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Studies on Origin, Intoxification, Detoxification and Metabolism of Marine Toxin (Tetrodotoxin and Paralytic Shellfish Toxin) and Their Molecular Biological Characteristics
|Contributors: ||NTOU:Department of Food Science|
|Issue Date: ||2011-06-28T05:52:47Z
|Abstract: ||摘要:近年來臺灣因河魨毒和麻痺性貝毒引起之食物中毒事件屢次發生，甚為國人所重 視，申請者以往對臺灣產河魨、熱帶性魚類、螺貝類和蟹類進行了一系列之毒性探討， 也建立了相當完整的魚貝類毒性資訊。本計畫第一年擬了解臺灣地區好氣性和厭氣性細 菌生產河魨毒和有毒藻類生產麻痺性貝毒之能力，探討河魨毒生產菌和產麻痺性貝毒藻 類來源之特殊生長需求條件，瞭解其生長特性及產毒最適條件，並量產河魨毒和麻痺性 貝毒，以利後續研究。並對菌種和藻種之長久保存進行探討。第二年計畫為探討螺貝類 和淡水河魨之蓄積河魨毒和麻痺性貝毒的能力，擬了解其體內結合毒素之高分子化合物 (結合蛋白質)，並比較不同有毒螺貝類和淡水河魨之結合蛋白質之生化特性及其種別差 異性，進行結合毒素蛋白質之活性部位分析。進一步探討河魨毒和麻痺性貝毒存在生物 體內之可能之蓄積機制，並對有毒藻種和產毒細菌進行基因鑑定。第三年計畫擬進行 in vivo 試驗，直接以口胃管灌食投予 Wistar 大白鼠不同劑量之河魨毒及麻痺性貝毒，觀 察並分析動物體之生理變異及行為變化。另利用高效能液相層析法/螢光衍生法及液相 層析串聯式質譜儀技術，對於毒素最初進入生物體，在體內血液中之毒含量、引發變異 之起始、作用時間之長短及最後離開生物體排出尿液中之代謝毒量作分析。另外，給予 大鼠不同飲用水包含逆滲透水、鹼性電解水、生理食鹽水及含硫胺基酸，經由不同時間 點收集其尿液及血液進行檢測分析毒素之濃度，以了解飲用水種類及含硫胺基酸是否會 加速毒素的排泄，以供醫療及檢驗分析參考之用途。其次擬進一步對同種不同毒生產性 之菌株和藻株，除了分析基因之差異外，進一步分析蛋白質體之差異性，以建立完整海 洋生物毒素來源之物種資料庫。|
Abstract:Food poisoning incidences by tetrodotoxin (TTX) and paralytic shellfish poison (PSP) in Taiwan occurred frequently in recent years and it was paid close attention by publics. In last two decades, government focused on prevention of TTX and PSP poisoning events. Some programs had been aimed at a series of investigations on toxin of Taiwan’s puffer fishes, coral reef fishes, mollusks as well as crabs, and the toxin profiles were established completely. Therefore, the reason how animals accumulated TTX/PSP and what the excreting pathways of those toxins are still under study. In this investigation, we will use animal models to test the accumulation mechanism and the excretion pathway. The first year is aimed to understand the producing ability of TTX from aerobic and anaerobic bacteria in toxic marine animals and PSP from toxic alga in Taiwan, focusing on optimal growth conditions and characteristics of TTX producing bacteria and PSP producing alga. The optimal growth and the best toxin producing conditions to mass production of TTX/PSP are also discussed in further study. Then, how to keep the species of TTX-producing bacteria and PSP-producing alga will be also studied. The next year, the toxin accumulating ability in mollusks and fresh water puffer fish will be emphasized, expect to separate the toxin-binding high molecular compound (protein) from those animals and to compare the biochemical and species differences of those derived compounds between toxic mollusks and fresh water puffer fish. The bioactive site of the compound in which responsible for binding toxins will be investigated, and the possible TTX/PSP accumulating mechanisms in toxic animals will be confirmed. The gene identification of toxic bacteria and alga will be also established. In third year, Wistar rats will be used for in vivo test, observing and analyzing their physical and behavior change during feeding period in different TTX/PSP doses by direct OG feeding. In addition, HPLC and LC-MS/MS will be applied to detect toxin content in blood and urine in different time course: TTX/PSP initially entering the animal body, the first syndrome appearance, time course of action and after the toxin excretion following urination. Finally, we will offer varied drinking water eg. RO water, alkaline water, normal saline and sulfur contained amino acid to rats, and collect their urine and blood in different periods. The concentration of TTX/PSP will be detected to find out whether the different drinking water or sulfur contained amino acid will accelerate toxin metabolism. The information will be applied for clinic and analysis application. Furthermore, different kinds of TTX-producing bacterial species and PSP-producing alga will be used to differentiate the DNA variations. Meanwhile, the variations of proteomics of those bacteria and alga are also undertaken to compare.
|Appears in Collections:||[食品科學系] 研究計畫|
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