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研究生:李承燊
研究生(外文):Li,Chen-Shen
論文名稱:酪胺對於南美白蝦免疫反應的影響
論文名稱(外文):Effects of tyramine on the immune responses of Litopenaeus vannamei
指導教授:鄭文騰鄭文騰引用關係
指導教授(外文):Cheng, Winton
口試委員:鄭文騰張欽泉黃榮富
口試委員(外文):Cheng, wintonChang, Chin-ChyuanHuang, Rong-Fu
口試日期:2017-01-16
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:水產養殖系所
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:2017
畢業學年度:105
語文別:中文
論文頁數:70
中文關鍵詞:南美白蝦酪胺溶藻弧菌
外文關鍵詞:Litopenaeus vannameiTyramineVibrio alginolyticus
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摘要
學號:M10413003
論文名稱:酪胺對於南美白蝦免疫反應的影響
總頁數:70
學校名稱:國立屏東科技大學      系(所)別:水產養殖所
畢業時間及摘要別:一百零五年學年度第一學期 碩士學位論文摘要
研究生:李承燊            指導教授:鄭文騰 博士

論文摘要內容:
在本篇的研究中,我們確定了酪胺 (tyramine) 對於南美白蝦免疫力的影響,測量了總血球數 (THC)、不同血球數 (DHC)、酚氧化酵素活性 (PO)、超氧陰離子含量 (O2-)、溶菌酶活性 (lysozyme)、以及感染溶藻弧菌 (Vibrio alginolyticus) 對於吞噬作用、清除能力之影響。並分別注射saline (控制組)、及注射酪胺2.5 × 10-6 mol shrimp-1、酪胺2.5 × 10-5 mol shrimp-1 (實驗組)。
結果表明,注射酪胺2.5 × 10-6 mol shrimp-1、酪胺2.5 × 10-5 mol shrimp-1後0.5、1 及 2小時,其總血球數、不同血球數顯著高於注射saline (控制組)。注射後0.5、1、2 及 4小時,其酚氧化酵素活性及超氧陰離子含量顯著較高。注射後0.5、1 及 4小時,其溶菌酶活性顯著較高。注射後2 及 4小時,其吞噬作用顯著較高。注射後2、4 及 8小時,其清除能力顯著較高。所有的免疫參數在注射酪胺後16小時恢復到對照值。注射酪胺後也顯著地降低病原體溶藻弧菌 (Vibrio alginolyticus) 攻擊的南美白對蝦的死亡率。這些結果表明,神經傳遞物質酪胺能促進免疫系統,並且能增強南美白蝦對於溶藻弧菌 (Vibrio alginolyticus) 的抵抗性。

關鍵字:南美白蝦、酪胺、溶藻弧菌
Abstract
Student ID:M10413003
Title of Thesis:Effects of tyramine on the immune responses of Litopenaeus vannamei
Total page:70
Name of Institute:Department of Aquaculture, National Pingtung University of Science and Technology     
Graduate date:Jan 16, 2017 Degree conferred:Master
Name of Student:Chen-Shen Li     Adviser:Winton Cheng
The Contents of Abstract in this thesis:
In this study, we determined the effect of tyramine (TA) on immunity of white shrimp, Litopenaeus vannamei. The total haemocyte count (THC), different haemocyte count (DHC), phenoloxidase activity, respiratory burst (release of superoxide anion), lysozyme activity, and phagocytic activity and clearance efficiency to the pathogen vibrio alginolyticus were measured when seawater shrimp Litopenaeus vannamei (11.18 ± 0.2) were individually injected with saline, or TA at 2.5 × 10-6 or 2.5 × 10-5 mol shrimp-1.Results showed that DHC and THC from 0.5 to 2 h, PO activity and respiratory burst from 0.5 to 4h, lysozyme activity from 0.5, 1 to 2 h, phagocytic activity from 2 to 4 h, clearance efficiency from 2 to 8 h significantly increased after TA injection at 2.5 × 10-6 or 2.5 × 10-5 mol shrimp-1. All of the immune parameters had returned to control values by 16 h after TA receiving. Injection of TA also significantly decreased the mortality of Litopenaeus vannamei challenged with the pathogen Vibrio alginolyticus. These results suggest that neurotransmitters TA promotes the immune resolution, which in turn enhance the resistance against to Vibrio alginolyticus of shrimp.

Keywords: Litopenaeus vannamei, Tyramine, Vibrio alginolyticus
目錄
摘要 Ⅰ
Abstract II
謝誌 Ⅳ
目錄 Ⅴ
圖表目錄 Ⅸ
第一章 前言 1
第二章 文獻回顧 7
2.1 南美白蝦之介紹 7
2.1.1南美白蝦之分類地位及生活習性 7
2.1.2 南美白蝦養殖概況 7
2.2甲殼類免疫系統介紹 8
2.2.1體液性免疫反應 8
2.2.2細胞性免疫反應 9
2.2.2.1 透明球 9
2.2.2.2半顆粒血球 9
2.2.2.3顆粒血球 10
2.2.2.4吞噬作用(Phagocytosis) 10
2.2.2.5包膜作用(Encapsulation) 10
2.2.2.6原酚氧化酵素系統(Prophenoloxidase, proPO) 11
2.2.2.7 76 kDa蛋白質(Peroxinectin) 12
2.2.2.8 β-glucan binding protein (BGBP) 12
2.2.2.9 反應氧族群(Radical Oxygen Intermediates, ROIs) 12
2.3 甲殼類緊迫神經內分泌機制 13
2.3.1酪胺 (Tyramine) 15
2.3.2 Tyramine 在無脊椎動物上的功能 15
第三章 材料與方法 18
3.1 實驗設計 18
3.1.1實驗動物 18
3.2 酪胺對南美白蝦免疫因子之影響分析 18
3.2.1 血淋巴之抽取 18
3.2.2 總血球數之計數 19
3.2.3 不同血球數之計數方法 19
3.2.4 酚氧化酵素活性之測定方法 19
3.2.5 超氧陰離子含量之測定方法 20
3.2.6 溶菌酶活性之測定方法 20
3.3 酪胺對南美白蝦抗病能力之影響分析 21
3.3.1 吞噬作用及清除能力試驗細菌之配製 21
3.3.2 吞噬作用 21
3.3.3 清除作用 21
3.3.4 攻試驗用病源菌 22
3.3.5攻擊試驗之實驗設計 22
3.4 酪胺對南美白蝦生理因子之影響分析 22
3.4.1 葡萄糖含量之測定 22
3.4.2血淋巴乳酸含量之測定 23
3.5 統計分析 24
第四章 結果 25
4.1 酪胺對南美白蝦免疫因子之影響 25
4.1.1 酪胺對南美白蝦總血球數之影響 25
4.1.2 酪胺對南美白蝦不同種類血球數之影響 27
4.1.3 酪胺對南美白蝦酚氧化酵素活性之影響 33
4.1.4 酪胺對南美白蝦超氧陰離子含量之影響 35
4.1.5 酪胺對南美白蝦溶菌酶活性之影響 37
4.2 酪胺對南美白蝦抗病能力之影響 39
4.2.1 酪胺對南美白蝦吞噬活性之影響 39
4.2.2 酪胺對南美白蝦清除能力之影響 41
4.2.3 攻擊試驗 43
4.3 酪胺對南美白蝦生理因子之影響 45
4.3.1 酪胺對南美白蝦血淋巴葡萄糖之影響 45
4.3.2 酪胺對南美白蝦血淋巴乳酸之影響 47
第五章 討論 49
5.1 酪胺對南美白蝦免疫因子之影響 49
5.1.1 酪胺對南美白蝦總血球數之影響 49
5.1.2 酪胺對南美白蝦不同種類血球數之影響 49
5.1.3 酪胺對南美白蝦酚氧化酵素活性之影響 50
5.1.4酪胺對南美白蝦超氧陰離子含量之影響 50
5.1.5 酪胺對南美白蝦溶菌酶活性之影響 50
5.2 酪胺對南美白蝦抗病能力之影響 51
5.3 酪胺對南美白蝦生理因子之影響 51
第六章 結論 53
參考文獻 54
附錄 66
作者介紹 70








圖表目錄
Fig. 1 Tyrosine代謝路徑 5
Fig. 2 實驗架構 6
Fig. 3 甲殼類緊迫神經內分泌機制 17
Fig. 4南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5、1、2、4、8、16及24小時測量總血球數,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 26
Fig. 5南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5、1、2、4、8、16及24小時測量透明球數,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 28
Fig. 6南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5、1、2、4、8、16及24小時測量顆粒球數,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 30
Fig. 7南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5、1、2、4、8、16及24小時測量半顆粒球數,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 32
Fig. 8南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5、1、2、4、8、16及24小時測量 (A) 總酚氧化酵素活性 (B) (顆粒血球 + 半顆粒血球) 之酚氧化酵素活性,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 34
Fig. 9南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5、1、2、4、8、16及24小時測量超氧陰離子含量,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 36
Fig. 10南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5、1、2、4、8、16及24小時測量溶菌酶活性,每個柱狀圖代表該組別的平均值並包含標準差在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 38
Fig. 11南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、2、4、8、16及24小時測量吞噬活性,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 40
Fig. 12南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、2、4、8、16及24小時測量清除能力,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05).. 42
Fig. 13南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5及1小時測量葡萄糖含量,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 46
Fig. 14南美白蝦分別注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1,並於處理後0、0.5及1小時測量乳酸含量,每個柱狀圖代表該組別的平均值並包含標準差。在相同注射時間點中出現的不同字母代表有顯著性差異 (p < 0.05). 48
Table. 1注射saline、酪胺2.5 × 10-6 mol shrimp-1及酪胺2.5 × 10-5 mol shrimp-1後,利用溶藻弧菌 (Vibrio alginolyticus) 進行攻擊實驗,並記錄7天累積死亡率. 44
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