臺灣博碩士論文加值系統

本實驗探討不同濃度及不同形式的硒對淡水長臂大蝦成長、抗氧化能力、免疫及抗病力之影響。將甲硫胺酸硒與硒酸鈉作為有機硒及無機硒之來源。五組實驗飼料分別為控制組 (未添加硒)、0.5和1 mg/kg的甲硫胺酸硒組、0.5和1 mg/kg的硒酸鈉組。實際所得的硒濃度在控制組、0.5、1 mg/kg的甲硫胺酸硒添加組與0.5、1 mg/kg的硒酸鈉添加組分別為1.17±0.07 mg/kg、1.53±1.36 mg/kg、2.19±0.86 mg/kg、1.59±0.21 mg/kg和2.22±0.45 mg/kg。以0.5和1 mg/kg的甲硫胺酸硒與硒酸鈉添加飼料餵養淡水長臂大蝦77天。在成長方面顯示控制組飼料中含有1.17 mg/kg的硒已足夠蝦子的成長所需。蝦子餵食硒添加飼料相對於控制組有較高的酵素活性，包括超氧歧化酶、麩胱苷肽過氧化酶、麩胱苷肽還原酶和麩胱苷肽硫轉移酶。在免疫因子方面，所有的硒滋養飼料組別的蝦子在總血球數、超氧歧化酶及清除作用，與控制組蝦子並無顯著性差異。然而，蝦子攝食含硒之飼料後其酚氧化酶活性、呼吸爆、麩胱苷肽過氧化酶活性、麩胱苷肽還原酶、麩胱苷肽硫轉移酶及吞噬能力皆明顯的提高。攝食0.5 mg/kg硒添加飼料之蝦子在注射真菌酵母菌後，死亡率明顯低於控制組蝦子。這意謂著於飼料中添加 0.5 mg/kg的硒，使硒之濃度使其達到1.53-1.59 mg/kg，有助於提高淡水長臂大蝦的抗氧化能力、免疫及抗病能力，而基礎飼料中含有1.17 mg/kg的硒已經足夠提供蝦子成長。

The objectives of this study were to investigate the effects of different levels and forms of selenium (Se) on the growth performance, antioxidant response, and immunity and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii. Sodium selenate (SSe) and seleno-L-methionine (MSe) were used as inorganic and organic Se source, respectively. Five experimental diets, including a control diet (without Se supplement), 0.5 and 1 mg/kg of MSe, 0.5 mg/kg and 1 mg/kg of SSe, were used. Providing the actual dietary values of control, 0.5, 1 mg/kg MSe supplied diet and 0.5, 1 mg/kg SSe supplied diet were 1.17±0.07 mg/kg, 1.59±0.21 mg/kg, 2.22±0.45 mg/kg, 1.53±1.36 mg/kg and 2.19±0.86 mg/kg, respectively. Prawns were fed with the experimental diets for 77 days. The growth performance trial showed that a basal diet containing about 1.17 mg/kg of Se is sufficient for growth. The prawn fed the Se supplied diets had greater antioxidant enzyme activities, including superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione S-transferase compared to those fed the control diet. In the trail of the immunity of prawn fed with Se supplied diets, no significant difference in total haemocyte count, superoxide dismutase and clearance efficiency were found among all treatments and control. However, phenoloxidase activity, respiratory burst, glutathione peroxidase, glutathione reductase, glutathione S-transferase and phagocytic activity of prawn fed with both forms of Se containing diets (organic Se and inorganic Se) were significant increased. For challenge test, prawn fed with two forms of Se containing diets at the level of 0.5 mg/kg has a significant lower mortality compared to the control prawn after being injected with Debaryomyces hansenii. It was suggest that feed contain with Se results in 1.5 mg/kg Se in diet may enhance prawn antioxidant status, immunity and disease resistance, and the basal diet containing 1 mg/kg of Se is enough for prawn growth.

目錄
中文摘要 Ⅰ
英文摘要… Ⅲ
目錄……… VI
圖目錄索引 XI
表目錄索引 XIII
第一章、前言 1
第二章、文獻回顧 5
2. 1 硒的特性 7
2. 2 硒對人體的影響 7
2. 2. 1 硒的吸收與代謝 10
2. 3 硒對水產動物的影響 13
2. 3. 1硒對魚類的影響 13
2. 3. 2 硒對蝦類的影響 14
2. 4 硒的毒性 14
2. 5 自由基與抗氧化系統 15
2. 6 甲殼類的免疫系統 20
第三章 材料與方法 25
3. 1 實驗動物 25
3. 2 實驗架構 25
3. 3 飼料配製 25
3. 4 飼料成分分析 27
3. 4. 1 水分的分析 27
3. 4. 2 灰分的分析 27
3. 4. 3 粗蛋白的分析 .... 27
3. 4. 4 粗脂肪的分析 28
3. 5 養成試驗 29
3. 6 水質檢測 29
3. 6. 1 溶氧的檢測 29
3. 6. 2 酸鹼值的檢測 29
3. 6. 3 溫度的檢測 30
3. 6. 4 氨氮的檢測 30
3. 6. 5 亞硝酸鹽的檢測 30
3. 7 成長指標 31
3. 8 硒含量分析 31
3. 9 抗氧化酵素分析 32
3. 9. 1 樣本的配製 32
3. 9. 2 蝦體組織麩胱苷肽過氧化酶之測定 32
3. 9. 3 蝦體組織麩胱苷肽還原酶之測定 33
3. 9. 4 蝦體組織麩胱苷肽硫轉移酶之測定 33
3. 9. 5 蝦體組織超氧歧化酵素之測定 34
3. 10. 抗病力及免疫因子試驗 35
3. 10. 1 病原菌之培養 35
3. 10. 2 攻擊試驗 36
3. 10. 3 免疫因子試驗 36
3. 10. 3. 1 血淋巴之抽取 36
3. 10. 3. 2 總血球之計數 36
3. 10. 3. 3 酚氧化酶之活性測定 37
3. 10. 3. 4 呼吸爆之活性測定 37
3. 10. 3. 5 血球均質液樣本的配製 38
3. 10. 3. 6 血球麩胱苷肽過氧化酶之檢測 38
3. 10. 3. 7 血球麩胱苷肽還原酶之檢測 38
3. 10. 3. 8 血球麩胱苷肽硫轉移酶之檢測 39
3. 10. 3. 9 血球超氧歧化酶之檢測 39
3. 11 蛋白質測定 39
3. 12 吞噬作用及清除效率之檢測 39
3. 12. 1 吞噬作用之測定 40
3. 12. 2 清除效率之測定 40
3. 13 統計分析 40
第四章、結果 41
4. 1 飼料成分分析 41
4. 2 水質檢測 41
4. 3 養成試驗 41
4. 4. 硒含量分析 44
4. 5. 抗氧化酵素分析 45
4. 5. 1 蝦體組織麩胱苷肽過氧化酶活性之分析 45
4. 5. 2 蝦體組織麩胱苷肽還原酶活性之分析 46
4. 5. 3 蝦體組織麩胱苷肽硫轉移酶活性之分析 47
4. 5. 4 蝦體組織超氧歧化酶之活性分析 48
4. 6 抗病力及免疫因子試驗 49
4. 6. 1 抗病力試驗 49
4. 6. 2總血球之數量 51
4.6. 3 血球酚氧化酶之活性 52
4. 6. 4 血球呼吸爆之活性 53
4. 6. 5 血球麩胱苷肽過氧化酶之活性 54
4. 6. 6 血球麩胱苷肽還原酶之活性 55
4. 6. 7 血球麩胱苷肽硫轉移酶之測定 56
4. 6. 8 血球超氧歧化酶之測定 57
4. 6. 9 吞噬作用之測定 58
4. 6. 10 清除作用之測定 59
第五章 討論 60
第六章 結論 66
第七章 參考文獻 67
作者簡介....... 94



















圖目錄索引
頁次
Fig. 1. Global aquaculture production of Macrobrach………………. 5
Fig. 2 Main Se forms in diet and human organism: Se ……………... 12
Fig. 3. Antioxidant action of Selenium as cofactor of the glutathione peroxidase…….......................................................................... 20
Fig. 4. Glutathione peroxidase activities of hepatopancreas, gills and muscle of prawn fed with control diet, and seleno-L- methionine and sodium selenate supplied………..................... 45
Fig. 5. Glutathione reducase activities of hepatopancreas, gills and muscle of prawn fed with control diet, and seleno-L -methionine and sodium selenate supplied…..……………...... 46
Fig. 6. Glutathione S-transferase activities of hepatopancreas, gills and muscle of prawn fed with control diet, and seleno-L -methionine and sodium selenate supplied…............................ 47
Fig. 7. Superoxide dismutase activities of hepatopancreas, gills and muscle of prawn fed with control diet, and seleno-L -methionine and sodium selenate supplied…………...………. 48
Fig. 8. Total haemocyte count of hemolymph of prawn fed with control diet, and seleno-L-methionine and sodium selenate supplied diets for ………………….………………………….. 51
Fig. 9. Phenoloxidase activities of hemolymph of prawn fed with control diet, and seleno-L-methionine- and sodium. .………... 52
Fig. 10. Respiratory burstof hemolymph of prawn fed with control diet, and seleno-L-methionine and sodium selenate supplied diets for 77 days………………………………………………. 53
Fig. 11. Glutathione peroxidase activities of hemolymph of prawn fed with control diet, and seleno-L-methionine and sodium selenate supplied diets for 77 days............................................. 54
Fig. 12. Glutathione reductase activities of hemolymph of prawn fed with control diet, seleno-L-methionine and sodium …….........
55
Fig. 13. Glutathione-S-transferase activities of hemolymph of prawn fed with control diet, seleno-L-methionine and sodium selenate supplied diets for 77 days..………………...……….... 56
Fig. 14. Superoxide dismutase activities of hemolymph of prawn fed with control diet, and seleno-L-methionine and sodium……... 57
Fig. 15. Phagocytosis rate of hemolymph of prawn fed with control diet, seleno-L-methionine and sodium selenate supplied…….. 58
Fig. 16. Clearance efficiency of hemolymph of prawn fed with control diet, seleno-L-methionine- and sodium supplied…………… 59

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