Μg STX Eq Research Articles

Contamination Status and Risk Assessment of Paralytic Shellfish Toxins in Shellfish along the Coastal Areas of China.

Paralytic shellfish toxins (PSTs) are widely distributed in shellfish along the coast of China, causing a serious threat to consumer health; however, there is still a lack of large-scale systematic investigations and risk assessments. Herein, 641 shellfish samples were collected from March to November 2020, and the PSTs' toxicity was detected via liquid chromatography-tandem mass spectrometry. Furthermore, the contamination status and potential dietary risks of PSTs were discussed. PSTs were detected in 241 shellfish samples with a detection rate of 37.60%. The average PST toxicities in mussels and ark shells were considerably higher than those in other shellfish. The PSTs mainly included N-sulfonylcarbamoyl toxins (class C) and carbamoyl toxins (class GTX), and the highest PST toxicity was 546.09 μg STX eq. kg-1. The PST toxicity in spring was significantly higher than those in summer and autumn (p < 0.05). Hebei Province had the highest average PST toxicity in spring. An acute exposure assessment showed that consumers in Hebei Province had a higher dietary risk, with mussels posing a significantly higher dietary risk to consumers. This research provides reference for the green and sustainable development of the shellfish industry and the establishment of a shellfish toxin prevention and control system.

Occurrence and spatial distribution of paralytic shellfish toxins in seawater and marine organisms in the coastal waters of Qinhuangdao, China

In recent years, paralytic shellfish toxins (PSTs) have been prevalent in the coastal waters of Qinhuangdao, the west coast of the Bohai Sea, China. The content of PSTs in shellfish often exceeded the regulatory limit of 800 μg STX equivalent (eq.) kg−1, which poses a serious threat to human health. In this study, two surveys were conducted in May 2021 and May 2022 to investigate the distribution of PSTs in the coastal waters of Qinhuangdao. Seawater, surface sediment, phytoplankton, zooplankton, and other marine organism samples were collected, and the composition and concentration of PSTs were analyzed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Results showed that multiple PST components were detected in all seawater samples collected at different depths, mainly including GTX1/4, GTX2/3, dcGTX2, STX and C1/2, and the highest concentration of PSTs reached 244 ng STX eq. L−1. The sediment samples also contained low levels of C1/2 and GTX2/3. Trace amounts of C1/2 and GTX1-4 were detected in phytoplankton and zooplankton. Moreover, all bivalve shellfish samples were found to contain PSTs, and the scallop Azumapecten farreri and the ark clam Anadara kagoshimensis showed relatively high concentrations of 607 and 497 μg STX eq. kg−1, respectively. In addition, low levels of PSTs were also found in some non-traditional PST vectors, including whelk Rapana venosa, octopus Amphioctopus ovulum, goby Ctenotrypauchen chinensis, and greenling Hexagrammos agrammus. Results of this study improve the understanding of the distribution of PSTs in seawater and marine organisms and the potential risk of persistent PSTs in seawater to marine ecosystems and human health.

Use of valvometry as an alert tool to signal the presence of toxic algae Alexandrium catenella by Mytilus edulis

Valvometry is a non-invasive technique used to continuously monitor gaping behaviour of bivalves at high frequency. In previous laboratory studies, valvometry has revealed a behavioural sensitivity of bivalves to the presence of toxic microalgae in seawater. However, the application of valvometry as an early-warning system for detecting natural occurrences of toxic microalgae and their resulting toxicity in bivalves remains largely unexplored. In this study, valvometry was used to characterise changes in blue mussels (Mytilus edulis) gaping behaviour during gradual exposure to the toxic dinoflagellate, Alexandrium catenella, which produces paralytic shellfish toxins (PST). Laboratory experiments were first performed to identify specific gaping behaviour changes and these responses were subsequently validated in natural seawater conditions in a second experiment. Under both laboratory and natural seawater conditions, mussels exposed to A. catenella tended to remain open (yawning) longer than non-exposed mussels. This change in gaping behaviour was observed at PST concentration as low as 30 μg STXeq 100 g–1 of mussel tissue. We suggest that increased opening is likely related to temporary muscular paralysis induced by toxic algae, as this mechanism has been previously reported in other bivalve species. Furthermore, we observed that biological rhythms of valve behaviour related to tidal and daily rhythms were modified when mussels were intoxicated by PSP (Paralytic Shellfish Poisoning). In conclusion, the effects of toxic algae on mussel gaping behaviour reveals that valvometry could be used as an early-warning tool for the presence of toxic Alexandrium sp. in the environment prior to mussels reaching the regulatory threshold (80 μg STXeq 100 g–1) for harvest interdiction.

Paralytic shellfish toxins in Alaskan Arctic food webs during the anomalously warm ocean conditions of 2019 and estimated toxin doses to Pacific walruses and bowhead whales

Climate change-related ocean warming and reduction in Arctic sea ice extent, duration and thickness increase the risk of toxic blooms of the dinoflagellate Alexandrium catenella in the Alaskan Arctic. This algal species produces neurotoxins that impact marine wildlife health and cause the human illness known as paralytic shellfish poisoning (PSP). This study reports Paralytic Shellfish Toxin (PST) concentrations quantified in Arctic food web samples that include phytoplankton, zooplankton, benthic clams, benthic worms, and pelagic fish collected throughout summer 2019 during anomalously warm ocean conditions. PSTs (saxitoxin equivalents, STX eq.) were detected in all trophic levels with concentrations above the seafood safety regulatory limit (80 μg STX eq. 100 g−1) in benthic clams collected offshore on the continental shelf in the Beaufort, Chukchi, and Bering Seas. Most notably, toxic benthic clams (Macoma calcarea) were found north of Saint Lawrence Island where Pacific walruses (Odobenus rosmarus) are known to forage for a variety of benthic species, including Macoma. Additionally, fecal samples collected from 13 walruses harvested for subsistence purposes near Saint Lawrence Island during March to May 2019, all contained detectable levels of STX, with fecal samples from two animals (78 and 72 μg STX eq. 100 g−1) near the seafood safety regulatory limit. In contrast, 64% of fecal samples from zooplankton-feeding bowhead whales (n = 9) harvested between March and September 2019 in coastal waters of the Beaufort Sea near Utqiaġvik (formerly Barrow) and Kaktovik were toxin-positive, and those levels were significantly lower than in walruses (max bowhead 8.5 μg STX eq. 100 g−1). This was consistent with the lower concentrations of PSTs found in regional zooplankton prey. Maximum ecologically-relevant daily toxin doses to walruses feeding on clams and bowhead whales feeding on zooplankton were estimated to be 21.5 and 0.7 μg STX eq. kg body weight−1 day−1, respectively, suggesting that walruses had higher PST exposures than bowhead whales. Average and maximum STX doses in walruses were in the range reported previously to cause illness and/or death in humans and humpback whales, while bowhead whale doses were well below those levels. These findings raise concerns regarding potential increases in PST/STX exposure risks and health impacts to Arctic marine mammals as ocean warming and sea ice reduction continue.

Paralytic shellfish toxins in Peruvian scallops associated with blooms of Alexandrium ostenfeldii (Paulsen) Balech & Tangen in Paracas Bay, Peru

In recent years, dense Alexandrium ostenfeldii blooms have been reported in different coastal areas. In this study, we report for the first time the occurrence of A. ostenfeldii blooms associated with the detection of paralytic shellfish toxins (PSTs) in the Peruvian scallop (Argopecten purpuratus) from Paracas Bay. Alexandrium ostenfeldii blooms occurred at the end of summer and early fall, after the increase of riverine input and under stratified conditions following a decrease in wind velocity. The highest abundances occurred during warm sea surface temperatures (18–27 °C). High PST concentrations that exceed the maximum permissible level (800 μg STX eq. kg−1) occurred even under low A. ostenfeldii abundances (20 × 103 cells l−1). Our results contribute to a better understanding of the dynamics of A. ostenfeldii in coastal systems influenced by riverine inputs and upwelling and can be used to improve monitoring programs and allow the implementation of mitigation measures along the Peruvian coast.

Effects of the toxic dinoflagellate Alexandrium catenella on the behaviour and physiology of the blue mussel Mytilus edulis

The effects of harmful algae on bivalve physiology are complex and involve both physiological and behavioural responses. Studying those responses is essential to better describe and predict their impact on shellfish aquaculture and health risk for humans. In this study we recorded for two months the physiological response of the blue mussel Mytilus edulis from Eastern Canada to a one-week exposure to a paralytic shellfish poisoning producing dinoflagellate strain of Alexandrium catenella, isolated from the St Lawrence estuary, Canada. Mussels in a ‘control’ treatment were fed continuously with a non-toxic diet, while mussels in a ‘starvation’ treatment were fed the same non-toxic diet the first week and subsequently starved for seven weeks. Mussels in a ‘toxic’ treatment received A. catenella for one week before being starved until the end of the experiment. Over a two-month experiment we monitored shell and tissue growth, filtration capacity, respiration rate, byssal attachment strength, valve opening behaviour, and toxin content in tissues. Mussels fed normally on the toxic dinoflagellate and accumulated an average of 51.6 µg STXeq 100 g–1 after one week of exposure. After seven weeks of depuration, about half of the specimen showed levels around 18 µg STXeq 100 g–1. The condition index of exposed mussels (‘toxic’ treatment) decreased rapidly from the start as compared to mussels that received a one-week non-toxic diet (‘starvation’ treatment). Oxygen consumption rates increased in the ‘toxic’ treatment before leveling out with that of mussels from the ‘starvation’ treatment. Valve opening amplitude was lower in the ‘toxic’ treatment during and following the exposure. Average valve closure duration was higher right after the exposure, during the peak of mussel tissue intoxication. No significant change in byssal thread strength was observed through time in each treatment but less force was required to detach mussels from the ‘toxic’ and ‘starvation’ treatments. The number of byssus threads produced by mussels exposed to the toxic dinoflagellate was also lower than in the control group. These results represent advancements in our understanding of the impacts of harmful algae on bivalves and contribute to the development of mitigation measures necessary to both the safety of consumers and the sustainability of aquaculture operations.

Chitosan performance during Paralytic Shellfish Toxins (PST) depuration of Mytilus chilensis exposed to Alexandrium catenella

Blooms of the dinoflagellate Alexandrium catenella, which produces Paralytic Shellfish Toxins (PST), generate serious socio-economic consequences for mariculture in Chile, especially for the production of Mytilus chilensis and other bivalves. Palliative strategies, such as the depuration of mussels in enriched water with chitosan offer encouraging prospects against the advance of contaminated areas and toxin persistence. Adult mussels were fed with A. catenella for 20 days and then were allowed to depurate using chitosan as facilitator, for the same period. Intoxicated mussels showed a reduction in feeding activity and rapid PST accumulation in 20 days (C = 451.5t + 1,673.6, R2 = 0.55 p = 0.008). Not enough evidence was found to indicate a positive effect of chitosan in mussel depuration after 20 days (C = −311.1t + 8,462.4, R2 = 0.8 p = 0.001). At the end of the study, toxicity was higher than 800 μg STX eq kg−1. C2 and GTX4 analogues were the most abundant in the dinoflagellate strain, while C2 and C1 were the most accumulated in mussels. The presence of C1 was notorious during depuration, as the persistence of GTX2,3. GTX5 was only detected in A. catenella, while STX was only present in mussels. Mussel sensitivity to the presence of the toxic dinoflagellate was observed in the present study. The biotransformation, selective elimination and epimerization processes were deduced from intoxication and depuration experiments.

Identifying the Source Organisms Producing Paralytic Shellfish Toxins in a Subtropical Bay in the South China Sea.

Identifying the exact phytoplanktonic sources of paralytic shellfish toxins (PSTs) is crucial for monitoring and preventing the buildup of toxin pollution, especially for causative species occurring at low levels. Phytoplankton and shellfish samples were simultaneously collected from representative mariculture zones in Daya Bay, China. Low concentration/low toxicity PSTs predominated with N-sulfocarbamoyl toxins 1, 2 (C1/2) being detected in phytoplankton (≤6.25 pmol L-1) and shellfish (≤0.21 μg STXeq g-1), which pose a potential risk of seafood poisoning. High-throughput sequencing of the phytoplankton samples based on 18S rDNA V4 regions identified 93 genera in 445 operational taxonomic units (OTUs). Five OTUs were assigned to Alexandrium hiranoi, Ambicodamus leei, Alexandrium pacificum, Alexandrium minutum, and an uncertain Alexandrium sp. A. pacificum and A. minutum are candidate PST producers and observed under the light microscope with densities of 66-972 cells L-1. Three strains of toxigenic species were successfully isolated and identified as A. pacificum and A. minutum based on their 18S rDNA V4 regions. The predominant toxins in A. pacificum were C1/2 (43.9-53.6 fmol cell-1) and resembled the toxins found in field samples predominated with C1/2. A. minutum produced only gonyautoxins 2/3 (8.03 fmol cell-1). Therefore, A. pacificum was identified as the predominant PST contributor in this area. This research makes a valuable contribution to the understanding of the traceability of phycotoxins in marine waters.

Experimental exposure of the mussel Mytilus platensis (d'Orbigny, 1842) to the dinoflagellate Alexandrium catenella from Argentine Patagonia.

Individuals of Mytilus platensis were exposed to Alexandrium catenella to evaluate the accumulation and metabolization of paralytic shellfish toxins (PST) over a period of 25 days. Mussels were collected from the intertidal zone of Cerro Avanzado, Argentine Patagonia. After 16 days, the toxins in the tissues of mussels were detected by the methods of mouse bioassay and high performance liquid chromatography with fluorometric detection (HPLC-FDL). The accumulation kinetics of PST toxins in M. platensis fed with A. catenella fitted to a linear function, in which the accumulation rate was 31.2 µg STX eq kg-1 day-1. After 16 days, the PST toxin level in tissues of mussels reached 1178 µg STX eq kg-1 exceeding the safety limit for human consumption (800 µg STX eq kg-1 tissue), whereas the highest PST toxin level was reached at the end of the experimentation (1613 µg STX eq kg-1) at 25 days. Differences in the toxin profile of the dinoflagellates and the tissues of the mussels confirmed biotransformation of PST in the mussel digestive system. The toxin profile of M. platensis was dominated by the gonyautoxins GTX1 and GTX4, while the toxin profile of A. catenella was dominated by the N-sulfocarbamoyl toxin C2. To our knowledge, this is the first experimentation on a laboratory scale of PST toxins accumulation in M. platensis with a native strain of A. catenella of Argentine Patagonia.

Paralytic shellfish poisoning due to ingestion of contaminated mussels: A 2018 case report in Caparica (Portugal).

In Portugal, the potent paralytic shellfish toxins (PSTs) have appeared irregularly since the onset of a national monitoring program for marine biotoxins in 1986. In years where high contamination levels were attained in bivalves, sporadic cases of human poisonings have been recorded, as in 1994 and 2007. The reappearance of high contamination levels led to the appearance of new cases during the autumn of 2018. This study details toxin ingestion, symptomatology and toxin elimination and metabolization in the fluids of two patients, who ingested mussels from the Portuguese southwest coast and required hospitalization due to the severity of symptoms. Toxin elimination was confirmed by ELISA in plasma and urine samples. In mussel samples, the toxin profile obtained by HPLC-FLD displayed a wide diversity of toxins, typical of Gymnodinum catenatum ingestion. However, in the urine samples, the toxin profile was reduced to B1 and dcSTX. Abundant compounds in mussels having an O-sulphate at C11, such as C1+2 and dcGTX2+3, were absent in urine. In plasma, PSTs were not detected by HPLC-FLD. Calculated toxin ingestion, resulting from consumption of an estimated 200-g portion, was in the range of 104–120 μg STX eq./kg b. w.

Paralytic Shellfish Toxin Uptake, Assimilation, Depuration, and Transformation in the Southeast Asian Green-Lipped Mussel (Perna viridis).

Bivalve molluscs represent an important food source within the Philippines, but the health of seafood consumers is compromised through the accumulation of harmful algal toxins in edible shellfish tissues. In order to assess the dynamics of toxin risk in shellfish, this study investigated the uptake, depuration, assimilation, and analogue changes of paralytic shellfish toxins in Perna viridis. Tank experiments were conducted where mussels were fed with the toxic dinoflagellate Alexandrium minutum. Water and shellfish were sampled over a six day period to determine toxin concentrations in the shellfish meat and water, as well as algal cell densities. The maximum summed toxin concentration determined was 367 µg STX eq./100 g shellfish tissue, more than six times higher than the regulatory action limit in the Philippines. Several uptake and depuration cycles were observed during the study, with the first observed within the first 24 h coinciding with high algal cell densities. Toxin burdens were assessed within different parts of the shellfish tissue, with the highest levels quantified in the mantle during the first 18 h period but shifting towards the gut thereafter. A comparison of toxin profile data evidenced the conversion of GTX1,4 in the source algae to the less potent GTX2,3 in the shellfish tissue. Overall, the study illustrated the temporal variability in Perna viridis toxin concentrations during a modelled algal bloom event, and the accumulation of toxin from the water even after toxic algae were removed.

Paralytic Shellfish Toxins and Ocean Warming: Bioaccumulation and Ecotoxicological Responses in Juvenile Gilthead Seabream (Sparus aurata).

Warmer seawater temperatures are expected to increase harmful algal blooms (HABs) occurrence, intensity, and distribution. Yet, the potential interactions between abiotic stressors and HABs are still poorly understood from ecological and seafood safety perspectives. The present study aimed to investigate, for the first time, the bioaccumulation/depuration mechanisms and ecotoxicological responses of juvenile gilthead seabream (Sparus aurata) exposed to paralytic shellfish toxins (PST) under different temperatures (18, 21, 24 °C). PST were detected in fish at the peak of the exposure period (day five, 0.22 µg g−1 N-sulfocarbamoylGonyautoxin-1-2 (C1 and C2), 0.08 µg g−1 Decarbamoylsaxitoxin (dcSTX) and 0.18 µg g−1 Gonyautoxin-5 (B1)), being rapidly eliminated (within the first 24 h of depuration), regardless of exposure temperature. Increased temperatures led to significantly higher PST contamination (275 µg STX eq. kg−1). During the trial, fish antioxidant enzyme activities (superoxide dismutase, SOD; catalase, CAT; glutathione S-transferase, GST) in both muscle and viscera were affected by temperature, whereas a significant induction of heat shock proteins (HSP70), Ubiquitin (Ub) activity (viscera), and lipid peroxidation (LPO; muscle) was observed under the combination of warming and PST exposure. The differential bioaccumulation and biomarker responses observed highlight the need to further understand the interactive effects between PST and abiotic stressors, to better estimate climate change impacts on HABs events, and to develop mitigation strategies to overcome the potential risks associated with seafood consumption.

Spatiotemporal distribution of paralytic shellfish poisoning (PSP) toxins in shellfish from Argentine Patagonian coast

Harmful algal blooms (HABs) have been recorded in the Chubut Province, Argentina, since 1980, mainly associated with the occurrence of paralytic shellfish poisoning (PSP) toxins produced by dinoflagellates of the genus Alexandrium. PSP events in this area impact on fisheries management and are also responsible for severe human intoxications by contaminated shellfish. Within the framework of a HAB monitoring program carried out at several coastal sites along the Chubut Province, we analyzed spatiotemporal patterns of PSP toxicity in shellfish during 2000–2011. The highest frequency of mouse bioassays exceeding the regulatory limit for human consumption was detected in spring and summer, with average values of up to ≈70% and 50%, respectively. By contrast, a lower percentage of positive bioassays (2–8%) or no toxicity at all was usually detected during autumn and winter. The most intense PSP events were usually observed between November and January, with values of up to 4,000 μg STX eq 100 g−1, and showed a marked interannual variability both in their magnitude and location. In addition, a severe PSP outbreak was recorded during autumn, 2009, at Camarones Bay, with toxicity values of up to 14,000 μg STX eq 100 g−1. The scallop Aequipecten tehuelchus showed significantly higher toxicity values compared to other shellfish species in SJG and SMG, suggesting a lower detoxification capacity. Our results contribute to the understanding of HABs dynamics on the Argentine Patagonian coast.

Paralytic Shellfish Toxins in Surf Clams Mesodesma donacium during a Large Bloom of Alexandrium catenella Dinoflagellates Associated to an Intense Shellfish Mass Mortality.

In late February 2016, a harmful algal bloom (HAB) of Alexandrium catenella was detected in southern Chiloé, leading to the banning of shellfish harvesting in an extended geographical area (~500 km). On April 24, 2016, this bloom produced a massive beaching (an accumulation on the beach surface of dead or impaired organisms which were drifted ashore) of surf clams Mesodesma donacium in Cucao Bay, Chiloé. To determine the effect of paralytic shellfish poisoning (PSP) toxins in M. donacium, samples were taken from Cucao during the third massive beaching detected on May 3, 2016. Whole tissue toxicity evidence a high interindividual variability with values which ranged from 1008 to 8763 μg STX eq 100 g−1 and with a toxin profile dominated by GTX3, GTX1, GTX2, GTX4, and neoSTX. Individuals were dissected into digestive gland (DG), foot (FT), adductor muscle (MU), and other body fractions (OBF), and histopathological and toxin analyses were carried out on the obtained fractions. Some pathological conditions were observed in gill and digestive gland of 40–50% of the individuals that correspond to hemocyte aggregation and haemocytic infiltration, respectively. The most toxic tissue was DG (2221 μg STX eq 100 g−1), followed by OBF (710 μg STX eq 100 g−1), FT (297 μg STX eq 100 g−1), and MU (314 μg STX eq 100 g−1). The observed surf clam mortality seems to have been mainly due to the desiccation caused by the incapability of the clams to burrow. Considering the available information of the monitoring program and taking into account that this episode was the first detected along the open coast of the Pacific Ocean in southern Chiloé, it is very likely that the M. donacium population from Cucao Bay has not had a recurrent exposition to A. catenella and, consequently, that it has not been subjected to high selective pressure for PSP resistance. However, more research is needed to determine the effects of PSP toxins on behavioral and physiological responses, nerve sensitivity, and genetic/molecular basis for the resistance or sensitivity of M. donacium.

Effect of modified clay on the transition of paralytic shellfish toxins within the bay scallop Argopecten irradians and sediments in laboratory trials

Shellfish aquaculture is an important marine industry that has been threatened by harmful algal blooms (HABs) and, in recent years, toxic HABs in particular. The modified clay (MC) treatment is the most promising method to mitigate HABs and has been frequently applied in the field. To determine how the application of MC affects the accumulation, transformation, and detoxification of paralytic shellfish toxins (PSTs) in the bay scallop Argopecten irradians and surrounding sediments, a series of experiments over 16 days in a laboratory simulated Alexandrium tamarense bloom were conducted. After 3 h of exposure to A. tamarense at 2070 (±120) cells mL−1, the toxicity level in the tissues of the scallops increased to 105.9 (±16.6) μg STX eq. 100 g−1 in the control group (group A, with no addition of MC), while the toxicity levels in the scallops of groups B and C (0.1 g L−1 and 0.5 g L−1 MC added, respectively) remained at relatively low levels. During the 3 h, the scallops in group A had incorporated 73.2 mol% of the toxin contained in the initial A. tamarense algal culture, the scallops in group B had incorporated 13.5 mol% of the toxin, and the scallops in group C showed almost no elevation in tissue toxin content. Meanwhile, 56.8 mol% and 107.1 mol% of the PSTs contained in the initial algal culture were settled in the sediments along with the MC-algal cells combination in group B and C, respectively. The results indicated that the application of MC can effectively prevent filter-feeding bivalves from ingesting toxic algal cells and decrease the accumulation of toxins by settling the toxic algal cells in the sediment. It was also indicated that the toxins accumulated within the sediments were more rapidly detoxified than that in the scallop tissues. Toxins accumulated by the scallops persisted in the tissue for approximately 16 days, while in the sediments, all of the PSTs contents were below the HPLC detection limits 4 days after the addition of MC.

Transformation and Depuration of Paralytic Shellfish Toxins in the Geoduck Clam Panopea globosa From the Northern Gulf of California

In January 2015 a harmful algae bloom (HAB) of the dinoflagellate Gymnodinium catenatum occurred in the Northern Gulf of California (NGC). This species produces paralytic shellfish toxins (PSTs) a group of potent neurotoxins. The harvesting and commercialization of geoduck Panopea globosa are important economic activities in this region. These activities were prohibited for several months due to the accumulation of PSTs in clam tissues. We analyzed PSTs concentrations in P. globosa collected on a weekly basis during 2015 near San Felipe, Baja California. The goal of the study was to evaluate the transformation and depuration characteristics of PSTs in the geoduck tissues. Three organisms from each sampling date were dissected and the PST content was evaluated in the visceral mass and siphon by high performance liquid chromatography with post column oxidation (HPLC-PCOX). Ten selected samples were also analyzed by hydrophilic interaction chromatography coupled to tandem mass spectrometry (HILIC-MS/MS). Toxicity in the siphon was lower than 800 µg STX eq kg-1 (the regulatory limit for PSTs) in all samples. In contrast, the maximum toxicity in the visceral mass exceeded 21 times this limit. It took 210 days to reach the regulatory limit after a maximum toxicity of 16,740 µg STXeq kg-1 was detected. Therefore, P. globosa is a slow detoxifier bivalve with a depuration rate of 4.26 day-1 (adjusted to an exponential decay model; r2=0.80). C1&2 were the most abundant analogues in siphon and viscera samples collected close to the HAB occurrence. The concentration of these analogues decreased and more toxic analogues as GTX5, dcGTX2 and dcSTX were detected. M-type analogs were detected by HILIC-MS/MS and represented up to 75% of total PSTs in some samples. M-type analogs contributed to 48% of toxicity estimated in the sample. We report for the first time the depuration rate, PSTs profile and its change over time in P. globlosa. This information is essential to characterize the metabolism of toxins in this economically important bivalve but also to develop management plans for the fishery if the organism is going to be recurrently exposed to PSTs producing blooms, as seems the case for the NGC.

Combined effects of warming and acidification on accumulation and elimination dynamics of paralytic shellfish toxins in mussels Mytilus galloprovincialis

Harmful algal blooms (HAB) have been increasing in frequency and intensity most likely due to changes on global conditions, which constitute a significant threat to wild shellfish and its commercial farming. This study evaluated the impact of increasing seawater temperature and acidification on the accumulation/elimination dynamics of HAB-toxins in shellfish. Mytilus galloprovincialis were acclimated to four environmental conditions simulating different climate change scenarios: i) current conditions, ii) warming, iii) acidification and iv) interaction of warming with acidification. Once acclimated, mussels were exposed to the paralytic shellfish toxins (PSTs) producing dinoflagellate Gymnodinium catenatum for 5 days and to non-toxic diet during the subsequent 10 days. High toxicity levels (1493 µg STX eq. kg−1) exceeding the safety limits were determined under current conditions at the end of the uptake period. Significantly lower PSP toxicity levels were registered for warming- and acidification-acclimated mussels (661 and 761 µg STX eq. kg−1). The combined effect of both warming and acidification resulted in PSP toxicity values slightly higher (856 μg STX eq. kg−1). A rapid decrease of toxicity was observed in mussels at the current conditions after shifting to a non-toxic diet, which was not noticed under the predicted climate change scenarios. Variability of each PST analogue, measured throughout the experiment, highlighted different mechanisms are associated with changes of each environmental factor, although both resulting in lower toxicity. Warming-acclimated mussels showed lower accumulation/elimination rates, while acidification-acclimated mussels showed higher capability to accumulate toxins, but also a higher elimination rate preventing high toxicity levels. As different mechanisms are triggered by warming and acidification, their combined effect not leads to a synergism of their individual effects. The present work is the first assessing the combined effect of climate change drivers on accumulation/elimination of PSTs, in mussels, indicating that warming and acidification may lead to lower toxicity values but longer toxic episodes. PSTs are responsible for the food poisoning syndrome, paralytic shellfish poisoning (PSP) in humans. This study can be considered as the first step to build models for predicting shellfish toxicity under climate change scenarios.

Exposure assessment to paralytic shellfish toxins through the shellfish consumption in Korea

Paralytic shellfish poisoning is caused by saxitoxin and its analogues. The paralytic shellfish toxins (PSTs) are produced by marine dinoflagellates and can be accumulated in filter feeding shellfish, such as mussel, clam, oyster and ark shell. The worldwide regulatory limits for PSTs in shellfish are set at 80 μg STX eq./100 g meat and this is widely accepted as providing adequate public health protection. In this study, we have determined five individual PSTs (STX, GTX1, GTX2, GTX3 and GTX4) in shellfish using LC-MS/MS and assessed the human acute and chronic exposures to PSTs through shellfish consumption. Food consumption data was obtained from the Korea National Health and Nutrition Examination Survey (KNHANES 2010–2015). The acute exposure using a large portion size of 88 g/day (95th percentile for consumers only) with maximum toxin level of 198.7 μg/kg was 0.30 μg/kg bw. Even though we estimated the acute exposure with a conservative manner, it was below the ARfDs (0.5 or 0.7 μg STX eq./kg bw) proposed by the international organizations, representing 43–60% of the ARfDs. The chronic exposures using mean consumption data for whole population with mean concentration of PSTs were ranged from 0.002 to 0.026 μg STX eq./kg bw/day. For consumers only, the chronic exposures were in the range of 0.012–0.128 μg STX eq./kg bw/day.

The role of a PSP-producing Alexandrium bloom in an unprecedented diamondback terrapin (Malaclemys terrapin) mortality event in Flanders Bay, New York, USA

Diamondback terrapins (Malaclemys terrapin) are a threatened or endangered species in much of their range along the U.S. Atlantic and Gulf coasts. Over an approximately three-week period from late April to mid-May 2015, hundreds of adult diamondback terrapins were found dead on the shores of Flanders Bay, Long Island, New York, USA. Concurrent with the mortality event, elevated densities of the paralytic shellfish toxin (PST)-producing dinoflagellate, Alexandrium fundyense (>104 cells L−1) and high levels of PST in bivalves (maximal levels = 540 μg STX eq. 100 g−1 shellfish tissue) were observed in the Flanders Bay region, resulting in shellfish bed closures in regional tributaries. Gross and histologic postmortem examinations of terrapins revealed no physical trauma to individuals or a common, underlying disease process to explain the deaths. PST compounds (0.2–12.5 μg STX eq. 100 g−1) were present in various M. terrapin tissues collected over the duration of the mortality event. High-throughput sequencing revealed that the ribbed mussel (Geukensia demissa, a PST vector) was present in the gastrointestinal tracks of all terrapin samples tested. While the potential of PST to cause mortality in chelonians has not been well-characterized, in the absence of other significant findings from necropsies and pathological analyses, we provide evidence that PST in shellfish was likely high enough to cause or contribute to the mortality in these small (<2.0 kg) animals.

Saxitoxin Level Comparation in Bali Sardine (Sardinella Lemuru) in Bali Strait in Different Monsoons

Sardinella fish ( Sardinella lemuru ) is a species of important pelagic fish from the family of Clupeidae found in Bali Strait. The dynamic condition of Bali Strait makes it fertile which is identical with plankton blooming. The objective of this research was to understand the potential of saxitoxin in sardinella fish because of dinoflagellates consumption in Bali Strait. The research was conducted in Southeast Monsoon (June - August 2015) and Northwest Monsoon (December 2015 – February 2016). The method used was in situ plankton sampling, counting and ELISA test to determine the level of saxitoxin in the fish. Hydro-oceanographic parameters measured were temperatures, salinity, DO (dissolved oxygen), pH level, phosphate level, nitrate level and transparency. They showed average optimum value for phytoplankton growth especially during Southeast Monsoon which its temperature lower and the nutrients (phosphate nitrate) higher than the Northwest Monsoon. This condition caused high richness of phytoplankton in the water and followed with dinoflagellates richness so they accumulated in sardinella fish. It was supported by the analysis of the fish gastric which showed positive correlation between dinoflagellates richness in the water and the fish gastric. ELISA test also showed that saxitoxin level of fish caught in Southeast Monsoon was higher than the one caught in Northwest Monsoon even though the level of saxitoxin was still in the safe range (less than 80 μg STXeq. per 100 g) but the danger of saxitoxin accumulation should be watched out for.