臺灣博碩士論文加值系統

腹足綱的軟體動物在全球的分佈大約有88,000種，台灣在近年來常有因為誤食有毒的苦螺而造成河魨毒素中毒的案件。金門四面環海有著豐富的海洋資源，當地人尤其喜愛食用海螺以及貝類等海產，其中螺類更是經常被用來烹煮成小菜食用，但是相較於台灣，金門對於河魨毒素相關研究仍非常有限，故本研究主要是採集金門金沙及金城潮間帶經常見到並且容易被採集誤食的苦螺來進行實驗，鑑定金門地區苦螺的物種及其體內河魨毒性與探討其腸道內容物物種。
螺類在傳統的物種鑑定上通常是以形態特徵來作為分類指標，但是如果螺類缺乏外殼，就會造成鑑定上的困難，因此在近幾年來多採用DNA來進行物種鑑定，能提高物種鑑定的準確性。本實驗搭配特定引子16S進行聚合酶連鎖反應對目標序列進行增幅，再進一步透過DNA膠體電泳進行確認，藉此了解此苦螺的物種與已知的有毒苦螺之間是否有關係。另外再以液相層析串聯式質譜儀分別對螺類的肌肉組織和腸道部位進行河魨毒素的檢測，了解螺體內是否含有河魨毒素。最後再利用膠體電泳進一步來檢測腸道內容物的物種。
藉由以上各項實驗結果可以發現金沙鎮所採集的苦螺其Query cover有91%以及Identification有98%與Thais clavigera最為符合，而金城地區採集的苦螺其Query cover有85%以及Identification有99%與T. clavigera最為符合，因為T. clavigera為肉食性的螺類，所以其腸道內容物物種經鑑定發現為Mancinella tuberosa、Purpura persica、Rapana bezoar、Thais luteostoma，皆為其他種的螺類，而透過液相層析串聯式質譜儀檢測後發現其並未含有河魨毒素，故可以推測金門地區的苦螺-T. clavigera為不含有河魨毒素的螺類。

Gastropod mollusk approximately 88,000 species distribute in the world. In recent years, many food poisons of tetrodotoxin (TTX) of marine snail in Taiwan has always happened. Kinmen is surrounded by the sea, have abundant marine resources. The locals like to edible oysters, marine snails, and much more seafood, among them, the marine snail has always cooking of dishes to eat. But compared to Taiwan, respect to tetrodotoxin information and studies are limited in Kinmen. In our study, primary to collection the marine snail has often seen and easy to ingestion along the coast in Kinmen to experiment, application the marine snail species and investigate the toxicity and the intestinal biological. The traditional identification method of marine snail species is identified by morphological characteristics, but it has difficult to identify if the marine snail without the shell. In recent years, identify species method is developed based on DNA to improve the accurate. In our study, using a 16S primer and coordinate to polymerase chain reaction (PCR) to amplified the sequence, and further to run electrophoresis to confirm the species, and take to understanding this species relationship with another marine snail has tetrodotoxin. In addition, using liquid chromatography tandem mass spectrometer (LC/MS/MS) to analysis tetrodotoxin in this marine snail species muscle and intestinal. Then take advantage of gel electrophoresis to understanding the intestinal biological.
The results show, Kinmen’s marine snail species is Thais clavigera, the T. clavigera is carnivorous marine snail, so we can find many another marine snail in the snail’s intestinal content, and we analysis TTX with LC/MS/MS can detection T. clavigera is non-toxic, from the results concluded that T. clavigera is a safety, non-toxic and can edible marine snail.

錄
授權書 i
論文考試審定書 ii
中文摘要 iii
英文摘要 iv
誌謝 v
目錄 vi
表目錄 vii
圖目錄 viii
壹、前言 1
貳、文獻整理 2
一、螺類引發河魨毒素之中毒事件 2
二、螺類分類與有毒螺類之種類 5
三、河魨毒素介紹 8
四、分子生物技術鑑定物種 12
五、利用粒線體DNA (mtDNA)鑑定物種之特性 14
六、河魨毒素成分分析方法 17
參、研究動機與目的 21
一、研究目的 21
二、實驗流程 21
肆、材料與方法 22
一、實驗材料 22
二、實驗方法 24
伍、結果 26
陸、討論 29
柒、結論 32
捌、參考文獻 33
附錄一、食品中海洋生物毒素之檢驗方法-河豚毒素之檢驗 84

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