臺灣博碩士論文加值系統

石斑魚是亞太地區重要的海水養殖物種。其中龍膽石斑具有高經濟價值、體型大及成長快速等優點，成為主推的養殖物種。但長期餵食商業飼料之龍膽石斑易出現脂肪肝及抗緊迫能力下降之問題，導致產量及經濟上損失。本研究擬探討不同濃度膽鹼飼料對龍膽石斑成長、油脂代謝及氨-氮耐受性之影響。
本研究共配製六種飼料分別為基礎魚粉飼料添加每公斤0 g (Control)、0.25 g (0.25CD)、0.5 g (0.5CD)、1 g (1CD)、2.5 g (2.5CD)及5 g (5CD)之氯化膽鹼，測得飼料膽鹼實際濃度分別為每公斤2.57 ±0.03 g、2.67 ±0.11 g、2.94 ±0.44 g、3.48 ±0.11 g、4.99 ±0.08 g及7.71 ±0.35 g膽鹼。以不同濃度膽鹼之飼料餵飼龍膽石斑56天與30天，各別觀察成長及油脂代謝與緊迫能力。在第一部分實驗，以不同濃度膽鹼飼料餵食龍膽石斑56天後，發現增重率、存活率及飼料效益沒有明顯的差異。然而，隨著膽鹼濃度增加，肝體比、肝臟中油脂、三酸甘油酯和膽固醇含量及血清中三酸甘油酯和膽固醇含量都出現下降的趨勢。此外，當龍膽石斑攝食膽鹼含量大於2.94 g/kg可增加背部肌肉油脂含量，亦可改善龍膽石斑在氨氮緊迫下能力，包括存活率及行為表現。由本研究證實，膽鹼對龍膽石斑之脂質代謝及緊迫的耐受性有很重要的改善作用。

Six fish meal basal diets supplied with 0 (control), 0.25, 0.5, 1, 2.5 and 5 g/kg of choline chloride, resulting in choline levels of 2.57, 2.67, 2.94, 3.84, 4.99 and 7.71 g/kg, respectively, were fed to giant grouper, Epinephelus lanceolatus, for 56 and 30 days to evaluate the growth and lipid metabolism, and stress tolerance respectively. In the first trial, fish fed different levels of choline containing diets for 56 days had no significant difference in weight gain, survival and feeding efficiency. Fish fed increased levels of dietary choline, however, tended to have decreases in the hepatic somatic index; lipids, triglycerides, and cholesterol in liver; and triglycerides and cholesterol in serum. A increase of lipid content in dorsal muscle was recorded in fish fed the diets containing choline >2.94 g/kg. Additionally, dietary choline improved the reactions of fish to ammonia stress, including survival and behavioural responses, in fish fed diets containing choline levels >2.94 g/kg. These findings indicate that choline plays important roles in lipid metabolism and stress tolerance in giant grouper.

中文摘要 I
英文摘要 III
目錄 V
圖目錄索引 VIII
表目錄索引 IX
第一章、前言 1
第二章、文獻回顧 4
2.1 龍膽石斑 4
2.1.1 龍膽石斑分類地位 4
2.1.2龍膽石斑形態特徵 4
2.1.3 生態習性 5
2.1.4龍膽石斑養殖與產業現況 5
2.1.5石斑養殖問題與解決 6
2.2膽鹼 7
2.2.1物理與化學性質 7
2.2.2膽鹼生理特性 7
2.2.2 代謝路徑 8
1. 乙醯化反應 (acetylation) 9
2. 氧化反應 (oxidation) 9
3. 磷酸化反應 (phosphorylation) 10
2.2.3膽鹼與油脂代謝 11
2.2.4膽鹼在水產養殖之抗緊迫 11
第三章、材料與方法 13
3.1飼料配製 13
3.2 成分分析 13
3.2.1 水分測定 13
3.2.2 灰份測定 14
3.2.3粗蛋白質測定 14
3.2.4 粗脂質測定 15
3.2.5 膽鹼分析 15
3.2.6 甲硫胺基酸分析 17
3.3實驗動物 19
3.4 成長試驗 20
3.4.1試水分析 20
3.4.1.2溫度 20
3.4.1.3 pH 20
3.4.1.4水中亞硝酸-氮分析 20
3.4.1.5 水中氨-氮 (Ammonia-N)分析 21
3.4.2 膽鹼對油脂代謝影響 22
3.4.2.1 肝臟與背部肌肉油脂分析 23
3.4.2.2 分析血清與肝臟三酸甘油酯含量 23
3.4.2.3 分析血清與肝臟膽固醇含量 24
3.4.2.4 分析血清高密度脂蛋白含量 24
3.4.2.5 分析血清低密度脂蛋白含量 25
3.4氨氮耐受性實驗 26
3.6統計分析 27
第四章、結果 28
4.1 成長表現 28
4.2 油脂代謝參數 29
4.2.1 肝體比與肝臟中三酸甘油酯、膽固醇及油脂含量 29
4.2.3背部肌肉油脂含量 29
4.3 氨-氮耐受性 30
第五章、討論 32
第六章、結論 37
參考文獻 52
作者簡介 60

孔垂永、陳瑛、陶得和、許開云 (2009)仔豬維生素B缺乏的診治。動物科學，24期，321頁。
古鎮均 (2007)箱網養殖龍膽石斑的管理。冉繁華，石斑，臺灣。60-70頁。
李雅雯 (1994)維生素C對餵食炸油之天竺鼠體內代謝的影響效應。中國文化大學生活應用科學系暨生活應用科學研究所碩士論文。
李克怡、洪祖培 (1997)維生素B1缺乏症 (腳氣病)─近年於日本再出現之多發性神經炎合併下肢水腫。當代醫學，46期，725-728頁。
邵廣昭 (2009)臺灣魚類資料庫。中央研究院，生物多樣性研究中心魚類生態與進化研究室。
林豐詔 (2003) 脂肪肝的診治及病例報告。中國中醫臨床醫學雜誌，9期，49-51頁。
林義福 (2011)維生素E之功能及對種雞之影響。臺灣農學會報，5期，487-499頁。
莊曉慧 (2007) 臺灣石斑魚產業競爭力與策略之分析。國立臺灣海洋大學環境生物與漁業科學學系碩士論文。
郭錦朱、郭誌明、沈昱徳、何碧月、周瑞良、陳紫媖 (2012)大蒜拮抗石斑腸炎弧菌症之效力。水產試驗東港生技研究中心。水試專訊，37期，26頁。
徐之勇、余燕、越恆章、馬金友、崔恆敏 (2010)飼料中添加膽鹼對雉雞脂肪肝的防治效果。河南農業科學，6期，128-131頁。
楊松昇、戴恭良、朱柏齡、林石化、林裕豐 (2001)尿毒症病患離子代謝之處理及維生素D之使用。腎臟與透析，4期，155-158頁。
楊惠郎、林翰佑、林青丘 (2011)石斑養殖科技。科學發展，462期 ，66-71頁。
鄭安倉 (2009)龍膽石斑生產概況。漁業推廣，275期 ，14-15頁。
戴揚 (2010) 臺灣龍膽石斑之養殖經濟分析。國立臺灣海洋大學水產養殖系碩士論文。
謝孟儒 (2010)飼料以廚餘粉取代魚粉對吳郭魚、龍膽石斑和點帶石斑及體組成的影響。國立海洋大學水產養殖系碩士論文。
簡宏達 (2002) 磷脂膽鹼及甜菜鹼對草蝦稚蝦膽鹼需求之影響。國立臺灣海洋大學食品科學系碩士論文。
蘇偉成、劉富光 (2005)專題報導水產養殖─臺灣水產養殖的永續經營。科學發展，385期，42-49頁。
AOAC (Association of Official Analytical Chemists) (1980) Methods of Anlysis, 13th edition.
Bloch, M. E. (1790) Naturgeschichte der ausl#westeur037#ndischen fische. German. p. 128.
Best, C. H., Maclean, D. L., Ridout, J. H. (1934) Choline and liver fat in phosphorus poisoning. The Journal of Physiology. 83: 275-284.
Bendschenider, K. and Robinson, R. J. (1952) A new spectrophotometric method for the de-termination of nitrite in sea water. Journal of Marine Research 11: 87-96.
Barlow, S. M., Pike, I. H. and Nixon, F. (1979) Choline contentof fish meals from various origins. Journal of the Science of Food and Agriculture 30: 89-92.
Chan, M. M. (1991) Choline. In: Handbook of Vitamins (2nd edn) (ed. by L. J. Machlin), Marcel Dekker, New York, NY. pp. 537-556.
Combs, G. F. (1992) The vitamins: fundamental aspects in nutrition and health. Academic Press, New York, NY.
Craig, S. R., Gatlin, D. M. (1996) Dietary choline requirement of juvenile red drum (Sciaenops ocellatus). Journal of Nutrition 126: 1696-1700.
Chou, R. L., Su, M. S., Chen, H. Y. (2001) Optimal dietary protein and lipid levels for juvenile cobia (Rachycentron canadum). Aquaculture 192, 81-89.
Cheung, O., Sanyal, A. J. (2010) Recent advances in nonalcoholic fatty liver disease. Current Opinion in Gastroenterology 26: 202-208.
Corbin, K. D. and Zeisel, S. H. (2012) Choline metabolism provides novel insights into non-alcoholic fatty liver disease and its progression. National Institutes of Health Public Access 28: 159-165.
Duan, Y., Zhu, X., Han, D., Yang, Y. and Xie, S. (2012) Dietary choline requirement in slight methionine-deficient diet for juvenile gibel carp (Carassius suratus gibelio). Aquaculture Nutrition 18: 620-627.
Food and Agriculture Organization of the United Nations, FAO (2012) The State of World Fisheries and Aquaculture 2012. pp. 3-40.
Gang, J., Jian, F. and Zhibiao, Q. (2006) Studies on the fatty liver diseases of Sciaenops ocellatus caused by differentether extract levels in diets. Frontiers of Biology in China. 1: 9-12.
Jiang, X., Jiang, Y., West, A. A., Perry, C. A., Malysheva, O. V., Devapatla, S., Pressman, E., Vermeylen, F. and Caudill, M. A. (2012)Maternal choline intake alters the epigeneticstate of fetal cortisol-regulating genes inhumans. The FASEB Journal 26: 3563-3574.
Jiang, G. Z., Wang, M., Liu, W. B., Li, G. F. and Qian, Y. (2013) Dietary choline requirement for juvenile blunt snoutbream, Megalobrama amblycephala. Aquaculture Nutition1 9: 499–505.
Kroening, H. G. and Pond, W. G. (1967) Methionine,choline and threonine interrelationships for growth and lipotropic action in the baby pig and rat. Journal of Animal Science 26:352-357.
Ketola, H. G. (1976) Choline metabolism and nutritional requirement of lake trout (Salvelinus namaycush). Journal of Animal Science 43: 474-477.
Kumar, N., Jadhao, S. B.,Chandan, N. K., Kumar, K., Jha, A.K., Bhushan, S., Kumar, S., Rana, R. S. (2012) Dietary choline, betaine and lecithin mitigates endosulfan-induced stress in Labeo rohita fingerlings. Fish Physiology and Biochemistry 38: 989-1000.
Lin, Y. H. and Shiau, S. Y. (2003) Dietary lipid requirement of grouper, Epinephelus malabaricus, and effects on immune responses. Aquaculture 225: 243-250.
Luo, Z., Liu, Y. J., Mai, K. S., Tian, L. X., Liu, D. H. and Tan, X. Y. (2004) Optimal dietary protein requirement of grouper Epinephelus coioidesjuveniles fed isoenergetic diets in floating net cages. Aquaculture Nutrition 10: 247-252.
Luo, Z., Liu, Y. J., Mai, K. S., Tian, L. X., Liu, D. H., Tan, X. Y. and Lin, H. Z. (2005a) Effect of dietary lipid level on growth performance, feed utilization and body composition of grouper Epinephelus coioides juveniles fed isonitrogenous diets in floating netcages. Aquaculture International 13: 257-269.
Luo, Z., Liu, Y. J., Mai, K. S., Tian, L. X., Yang, H. J., Tan, X. Y. and Liu, D. H. (2005b) Dietary L-methionine requirement of juvenile grouper Epinephelus coioides at a constant dietary cystine level. Aquaculture 249: 409-418.
McDowell, L. R. (1989) Vitamins in animal nutrition. Academic Press, New York. pp. 347-364.
Menten, J. F. M., Pesti, G. M. and Bakalli, R. I. (1997) A newmethod for determining the availability of choline insoybean meal. Poultry Science 76: 1292-1297.
Mai, K., Xiao, L., Ai, Q., Wang, X., Xu, W., Zhang, W., Liufu, Z., Ren, M. (2009) Dietary choline requirement for juvenile cobia, Rachycentron canadum. Aquaculture 289: 124-128.
Mustafa, I. H., Elkamel, A., Ibrahim, G., Chen, P., Elnashaie, S. S. E. H. (2009) Effect of choline and acetate substrates on bifurcation and chaotic behavior of acetylcholine neurocycle and Alzheimer's and Parkinson's diseases. Chemical Engineering Science 64: 2096-2112.
Marcolin, #westeur010#., Forgiarini, L. F., Tieppo, J., Dias, A. S., Freitas, L. A. R., Marroni, N. P. (2011) Methione- and choline-deficient diet induces hepatic changes characteristic of non-alcoholic steatohepatitis. Arquivos de Gastroenterologia 48 :72-79.
Michael, F. R. and Koshio, S. (2013) Dietary choline improvesthe stress tolerance against freshwater and formaline inpost larval Kuruma shrimp, Marsupenaeus japonicas. Aquaculture 2013-Meeting Abstract (n.d.) from the World Wide Web: https://www.was.org/meetings/ShowAbstract.aspx?Id=28390.
National Research Council (2011) Nutrient Requirements of fish. National Academic Press, Washington D. C.
Network of Aquaculture Centres in Asia-Pacific, NACA (2011) Regional review on status and trends in aquaculture development in asia-pacific 2010. Food and Agriculture Organization of the United Nations, FAO. pp. 21-23.
Patterson, K. Y., Bhagwat, S. A., Williams, J. R., Howe J. C., Holden J. M. (2008) USDA database for the choline content of common foods. U.S. Department of Agriculture, Agricultural Research Service, Beltsville Human Nutrition Research Center, Nutrient Data Laboratory.
Pankhurst, N. W. (2011) The endocrinology of stress infish: an environmental perspetive. General and Comparative Endocrinology 170: 265-275.
Poston, H. A., (199l) Choline requirement of swim-up rainbow trout fry. Fish-Culturist 53: 220-223.
Roem, A. J., Stickney, C. C., Kohler, C. C. (1990) Vitamin requirements of blue tilapia in a recirculating water system. Fish-Culturist 52: 15-18.
Rumsey, G. L., (1991) Choline-betaine requirements of rainbow trout (Oncorhynchus mykiss). Aquaculture 95: 107-l 16.
Rosa, J. G. S., Koakoski, G., Piato, A. L., Kreutz, L. C., Fagundes, M., Marqueze, A. and Barcellos, L. J. G. (2013) Impairment of cortisol response to stress in zebrafish acutelyexposed to methyl-parathion. Journal of Environmental Science and Technology 6: 57-62.
Solorzano, L. (1969) Determination of ammonia in natural waters by the phenolhypochlorite method. Limnol. Oceanogr 14: 799-801.
Shiau, S. Y. and Lan, C. W. (1996) Optimum dietary proteinlevel and protein to energy ratio for growth of grouper (Epinephelus malabaricus). Aquaculture 145: 259-266.
Shiau, S. Y., Lo, P. S. (2000) Dietary choline requirements of juvenile hybrid tilapia, Oreochromis niloticus #westeur024# O. aureus. The Journal of Nutrition 130: 100-103.
Tsai, C. E. and Chiang, C. (1996) Effect of methionine and choline, the precursors of lecithin on plasma and liver lipids in rats. Nutritional Sciences Journal 21:13-25.
Twibell, R. G. and Brown, P. B. (2000) Dietary choline equirement of juvenile yellow perch (Perca flavescens). Journal of Nutrition 130: 95-99.
Tienboon, P. and Wangpakapattanawong, P. (2007) Vitamin A status of the minority ethnic group of Karenhill tribe children aged 1-6 years in Northern Thailand. Asia Pacific Journal of Clinical Nutrition. 16: 158-162.
Teodoro, J. S., Rolo, A. P., Duarte, F. V., Sim#westeur054#es, A. M., Palmeira, C. M. (2008) Differential alterations in mitochondrial function induced by a choline-deficient diet: understanding fatty liver disease progression. Mitochondrion 8: 367-376.
Wilson, R. P. and Poe, W. E. (1988) Choline nutrition of fingerling channel catfish. Aquaculture 68: 65-71.
Woodward, B. (1994) Dietary vitamin requirements of cultured young fish, with emphasis on quantitative estimates for salmonids. Aquaculture 124: 133-168.
Wang, D. Z., Zhao, L. and Tan, Y. J. (1995) Requirement of the fingerling grass carp (Ctenopharyngodon idellus) for choline. Journal of Fisheries of China 19: 133-139.
Woollard, D. C. (2000) Determination of choline in milk and infant formulas byenzymatic analysis: collaborative study. Journal of AOAC International 83: 131-138.
Wu, P., Feng, L., Kuang, S. Y., Liu, Y., Jiang, J., Kai, H., Jiang, W. D., Li, S. H., Tang, L., Zhou, X. Q. (2011) Effect of dietary choline on growth, intestinal enzyme activities and relative expressions of target of rapamycin and eIF4E-binding protein 2 gene in muscle, hepatopancreas and intestine of juvenile Jian carp (Cyprinus carpio var. Jian). Aquaculture 317: 107-116.
Wahlqvist, M. L., Tanaka, K., Tzeng, B. H. (2013) Clinical decision-making for vitamin K-1 and K-2 deficiency and coronary artery calcification with warfarin therapy: are diet, factor Xa inhibitors or both the answer? Asia Pacific Journal of Clinical Nutrition 22: 492-496.
Wang, M., Jiang, G. Z., Liu, W. B., Qian, Y. Zhu, J. (2014) Utilization of choline in different levels and dietary choline availability values in seven common feed ingredients for juvenile blunt snout snout bream, megalobrama amblycephala. Acta Hydrobiologica Sinica 38: 51-57.
Yao, Z. and Vance, D. E. (1988) The active synthesis of phosphatidylcholine is required for very low density lipoprotein secretion from rat hepatocytes. The Journal of Biological Chemistry 263: 2998-3004.
Yang, D. Q., Chen, F., Ruan, G. L. (2006) Effect of dietary choline on the growth, tissue lipid content and activities of digestive enzymes of Monopterus albus. Journal of Fisheries of China 30: 676-682.
Yeh, S. P., Shiu, P. J., Guei, W. C., Lin, Y. H., Liu, C. H. (2013) Improvement in lipid metabolism and stresstolerance of juvenile giant grouper, Epinephelus lanceolatus (Bloch), fed supplemental choline. Aquaculture Research 2013: 1-12
Zhang, Z. and Wilson, R. P. (1999) Reevaluation of the choline requirement of fingerling channel catfish (Ictalurus punctatus) and determination of the availability of choline in common feed ingredients. Aquaculture 180: 89-98.