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研究生:曾怡婷
研究生(外文):Tseng I Ting
論文名稱:內因性因子與外因性因子-氨、亞硝酸及硫酸銅影響白蝦免疫反應
論文名稱(外文):Effects of intrinsic factors and extrinsic factors - ammonia, nitrite and copper sulfate on the immune response of white shrimp Litopenaeus vannamei
指導教授:陳鴻鳴陳鴻鳴引用關係
指導教授(外文):Chen Hung Min
學位類別:碩士
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:2002
畢業學年度:90
語文別:中文
論文頁數:83
中文關鍵詞:白蝦總血球數透明血球顆粒血球酚氧化酵素超氧離子超氧歧化酵素Vibrio alginolyticus
外文關鍵詞:Litopenaeus vannameitotal haemocyte counthyaline cellgranular cellphenoloxidase activityrespiratory burstsuperoxide dismutaseVibrio alginolyticus
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摘要
研究白蝦Litopenaeus vannamei內因性因子-性別、體重、脫殼週期及攝餌率與其免疫因子之關係,研究外因性因子-氨、亞硝酸及硫酸銅對於白蝦免疫因子的影響,以及白蝦感染Vibrio alginolyticus,曝露於氨、亞硝酸及硫酸銅濃度下之抵抗力。免疫因子分析包括總血球數、透明血球數、顆粒血球數、酚氧化酵素、超氧離子及超氧歧化酵素。
白蝦之總血球數、透明血球數、顆粒血球數、酚氧化酵素及超氧離子並不受性別與體重(3.68~30.38 g)影響。白蝦在不同脫殼週期(A、B、C、D0/D1及D2/D3)中,其總血球數、透明血球數、顆粒血球數、酚氧化酵素及超氧離子均以在脫殼間期(C期)顯著高於脫殼後期(A期),總血球數、透明血球數及顆粒血球數在C期及脫殼前期(D0/D1)無顯著差異,酚氧化酵素及超氧離子在C期及脫殼前期(D0/D1及D2/D3)無顯著性差異。白蝦在不同攝餌率(0﹪、0.1﹪、0.5﹪、1.0﹪及2.0﹪)下,以攝餌率0﹪及0.1﹪經7天後,其總血球數及酚氧化酵素活性明顯下降,而超氧離子在各組間則無顯著性差異。
白蝦感染V. alginolyticus(1 ×106 cfu shrimp-1)後,曝露在5.5 mg l-1氨-氮或以上濃度,經168小時後,其累積死亡率均顯著高於對照組。白蝦曝露於11.17及22.32 mg l-1氨-氮,經2天後,其超氧離子濃度上升,而超氧歧化酵素下降,經7天後,酚氧化酵素活性下降,而白蝦之總血球數、透明血球數及顆粒血球數曝露在0、1.07、5.40、11.17及22.32 mg l-1氨-氮組之間沒有顯著差異。
白蝦感染V. alginolyticus(1 ×106 cfu shrimp-1)後,曝露於5.5 mg l-1亞硝酸-氮或以上濃度,經168小時後,其累積死亡率均顯著高於對照組。白蝦曝露於4.94 mg l-1亞硝酸-氮或以上濃度,經2天後,其總血球數及酚氧化酵素活性下降,經7天後,超氧離子濃度上升。
白蝦感染V. alginolyticus(3 ×105 cfu shrimp-1)後,曝露於5 mg l-1 Cu2+或以上濃度,經168小時後,其累積死亡率均顯著高於對照組。白蝦曝露於5 mg l-1 Cu2+或以上濃度,12及24小時,其總血球數及酚氧化酵素活性下降,超氧離子濃度上升。
本實驗證實白蝦免疫能力與性別及體重(3.68~30.38 g)無關,白蝦在不同脫殼週期,以脫殼間期(C期)時免疫能力較好,而投餌率在0及0.1﹪時,白蝦免疫能力下降。白蝦感染V. alginolyticus,曝露在5.5 mg l-1氨-氮、5.5 mg l-1亞硝酸-氮或5 mg l-1 Cu2+,其抵抗力下降,白蝦曝露於氨-氮11.17 mg l-1、4.94 mg l-1亞硝酸-氮或5 mg l-1 Cu2+,其酚氧化酵素活性下降。酚氧化酵素可以當作白蝦免疫能力指標。養殖過程中應注意水質的變化及白蝦的免疫力而做適當管理。
關鍵字:白蝦、性別、體重、脫殼週期、攝餌率、氨、亞硝酸、硫酸銅、總血球數、透明血球、顆粒血球、酚氧化酵素、超氧離子、超氧歧化酵素、Vibrio alginolyticus。
Abstract
This paper deals with the effects of intrinsic factors like gender, size, molt stages and feeding rate, and the effect of extrinsic factors like ammonia, nitrite and copper sulfate on the immune response of Litopenaeus vannamei. Immune tools used are total haemocyte count (THC), hyaline cell, granular cell, phenoloxidase, respiratory burst and superoxide dismutase (SOD). L. vannamei challenged with Vibrio alginolyticus were placed in water containg different concentrations of ammonia, nitrite and copper sulfate, and the mortality of shrimp was also studied.
No significant differences in THC, hyaline cell, granular cell, phenoloxidase and respiratory burst were observed between male and female shrimp and among shrimp weightd from 3.68 to 30.38 g. Among molt stages of A, B, C, D0/D1 and D2/D3, the THC, hyaline cell, granular cell, phenoloxidase and respiratory burst in C stage are higher than those in A stage. The shrimp with no feeding or feeding rate at 0.1﹪showed significantly lower THC and phenoloxidase activity after 7 days.
The mortality of L. vannamei challenged with V. alginolyticus at 1 x 106 cfu shrimp-1 and exposed to 5.5 mg l-1 ammonia-N was significantly higher than that of shrimp challenged with V. alginolyticus at same dose and exposed in the control solution. L. vannamei following 2 days exposure to 11.17 and 22.32 mg l-1 ammonia-N decreased its SOD, but increased its respiratory burst, and the shrimp following 7 days exposure to 11.17 and 22.32 mg l-1 ammonia-N decreased its phenoloxidase activity. However, no significant differences in THC, hyaline cells and granular cells were observed among the shrimp exposed to 0, 1.07, 5.40, 11.17 and 22.32 mg l-1 ammonia-N.
The mortality of L. vannamei challenged with V. alginolyticus at 1 x 106 cfu shrimp-1 and exposed to 5.5 mg l-1 nitrite-N was significantly higher than that of shrimp challenged with V. alginolyticus at same dose and exposed in the control solution. L. vannamei following 2 days exposure to 4.94 mg l-1 nitrite-N decreased its THC and phenoloxidase activity, and the shrimp following 7 days exposure to 4.94 mg l-1 nitrite-N increased its respiratory burst.
The mortality of L. vannamei challenged with V. alginolyticus at 3 x 105 cfu shrimp-1 and exposed to 5 mg l-1 Cu2+ was significantly higher than that of shrimp challenged with V. alginolyticus at same dose and exposed in the control solution. L. vannamei following 24 h exposure to 5 mg l-1 Cu2+ decreased its THC and phenoloxidase activity, but increased its respiratory burst.
The present study documented that no significant difference in immune ability was observed between male and female L. vannamei, and among the shrimp weighed from 3.68 g to 30.38 g. L. vannamei with no feeding or the shrimp at feeding rate of 0.1 % decreased its immune ability. L. vannamei challenged with V. alginolyticus decreased its resistance when exposed to 5.5 mg l-1 ammonia-N, 5.5 mg l-1 nitrite or 5 mg l-1 Cu2+ .It is concluded that L. vannamei exposed to 11.17 mg l-1 ammonia-N, 4.94 mg l-1 nitrite-N or 5 mg l-1 Cu2+ decreased its immune ability by reducing its phenoloxidase activity. Management of water quality and maintaining better immune ability of cultured animals are suggested during shrimp farming.
Keyword: Litopenaeus vannamei, gender, size, molt cycle, feeding rate, ammonia, nitrite, copper sulfate, total haemocyte count, hyaline cell, granular cell, phenoloxidase activity, respiratory burst, superoxide dismutase, Vibrio alginolyticus.
目錄
目錄………………………………………………………………………..I
中文摘要…………………………………………………………………III
英文摘要…………………………………………………………………VI
第一章 前言 1
第二章 文獻整理 4
1. 白蝦簡介 4
2. 甲殼類的防禦機制 4
2.1 體液性免疫 4
2.2 細胞性免疫 6
3. 影響甲殼類免疫防禦機制因子 10
3.1 內因性因子對甲殼類免疫因子之影響 11
3.2 外因性因子對甲殼類免疫防禦機制之影響 12
第三章 材料與方法 18
1. 實驗用蝦 18
2. 實驗用水 18
3. 血淋巴之抽取 18
4. 實驗設計 18
5. 血淋巴免疫因子之分析 22
6. 感染實驗 24
7. 統計分析 26
第四章 結果 27
1. 內因性因子對白蝦免疫防禦機制之影響 27
1.1白蝦性別與免疫因子之關係 27
1.2白蝦體重與免疫因子之關係 27
1.3白蝦脫殼週期與免疫因子之關係 28
1.4白蝦攝餌率與免疫因子之關係 29
2. 外因性因子對白蝦免疫防禦機制之影響 29
2.1 氨對白蝦免疫因子之影響 29
2.2 亞硝酸對白蝦免疫因子之影響 31
2.3 硫酸銅對白蝦免疫因子之影響 32
第五章 討論 34
1. 內因性因子對白蝦免疫防禦機制之影響 34
1.1白蝦性別及體重與免疫因子之關係 34
1.2白蝦脫殼週期與免疫因子之關係 35
1.3白蝦攝餌率與免疫因子之關係 36
2. 外因性因子對白蝦免疫防禦機制之影響 37
2.1 氨對白蝦免疫因子之影響 37
2.2 亞硝酸對白蝦免疫因子之影響 39
2.3 硫酸銅濃度對白蝦免疫因子之影響 40
參考文獻 43
表………………………………………………………………………….57
圖………………………………………………………………………….62
附錄……………………………………………………………………….80
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