臺灣博碩士論文加值系統

本研究探討飼料中添加不同濃度雨生紅球藻 (Haematococcus pluvialis) 藻粉及萃取之蝦紅素，對人工繁殖眼斑海葵魚 (Amphiprion ocellaris) 活存、成長及體表呈色之影響。實驗分成三個部分，首先大量培養雨生紅球藻，製作成凍乾藻粉後，以最適化學萃取方法進行蝦紅素萃取。分別在飼料中添加雨生紅球藻凍乾藻粉及萃取之蝦紅素，投餵眼斑海葵魚，觀察有無萃取處理之雨生紅球藻對眼斑海葵魚活存、成長及呈色影響，確定眼斑海葵魚可吸收雨生紅球藻萃取之蝦紅素。最後，以飼料中添加雨生紅球藻萃取之蝦紅素，觀察不同濃度對於眼斑海葵魚的活存、成長及呈色影響，與市售市售合成蝦紅素比較差異。
實驗一、探討商業水耕培養液─花寶 4 號對雨生紅球藻之最大成長生物量影響，並與 Jaworski’s medium (JM) 比較。雨生紅球藻粉以兩種萃取法，醋酸-DMSO 萃取法及 HCl-丙酮萃取法檢測雨生紅球藻粉破壁及萃取效果。結果顯示，花寶 4 號培養液濃度 10 mg/L、20 mg/L、40 mg/L 及 60 mg/L 皆對雨生紅球藻生物量有提高之影響，其中濃度 20 mg/L 及 60 mg/L，在短時間內有顯著生物量成長，濃度 40 mg/L 於培養第 26 天有最高生物量 1.16 ± 0.81 × 105 cells/mL。醋酸-DMSO 萃取法及 HCl-丙酮萃取法對雨生紅球藻之蝦紅素萃取量分別為 19.20 ± 0.49 mg/g cell 及 21.99 ± 0.52 mg/g cell，兩種萃取法進行葉綠素去除步驟之萃取量分別為 15.80 ± 0.98 mg/g cell 及 20.41 ± 0.52 mg/g cell。HCl-丙酮萃取法對於雨生紅球藻之蝦紅素萃取量較高，且萃取量不受葉綠素去除步驟之影響。
實驗二、探討飼料中添加雨生紅球藻粉，投餵 0.19 ± 0.05 g眼斑海葵魚，觀察 10 週後，記錄其活存、成長及呈色狀況，並與市售合成蝦紅素比較差異。結果顯示，投餵添加藻粉組別，其末重為 0.22 ~ 0.26 g，增重率為 18.86 ~ 30.56 %，飼料轉換率為 3.24 ~ 5.01，特殊成長率為 0.21 ~ 0.33 %，活存率為 84 ~ 98 %；餵食市售合成蝦紅素組，其末重為 0.23 g，增重率為 24.42 %，飼料轉換率為 4.28，特殊成長率為 0.27 %，活存率為 94 %，結果顯示，飼料中添加藻粉，對眼斑海葵魚的活存、成長及體表增豔無顯著效果。
接續實驗觀察 8 週，檢測雨生紅球藻萃取之蝦紅素，對眼斑海葵魚體表增豔觀察，結果顯示，投餵萃取蝦紅素，隨投餵濃度增加有顯著增豔效果，色彩指標 L* 值為 43.17 ~ 48.57，a* 值為 49.97 ~ 58.67，b* 值為 56.37 ~ 59.93，C* 值為 77.32 ~ 83.85，H* 值為 43.88 ~ 49.80。
實驗三、探討飼料中添加不同濃度雨生紅球藻萃取之蝦紅素，對眼斑海葵魚活存、成長及呈色觀察，並與市售合成蝦紅素比較差異。投餵萃取之蝦紅素 8 週，其末重為 0.36 ~ 0.39 g，增重率為 32.61 ~ 38.99 %，飼料轉換率為 2.10 ~ 2.46，特殊成長率為 0.35 ~ 0.41 %，活存率為 100 %。結果顯示，萃取之蝦紅素100 mg/kg 對眼斑海葵魚，有較佳增重率及飼料轉換率，添加量 200 mg/kg 有較佳的增豔效果。

The purpose of the experiment is to investigate the effect of dietary supplementation with microalgae Haematococcus pluvialis on survival, growth and pigmentation of Amphiprion ocellaris. The experiment was divided into three parts. First, Haematococcus pluvialis was cultured in a great amount, and was lyophilized into algae powder. Optimal extraction method was adopted to extract astaxanthin from Haematococcus pluvialis. Haematococcus pluvialis algae powder and extracted astaxanthin was added separately to the diets to feed Amphiprion ocellaris as to know whether dietary supplementation with either algae powder or extracted astaxanthin had effect on survival, growth and pigmentation. Amphiprion ocellaris was made sure to absorb the astaxanthin extracted from Haematococcus pluvialis. Finally, Amphiprion ocellaris was fed with the diet supplementations with different concentrations of extracted astaxanthin. By doing so, the effect on survival, growth and pigmentation of Amphiprion ocellaris can be observed. The result was used to compare with commercially available synthetic astaxanthan.
Experiment I aimed to know the effect of commercially available hydroponic medium─Hyponex Precious the 4th on the maximum biomass of Haematococcus pluvialis, and to compare the result with the JM medium. In order to investigate the astaxanthin extraction yield, the algae powder was extracted in two different ways, acetic acid-DMSO method and HCl-acetone method. The biomass of Hyponex Precious the 4th medium at concentrations 10 mg/L, 20 mg/L, 40 mg/L, 60 mg/L were all increased in a short period of time, among the all, concentration 20 mg/L, 60 mg/L yield the most significant results. The maximum biomass reached at the concentration of 40 mg/L on the 26th day (1.16 ± 0.81 × 105 cells/mL). The astaxanthin yield of acetic acid-DMSO method and HCl-acetone method was 19.20 ± 0.49 mg/g cell and 21.99 ± 0.52 mg/g cell separately. With the procedure of removing chlorophylls, the astaxanthin yield was 15.80 ± 0.98 mg/g cell and 20.41 ± 0.52 mg/g cell separately, while the astaxanthin yield of HCl-acetone method remained the same.
Experiment II aimed to observe the effect of dietary supplementation with Haematococcus pluvialis algae powder on survival, growth and pigmentation of Amphiprion ocellaris for 10 weeks with initial weight of 0.19 ± 0.05 g, and to compare the result with commercially available synthetic astaxanthan. The final weight of Amphiprion ocellaris with diet containing algae powder was 0.22 ~ 0.26 g, weight gain was 18.86 ~ 30.56 %, feed conversion rate was 3.24 ~ 5.01, specific growth rate was 0.21 ~ 0.33 %, the survival rate was 84 ~ 98 %. The final weight of Amphiprion ocellaris with diet containing synthetic astaxanthin was 0.23 ± 0.06 g, weight gain was 24.42 ± 4.76 %, feed conversion rate was 4.28 ± 0.70, specific growth rate was 0.27 ± 0.05 %, and survival rate was 94 ± 2.83 %. The result did not show significant effect on dietary supplementation with Haematococcus pluvialis algae powder on survival and growth. No significant effect was observed on pigmentation for dietary supplementation with Haematococcus pluvialis algae powder as well.
Experimental observation proceeded to detect dietary supplementation with high concentrations of astaxanthin extracted from Haematococcus pluvialis for 8 weeks. The result showed an increase of the skin pigmentation as the concentration of extracted astaxanthin in diet increased. The color parameter L* value was 43.17 ~ 48.57, a* value was 49.97 ~ 58.67, b* value was 56.37 ~ 59.93, C* value was 77.32 ~ 83.85, and H* value was 43.88 ~ 49.80.
Experiment III aimed to examine the effect of dietary supplementation with various concentrations of astaxanthin extracted from Haematococcus pluvialis on survival, growth and pigmentation of Amphiprion ocellaris for 8 weeks with initial weight of 0.28 ± 0.06 g, and to compare the result with commercially available synthetic astaxanthan. The final weight of Amphiprion ocellaris with diet containing extracted astaxanthin was 0.36 ~ 0.39 g, weight gain was 32.61 ~ 38.99 %, feed conversion rate was 2.10 ~ 2.46, specific growth rate was 0.35 ~ 0.41 %, the survival rate was 100 %. The result show that Amphiprion ocellaris fed with extracted astaxanthin 100 mg/kg had better weight gain and feed conversion rate. As for the pigmentation, the treatment of 200 mg/kg showed a better result.

謝辭 I
摘要 II
Abstract III
目錄 V
表目錄 VII
圖目錄 IX
附錄 X
第一章、前言 1
第二章、文獻整理 3
一、雨生紅球藻 3
二、眼斑海葵魚 7
第三章、材料與方法 10
一、實驗設計 10
二、實驗藻種 11
三、藻種培養條件及誘導條件 11
四、實驗動物 11
五、實驗動物養殖系統 11
六、實驗養殖條件 12
七、實驗飼料配製 12
八、實驗分析項目 12
九、體表比色方法 14
十、統計分析 15
第四章、結果 16
一、雨生紅球藻之培養及蝦紅素萃取定量 16
二、飼料添加雨生紅球藻粉及萃取之蝦紅素對眼斑海葵魚活存、成長及呈色之影響 18
三、飼料添加不同濃度雨生紅球藻萃取之蝦紅素對眼斑海葵魚活存、成長及呈色之影響 25
第五章、討論 30
一、雨生紅球藻之培養及蝦紅素萃取定量 30
二、飼料添加雨生紅球藻粉及萃取之蝦紅素對眼斑海葵魚活存、成長及呈色之影響 32
第六章、結論 39
參考文獻 40

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