• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.1
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• pp.158-160
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• 1997

Food poisoning accident occurred on May, 1996 at Oepo, Geoje County, Kyeongnam Province, Korea, and two persons were died within a few hours after ingestion of the soup prepared with wild mussel, Mytilus edulis, harvested on the sea rock. Paralytic shellfish poisons (PSP) were elucidated as the responsible toxins for the food poisons accident because the wild mussels caught after three days at the near place from the accident contained high toxicity of PSP ranged $650\~1000MU/g$ of edible meat by mouse bioassay. Gonyautoxin-1+4 $(42.7\%)$ and C1+C2 $(40.0\%)$ were detected as the major toxins in the mussels by fluorometric HPLC method. Although, the poison extracted out with drip during freezing and thawing, and the toxicity gradually decreased by boiling for 20 minutes, over 30 MU/g of toxins remained in the soup and meat, which indicated that they could be able to make food poisoning.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.2
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• pp.147-153
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• 2013

To prevent paralytic shellfish poisoning (PSP) due to the consumption of shellfish contaminated with PSP toxins, the quantitative analysis of these toxins is very crucial. The AOAC International mouse bioassay (MBA) has been used widely for the routine monitoring of PSP toxins for more than 50 years. However, this method has low sensitivity and high limit of quantification (LOQ) and interferences from other components in the extract, and it cannot determine toxic profiles. Ethical problems also exist with the continued use of this live mouse assay. To establish an alternative method to the MBA used for PSP toxins analysis, we attempted to optimize the analysis conditions of a precolumn high-performance liquid chromatography (HPLC) oxidation method and succeeded in validating its accuracy and precision in quantifying PSP toxins. A clear peak and the isolation of PSP toxins were obtained by injecting the working standards of Certified Reference Materials using HPLC. The LOQ of the precolumn HPLC oxidation method for PSP toxins was about $0.1002{\mu}g/g$, which represented an approximately fourfold improvement in detection capability versus the AOAC MBA. The intra-accuracy and precision for PSP toxins in oysters were 77.0-103.3% and 2.0-5.7%, respectively, while the respective inter-accuracy and precision were 77.3-100.7% and 2.4-6.0%. The mean recoveries of PSP toxins from oysters were 75.2-112.1%. The results of a comparison study showed good correlation between the results of the precolumn HPLC oxidation method and those of MBA, with a correlation factor of 0.9291 for mussels. The precolumn HPLC oxidation method may be used as an alternative to, or supplementary method with, MBA to monitor the occurrence of PSP toxins and to analyze the profiles of these toxins in shellfish.

• Fisheries and Aquatic Sciences
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• v.16 no.3
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• pp.137-142
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• 2013

To understand critical aspects of paralytic shellfish poisoning (PSP) in a chief area of bivalve production in Korea, seasonal variation in PSP toxins in bivalves collected from Jinhae Bay, Korea in 2009 was surveyed by the pre-column high-performance liquid chromatography oxidation method. We also confirmed the profiles of major bivalves such as oysters Crassostrea gigas and mussels Mytilus galloprovincialis in Jinhae Bay. PSP toxins in the bivalves showed remarkable seasonal variation. PSP toxin levels were detected from April to May in 2009, and the highest total toxin levels at all stations were recorded in May. The major toxins in bivalves were gonyautoxin [GTX] 1&4 and C 1&2; in oysters GTX 2&3 were also detected as major components. GTX 1&4, which showed the highest PSP toxin levels at each station, accounted for the highest proportions of toxin components in mussels and oysters (64.5-71.3% and 41.4-42.4%, respectively). It was also confirmed that the highest toxicity (in ${\mu}g$ saxitoxin [STX] eq/g) was derived from GTX 1&4. The highest total toxicity (in ${\mu}g$ STX eq/g) was approximately 2-8-fold higher in mussels than in oysters collected from the same station. PSP toxin levels in bivalves differed significantly according to the sample collection station. However, the profiles of toxins in the bivalves did not show significant differences during the survey period according to sample collection station. This study shows that PSP toxin levels in some samples from Jinhae Bay were above the regulatory limit in Korea during a specific period in spring.

• Fisheries and Aquatic Sciences
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• v.8 no.2
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• pp.76-82
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• 2005

We analyzed the paralytic shellfish poisoning (PSP) toxins of the toxic marine dinoflagellate Alexandrium tamarense collected from Dadaepo and Gaduck-do in Busan and from Sujeong-ri in Jinhae Bay, Korea, in April 2003. We also analyzed the PSP toxin of mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) collected around Busan and Jinhae Bay. PSP toxin analyses were conducted by high performance liquid chromatography (HPLC). Fifteen cultured A. tamarense isolates contained 2.78 to 57.47 fmol/cell, with nearly identical toxin profiles: major components C2, GTX4; minor components C1, GTX1, NEO; and trace components GTX2, GTX3, STX. PSP toxin contents were 0 to $492\;\mu{g}$ STXeq/100 g in mussels and 0 to $48\;\mu{g}$ STXeq/100 g in oysters. Mussels at Gijang and Sujeong-ri contained the most PSP toxin contents ($492\;\mu{g}\;STXeq/100\;g\;and\;252\;\mu{g}\;STXeq/100\;g,\;respectively$), exceeding the quarantine level ($80\;\mu{g}$ STXeq/100 g). Their dominant toxin components were C2, C1, GTX2, and GTX3; the minor components GTX1, GTX4, GTX5, and NEO were sporadically detected. Phytoplankton contained 0.774 fmol/L seawater and 1.228 fmol/L seawater at Gijang and Sujeong-ri in April. At that time, Alexandrium cells were present in the water column at Gijang at 2,577 cells/mL and at Sujeong-ri at 6,750 cells/mL. Overall, we found the high and similar PSP toxin contents in AZexandrium isolates and mussels, and a correlation between occurrence of toxic Alexandrium cells in the water column and mussel intoxification. High densities of toxic Alexandrium cells in the water column immediately preceded shellfish intoxification at Gijang and Sujeong-ri in April.

• Journal of Environmental Science International
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• v.18 no.12
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• pp.1331-1338
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• 2009

Marine dinoflagellate Alexandrium minutum producing paralytic shellfish toxins is responsible for paralytic shellfish poisoning (PSP). To investigate its temporal distributions in Chinhae Bay where PSP occurs annually, SYBR Green I based A. minutum-specific real-time PCR probe was developed on the LSU rDNA region. Assay specificity and sensitivity were tested against related species, and its specificity was further confirmed by sequencing of field-derived samples. Ten months field survey in 2008 (a total 100 surface water samples) by using the real-time PCR probe showed that A. minutum was detected at very low densities of 1-4 cells $L^{-1}$ in May and June being spring in Chinhae Bay, Korea.

• Fisheries and Aquatic Sciences
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• v.12 no.3
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• pp.227-235
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• 2009

The marine toxic dinoflagellate Alexandrium catenella has been implicated in numerous paralytic shellfish poisoning (PSP) events in many countries. Due to difficulties in rapidly identifying A. catenella, field-based study of this species has been problematic. The present study developed a TaqMan format A. catenella-specific probe for real-time PCR assay (specific to Korean genotype) based on LSU rDNA sequence information for studying geographic and temporal distribution of the species in surface sediments and water columns of Jinhae Bay, Korea. The field survey from 2007 to 2008 revealed that A. catenella occurred in most seasons at low densities, mostly below 1 cell $mL^{-1}$, and was more abundant in spring (maximum cell density of 2 cells $mL^{-1}$) when shellfish exceed the quarantine toxin level for PSP toxins in Jinhae Bay.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.39 no.1
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• pp.49-54
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• 2006

Cultured cells of the toxic Alexandrium tamarense were fed to four mussel species, Mytilus coruscus, M. edulis, M. galloprovincialis and Septifer vulgatus, to examine the interspecies and interlocality differences in the ability to accumulate paralytic shellfish poisoning (PSP) toxins. Toxin content of A. tamarense cells varied during culture period. In contrast, toxin composition in the cell (C1,2, GTX1-4 and neoSTX) was constantly stable. In feeding experiment, the four mussel species collected from Geoje intoxicated after uptake of A. tamarense. Toxin content ($average{\pm}SD\;{\mu}g$ STXeq/100 g) of M. coruscus, M. edulis, M. galloprovincialis and Septifer vulgatus were $1,660{\pm}79,\;3,914{\pm}2,242,\;5,626{\pm}1,620\;and\;958{\pm}163$, respectively. Toxin profiles included C1,2, GTX1,4 and neoSTX as the major components, and dcGTX2,3, GTX2,3, neoSTX and STX as the minor ones. Toxin accumulation of three mussel species collected from Pohang, Geoje and Anmyon-do showed interspecies and interlocality differences. Toxin content ($average{\pm}SD\;{\mu}g$ STXeq/100 g) were $91{\pm}4,\;151{\pm}14,\;39{\pm}3$ in M coruscus, $189{\pm}1,\;231{\pm}11,\;206{\pm}15$ in M edu/is and $214{\pm}28,\;326{\pm}30,\;291{\pm}26$ in M. galloprovincialis in order of Anmyon-do, Geoje and Pohang.

• Fisheries and Aquatic Sciences
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• v.16 no.3
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• pp.159-164
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• 2013

The mouse bioassay and high performance liquid chromatography (HPLC) post-column oxidation method are different methods of quantifying paralytic shellfish poisoning toxins. In this study, we compared their ability to accurately quantify the toxicity levels in two types of field sample (oysters and mussels) with different toxin profiles for routine regulatory monitoring. A total of 72 samples were analyzed by both methods, 44 of which gave negative results, with readings under the limit of detection of the mouse bioassay ($40{\mu}g/100g$ saxitoxin [STX] eq). In 14 oysters, the major toxin components were gonyautoxin (GTX) 1, -2, -3, -4, -5, decarbamoylgonyautoxin-2 (dcGTX2), and decarbamoylsaxitoxin (dcSTX), while 14 mussels tested positive for dcSTX, GTX2, -3, -4, -5, dcGTX2, neosaxitoxin (NEO), STX, and dcSTX. When the results obtained by both methods were compared in two matrices, a better correlation ($r^2=0.9478$) was obtained for mussels than for oysters ($r^2=0.8244$). Additional studies are therefore needed in oysters to investigate the differences in the results obtained by both methods. Importantly, some samples with toxin levels around the legal limit gave inconsistent results using HPLC-based techniques, which could have a strong economic impact due to enforced harvest area closure. It should therefore be determined if all paralytic shellfish poisoning toxins can be quantified accurately by HPLC, and if the uncertainties of the method lead to doubts regarding regulatory limits.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.3
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• pp.364-372
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• 1995

On the outbreak of paralytic shellfish poisoning in April 1993 in most of shellfish harvesting areas in Jinhae Bay, Korea, to clarify the toxin production of causative organism Alexandrium species, 19 axenic clonal isolates established from the benthic resting cysts in three different stations of those culture grounds were subjected to PSP toxin analysis by HPLC. Individual toxin content per cell was highly variable among the strains isolated from a sampling area and originated from an individual cyst. Average toxin contents in those areas revealed higher values of 54-70 fmol/cell. Toxin profiles included C1/C2(epiGTX8/GTX8), GTX1/GTX4 and neoSTX as the major components, and GTX2/GTX3, GTX5, C4, dcSTX and STX as the minor or sporadic ones. neoSTX on the dominant toxins showed not only most diverse compositional changes comprising $5-54 mol\%$ ranges but also no detection on the half of the strains examined, which were implicated in arising of heterogeneity with a genetic trait within a geographical region. When average toxin composition was compared, carbamate toxins comprised large proportions of $57\%,\;54\%\;and\;67\%$ as total toxin in St. 1, St. 2 and St. 4, respectively. These results suggested that an extensive paralytic shellfish toxification in Jinhae Bay could be largely due to the production of highly potent carbamate toxins in the causative dinoflagellate Alexandrium species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.4
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• pp.293-299
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• 1987

At various times and places all over the world men have become ill and some have died after eating shellfish that were intoxicated with paralytic shellfish poison(PSP) caused by Protogonyaulax spp. In late March, 1986, two persons were dead by ingesting wild sea mussels, Mytilur edulis, grown at bottom of an anchored waste ship to be dismantled at Gamchun Bay, Pusan, Korea. The samples were collected from the bottom of the ship during April $1\~April$ 8 of the year to find the cause of the food poisoning accident. The toxicity was estimated by bioassay with ICR male mouse, while the toxins were extracted and characterized. The toxins were extracted with acidified $80\%$ ethanol. The extract was defatted three times with dichloromethane, treated with activated charcoal, and then purified by chromatography on Bio-Gel P-2 and Bio-Rex 70. The toxic fractions obtained were analysed by cellulose acetate membrane electrophoresis, thin layer chromatography and high performance liquid chromatogaphy. The range and the average of PSP-toxicity of the samples were $132\~295\;MU/g$, 203 MU/g respectively. The amount of PSP was $26.4\~58.9{\mu}g/g$ of whole meat in range and $40.6{\mu}g/g$ in average. The toxicity of the digestive gland of the samples was 9 times higher than that of edible meat (except digestive gland) as $439\~979MU/g$, and it was about $70\%$ in total toxin. The compositional analytical results of the paralytic shellfish toxin, Gonyautoxin $1\~4$ were the major part of the PSP and Saxitoxin and neosaxitoxin were detected as the minor component. It was concluded that the food poisoning accident was caused not by Saxitoxins but by Gonyautoxins.