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研究生:潘志弘
研究生(外文):Chih-Hung Pan
論文名稱:影響草蝦類胡蘿蔔素分佈因素及蝦紅素在草蝦生物功能之研究
論文名稱(外文):Factors Affecting Carotenoids Distribution and Biological Functions of Aataaxanthin in Giant Tiger Penaeus monodon
指導教授:陳瑤湖陳瑤湖引用關係
指導教授(外文):Yew-Hu Chien
學位類別:博士
校院名稱:國立海洋大學
系所名稱:水產養殖學系
學門:農業科學學門
學類:漁業學類
論文種類:學術論文
論文出版年:2001
畢業學年度:90
語文別:中文
中文關鍵詞:呈色生物功能
外文關鍵詞:pigmentationbiological function
相關次數:
  • 被引用被引用:10
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中文摘要
甲殼類無法自行合成類胡蘿蔔素。然而對蝦類主要的類胡蘿蔔素,蝦紅素,不僅在不同組織的色澤分佈扮演重要的角色,同時也具有生物功能,如營養需求、生殖表現、蛋白質的穩定及酵素的活性、抗氧化能力及免疫功用。本研究以五項試驗分別針對影響草蝦類胡蘿蔔素分佈的內因性與外因性的因子及蝦紅素的一些生物功能進行探討:
第一項研究為大小、性別及不同脫殼週期對養殖草蝦在不同組織的類胡蘿蔔素含量之研究。性別間肝胰臟、頭、肌肉及殼的類胡蘿蔔素含量沒有差異。在 0.4-2.8 g的體重範圍,類胡蘿蔔素及蝦紅素的含量隨蝦體重而降低。頭、肌肉及殼的類胡蘿蔔素和蝦紅素含量在脫殼週期之 D期最高,B期最低。肌肉有最低的蝦紅素濃度。在各個脫殼週期, 頭的蝦紅素含量比殼高。頭及肌肉的類胡蘿蔔素含量不僅受脫殼週期的影響也受水份含量的影響,殼的類胡蘿蔔素只受脫殼週期的影響。
第二項研究為飼養冷凍豐年蝦無節幼蟲及臺灣毛蝦對草蝦後期幼蟲成長、活存、蝦紅素濃度及色素組成之影響。豐年蝦無節幼蟲主要含裸藻酮素、海膽酮素及β-胡蘿蔔素,但沒有蝦紅素。臺灣毛蝦的類胡蘿蔔素主要為蝦紅素。豐年蝦無節幼蟲比臺灣毛蝦含有較高的總類胡蘿蔔素。在飼養四週後,飼養豐年蝦的草蝦後期幼蟲其活存率比飼養臺灣毛蝦的高。飼養豐年蝦與飼養臺灣毛蝦的草蝦後期幼蟲其成長沒有差異。飼養豐年蝦的草蝦後期幼蟲其總蝦紅素、游離蝦紅素、單酯化蝦紅素及雙酯化蝦紅素的濃度均高於飼養臺灣毛蝦者。除了游離蝦紅素之外,飼養豐年蝦的草蝦後期幼蟲其總蝦紅素、單酯化蝦紅素、雙酯化蝦紅素並未隨飼育時間而下降。但飼養臺灣毛蝦的草蝦後期幼蟲其總蝦紅素、游離蝦紅素、單酯化蝦紅素或雙酯化蝦紅素均隨飼育期間而下降。餵食這兩種蝦的草蝦後期幼蟲其活存率均與組織中的游離蝦紅素呈正相關。餌料中的總類胡蘿蔔素含量比其色素組成較會影響草蝦後期幼蟲的蝦紅素的濃度及組成。
第三項研究為光照、藻類、色素濃度對草蝦後期幼蟲成長、活存及呈色之影響。在飼養第四週之後隨著蝦的成長體蝦紅素濃度明顯下降。僅在第四週光照(24小時光照比黑暗)會明顯的防止體蝦紅素的降低,但藻水(Isochrysis galbana,3.36-8.70 mg/L)及飼糧蝦紅素(80 mg/kg)從第一週開始即明顯防止蝦體蝦紅素的下降。光照、藻類及飼糧蝦紅素對草蝦成長沒有顯著的影響。整個試驗中,飼養在光環境的蝦比在暗環境有較高的活存率。飼糧蝦紅素僅在第一週改善活存率。僅在第四週蝦的活存率才和體蝦紅素濃度成正相關。當草蝦成長時體內的蝦紅素會降低,為了要維持更好的活存率,維持草蝦後期幼蟲體蝦紅素的一定含量是必須的。
第四項研究為餵食過高量蝦紅素之幼草蝦在缺氧緊迫下的死亡率及其致死過程。在缺氧緊迫下(溶氧<1.0 mg/l,4 h),已餵食高量蝦紅素(360 mg/kg)一週的處理組,其活存率較控制組的蝦高。在11%的顯著水準下, 處理組蝦的致死溶氧量較控制組為低。蝦紅素不僅能幫助草蝦容忍低溶氧,而且會依環境的溶氧條件扮演緩衝氧的角色。
第五項研究為草蝦稚蝦飼養蝦紅素對環境緊迫及病原攻擊的忍受性。本研究以攝取蝦紅素來增加蝦體蝦紅素含量,增強蝦的抗氧化能力,以忍受物理緊迫(5分鐘內水溫急速變化 27℃至 5℃及/或鹽度變化 32 至 0 ppt)、化學緊迫(72 小時氨在0,0.02, 0.2, 2, 20 mg/L的毒性測驗)及病原攻擊(持續5天的弧菌水浴)。在物理緊迫下,處理組比控制組有較高的恢復率(56-55% vs 48%),此結果顯示提供蝦紅素可以改善對溫度及滲透壓緊迫的抵抗力。由於蝦紅素的存在,血淋巴總抗氧化狀態增強,氧化歧化酵素降低。攝取蝦紅素可以改善肝胰臟的功能,因為攝食240 mg/kg的蝦紅素其天門冬氨酸轉氨酵素顯著高於攝食80 及 0 mg/kg的蝦。但是血淋巴的丙氨酸轉氨酵素及天門冬氨酸轉氨酵素含量無法反映在溫度及鹽度緊迫後健康的改善。飼料內提供 80 mg/kg 的蝦紅素,當面臨氨緊迫時會改善活存率。天門冬氨酸轉氨酵素是四指標中唯一能適當的反映蝦面臨氨緊迫時的生理反應及活存率。在飼料裡添加80 mg/kg 的蝦紅素會改善蝦對病原的抵抗力,但僅顯現在第 48 小時的觀察。蝦紅素的保護時間與血淋巴的過氧化抑制效果變成有效的時間一致。本研究結果顯示蝦紅素對草蝦是一種”半基本”營養素;當草蝦面臨物理、化學緊迫及病原攻擊時可能變成一相當關鍵性的營養素。
英文摘要
Crustaceans are unable to synthesize carotenoids de novo. However, astaxanthin, the predominant carotenoid in penaeids, plays important roles not only on color distribution in various tissues but also on biological functions, such as nutrition requirement, reproduction performance, stabilization of protein and enzyme activity, antioxidative ability, and immunity. This study was aimed to find out the intrinsic and extrinsic factors that affected carotenoids distribution and some of the biological functions of astaxanthin in giant tiger prawn, Penaeus monodon through five studies:
Study 1- Carotenoid content in various tissues of cultured P. monodon by their sexes, sizes, and molting stages. There were no differences between sexes in carotenoid content in hepatopancreas, head, flesh, and shell. Carotenoid content decreased with prawn weight in range of 0.4-2.8 g. Carotenoid and astaxanthin contents in head, flesh, and shell were highest at D stage and lowest at B stage. Flesh had the lowest astaxanthin concentration. Head had higher astaxanthin content than shell at all stages. Carotenoid contents in head and flesh were affected by not only the molting stages but also the water content and carotenoid contents in shell were affected by mainly the molting stages.
Study 2 - Astaxanthin concentration and composition, survival, and growth of postlarvae P. monodon fed with frozen brine shrimp Artemia sp. nauplii and minced Taiwan mauxia shrimp Acetes intermedius. Diet B (brine shrimp) contained mainly canthaxanthin, echinenone, and β-carotene, but no astaxanthin. Carotenoids in Diet M (mauxia shrimp) were mainly astaxanthin. Diet B had higher total carotenoid (TC) than Diet M. After 4 weeks’ feeding, B-prawn had higher survival than M-prawn. No difference in growth was found between B-prawn and M-prawn. The concentrations of total astaxanthin (TA), free astaxanthin (FA), astaxanthin monoester (MA), and astaxanthin diester (DA) in B-prawn were all higher than those in M-prawn. Except for FA, no reduction of TA, MA, or DA concentration during the feeding interval was found in B-prawn. However, concentration of TA, FA, MA, and DA in M-prawn all decreased. Survivals of both prawns were positively correlated to tissue FA concentration. TC content of the diets had a greater influence on resulting prawn astaxanthin concentration and composition than carotenoid fractions of the diets.
Study 3 - Effects of light regime, algae in the water, and dietary astaxanthin on pigmentation, growth, and survival of P. monodon postlarvae. After 4 weeks’ rearing, body astaxanthin concentrations decreased significantly as prawns grew. Light (24-h light vs. dark) had significant effects in preventing body astaxanthin reduction only during the 4th week, but both algae in the water (Isochrysis galbana, 3.63-8.70 μg/L vs. none) and dietary astaxanthin (80 mg astaxanthin/kg vs. none) had significant effects in preventing body astaxanthin reduction from the 1st week. There were no significant effects of light regime, algae in the water, or dietary astaxanthin on prawn growth. Throughout the experiment, the survival of prawn reared in the light was higher than that for prawn reared in the dark. The survival of prawn reared in algae-containing water was higher than that of prawn reared in clear water except in the 3rd week. Dietary astaxanthin improved prawn survival only in the 1st week. Only in the 4th week did survival have a positive correlation with prawn body astaxanthin concentration. It is essential to maintain a certain level of astaxanthin in tiger prawn postlarvae for better survival while the prawn are growing and their astaxanthin is decreasing.
Study 4 - Oxygen depletion stress on mortality and lethal course of juvenile P. monodon fed high level of dietary astaxanthin. Under oxygen depletion stress (dissolved oxygen < 1.0 mg/l for 4 h), P-prawn, which had fed high level of dietary astaxanthin (360 mg/kg) for 1 week had higher survival rate than C-prawn, the control. Lethal dissolved oxygen level of P-prawn was lower than C-prawn at a significant level of 11%. Astaxanthin not only helped the prawn tolerate lower dissolved oxygen but also may play a buffer role on oxygen intake according to the environment dissolved oxygen conditions.
Study 5 - The resistance to environmental stresses and pathogen challenge by P. monodon juvenile fed diets supplemented with astaxanthin. This study was aimed to determine if increasing body astaxanthin content through dietary supplementation (80 mg/kg and 240 mg/kg astaxanthin for 8 weeks) could enhance prawn’s antioxidant defense capability and resistance to physical stress (a rapid change in water temperature from 27 C to 5 C and/or salinity from 32 ppt to 0 ppt for 5 minutes), chemical stress (ammonia toxicity at 0, 0.02, 0.2, 2, and 20 mg/L ammonia-N for 72 h), and pathogen challenge (bath challenge of Vibrio damsela for 5 days). Significantly higher recovery rates in treated prawn than control prawn (56-55% vs. 48%) suggested that astaxanthin supplement had improved the resistance against thermal and osmotic stress. Haemolymph total antioxidant status (TAS) was improved and haemolymph superoxide dismutase (SOD) was reduced by the presence of dietary astaxanthin. Shrimp hepatopancreatic function may have been improved by dietary astaxanthin since haemolymph astpartate aminotransferase (AST) of prawn fed 240 mg/kg astaxanthin was significantly higher than those of prawn fed 80 mg/kg and 0 mg/kg astaxanthin. However, haemolymph alanine aminotransferase (ALT) and AST content of prawn did not reflect improvement in health following thermal and osmotic stresses. Astaxanthin supplement in diet at 80 ppm improved survival rate of shrimp subjected to ammonia stress. AST was the only indicator among the four to adequately reflect the physiological response or survival rate of prawn subjected to ammonia stress. Astaxanthin supplement in diet at 80 ppm improved the shrimp against pathogen challenge, however, only at 48 h after the contamination. The timing of astaxanthin protection coincided with the time when hydroperoxide inhibition in haemolymph became effective. This study has demonstrated that for P. monodon, astaxanthin is a ‘semi-essential’ nutrient, which can become critical when the animal is under the physical and chemical stress and the pathogen challenge.
目錄
目錄I
謝辭V
中文摘要VI
英文摘要VIII
第一章 前言1
1甲殼類色素研究簡史因子2
2-1影響類胡蘿素的內因性因子2
2-2影響類胡蘿素的外因性因子3
3甲殼類色素的生物功能4
3-1抗氧化能力5
3-2營養需求5
3-3免疫或抗病源感7
4研究方向7
第二章文獻整理8
1甲殼類色素的特性、結構與種類8
2甲殼類色素的分布10
2-1內因性因子的影響12
2-2外因性因子的影響13
3甲殼類色素的生物功能13
3-1營養需求(維生素A)14
3-2生殖功能14
3-3穩定蛋白質及酵素活性15
3-4氧的貯備能力15
3-5抗氧化特性16
3-6免疫或抗病原感染20
I.內因性因子之研究
第三章養殖之草蝦在性別、大小對不同脫殼週期與類胡蘿蔔素的含量的關係26
1 摘要26
2前言27
3材料與方法28
3-1大小試驗28
3-2性別試驗28
3-3不同脫殼週期試驗28
3-4脫殼期數及性別的判別28
3-5色素的萃取及分析29
3-5-1重要化學藥品及標準品29
3-5-2蝦紅素標準品的萃取及分析29
3-5-3樣本類胡蘿蔔素及蝦紅素的萃取及分析29
3-5-4分析程序30
3-5-5統計測定30
4結果30
4-1類胡蘿蔔素之含量與分佈31
4-2蝦紅素之含量與分佈31
5討論32
5-1類胡蘿蔔素之含量與分佈32
5-2蝦紅素之含量與分佈34
II.外因性因子之研究
第四章餵飼冷凍豐年蝦(Artemia sp.)無節幼蟲及絞碎之台灣毛蝦(Acetes intermedius)對草蝦後期幼蟲成長、活存、呈色及其色素組成之研究45
1摘要45
2前言46
3材料及方法47
3-1試驗設計47
3-2餌料及飼育方法47
3-3飼育48
3-4蝦紅素分析48
3-5統計分析48
4結果48
4-1活存與成長48
4-2呈色49
5討論50
5-1活存50
5-2成長51
5-3呈色51
第五章光照、藻類、飼料中的色素添加對草蝦呈色、成長和活存的影響61
1摘要62
2前言62
3材料與方法62
3-1試驗設計62
3-2飼料準備63
3-3藻水63
3-4光照63
3-5蝦的飼養63
3-6藻類中類胡蘿蔔素之分析64
3-7蝦紅素分析64
3-8統計分析64
4結果64
4-1呈色64
4-2成長65
4-3活存66
5討論66
5-1呈色66
5-2成長69
5-3活存(Survival)69
III.生物功能之研究
第六章餵食高量蝦紅素對幼草蝦在缺氧緊迫下的死亡率及致死的影響85
1摘要85
2前言86
3材料與方法86
3-1飼料準備86
3-2缺氧緊迫試驗87
3-3致死氧試驗88
3-4蝦紅素分析88
4結果與討論88
第七章草蝦體內蝦紅素含量對環境緊迫與抗病原之影響95
1摘要95
2前言96
3材料與方法99
3-1試驗設計99
3-2養殖方法100
3-3飼料配方100
3-4耐緊迫試驗100
3-4-1試驗(I):溫、鹽度之緊迫試驗100
3-4-2試驗(II):氨的緊迫試驗101
3-4-2-1氨測試溶液101
3-4-2-2試驗設計101
3-4-3試驗(III):抗病原菌的試驗101
3-5樣本處理102
3-5-1試藥102
3-6酵素的活性的測定103
3-6-1酵素的活性103
3-6-1-1TAS測定103
3-6-1-2SOD測定104
3-6-1-3AST、ALT測定105
3-6-2血淋巴蛋白質105
3-6-3LPO(Lipid peroxides)的測定105
3-7蝦紅素的萃取及分析106
3-8統計分析106
4結果106
4-1飼養後草蝦蝦紅素含量、成長與活存率107
4-1-1飼育後草蝦體內血液抗氧化酵素107
4-1-2過氧化物抑制狀態107
4-2溫度、鹽度緊迫108
4-3氨緊迫109
4-4抗病原菌緊迫110
4-4-1抗病原菌緊迫草蝦體內血液抗氧化酵素110
5討論111
5-1呈色111
5-2活存與成長111
5-3物理緊迫-溫、鹽度激烈下降112
5-3-1恢復率112
5-3-2TAS與SOD113
5-3-3AST與ALT115
5-4化學緊迫-氨毒性116
5-5毒性試驗118
第八章結論140
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