 L, at the salinity of 25 and 1/4 times of the f/2 medium concentration. <italic>A</italic>.<italic>tamarense</italic> blooms with high PSP toxicity may occur in marine environments with high salinity, sufficient space and high eutrophication. Thus, in order to reduce the risk of harmful <italic>A</italic>.<italic>tamarense</italic> blooms with high PSP toxicity, the supervision of sea areas that meet the above conditions should be strengthened carefully.

Sulfitobacter alexandrii sp. nov., a new microalgae growth-promoting bacterium with exopolysaccharides bioflocculanting potential isolated from marine phycosphere.

Marine phycosphere harbors unique cross-kingdom associations with enormous ecological significance in aquatic ecosystems as well as relevance for algal biotechnology industry. During our investigating the microbial composition and bioactivity of marine phycosphere microbiota (PM), a novel lightly yellowish and versatile bacterium designated strain AM1-D1T was isolated from cultivable PM of marine dinoflagellate Alexandrium minutum amtk4 that produces high levels of paralytic shellfish poisoning toxins (PSTs). Strain AM1-D1T demonstrates notable bioflocculanting bioactivity with bacterial exopolysaccharides (EPS), and microalgae growth-promoting (MGP) potential toward its algal host. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain AM1-D1T was affiliated to the members of genus Sulfitobacter within the family Rhodobacteraceae, showing the highest sequence similarity of 97.9% with Sulfitobacter noctilucae NB-68T, and below 97.8% with other type strains. The complete genome of strain AM1-D1T consisted of a circular 3.84-Mb chromosome and five circular plasmids (185, 95, 15, 205 and 348Kb, respectively) with the G+C content of 64.6%. Low values obtained by phylogenomic calculations on the average nucleotide identity (ANI, 77.2%), average amino acid identity (AAI, 74.7%) and digital DNA-DNA hybridization (dDDH, 18.6%) unequivocally separated strain AM1-D1T from its closest relative. The main polar lipids were identified as phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol, one unidentified phospholipid and one unidentified lipid. The predominant fatty acids (> 10%) were C18:1 ω7c, C19:0 cyclo ω8c and C16:0. The respiratory quinone was Q-10. The genome of strain AM1-D1T was predicted to encode series ofgene clusters responsible for sulfur oxidation (sox) and utilization of dissolved organic sulfur exometabolites from marine dinoflagellates, taurine(tau) and dimethylsulfoniopropionate (DMSP) (dmd), as well as supplementary vitamin B12 (cob), photosynthesis carotenoids (crt) which are pivotal components during algae-bacteria interactions. Based on the evidences by the polyphasic characterizations, strain AM1-D1T represents a novel species of the genus Sulfitobacter, for which the name Sulfitobacter alexandrii sp. nov. is proposed. The type strain is AM1-D1T (= CCTCC 2017277T = KCTC 62491T).

Algal blooms of Alexandrium spp. and Paralytic Shellfish Poisoning toxicity events in mussels farmed in Sicily.

Marine biotoxins can accumulate in filter- feeders bivalve molluscs, that may represent a source of potential health problems being vectors of toxins, that are transferred to humans through their consumption. Harmful Algal Blooms impact on aquaculture may give also economic losses due to temporary closures of contaminated shellfish harvest and marketing. The presence of toxic algae for Paralytic Shellfish Poisoning (PSP), with recurrent toxic blooms of dinoflagellates, such as several Alexandrium species, been known since 2000 in the waters of an Ionian bay of Sicily, the Syracuse harbour, where shellfish farms are located. Our previous works reported in this area the positivity for PSP toxin in mussels (Mytilus galloprovincialis) with saxitoxin concentrations above the limit of the law and the simultaneous presence of toxic species of the genus Alexandrium in the waters. This work reports new recent episodes of algal blooms of Alexandrium minutum in the waters of the Syracuse harbour and PSP toxin contamination in farmed mussels, with values beyond the limits established by law, with the consequent immediate closure of the production area. PSP toxicity was detected with the MBA (Mouse Bioassay) with the confirm carried out with Lawrence method to quantify the total saxitoxin equivalents and characterize the toxic profile. Regular application of the implemented health plan is very important in order to prevent any risk and protect consumer health.

Application of Six Detection Methods for Analysis of Paralytic Shellfish Toxins in Shellfish from Four Regions within Latin America.

With the move away from use of mouse bioassay (MBA) to test bivalve mollusc shellfish for paralytic shellfish poisoning (PSP) toxins, countries around the world are having to adopt non-animal-based alternatives that fulfil ethical and legal requirements. Various assays have been developed which have been subjected to single-laboratory and multi-laboratory validation studies, gaining acceptance as official methods of analysis and approval for use in some countries as official control testing methods. The majority of validation studies conducted to date do not, however, incorporate shellfish species sourced from Latin America. Consequently, this study sought to investigate the performance of five alternative PSP testing methods together with the MBA, comparing the PSP toxin data generated both qualitatively and quantitatively. The methods included a receptor binding assay (RBA), two liquid chromatography with fluorescence detection (LC-FLD) methods including both pre-column and post-column oxidation, liquid chromatography with tandem mass spectrometry (LC-MS/MS) and a commercial lateral flow assay (LFA) from Scotia. A total of three hundred and forty-nine shellfish samples from Argentina, Mexico, Chile and Uruguay were assessed. For the majority of samples, qualitative results compared well between methods. Good statistical correlations were demonstrated between the majority of quantitative results, with a notably excellent correlation between the current EU reference method using pre-column oxidation LC-FLD and LC-MS/MS. The LFA showed great potential for qualitative determination of PSP toxins, although the findings of high numbers of false-positive results and two false negatives highlighted that some caution is still needed when interpreting results. This study demonstrated that effective replacement methods are available for countries that no longer wish to use the MBA, but highlighted the importance of comparing toxin data from the replacement method using local shellfish species of concern before implementing new methods in official control testing programs.

Random forest classification to determine environmental drivers and forecast paralytic shellfish toxins in Southeast Alaska with high temporal resolution

Paralytic shellfish poison toxins (PSTs) produced by the dinoflagellate in the genus Alexandrium are a threat to human health and subsistence lifestyles in Southeast Alaska. It is important to understand the drivers of Alexandrium blooms to inform shellfish management and aquaculture, as well as to predict trends of PST in a changing climate. In this study, we aggregate environmental data sets from multiple agencies and tribal partners to model and predict concentrations of PSTs in Southeast Alaska from 2016 to 2019. We used daily PST concentrations interpolated from regularly sampled blue mussels (Mytilus trossulus) analyzed for total PSTs using a receptor binding assay. We then created random forest models to classify shellfish above and below a threshold of toxicity (80 µg 100 g−1) and used two methods to determine variable importance. We obtained a multivariate model with key variables being sea surface temperature, salinity, freshwater discharge, and air temperature. We then used a similar model trained using lagged environmental variables to hindcast out-of-sample (OOS) shellfish toxicities during April-October in 2017, 2018, and 2019. Hindcast OOS accuracies were low (37–50%); however, we found forecasting using environmental variables may be useful in predicting the timing of early summer blooms. This study reinforces the efficacy of machine learning to determine important drivers of harmful algal blooms, although more complex models incorporating other parameters such as toxicokinetics are likely needed for accurate regional forecasts.

Shellfish contamination with marine biotoxins in Portugal and spring tides: a dangerous health coincidence.

Bivalve molluscs can acquire marine biotoxins by filter-feeding upon certain toxin-producing microalgae. The two most common syndromes observed in temperate coastal waters have been diarrhetic shellfish poisoning (DSP) and paralytic shellfish poisoning (PSP). While DSP is a non-fatal gastrointestinal syndrome, PSP is a neurological syndrome which can lead to death by respiratory paralysis in high intoxication scenarios. In Portugal, the presence of both DSP and PSP toxins leads to recurrent seasonal bans of bivalve harvesting. On a few occasions, the bans were not placed in time, not properly disseminated to the public or were disregarded by recreational harvesters. Several cases of poisonings have been studied in collaboration between health authorities and the laboratory in charge of the biotoxin monitoring programme. Some of the outbreaks have even called the attention of the local media. In several of these recorded cases, a common trait has emerged throughout the years: bivalve harvest had often been done during very low tides attributed to either new or full moons. These tides expose intertidal bivalves more widely, increase harvesting time, and allow picking of larger-sized specimens. In some occasions, the consumers were extremely unfortunate because a noxious coincidence had occurred: larger-sized specimens were available but had attained the highest toxin content of the toxic season. This review alerts that despite costly monitoring programmes have been perfected through the years, human poisonings still take place due to the rapid increase in toxin levels and/or disrespect of harvest bans.

Wind-driven development and transport of Gymnodinium catenatum blooms along the coast of Fujian, China

Gymnodinium catenatum is a cosmopolitan, bloom-forming dinoflagellate known to produce a suite of potent paralytic shellfish poisoning (PSP) toxins. Here, we revisit two major blooms of G. catenatum along the Fujianese Coast, China, in 2017 and 2018. The impact area of the 2017 bloom was larger than that of the 2018 event. Field sampling and remote satellite sensing revealed that alongshore transport driven by the southwest wind, as well as physical accumulation driven by the northeast wind, played important roles in the development and distribution of the two bloom events. The relationship between wind-induced hydrodynamic conditions and the unprecedented HAB events established in this study adds greatly to our understanding of algal bloom dynamics along the Fujianese coast. These results improve our ability to detect, track, and forecast G. catenatum blooms, thereby potentially minimizing the negative impacts of future HAB events.

First determination of extracellular paralytic shellfish poisoning toxins in the culture medium of toxigenic dinoflagellates by HILIC–HRMS

Paralytic shellfish poisoning (PSP) toxins have received considerable attention in recent years because of their adverse effects on marine breeding industries and human health. In this study, a reliable method for the analysis of extracellular PSP toxins in the culture medium of marine toxic dinoflagellates was developed for the first time using graphitized carbon black–solid-phase extraction and hydrophilic interaction liquid chromatography–high-resolution mass spectrometry. The limit of quantification of typical PSP toxins in algal culture medium ranged from 0.072 μg/L to 0.151 μg/L under optimal conditions. Satisfactory absolute recoveries (87.5%–102.4%), precision (relative standard deviation ≤ 7.6%), and linearity (R2 ≥ 0.9998) were also achieved. In addition, the proposed method was applied to screen and determine the extracellular PSP toxins of two typical toxigenic dinoflagellates, Alexandrium minutum and Alexandrium tamarense. The total concentrations of the extracellular PSP toxins in A. minutum and A. tamarense over the whole growth period were within 2.0–735.5 and 2.0–19.2 μg/L, respectively. The concentrations of extracellular PSP toxins varied remarkably in the different growth stages of A. minutum and A. tamarense, and the contents of some extracellular PSP toxins were substantially higher than those of intracellular PSP toxins. Therefore, the extracellular PSP toxins released by toxigenic red tide algae cannot be ignored, and their environmental fate, bioavailability, and potential harm to aquatic environment need to be investigated in future studies.