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研究生:張宏揚
研究生(外文):Chang, Hung-Yang
論文名稱:飼糧類胡蘿蔔素及大蒜粉對金目鱸的成長、活存率、抗氧化及非特異免疫能力的影響
論文名稱(外文):Effects of dietary carotenoids and garlic powder on growth、survival rate、antioxidant and non-specific immune capacity of Lates calcarifer
指導教授:潘志弘潘志弘引用關係
指導教授(外文):Pan, Chih-Hung
口試委員:鄭文騰黃貴民
口試委員(外文):Cheng, WintonHuang, Kuei-Min
口試日期:2011-06-28
學位類別:碩士
校院名稱:國立高雄海洋科技大學
系所名稱:水產養殖研究所
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:2011
畢業學年度:99
語文別:中文
論文頁數:117
中文關鍵詞:金目鱸β-胡盧蔔素蝦紅素大蒜粉抗氧化非特異免疫
外文關鍵詞:barramundiβ-caroteneAstaxanthingarlic powderantioxidantnon-specific immune
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本研究主要目的在於探討飼糧類胡蘿蔔素 (Carotenoids) 及大蒜粉 (Garlic powder) 對金目鱸 (Late calcarifer) 的成長、活存率、抗氧化,及非特異免疫能力的影響,整個實驗共有11個處理組,並分成類胡蘿蔔素及大蒜粉兩種添加物,分別為 β-胡蘿蔔素 (β-carotene)、蝦紅素 (Astaxanthin) 及大蒜粉各0、80、160及320 mg/kg四種濃度,並投餵金目鱸12週,經過12週的飼養週期後,在各處理組間的成長及活存率沒有顯著的差異 (p>0.05),而在實驗當中血漿中的總抗氧化狀態 (Total antioxidative status, TAS)、抗氧化酵素 [超氧歧化酵素 (Superoxide dismutase, SOD)、麩胱甘肽過氧化酵素 (Glutathioine peroxidase, GPx)、麩胱甘肽還原酵素 (Glutathione reductase, GR) ] 以及轉氨酵素 [天門冬氨酸轉氨酵素 (Aspartate aminotransferase, AST)、丙氨酸轉氨酵素 (Alanine aminotransferase, ALT) ] 被用來當作抗氧化能力的評估指標,實驗結果顯示,TAS 及 GR 的活性與控制組間沒有顯著的差異 (p>0.05),然而類胡蘿蔔素的處理組與控制組相比較,有較低的AST、ALT 及 SOD 的活性表現。在 GPx 的活性以飼糧160及320 mg/kg的蝦紅素顯著高於 β-胡蘿蔔素、大蒜粉以及控制組。
非特異免疫防禦以細胞性 (吞噬活性、呼吸爆)、體液性 (溶菌酵素)、SOD 及 GPx 做為評估的指標。在呼吸爆的活性上,類胡蘿蔔素及未氧化脂質的處理組與氧化脂質組有顯著的差異 (p<0.05)。在頭腎白血球的吞噬活性以飼糧80 mg/kg 的 β-胡盧蔔素處理組顯著高於蝦紅素、大蒜粉及控制組3組處理組,然而在吞噬活性上並沒有觀察到魚類飼糧蝦紅素、β-胡盧蔔素及大蒜粉有顯著的差異 (p>0.05)。血清中溶菌酵素的活性以飼糧大蒜粉比氧化脂質組有較高的活性表現,此外也觀察到 SOD 的活性隨著飼糧中蝦紅素及 β-胡盧蔔素濃度的增加其活性也隨之降低,並與氧化脂質組有顯著的差異 (p<0.05)。但在大蒜粉的處理組與氧化脂質組的 SOD 活性則是沒有顯著差異 (p>0.05)。GPx 以飼糧蝦紅素及 β-胡盧蔔素比氧化脂質組有較高的活性表現,並有顯著差異 (p<0.05)。然而大蒜粉與氧化脂質處理組間的 GPx 活性則是沒有顯著差異 (p>0.05)。

This study was aimed to find out the effects of dietary supplemented carotenoids and garlic powder on growth, survival rate, antioxidant capacity and non-specific immune of barramundi, Late calcarifer. Eleven experimental diets supplemented with two types of carotenoid and garlic powder : astaxanthin, β-carotene, and garlic powder, at four concentrations : 0, 80, 160, and 320 mg/kg were feed to the fish for twelve weeks. No differences in growth and survival rate of the fish among treatments were found after 12-week rearing. Serum total antioxidant status (TAS). Serum antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidases (GPx), glutathione reductase (GR)] and serum transaminases [alanine aminotrans- ferase (ALT). asparatate aminotransferase (AST)] were chosen as indices of fish antioxidant capacity. There were no differences in TAS and GR between treatments and the control fish. The CD-fed fish had lower AST, ALT, and SOD activities than the control fish. The GPx of fish fed 160 and 320 mg/kg astaxanthin diets were significantly higher than those of fish fed β-carotene , garlic powder and the control diet. There were significantly different in GPx between astaxanthin treatment and the control diet.
Non-specific immune response (phagocytic activity, respiratory burst , lysozyme activity), superoxide dismutase and glutathione peroxidase activity were evaluated. Carotenoid and non-oxidized oil treated fish showed significantly higher respiratory burst activity compared with the oxidized oil groups. The phagocytic activity of head kidney leukocytes of fish fed 80 mg/kg β-carotene (BI) containing diets were significantly higher than those of fish fed astaxanthin, garlic powder and the control diet. However, no significant differences in phagocytic activity were observed between fish fed the astaxanthin, β-carotene and garlic powder supplemented diets. The serum lysozyme activities of fish fed garlic powder diets were significantly higher than those of fish fed the oxidized oil groups. Significant differences in lysozmye activities were found between fish fed garlic powder and the oxidized oil groups. Decreasing trends in SOD activities with increasing dietary astaxanthin and β-carotene were also observed. There were significant differences in SOD activity between fish fed the astaxanthin, β-carotene and the oxidized oil group. However, no significant differences in SOD activity were observed between fish fed garlic powder supplemented diets and the oxidized oil group. GPx activities of fish fed astaxanthin and β-carotene were significantly higher than those of fish fed the oxidized oil group. There were not significantly different in GPx activities between fish fed garlic powder supplemented diets and. the oxidized oil group.

目錄

摘要..................................................... Ⅰ
Abstract................................................. Ⅲ
誌謝..................................................... Ⅴ
目錄..................................................... Ⅵ
表目錄................................................... Ⅸ
圖目錄................................................... Ⅹ
略語表................................................... ⅩⅠ
第1章 前言.............................................. 1
第2章 文獻整理.......................................... 3
2.1 金目鱸的生態及分類地位介紹........................... 3
2.2 脂質的氧化........................................... 4
2.3 動物體內的氧化與抗氧化................................ 5
2.4 抗氧化物在體內預防氧化緊迫的防禦關聯................. 8
2.5 魚類的免疫系統....................................... 10
2.6 特異性免疫系統....................................... 10
2.7 非特異性免疫系統..................................... 11
2.8 類胡蘿蔔素簡介........................................ 13
2.9 β-胡蘿蔔素簡介.......................................... 16
2.10 蝦紅素簡介............................................ 17
2.11 大蒜簡介.............................................. 18
第3章 飼糧類胡蘿蔔素及大蒜粉對金目鱸的成長、活存率與抗氧化能力的影 響......................................................... 21
3.1 實驗設計................................................ 21
3.2 飼糧準備................................................ 22
3.3 魚的飼養................................................ 22
3.4 脂質過氧化物價的測定 (Peroxide value, POV)............... 23
3.5 抗氧化酵素活性的測定.................................. .... 24
3.5.1 樣本處理 (粗酵素製備).................................. 24
3.5.2 總抗氧化狀態 (Total antioxidative status, TAS)........ 24
3.5.3 天門冬氨酸轉氨酵素 (Aspartate aminotransferase, AST).. 25
3.5.4 丙氨酸轉氨酵素 (Alanie aminotransferase, ALT)......... 25
3.5.5 麩胱甘肽過氧化酵素 (Glutathione peroxidase, GPx)...... 26
3.5.6 麩胱甘肽還原酵素 (Glutathione reductase, GR).......... 27
3.5.7 超氧歧化酵素 (Superoxide dismutase, SOD)............. 27
3.5.8 血球溶破液中蛋白質................................... 28
3.6 統計分析............................................... 29
3.7 結果................................................... 30
3.8 討論................................................... 35
第4章 飼糧類胡蘿蔔素及大蒜粉對金目鱸非特異免疫反應之影響.......... 61
4.1 實驗設計............................................... 61
4.2 飼糧準備............................................... 62
4.3 魚的飼養............................................... 62
4.4 免疫能力分析............................................ 62
4.4.1 血清採集.......................................... 62
4.4.2 頭腎巨噬細胞的分離................................ ... 62
4.4.3 細胞的粉碎........................................ 63
4.4.4 巨噬細胞呼吸爆 (Respiratory burst)................... 63
4.4.5 溶菌酵素活性測定 (Lysozyme activity assay)............ 64
4.4.6 吞噬率 (Phagocytic rate)............................. 64
4.4.7 SOD 測定......................................... 65
4.4.8 GPx 測定.......................................... 65
4.4.9 血球溶破液中蛋白質................................ 65
4.5 統計分析............................................. 65
4.6 結果................................................. 66
4.7 討論................................................. 70
結論..................................................... 86
參考文獻................................................. 87
表目錄
表一. 基礎飼料成分.......................................... 46
表二、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 TAS (m/mole serum) 之活性表現.......................................................... 47
表三、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 AST (unit/mg protein) 之活性表現......................................................... 48
表四、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 ALT (unit/mg protein) 之活性表現........................................................ 49
表五、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 SOD (unit/mg protein) 之活性表現........................................................ 50
表六、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 GPx (unit/mg protein) 之活性表現......................................................... 51
表七、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 GR (unit/mg protein)之活性表現............................................................ 52
表八、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸呼吸爆 (O.D.630) 之活性表現............................................................ 76
表九、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸溶菌酵素 (μg/mL) 之活性表現............................................................ 77
表十、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸吞噬率 (%) 之活性表現............................................................. 78
表十一、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 SOD (unit/mg protein) 之活性表現............................................................. 79
表十二、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 GPx (unit/mg protein) 之活性表現.............................................................. 80
圖目錄
圖一、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸成長之表現................. 53
圖二、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸活存率之表現............... 54
圖三、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 TAS 之活性表現............. 55
圖四、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 AST 之活性表現............. 56
圖五、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 ALT 之活性表現............. 57
圖六、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 SOD 之活性表現............. 58
圖七、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 GPx 之活性表現............. 59
圖八、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 GR 之活性表現.............. 60
圖九、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸呼吸爆之活性表現............. 81
圖十、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸溶菌酵素之活性表現........... 82
圖十一、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸吞噬率之活性表現........... 83
圖十二、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 SOD 之活性表現........... 84
圖十三、飼糧類胡蘿蔔素及大蒜粉12週,金目鱸 GPx 之活性表現........... 85



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