Shellfish Toxin Profiles Research Articles

Variability in Paralytic Shellfish Toxin Profiles and Dinoflagellate Diversity in Mussels and Seawater Collected during Spring in Korean Coastal Seawater.

Paralytic shellfish toxins (PSTs) are potent neurotoxins produced by certain microalgae, particularly dinoflagellates, and they can accumulate in shellfish in coastal seawater and thus pose significant health risks to humans. To explore the relationship between toxicity and PST profiles in seawater and mussels, the spatiotemporal variations in PST concentrations and profiles were investigated along the southern coast of Korea under peak PST levels during spring. Seawater and mussel samples were collected biweekly from multiple stations, and the toxin concentrations in the samples were measured. Moreover, the dinoflagellate community composition was analyzed using next-generation sequencing to identify potential PST-producing species. The PST concentrations and toxin profiles showed substantial spatiotemporal variability, with GTX1 and GTX4 representing the dominant toxins in both samples, and C1/2 tending to be higher in seawater. Alexandrium species were identified as the primary sources of PSTs. Environmental factors such as water temperature and salinity influenced PST production. This study demonstrates that variability in the amount and composition of PSTs is due to intricate ecological interactions. To mitigate shellfish poisoning, continuous monitoring must be conducted to gain a deeper understanding of these interactions.

Interannual variability in mesoscale distribution of Dinophysis acuminata and D. acuta in Northwestern Patagonian fjords

Dinophysis acuminata and D. acuta, which produce diarrheogenic toxins and pectenotoxins in southern Chile, display site-specific differences in interannual variability (2006 – 2018) in Reloncaví, Pitipalena and Puyuhuapi fjords (41 – 46 °S), Chilean Patagonia. Linear Models show decreasing trends in rainfall and river discharge. Latitudinal decreasing gradients in SST temperature and vertical salinity gradients were observed. A brackish water layer (FW salinity <11 psu), permanently present in Reloncaví, decreased in thickness with time in Pitipalena and was usually absent in Puyuhuapi, the only fjord where D. acuta reached bloom (>103 cells L‒1) densities every season. Dinophysis acuminata, associated with toxin profiles in shellfish that include only pectenotoxins, bloomed everywhere with a poleward increasing gradient. Absence of the FW layer provides a possible index of risk for D. acuta blooms. An apparent poleward shift of D. acuta populations, responsible for DSP outbreaks in Reloncaví in the 1970s, and the recent EU deregulation of pectenotoxins will have a positive impact on the mussel industry in Los Lagos Region. Changes to ongoing monitoring protocols to improve risk assessment capabilities are suggested.

Paralytic shellfish toxin profiles in mussel, cockle and razor shell under post-bloom natural conditions: Evidence of higher biotransformation in razor shells and cockles

Concentrations of the paralytic shellfish toxins GTX6, C1+2, GTX5, C3+4, dcSTX, dcNEO and dcGTX2+3 were determined by LC-FLD in composite samples of whole soft tissues of mussels (Mytilus galloprovincialis), cockles (Cerastoderma edule) and razor shells (Solen marginatus) after exposure to a Gymnodinium catenatum bloom. Specimens were harvested weekly during three months under natural depuration conditions in the Mira branch of Aveiro lagoon, Portugal. Under the decline of G. catenatum cell densities, elimination or transformation of the uptake toxins associated with the ingestion of toxic cells differed among the surveyed species. Ratio between the toxins dcSTX plus dcGTX2+3 plus dcNEO and toxins GTX6 plus GTX5 plus C1+2 plus C3+4 was used to illustrate the biotransformation occurring in the bivalves. Enhancement of the ratios was observed for razor shells and cockles seven weeks after the peak of the algal bloom. Most likely it reflects more intense biotransformation in razor shells and cockles than in mussels. Conversion into toxins of higher toxicity may prolong the bivalve toxicity. These results show the complexity of toxin elimination in bivalves under post-bloom conditions and emphasize the pertinence of monitoring programs of bivalve toxicity in order to protect human health.

Insertions within the Saxitoxin Biosynthetic Gene Cluster Result in Differential Toxin Profiles.

The neurotoxin saxitoxin and related paralytic shellfish toxins are produced by multiple species of cyanobacteria and dinoflagellates. This study investigates the two saxitoxin-producing strains of Scytonema crispum, CAWBG524 and CAWBG72, isolated in New Zealand. Each strain was previously reported to have a distinct paralytic shellfish toxin profile, a rare observation between strains within the same species. Sequencing of the saxitoxin biosynthetic clusters ( sxt) from S. crispum CAWBG524 and S. crispum CAWBG72 revealed the largest sxt gene clusters described to date. The distinct toxin profiles of each strain were correlated to genetic differences in sxt tailoring enzymes, specifically the open-reading frame disruption of the N-21 sulfotransferase sxtN, adenylylsulfate kinase sxtO, and the C-11 dioxygenase sxtDIOX within S. crispum CAWBG524 via genetic insertions. Heterologous overexpression of SxtN allowed for the proposal of saxitoxin and 3'-phosphoadenosine 5'-phosphosulfate as substrate and cofactor, respectively, using florescence binding assays. Further, catalytic activity of SxtN was confirmed by the in vitro conversion of saxitoxin to the N-21 sulfonated analog gonyautoxin 5, making this the first known report to biochemically confirm the function of a sxt tailoring enzyme. Further, SxtN could not convert neosaxitoxin to its N-21 sulfonated analog gonyautoxin 6, indicating paralytic shellfish toxin biosynthesis most likely occurs along a predefined route. In this study, we identified key steps toward the biosynthetic conversation of saxitoxin to other paralytic shellfish toxins.

Dinophysis acuta in Scottish Coastal Waters and Its Influence on Diarrhetic Shellfish Toxin Profiles.

Diarrhetic shellfish toxins produced by the dinoflagellate genus Dinophysis are a major problem for the shellfish industry worldwide. Separate species of the genus have been associated with the production of different analogues of the okadaic acid group of toxins. To evaluate the spatial and temporal variability of Dinophysis species and toxins in the important shellfish-harvesting region of the Scottish west coast, we analysed data collected from 1996 to 2017 in two contrasting locations: Loch Ewe and the Clyde Sea. Seasonal studies were also undertaken, in Loch Ewe in both 2001 and 2002, and in the Clyde in 2015. Dinophysis acuminata was present throughout the growing season during every year of the study, with blooms typically occurring between May and September at both locations. The appearance of D. acuta was interannually sporadic and, when present, was most abundant in the late summer and autumn. The Clyde field study in 2015 indicated the importance of a temperature front in the formation of a D. acuta bloom. A shift in toxin profiles of common mussels (Mytilus edulis) tested during regulatory monitoring was evident, with a proportional decrease in okadaic acid (OA) and dinophysistoxin-1 (DTX1) and an increase in dinophysistoxin-2 (DTX2) occurring when D. acuta became dominant. Routine enumeration of Dinophysis to species level could provide early warning of potential contamination of shellfish with DTX2 and thus determine the choice of the most suitable kit for effective end-product testing.

A review of the global distribution of Alexandrium minutum (Dinophyceae) and comments on ecology and associated paralytic shellfish toxin profiles, with a focus on Northern Europe.

Alexandrium minutum is a globally distributed harmful algal bloom species with many strains that areknown to produce paralytic shellfish toxins (PSTs) and consequently represent a concern to human andecosystem health. This review highlightsthat A.minutum typically occurs in sheltered locations, with cell growth occurring during periods of stable water conditions. Sediment characteristics are important in the persistence of this species within a location, with fine sediments providing cyst deposits for ongoing inoculation to the water column. Toxic strains of A.minutum do not produce a consistent toxin profile, different populations produce a range of PSTs in differing quantities. Novel cluster analysis of published A.minutum toxin profiles indicates five PST profile clusters globally. Some clusters are grouped geographically (Northern Europe) while others are widely spread. Isolates from Taiwan have arange of toxin profile clusters and this area appears to have the most diverse set of PST producing A.minutum populations. These toxin profiles indicate that within the United Kingdom there are two populations of A.minutum grouping with strains from Northern France and Southern Ireland. There is a degree of interconnectivity in this region due tooceanic circulation and a high level of shipping andrecreational boating. Further research into theinterrelationships between the A.minutum populations in this global region would be of value.

Domoic acid--a new toxin in the Croatian Adriatic shellfish toxin profile.

This is the first study that presents concentrations of domoic acid detected in the whole shellfish tissue from breeding and harvesting areas along the Croatian coast of the Adriatic Sea during the period 2006 to 2008. Shellfish sample analyses after SAX cleaning procedures, using a UV-DAD-HPLC system, showed the presence of domoic acid in four species. The most prevalent of those species were the blue mussel (Mytilus galloprovincialis), followed by European flat oyster (Ostrea edulis), Mediterranean scallop(Pecten jacobaeus) and proteus scallop (Flexopecten proteus). Domoic acid, a potentially lethal phycotoxin that causes amnesic shellfish poisoning (ASP), was detected for the first time in January 2006 with the highest value of 6.5486 μg g-1 in whole shellfish tissue. Pseudo-nitzschia spp. bloom events preceded these high domoic acid concentrations. According to this study, retention of domoic acid in the blue mussel M. galloprovincialis is more than 42 days. This investigation indicates the first presence of domoic acid in Croatian shellfish, but in concentrations under the regulatory limit (20 μg g-1), therefore shellfish consumption was not found to endanger human health.

A convenient and cost-effective method for monitoring marine algal toxins with passive samplers

Passive sampling disks were developed based on the method of MacKenzie, L., Beuzenberg, V., Holland, P., McNabb, P., Selwood, A. [2004. Solid phase adsorption toxin tracking (SPATT): a new monitoring tool that simulates the biotoxin contamination of filter feeding bivalves. Toxicon 44, 901–918] and protocols were formulated for recovering toxins from the adsorbent resin via elution from small columns. The disks were used in field studies to monitor in situ toxin dynamics during mixed algal blooms at Flødevigen in Norway. Examples are given from time-integrated sampling using the disks followed by extraction and high performance liquid chromatography–mass spectrometry (HPLC–MS) analysis for azaspiracids, okadaic acid analogues, pectenotoxins, yessotoxins and spirolides. Profiles of accumulated toxins in the disks and toxin profiles in blue mussels ( Mytilus edulis) were compared with the relative abundance of toxin-producing algal species. Results obtained showed that passive sampling disks correlate with the toxin profiles in shellfish. The passive sampling disks were cheap to produce and convenient to use and, when combined with HPLC–MS or enzyme-linked immunosorbent assay (ELISA) analysis, provide detailed time-averaged information on the profile of lipophilic toxin analogues in the water. Passive sampling is therefore a useful tool for monitoring the exposure of shellfish to the toxigenic algae of concern in northern Europe.

Toxic Alexandrium minutum (Dinophyceae) from Vietnam with new gonyautoxin analogue

Clonal cultures of Alexandrium species collected from a shrimp pond on the northern coast of Vietnam were established and morphologically identified as Alexandrium minutum. Nucleotide sequences of domains 1 and 2 of the large subunit ribosomal (LSU) rRNA gene showed high sequence similarity to A. minutum isolates from Malaysia. Paralytic shellfish toxin profile of the clones was characterized by the dominance of GTX4, GTX1, and NEO. GTX3, GTX2, and dcSTX were also present in trace amount. Toxin content varied among the strains and growth stages, ranged from 3.0 to 12.5 fmol cell −1. In addition to these known toxin components, a new gonyautoxin derivative was detected by HPLC, eluting between GTX4 and GTX1. The peak of this compound disappeared under non-oxidizing HPLC condition but unchanged either after treated with 0.05 M ammonium phosphate/10% mercaptoethanol or 0.1N HCl hydrolysis. LCMS ion scanning showed a parental ion of [ M + H] + at m/ z 396, [ M − SO 3] + at m/ z 316, and [ M − SO 4] + at m/ z 298. Based on these results, the derivative was identified as deoxy-GTX4-12ol, and this represents the first report of this toxin analogue.

Paralytic shellfish toxin profiles and toxin variability of the genus Alexandrium (Dinophyceae) isolated from the Southeast China Sea

Paralytic shellfish toxin (PST) profiles of 16 Alexandrium isolates from the Southeast China Sea were analyzed by high-pressure liquid chromatography. Toxin content and composition of three A. tamarense isolates, ATDH01, ATGX02 and ATMJ02, were also investigated at different growth phases and under various culture conditions. Our results showed that six strains of A. affine were non-toxic, while 10 strains of A. tamarense and A. catenella were toxic. These toxic isolates grown in the same culture conditions consistently produced an unusually high proportion of the N-sulfocarbamoyl toxin C1/2 (around 60–80% of total toxins) and medium amounts of gonyautoxin GTX5 (around 15–30% of total) with only trace quantities (<5% of total) of other saxitoxin derivatives (i.e. GTX1, GTX3, GTX4 and neoSTX). The toxin composition of three A. tamarense isolates did not vary with the growth phases, although higher toxin contents (Qt, fmol cell −1) were found in the exponential phase. Variations in temperature, salinity and nutrient levels affected toxin content of three A. tamarense isolates but they did not have pronounced effects on the toxin composition (mole %). These results indicate that toxin composition remained relatively constant under various culture conditions, suggesting that toxin composition could be used as a stable biomarker for the Alexandrium species in this region. However, comparison of toxin profiles between isolates from different localities require special caution since isolates even from the same region can have distinct toxin profiles.

Paralytic shellfish toxin profiles in net phytoplankton samples from Bahía Concepción, Gulf of California, Mexico

a Centro Interdisciplinario de Ciencias Marinas (CICIMAR-IPN), Avenida Politecnico s/n, Col. Playa Palo de Santa Rita, La Paz, B.C.S. 23090, Mexico b Apdo. Postal 592, Col. Centro, La Paz, B.C.S. 23000, Mexico c Centro de Investigaciones Biologicas del Noroeste (CIBNOR), Mar Bermejo 195, Col. Playa Palo de Santa Rita, La Paz, B.C.S. 23090, Mexico d Friedrich-Schiller University, Faculty of Biology and Pharmacy, Dornburgerstrase 25, 07743 Jena, Germany

Comparative paralytic shellfish toxin profiles in the strains of Gymnodinium catenatum Graham from the Gulf of California, Mexico

Comparative paralytic shellfish toxin profiles in the strains of Gymnodinium catenatum Graham from the Gulf of California, Mexico

Paralytic shellfish toxin profiles of different geographic populations of Gymnodinium catenatum (Dinophyceae) in Korean coastal waters

Paralytic shellfish toxin profiles of different geographic populations of Gymnodinium catenatum (Dinophyceae) in Korean coastal waters

Paralytic shellfish toxin profiles of different geographic populations of Gymnodinium catenatum (Dinophyceae) in Korean coastal waters

SUMMARY Paralytic shellfish poisoning toxin profiles of dinoflagellate cultures of Gymnodinium catenatum Graham from the Yellow and South Seas in Korea were investigated by high performance liquid chromatography fluorometric detection. Strains from the Yellow Sea had predominantly carbamate toxins, while strains from Sujeongri and Chindong in the South Sea contained the N-sulfocarbamoyl toxins, Cl,2, as major components including the presence of GTX5 and dcSTX in some strains. Toxin profiles from St. Deukryang Bay strains (South Sea) showed both characteristics of those in the South Sea and those in the Yellow Sea. Thirty strains could be divided into three groups based on cluster analysis of toxin compositions. Group I (Yellow Sea strains) was distinguished from Group II (Sujeongri and Chindong strains) by the absence of GTX5 and dcSTX. Group III comprised Deukryang Bay strains. In conclusion, the Yellow Sea and the South Sea were found to have different dinoflagellate populations with different toxin compositions.

Comparative paralytic shellfish toxin profiles in two marine bivalves during outbreaks of Gymnodinium catenatum (Dinophyceae) in the Gulf of California

Comparative paralytic shellfish toxin profiles in two marine bivalves during outbreaks of Gymnodinium catenatum (Dinophyceae) in the Gulf of California

Comparative study on paralytic shellfish toxin profiles of the dinoflagellate Gymnodinium catenatum from three different countries

Paralytic shellfish toxin profiles of the dinoflagellate Gymnodinium catenatum Graham were investigated as a possible biochemical marker to distinguish different geographic populations of this species. Isolates obtained between 1986 and 1988 from Japan, Tasmania (Australia) and Galicia (Spain) were cultured under similar conditions and the toxins produced were analyzed using HPLC. Variations in temperature, salinity, and nitrate and phosphate levels in the culture medium had no significant effect on the toxin profile, suggesting that toxins can be used as a stable biochemical marker for this dinoflagellate. All the isolates produced mainly toxins of the N-sulfocarbamoyl group (C1–C4, gonyautoxins 5 and 6) but their relative abundance differed according to their geographic origin. Furthermore, only the Australian population produced the newly found 13-deoxydecarbamoyl toxins, and these could readily be used to distinguish the Australian populations from those of the other two countries.

Quantification of diarrhetic shellfish toxins and identification of novel protein phosphatase inhibitors in marine phytoplankton and mussels

Liquid chromatography (LC)-linked protein phosphatase 1/2A (PP-1/PP2A) bioassay was used to quantitatively identify diarrhetic shellfish toxins in marine phytoplankton (cultured and natural assemblages) and commercially available mussels. Using this approach, multiple protein phosphatase inhibitor profiles of varying composition were found in diarrhetic mussels from Holland and Canada. Based on LC elution positions and relative activity versus PP-1 and PP-2A, at least six inhibitors distinct from known diarrhetic shellfish toxins were identified and termed mussel phosphatase inhibitor (MPI) 19,22,23,25,33 and 42. The levels of these inhibitors, in okadaic acid equivalent units, varied from 100 pg to 3350 ng per g shellfish tissue. The combined levels of PP-1/2A inhibitors in all instances superseded that of okadaic acid/dinophysistoxin-1 and may contribute to the diarrhetic shellfish toxin profile of the contaminated mussels. The efficacy of LC-protein phosphatase bioassay was established for cultured phytoplankton where picogram levels of okadaic acid could be detected from microgram extracts of Prorocentrum lima. Analyses of plankton net tows from estuarine mussel culture sites in Eastern Canada revealed a heterogeneous population of protein phosphatase inhibitors, with dinophysistoxin-1 being most prevalent. This toxin was predominant for at least 2 months in mussel populations in the immediate vicinity of plankton sampling sites. The results are consistent with a hypothetical model in which marine bacteria, cyanobacteria and dinoflagellates combine to produce a variety of protein phosphatase inhibitors effective against signal transduction pathways in higher eukaryotes.

Toxicity and paralytic shellfish toxin profiles of the xanthid crabs, Lophozozymus pictor and Zosimus aeneus, collected from some Australian coral reefs

—Purification of toxic aqueous extracts from the xanthid crabs Zosimus aeneus and Lophozozymus pictor, collected from Australian waters, yielded paralytic shellfish toxins, including saxitoxin (STX), neosaxitoxin (neoSTX) and gonyautoxins 1, 2 and 4 (GTX 1,2,4). No more than two paralytic shellfish toxins were found in any of the purified extracts from any specimen. Four specimens of Z. aeneus and one specimen of L. pictor each contained more toxic material than the suggested human oral lethal dose. The moult of a specimen of L. pictor was toxic, which may indicate a route in crabs for toxin removal.