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研究生:何佩蒨
研究生(外文):Ho, Pei-Chien
論文名稱:龍虎石斑之基因魚種鑑定及龍虎石斑和龍膽石斑肝臟中熱帶性海魚毒、維生素 A 和脂肪酸含量之探討
論文名稱(外文):Studies on Genetic Identification of Epinephelus fuscoguttatus × Epinephelus lanceolatus and Level of Ciguatera Toxins, Vitamin A and Fatty Acids in Liver of E. fuscoguttatus × E. lanceolatus and E. lanceolatus
指導教授:黃登福黃登福引用關係
指導教授(外文):Hwang, Deng-Fwu
口試委員:蕭泉源林欣榮陳泰源
口試委員(外文):Shiau, Chyuan-YuanLin, Shin-JungChen, Tai-Yuan
口試日期:2020-06-11
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2020
畢業學年度:108
語文別:中文
論文頁數:95
中文關鍵詞:龍虎石斑龍膽石斑基因鑑定維生素 A 中毒熱帶性海魚毒脂肪酸
外文關鍵詞:Epinephelus fuscoguttatus × E. lanceolatusE. lanceolatusgenetic identificationvitamin A poisoningciguatera toxinsfatty acids
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石斑魚 (groupers) 泛指石斑魚屬 (Epinephelus) 或喙鱸屬 (Mycteroperca) 的魚類,分布於全球熱帶及亞熱帶海域。而由龍膽石斑 (Epinephelus lanceolatus) 及老虎石斑 (Epinephelus fuscoguttatus) 雜交而來的龍虎石斑 (E. fuscoguttatus × E. lanceolatus) 為近年來台灣新興的養殖石斑魚種。石斑魚為高經濟價值魚類,常被與其相似且較便宜的魚種替代,且加工處理過的石斑魚更難利用外觀特徵來辨識其種類。此外,石斑魚為台灣可能含有熱帶性海魚毒的珊瑚礁魚種之一,其毒素會經由食物鏈積累在魚體中,尤其是肝臟和內臟部位,且魚體越大其所累積的毒素含量越多。而肝臟為人類和動物主要儲存維生素 A 的器官,研究指出,長期食用大量動物肝臟、魚肝油或服用高劑量補充劑是造成維生素 A 中毒之主因,攝取過量可能會導致急性中毒或慢性中毒。本研究第一章將龍虎石斑之 DNA 經聚合酶連鎖反應 (polymerase chain reaction, PCR) 擴增核基因 RYR3 並定序,獲得其 RYR3 的等位基因序列,與 NCBI 資料庫進行同源性比對,發現與 genbank 上之具有龍膽石斑及老虎石斑之雜交種及老虎石斑魚這兩種魚之基因親緣性最高 (> 99%),再结合母系遺傳的粒線體 DNA 基因的同源比對结果,則可確定其母系為老虎石斑,父系為龍膽石斑。第二章為分析龍虎石斑及龍膽石斑肝臟的一般成分、熱帶性海魚毒、維生素 A 含量及脂肪酸組成。結果顯示,養殖龍虎石斑肝臟之水分含量為 52.47±4.66%,粗蛋白為 5.90±1.31%,粗脂肪為 16.50±4.47%,灰分為 0.82±0.08%,碳水化合物為 24.30±3.27%,其脂肪酸組成以肉豆蔻酸 (C14:0)、油酸 (C18:1) 與亞麻油酸 (C18:2) 為主,含量分別為 29.68±3.91 mg/g lipid、10.23±6.61 mg/g lipid 及6.78±5.17 mg/g lipid;而養殖龍虎石斑肝臟之維生素 A 含量平均為 988.55 ± 692.82 IU/g,其魚體大小、肝臟大小與肝臟中維生素 A 之含量並無相關性,此外,所有養殖龍虎石斑之肝臟皆無熱帶性海魚毒檢出。另外,市售龍膽石斑肝臟之水分含量為 53.91±9.48%,粗蛋白為 8.68±2.48%,粗脂肪為 15.73±7.52%,灰分為 0.75±0.21%,碳水化合物為 19.32±8.91%,其脂肪酸組成以肉豆蔻酸 (C14:0)、棕櫚酸 (C16:0) 與 DHA (C22:6) 為主,含量分別為 34.20±8.85 mg/g lipid、10.34±2.29 mg/g lipid 及10.53±2.13 mg/g lipid,而所有龍膽石斑之肝臟皆不具熱帶性海魚毒,此外,龍膽石斑肝臟之維生素 A 含量平均為 1,544.03±2,534.87 IU/g,但其中有些體重大於 20 kg 的檢體,其魚體及肝臟愈大,其肝臟所累積之維生素 A 含量愈高,因此建議避免攝食大型龍膽石斑 (≥ 20 kg) 之肝臟,以減少中毒之風險。
Groupers (family Epinephelidae) inhabit in coral reef area of the tropics and subtropics around the world. Hybrid grouper (Epinephelus lanceolatus♂ × E. fuscoguttatus♀) has been widely cultured in the southeast coast of Taiwan due to its fast growth, popular taste, and high nutritional and economic value. They are economically valuable and are substituted by cheaper fish sometimes due to economic profit. In groupers, color patterns may change substantially within a species. Even the same species may have different color patterns in various living environments and different developmental stages. Furthermore, it is difficult to identify clearly the species of processed products based on their morphology. Grouper is one of the coral reef species that may contain ciguatera toxins in Taiwan. Ciguatera toxins are concentrated in fish via the food chain. Contaminated fish accumulate ciguatera toxins in the flesh, muscle, skin, and viscera, especially in the liver and viscera. The larger the fish body, the more toxins it accumulates. On the other hand, the liver is the main organ for storing vitamin A in humans and animals. Studies have shown that long-term consumption of large amounts of animal liver, cod liver oil or high-dose vitamin A supplements is the main cause of vitamin A poisoning. Excessive intake of vitamin A may cause acute or chronic poisoning. In the first chapter of this study, E. fuscoguttatus × E. lanceolatus was tested for the PCR amplification of RYR3 gene. The identity of the species was checked by BLAST analysis. The two alleles of RYR3 shared the highest identity with those of E. fuscoguttatus and E. lanceolatus (>99%). Nuclear gene combined with the mitochondrial DNA gene sequencing results. Therefore, we could determine that E. fuscoguttatus was the female parent, while E. lanceolatus was the male parent. The second chapter analyzed the level of proximate, fatty acids composition, ciguatera toxins and vitamin A in liver of cultured E. fuscoguttatus × E. lanceolatus and commercial E. lanceolatus. The results show the average moisture, crude protein, crude lipid, ash and carbohydrate of the liver in cultured grouper were 52.47±4.66, 5.90±1.31, 16.50±4.47, 0.82±0.08 and 24.30±3.27%, respectively. The major fatty acids were myristic acid (C14:0), oleic acid (C18:1) and linoleic acid (C18:2). The amount of the major fatty acids were 29.68±3.91, 10.23±6.61 and 6.78±5.17 mg/g lipid, respectively. The average level of vitamin A in cultured E. fuscoguttatus × E. lanceolatus liver was 988.55 ± 692.82 IU/g. Regression model shows that there is no relationship between body weight, liver weight and vitamin A level. The ciguatera toxin of liver in cultured E. fuscoguttatus × E. lanceolatus was not detected. The average moisture, crude protein, crude lipid, ash and carbohydrate of the liver in commercial E. lanceolatus were 53.91±9.48, 8.68±2.48, 15.73±7.52, 0.75±0.21 and 19.32±8.91%, respectively. The dominant fatty acids were myristic acid (C14:0), palmitic acid (C16:0) and docosahexaenoic acid (C22:6). The amount of the dominant fatty acids were 34.20±8.85, 10.34±2.29 and 10.53±2.13 mg/g lipid, respectively. The average level of vitamin A in commercial E. lanceolatus liver was 1,544.03 ± 2,534.87 IU/g. Regression model shows that the more body weight and liver weight, the higher vitamin A level in liver of commercial E. lanceolatus. The ciguatera toxin of liver in commercial E. lanceolatus was not detected. Acute vitamin A poisoning and ciguatera poisoning may occur simultaneously due to ingestion the liver of large-sized E. lanceolatus (body weight ≥ 20 kg).
摘要 I
Abstract II
目錄 IV
壹、文獻整理 1
一、石斑魚之簡介 2
二、魚種鑑定 4
三、熱帶性海魚毒之簡介 9
四、維生素 A 之簡介 13
五、脂肪酸之簡介 18
貳、研究內容 19
第一章、龍虎石斑之基因鑑定 20
一、前言 20
二、材料與方法 21
三、結果 24
四、討論 26
五、圖表 28
第二章、龍虎石斑和龍膽石斑肝臟中維生素A含量、熱帶性海魚毒性及脂肪酸之分析 42
一、前言 42
二、材料與方法 43
三、結果 48
四、討論 51
五、圖表 54
參、總結論 76
肆、參考文獻 77
謝辭 95
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