• 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.

• Journal of Preventive Medicine and Public Health
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• v.21 no.1 s.23
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• pp.163-171
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• 1988

The paralytic shellfish poisoning was occurred among 25 laborers who worked at breaking-up of ships in Pusan for 5 days from March 29 to April 2 of 1956. For the purpose of accurately defining the paralytic shellfish poison(PSP) , the authors carried out mouse bioassay and chemical analysis. The results were summarized as follows: 1. The mean amount of Paralytic shellfish toxin was $1,207.8{\mu}g$ Per 100gm meat, and the mean death time of mouse was 5 minutes 16 second. 2. The properties of the PSP were mainly gonyautoxin group by chemical analysis(TLC, IR, $^{1}H-NMR$).

• Journal of Food Hygiene and Safety
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• v.14 no.3
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• pp.265-269
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• 1999

Paralytic shellfish poison threatens public health most seriously from April to early May every year and gives adverse effects on export of these products. Major shellfish products exported were canned oyster, Crassostrea gigas and blue mussel, Mytilus edulis. Toxicities of canned shellfishes with toxin of low levels were mostly inactivated during the processing; in contrast, residual toxicity was of great concern with canned products from highly toxic shellfishes. This study was to provide basic data to establish food safety measure by evaluating the changes of toxicity during 2 year storage of canned products with toxic blue mussel and oyster. Any significant difference was not observed between two samples. Boiled can and smoked can showed inactivation of toxicity to some extent, whereas acidified can did not show reduction of toxicity even after 2 year storage. In case the initial toxicity of shellfish was high long term storage could not inactivate the toxicity of the canned product.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.6
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• pp.546-549
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• 2000

For the establishment of biodetoxification method which can be acceptable for live bivalves, paralytic shellfish poison (PSP) detoxification bacteria were isolated from sea water and bivalves, and PSP detoxification activity and optimal growth condition of the isolated strains were investigated. from the bivalve and sea water samples, 8 strains of PSP detoxification bacteria were isolated. Of the isolated strains, CW-6 isolated from sea water shown strong PSP detoxification activity and decomposed completely 18 nmole/g of GTX2 after 3 days incubation in artificial medium. The selected stain CW-6 shown typical characteristics of the Enterobacter sp. and identified as Enterobacter sp, CW-6. Optimal growth condition of the Enterobacter sp. CW-6 were $35^{\circ}C$, pH 7 and $NaCl 1{\%}$, respectively.

• Korean Journal of Food Science and Technology
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• v.44 no.4
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• pp.390-392
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• 2012

Diarrhetic shellfish poisoning toxins were investigated by liquid chromatography-electrospray ionization mass spectrometry (LC-MS/MS). Okadaic acid (OA), Dinophysistoxin-1 (DTX1), Pectonotoxin2, (PTX2) and Yessotoxin (YTX) in bivalves were quantified. OA were found in four samples; mussel Mytilus edulis (0.001 ${\mu}g/g$), Oyster Crassostrea gigas (0.004 and 0.001 ${\mu}g/g$) and manila clam Ruditapes philippinarum (0.001 ${\mu}g/g$). DTX1, PTX2, and YTX were not detected from all of the samples examined.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.18-25
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• 1990

Paralytic Shellfish Poison (PSP) is mainly produced by marine dinoflagellates such as Protogonyaulax sp. and Pyrodinium sp.. The PSP was known to be accumulated in digestive gland of shellfish as result of feeding toxic dinoflagellates. PSP illness when occurs when one eats PSP intoxicated shellfish. Therefore PSP is becoming as serious problem in food hygiene and shellfish cultivation industry. The purpose of this study was to develop detoxification method for utilization of PSP intoxicated sea mussel and prevent from PSP illness. The PSP was extracted with 0.1 N HCl solution from the submitted sea mussel, then the toxicity was measured by mouse assay according to Official Methods of Analysis of the Association of Official Analytical Chemists. No detoxification effect was observed by adding extracted juice of garlic and ginger. When the sea mussel homogenate was heated at various temperatures, the PSP toxicity was not changed significantly at below $70^{\circ}C$ for 60 minutes but it was decreased as the heating temperature was increased. For example, when the sea mussel homogenate was heated at 100, $121^{\circ}C$ for 10 minutes, the toxicity was decreased about 67 and 90%, respectively. When the sea mussel containing 645 $\mu$g PSP per 100g of edible meat was processed according to general shellfish canning procedure, the toxicity was decreased as the level of PSP undetected by mouse assay.

• Journal of Marine Life Science
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• v.9 no.1
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• pp.9-21
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• 2024

Paralytic shellfish poisoning (PSP) including Saxitoxin (STX) is caused by harmful algae, and poisoning occurs when the contaminated seafood is consumed. The mouse bioassay (MBA), a standard test method for detecting PSP, is being sanctioned in many countries due to its low detection limit and the animal concerns. An alternative to the MBA is the Neuro-2a cell-based assay. This study aimed to establish various test conditions for Neuro-2a assay, including cell density, culture conditions, and STX treatment conditions, to suit the domestic laboratory environment. As a result, the initial cell density was set to 40,000 cells/well and the incubation time to 24 hours. Additionally, the concentration of Ouabain and Veratridine (O/V) was set to 500/50 μM, at which most cells died. In this study, we identified eight concentrations of STX, ranging from 368 to 47,056 fg/μl, which produced an S-shaped dose-response curve when treated with O/V. Through inter-laboratory variability comparison of the Neuro-2a assay, we established five Quality Control Criteria to verify the appropriateness of the experiments and six Data Criteria (Top and Bottom OD, EC₅₀, EC₂₀, Hill slop, and R² of graph) to determine the reliability of the experimental data. The Neuro-2a assay conducted under the established conditions showed an EC₅₀ value of approximately 1,800~3,500 fg/μl. The intra- & inter-lab variability comparison results showed that the coefficients of variation (CVs) for the Quality Control and Data values ranged from 1.98% to 29.15%, confirming the reproducibility of the experiments. This study presented Quality Control Criteria and Data Criteria to assess the appropriateness of the experiments and confirmed the excellent repeatability and reproducibility of the Neuro-2a assay. To apply the Neuro-2a assay as an alternative method for detecting PSP in domestic seafood, it is essential to establish a toxin extraction method from seafood and toxin quantification methods, and perform correlation analysis with MBA and instrumental analysis methods.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.2
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• pp.174-180
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• 2020

The mouse bioassay has been used widely for the monitoring of paralytic shellfish toxins (PSTs) in many countries. However, this method shows low sensitivity and high limit of detection (LOD), as well as it cannot confirm toxic profiles. Recently, LC-MS/MS method was studied for the quantitative of PSTs, however, the method has any problems with unstable retention times by ionization suppression caused by high salt concentration in shellfish extracts. To establish an alternative method for PSTs analysis, we tried to original LC-MS/MS methods adding desalting operation using amorphous graphitized polymer carbon solid-phase extraction cartridges. The method validation was conducted to determine linearity, limit of detection, limit of quantification (LOQ), accuracy, and precision in quantifying PSTs. The correlation coefficients for all tested PSTs maintained over 0.999. The LODs and LOQs for all PSTs were about 0.19-1.05 ㎍/kg and 0.58-3.18 ㎍/kg, respectively. The accuracies for PSTs were 95.4-107.7% for saxitoxin group, 97.1-100.9% for gonyautoxin group, 99.0-100.8% for N-sulfocarbamoyl toxin group, and 96.8-104.6% for decarbamoyl toxin group. These results indicate that the modified LC-MS/MS method was appropriate for analyzing the PSTs in shellfish and tunicates.

• 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.45 no.5
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• pp.465-471
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• 2012

To compare the accumulation of paralytic shellfish poison (PSP) in different marine organisms, the occurrence and variation of PSP were surveyed in blue mussel Mytilus edulis, oyster Crassostrea gigas, short neck clam Ruditapes philippinarum, bay scallop Argopecten irradians, and warty sea squirt Styela clava collected from Jinhae Bay, Korea, in 2005 and 2006 year. We also investigated the ability of the blue mussel to detoxify PSP by relaying and depuration (via the water flow or water circulation system). In the marine organisms examined, PSP levels were the highest in blue mussel, followed in order by bay scallop, oyster, short neck clam, and warty sea squirt. Comparing the maximum PSP levels in the bivalve species examined in 2005 and 2006, PSP in blue mussel was 1.6-2.0, 4.0-5.9, and 5.1-6.0 times higher than in bay scallop, oyster, and short neck clam, respectively. Therefore, blue mussel could be useful as a bioindicator for PSP monitoring. With the increasing PSP levels in blue mussel in 2006, the proportion of PSP in its digestive gland increased to 95.1% when the maximum level was detected from the whole tissues of blue mussel on May 29. Subsequently, the PSP proportion in the digestive gland decreased as the PSP level in whole tissue decreased. The detoxification of PSP in blue mussel was greatest with relaying, followed by the water flow, and water circulation systems. Relaying decreased the PSP level below the regulatory limit of $80{\mu}g$/100 g after 2 days in low toxic sample with $124{\mu}g$/100 g, and after 7 days in high toxic sample with $401{\mu}g$/100 g. During depuration in the blue mussel with $401{\mu}g$/100 g via the water flow system, the PSP amounts in the digestive gland decreased by about 50% after 1 day, and about 77% after 7 days. In contrast, the PSP amounts in the soft body, gill, and mantle did not change significantly with depuration.