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研究生(外文):Luis Fernando Valenzuela Munguia
論文名稱(外文):Effects of Dietary Supplement of Algae Extract on Growth, Immunity, Antioxidant Capacity and Oxidative Stress Response of Pacific White Shrimp (Litopenaeus vannamei)
指導教授(外文):Yew-Hu Chien
口試委員(外文):Huang, Chen-HueiLai, Hong-ThihCheng, Da-ChihYew-Hu Chien
外文關鍵詞:Litopenaeus vannameiantioxidant extractsantioxidant responseimmune responsemetabolic response
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此研究的目的是要評估飼糧中添加藻類(Spirulina sp., Chlorella sp., Nannochloropsis sp., Sargassum wightii及 Ulva lactuca)萃取物對太平洋白蝦(Litopenaeus vannamei)的成長、免疫、抗氧化能力及氧化緊迫反應的影響。十二個處理包括控制組(C)、蝦紅素標準控制組(AX)及使用藻類的10組,其中5組為單倍濃度Chlorella (Cx1)、Spirulina (Sx1)、Nannochloropsis (Nx1)、Sargassum(Sx1)及Ulva (Ux1);相對兩倍濃度的是Cx2、Sx2、Nx2、Sx2及Ux2。均衡組的比較顥示含萃取物的組及蝦紅素比控制組有顯著較高的平均終重、平均增重及平均比成長。總血球數(THC)在正常及低溶氧狀況下,有添加組皆高於控制組。從最高至最低在無緊迫下,紅血球的排序為:(Cx2 = Sx2 = Wx2 = Ux2 = Nx2) > (Sx1 = Cx1 = Wx1 =Ux1 = Nx1) > AX > C;在有緊迫下排序為:(Ux2 = Nx2 = Wx2 = Ux2 = Nx2) > (Cx1 = Sx1 = Nx1 =Wx1 = Ux1) > AX > C。SOD在不同處理及濃度下呈現差異且增加不同。在緊迫下,GPx在添加組顯著的不同於控制組。然而儘管來源或濃度不同,GPx在添加組之間並無差異。類似於GPx,GR在緊迫下,添加組比蝦紅素組及控制組高,代謝反應進行均衡組分析顯示在無緊迫與有緊迫下Gluc及Trigs有三種比較呈現顯著:控制組比處理組、蝦紅素組比藻萃取物組、以及藻萃取物中單倍濃度比雙倍濃度在低溶氧的緊迫下,雙倍濃度組的蝦比單倍濃度組的蝦活存較久。整體而言,白蝦餵食添加藻類萃取物的飼糧可增進其免疫、抗氧化及代謝之反應。
The aim of this study was to evaluate the effects of dietary antioxidant supplements extracted from algae (Spirulina sp., Chlorella sp., Nannochloropsis sp., Sargassum wightii and Ulva lactuca) on growth, immunity, antioxidant capacity and oxidative stress response of pacific white Shrimp (Litopenaeus vannamei). Twelve treatments were made including a Control (C), standard control astaxanthin (AX) and 10 diets using algae, five in single concentration Chlorella (Cx1), Spirulina (Sx1), Nannochloropsis (Nx1), Sargassum(Sx1) and Ulva (Ux1), and five with double concentration Chlorella (Cx2), Spirulina (Sx2), Nannochloropsis (Nx2), Sargassum(Sx2) and Ulva (Ux2). After orthogonal comparison, the treatments supplemented with raw extract and astaxanthin compared to control obtained significantly higher averages of MFW, MWG and SGR. Total hemocytes count (THC) showed to be significantly higher in supplemented diets compared to control under normal and hypoxia conditions. Analysis of non-stressed and stressed shrimp immune parameters ranked the greatest to the least as follow: RB under no stress: (Cx2 = Sx2 = Wx2 = Ux2 = Nx2) > (Sx1 = Cx1 = Wx1 =Ux1 = Nx1) > AX > C. RB under stress: (Ux2 = Nx2 = Wx2 = Ux2 = Nx2) > (Cx1 = Sx1 = Nx1 =Wx1 = Ux1) > AX > C. Superoxide dismutase (SOD), was very different and increased differentially under varying concentration of treatment supplementation. Glutathione peroxide (GPx), under stress was significantly different when comparing the supplemented diets against control. However, supplemented diets were no significantly different, despite, source or concentration. Glutathione reductase (GR), similarly to GPx under stress was significantly higher when comparing the supplemented diets against control and astaxanthin. The orthogonal analysis for the metabolic response showed, Gluc and Trigs non-stressed and stressed condition had significant difference in three levels of comparison control vs. treatments, astaxanthin vs. algae extracts and algae extracts x1 vs. x2. Hypoxia test revealed that double concentration diets lasted for longer time under stress conditions and were significantly higher than single concentration of supplemented diets. Overall, results in this study showed that immune response, antioxidant response and metabolic response of L. vannamei, were enhanced significantly by the supplementation of algae-derived antioxidant in their diets.
Acknowledgements iii
摘要 iv
Abstract v
Tables of Contents vi
List of Tables ix
List of Figures xi
1. Introduction 1
1.1 Importance of Shrimp in aquaculture 2
1.2 Reactive Oxygen Species 2
1.3 Antioxidants 2
1.4 Importance of Micro and Macroalgae 3
1.5 Polyphenols 3
1.6 General Objective 3
2. Literature Review 4
2.1 White Shrimp 5
2.1.1 White Shrimp Taxonomy 5
2.1.2 Shrimp Immune Capacity 5
2.1.3 Shrimp Immune Capacity Assessment 6
2.2 Algae 7
2.2.1 Microalgae 7
2.2.2 Macroalgae 8
2.3 Antioxidants 9
2.4 Polyphenols 9
2.4.1 Role of Polyphenols in Algae 10
2.5 Reactive Oxygen Species 10
3. Material and Methods 11
3.1 Algae Source 12
3.2 Algae Extraction Method 12
3.3 Total Phenolic Content Estimation 12
3.4 Antioxidant Essays 13
3.4.1 Total Antioxidant Status (TAS) 13
3.4.2 Diphenyl-2-picrylhydrazyl (DPPH) 13
3.4.3 Ferric Reducing Antioxidant Power (FRAP) 14
3.5 In vitro Antimicrobial Activity 14
3.5.1 Microbes 14
3.5.2 Antibacterial Disk Diffusion Test 14
3.6 Experimental Design 14
3.7 Experimental System 15
3.8 Shrimp Source 15
3.9 Experimental Diets 15
3.10 Sample Collection 16
3.11 Oxygen Depletion Stress Test 16
3.12 Biochemical Analysis 16
3.12.1 Antioxidant Capacity 16
3.12.2 Immune Response 16
3.12.3 Metabolic Response 17
3.13 Growth Parameters Measured 17
3.13.1 Survival and Growth Performance 17
3.14 Proximate Analysis 17
3.15 Statistical Analysis 17
4. Results 19
4.1 Total phenolic content 20
4.2 Total Antioxidant Status Determination (ABTS) 20
4.3 Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay 20
4.4 Ferric Reducing Antioxidant Power (FRAP) Antioxidant Assay 21
4.5 Statistical Correlation of Different Antioxidant Assays Against TPC 21
4.6 Disk Diffusion Test Antibacterial Assay 22
4.7 Shrimp Growth Performance and Muscle Proximate Analysis 22
4.8 Shrimp Immune Response 23
4.9 Shrimp Antioxidant Response 24
4.10 Shrimp Metabolic Response 25
4.11 Oxygen Depletion Stress Test 27
4.12 Correlation Among Parameters 27
5. Discussion 28
5.1 Algae Total Phenolic Content 29
5.2 Algae Total Antioxidant Assay 30
5.3 Algae Diphenyl-2-picrylhydrazyl 31
5.4 Algae Ferric Reducing Antioxidant Power 32
5.5 Statistical Correlation of Different Antioxidant Assays Against TPC 32
5.6 Disk Diffusion Test Antibacterial Assay 33
5.7 Shrimp in vivo Experiment 33
6. Conclusion and Recommendations 37
References 40
Tables 51
Figures 70
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