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Development of An ICR mouse bioassay for Toxicity Evaluation in Neurotoxic Poisoning Toxins-Contaminated Shellfish

Biomedical and Environmental Sciences 2013年第5期26卷 346-364页

作者： WONG Chun Kwan HUNG Patricia KAM Kai Man Biotoxin Laboratory Microbiology DivisionPublic Health Laboratory Services BranchCentre for Health ProtectionDepartment of HealthShek Kip MeiKowloon

Objective To develop an ICR （female） mouse bioassay （MBA） for toxicity confirmation and evaluation of neurotoxins （brevetoxins）-contaminated shellfish. Methods Brevetoxins （BTX-B） as a causative agent of neurotoxic shellfish poisoning （NSP） under different shellfish matrices were intraperitoneally injected at different doses into mice to study their toxic effects and to differentiate the range of lethal and sublethal dosages. Their sensitivity and specificity were analyzed with 2 competitive ELISA kits for quantitative determination of standard BTX-B and dihydroBTX-B under different shellfish matrix-diluent combinations. Detection rates of MBA and two antibody-based assays for BTX-B from field NSP-positive shellfish samples were compared. Results BTX-B could be detected in shellfish tissues at concentration of 50-400 μg/100 g under shellfish matrix-Tween-saline media, which were appropriate to identify toxic shellfish at or above the regulatory limit （80 μg/100 g shellfish tissues）. The LD 50 identified was 455 g/kg for BTX-B under general shellfish matrices （excluding oyster matrices） dissolved in Tween-saline. The presence of shellfish matrices, of oyster matrices in particular, retarded the occurrence of death and toxicity presentation in mice. Two antibody-based assays, even in the presence of different shellfish matrix-diluent combinations, showed acceptable results in quantifying BTX-B and dihydroBTX-B well below the regulatory limit. Conclusion The two ELISA analyses agree favorably （correlation coefficient, r 0.96; Student＇s t-tests, P〉0.05） with the developed bioassay.

关键词： Antibody-based assay Brevetoxin ICR female mice LD 50 mouse bioassay Neurotoxic shellfish poisoning

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Features of Ciguatera Fish Poisoning Cases in Hong Kong 2004-2007

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Biomedical and Environmental Sciences 2008年第6期21卷 521-527页

作者： CHUN-KWAN WONG PATRICIA HUNG KELLIE L. H. LEE TINA MOK THOMAS CHUNG KAI-MAN KAM Biotoxin Laboratory Public Health Laboratory Services Branch Centre for Health Protection Department of Health Shek Kip Mei Kowloon Food and Environmental Hygiene Department Queensway Government Offices

Ohjective To review the clinical features and laboratory investigations of ciguatera patients in Hong Kong between 2004 and 2007 in order to show the timely sampling of implicated fish from ciguatera victims and application of validated mouse bioassay for confirming suspected clinical cases of ciguatera. Methods Diagnosis of the ciguatera victims was based on history of coral fish consumption and clinical presentations stated in official guidelines for clinical diagnosis of ciguatera fish poisoning in Hong Kong. Food remnants of coral fish samples were collected swiftly from ciguatera victims between 2004 and 2007 for ciguatoxins （CTXs） analysis. Results Major clinical symptoms in ciguatera patients included gastrointestinal and neurological effects including limb numbness and diarrhoea, which developed at 0.5 to 15 hours after consumption of fish. In most cases, neurological symptoms were more common than gastrointestinal symptoms. A broad range of attack rate （10%-100%） was observed in each ciguatera outbreak. Validated mouse bioassay on ether extracts of the food remnant samples confirmed that all were CTXs-positive （〈0.5 - 4.3 MU/20 mg ether extract） and directly linked to the corresponding ciguatera cases. Conclusion Consistency between clinical and laboratory analysis for ciguatera poisoning illustrates the application of laboratory mouse bioassay in a timely fashion for confirming ciguatera poisoning cases and implementing effective public health measures. With further improvement in laboratory techniques, features of ciguatera fish poisoning cases can be better defined, Further studies are needed to determine the risk of each class of CTXs （Pacific-, Indian- and Caribbean-CTXs） in Hong Kong.

关键词： Ciguatera poisoning Ciguatoxin Food remnants Hong Kong mouse bioassay

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Toxicity assessment and screening of tetrodotoxin in the oblong blowfish(Takifugu oblongus)from the Tamil Nadu Coast of Bay of Bengal,India

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Asian Pacific Journal of Tropical Medicine 2017年第3期10卷 263-268页

作者： Indumathi SM Khora SS Department of Integrative Biology School of Biosciences and TechnologyVIT UniversityVellore-632014Tamil NaduIndia

Objective:To assess the toxicity of the pufferfish Takifugu oblongus,from Chennai coast,Tamil Nadu,India and to detect the presence of Tetrodotoxin(TTX).Methods:The toxicity was evaluated by mouse bioassay using Swiss Albino mice which were expressed in mouse units(MU).Gross anatomical features were observed which is followed by histopathology of the dead mice tissues to establish the toxicity.Instrumental analysis for the presence of tetrodotoxin was also performed through GC-MS and HPLC.Results:The toxicity of ovary was the maximum with 163 MU/g and lowest toxicity was observed in skin with 75.88 MU/g.Histopathological analyses of the dead mice showed various cellular degenerations and inflammations.The amount of Tetrodotoxin detected through GC-MS and HPLC was more reliable and sensitive than the customary mouse bioassay as instrumental analyses were able to detect even nanograms of the toxin.Conclusions:The present study evidently proved that Takifugu oblongus is highly toxic and consumption of the same can pose serious threat for health and possible lethality to humans.

关键词： Food intoxications Takifugu oblongus mouse bioassay Tetrodo toxin GC-MS HPLC

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