臺灣博碩士論文加值系統

大花咸豐草 (Bidens alba) 與左手香 (Plectranthus amboinicus) 是臺灣常見的藥用植物，已被證實有提升生物免疫力的效果，因此本論文研究目的在探討經熱水萃取後的大花咸豐草或左手香對於南美白對蝦的非特異性免疫之影響。
首先以大花咸豐草或左手香萃取液進行體外實驗，確認大花咸豐草或左手香萃取液對白蝦血球具有良好細胞相容性，並增加吞噬作用 (phagocytosis) 、超氧陰離子 (Superoxide anion, O2-) 產生率及酚氧化酵素活性 (phenoloxidase, PO) 。接著進行連續投餵28天活體實驗結果顯示，以10、20及40 g/kg的大花咸豐草與左手香萃取物分別添加於飼料中連續投餵白蝦28天，結果發現各實驗組均能提升白蝦的總血球數、吞噬作用、超氧陰離子產生率與酚氧化酵素活性。接著連續投餵實驗飼料56天，結果發現20 g/kg大花咸豐草萃取物組與20 g/kg左手香萃取物組顯著提升增重率、特殊成長率並降低飼料轉換率。
綜合上述研究結果，經過熱水萃取後的大花咸豐草或左手香添加於飼料中投餵，均可提升白蝦的非特異性免疫反應以及成長能力，極具潛力開發為水產飼料添加劑，促進未來水產養殖產業發展。

The present study is conducted to investigate the effects of dietary supplemented with Bidens alba (BAE) and Plectranthus amboinicus (PAE) on non-specific immune responses and growth of white shrimp (Litopenaeus vannamei).
As a first step, we determined the effect of the BAE and PAE on whire shrimps heamocytes in vitro test. The results found that BAE and PAE are non-toxicity to white shrimp heamocytes, and increased phagocytosis activity, superoxide anion production rate and phenoloxidase activity.
Following the result from in vivo experiment, we designed four diets containing 10, 20 and 40 g/kg of BAE and PAE for 28 days feeding white shrimps. The result showed both of the BAE and PAE enhanced the non-specific immune responses, such as total haemocyte counts (THC), phagocytosis activity, superoxide anion (O2-) production rate and phenoloxidase activity. In another experiment, white shrimps fed the diet concluding the doses of extracts similarly to the in vivo experiment. Results showed that the shrimps received 20 g/kg of BAE and 20 g/kg of PAE can significantly inproved the growth performance, such as weight gain, specific growth rate and feed coversion ratio after 56 days for feeding trial.
Based on the present results, the administration of Bidens alba or Plectranthus amboinicus extracts supplemented diets can enhanced the non-specific immune responses and improve growth performance of white shrimp. It has great potential to be developed as an aquatic feed additive to promote future development of aquaculture industry.

謝辭 I
摘要 IV
Abstract V
目錄 VI
表目錄 X
圖目錄 XI
第一章、前言 1
第二章、文獻整理 2
一、 南美白對蝦簡介 2
1. 分類與簡史 2
二、 甲殼類免疫系統 2
1. 體液性免疫 (humoral immune response) 3
2. 細胞性免疫 (cellular immune response) 4
三、 藥用植物研究 7
1. 藥用植物於水產養殖上應用 7
2. 大花咸豐草 9
2.1 分類與簡史 9
3. 左手香 10
3.1 分類與簡史 10
第三章、材料與方法 13
ㄧ、實驗設計 13
實驗一：大花咸豐草或左手香萃取液對白蝦血球細胞安全性測試及體外非特異性免疫之影響 13
實驗二：飼料中添加大花咸豐草或左手香萃取液對白蝦非特異性免疫之影響….. 13
實驗三：飼料中添加大花咸豐草或左手香萃取液對白蝦成長之影響 14
二、實驗動物 15
三、實驗環境 15
四、實驗用海水 15
五、實驗用添加物製備 15
六、體外實驗溶液配置 15
七、實驗用飼料配製 16
八、非特異性免疫反應分析方法 16
1. 蝦血淋巴液之抽取 16
2. 總血球數計數 (Total haemocyte counts, THC) 16
3. 血球細胞安全性試驗 16
4. 吞噬作用 (Phagocytosis) 之測定 16
5. 超氧陰離子(Superoxide anion, O2-)產生率之測定 17
6. 酚氧化酵素 (Phenoloxidasse system, PO system)活性之測定 17
九、成長分析方法 18
十、統計分析 18
第四章、結果 19
一、 大花咸豐草與左手香萃取液對白蝦血球細胞安全性測試與體外非特異性免疫之影響 19
1. 大花咸豐草與左手香萃取液對白蝦血球細胞安全性之影響 19
2. 大花咸豐草與左手香萃取液體外培養白蝦血球細胞對吞噬率及吞噬指數之影響 19
3. 大花咸豐草與左手香萃取液體外培養白蝦血球細胞對超氧陰離子產生率之影響 19
4. 大花咸豐草與左手香萃取液體外培養白蝦血球細胞對酚氧化酵素活性之影響 19
二、 飼料中添加大花咸豐草或左手香萃取液對白蝦非特異性免疫之影響 20
1. 飼料中添加大花咸豐草與左手香萃取物對白蝦總血球數之影響 20
2. 飼料中添加大花咸豐草與左手香萃取物對白蝦吞噬率之影響 20
3. 飼料中添加大花咸豐草與左手香萃取物對白蝦吞噬指數之影響 20
4. 飼料中添加大花咸豐草與左手香萃取物對白蝦超氧陰離子之影響 20
5. 飼料中添加大花咸豐草與左手香萃取物對白蝦酚氧化酵素活性之影響…… 21
三、 飼料中添加大花咸豐草或左手香萃取液對白蝦成長表現之影響 21
第五章、討論 22
一、 大花咸豐草或左手香萃取液對白蝦血球細胞安全性測試及體外非特異性免疫之影響 22
1. 細胞安全性測試 22
2. 白蝦血球細胞體外培養中添加不同濃度大花咸豐草或左手香萃取液對非特異性免疫之影響 23
二、 飼料中添加大花咸豐草或左手香萃取液對白蝦非特異性免疫之影響 23
1. 飼料中添加大花咸豐草或左手香萃取物對白蝦總血球數之影響 23
2. 飼料中添加大花咸豐草或左手香萃取液對白蝦吞噬率 (Phagocytic rate) 及吞噬指數 (Phagocytic index) 之影響 24
3. 飼料中添加大花咸豐草或左手香萃取液對白蝦超氧陰離子之影響 24
4. 飼料中添加大花咸豐草或左手香萃取液對白蝦酚氧化酵素活性之影響……. 25
三、 飼料中添加大花咸豐草或左手香萃取液對白蝦成長能力之影響 26
第六章、結論 27
參考文獻 28
附錄 53
附錄一、南美白對蝦血球細胞類型 53
附錄二、白蝦血球細胞在不同染色法所佔比例 54
附錄三、血球細胞型態 54
附錄四、大花咸豐草 (Bidens alba) 54
附錄五、左手香(Plectranthus amboinicus) 55
附錄六、MTT asay化學反應式 55
附錄七、NBT還原超氧陰離子化學式 55
附錄八、吞噬作用-箭頭處為白蝦血球細胞吞噬乳珠 56
附錄九、連續投餵白蝦實驗飼料28天成長表現 57
附錄十、免疫藥品配方 58

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