本研究探討枯草芽孢桿菌孢子能否加快完整蛋白質飼料中結合型胺基酸在點帶石斑魚腸道中的消化速度，縮短與飼料中額外添加之結晶甲硫胺酸被魚體吸收的時間差，進而改善在高植物蛋白質情況下點帶石斑魚對結晶胺基酸的利用度，達到有效率合成體蛋白質並促進成長表現。實驗分為兩部分進行，一為14週之成長實驗，評估投餵魚粉飼料、高植物蛋白質飼料以及枯草芽孢桿菌孢子加在有/無添加結晶甲硫胺酸之高植物蛋白質飼料後，對點帶石斑魚稚魚之成長表現、蛋白質效率、蛋白質及胺基酸消化率、非專一性免疫以及血清和肌肉中游離胺基酸含量的影響。二為灌食實驗，藉由強制灌食後點帶石斑魚血液游離胺基酸濃度變化，瞭解飼料中胺基酸的吸收時間，反映出枯草芽孢桿菌添加對植物蛋白質消化的影響。結果顯示，高植物蛋白質飼料添加結晶甲硫胺酸顯著 (P<0.05)改善點帶石斑魚成長表現，但B. subtilis 孢子添加在有/無添加結晶甲硫胺酸之高植物蛋白質飼料中，對點帶石斑魚成長表現、蛋白質效率、蛋白質與胺基酸消化率、肌肉游離胺基酸濃度和非專一性免疫反應的影響並不顯著 (P>0.05)，且大多無正面效果。點帶石斑魚經灌食高植物蛋白質飼料後，血清甲硫胺酸濃度比灌食魚粉飼料者較慢達到高峰值，並在達到高峰值之後急遽下降；但灌食添加結晶甲硫胺酸的高植物蛋白質飼料時，血清甲硫胺酸濃度急遽下降的情況獲得改善；進一步加入B. subtilis孢子，血清中各種游離必需胺基酸 (含甲硫胺酸)濃度又會在達到高峰值之後急遽下降。由本研究得知，在高比例植物蛋白質飼料中添加B. subtilis孢子，不會提升點帶石斑魚利用結晶甲硫胺酸的能力，也不影響石斑魚的成長；但添加結晶甲硫胺酸則顯著改善成長。

With the aim to enhance the efficiency of utilization of crystalline methionine supplemented in the high plant-protein diet for grouper (Epinephelus coioides), this study used Bacillus subtitlis spore as a probiotic additive in the diet to shorten the absorption time difference between protein-bound amino acid and crystalline methionine. The study was conducted in two parts. In the first part, juvenile groupers were fed for 14 weeks with 5 experimental diets: fish meal diet, high plant-protein diet with/without crystalline methionine, as well as with/without B. subtitlis spore separately. Growth performance, PER, protein digestibility, amino acid digestibility, non-specific immune responses, and free amino acid concentration in both muscle and serum were assessed. The second part was a time-series study on serum free amino acids concentration after a force-feeding experiment. The results showed that crystalline methionine supplementation in the high plant-protein diet effectively improved the growth of E. coioides (P<0.05). However, B. subtitlis spore supplementation did not affect fish growth performance significantly (P>0.05). A delay in the appearance of peak serum amino acid concentration was observed when fishmeal was partially replaced by soy protein. On the other hand, the force-feeding experiment showed that serum essential amino acid (include methionine) concentrations droped drastically after they reached the peak concentrations from being forced-fed with the B. subtilis containing diet. Supplementation of crystalline methionine seemed to ease the drop of serum methionine concentration. Based on these results, it is concluded that addition of B. subtitlis spore in high plant-protein diet for the grouper does not enhance the utilization of crystalline methionine, but supplementation of crystalline methionine significantly improve the growth performance of the grouper.

章次 頁數
中文摘要 i
英文摘要 ii
目錄 iv
圖目錄 vi
表目錄 vii
文獻回顧 1
一. 石斑魚簡介/石斑魚養殖近況 1
二. 蛋白質 1
三. 胺基酸 4
四. 飼料中蛋白質主要來源 7
五. &#63847;同蛋白質來源取代魚粉之研究 7
六. 魚類之結晶胺基酸營養 9
七. 蛋白質和胺基酸對魚類免疫的影響 12
八. 微生物與魚類的關係 13
前言 18
材料與方法 20
一. 益生菌菌株來源及培養 21
二. 實驗飼料 21
三. 實驗動物及飼育方式 22
四. B. subtilis在飼料及養殖水體中的活存 24
五. 成長指標 25
六. 魚體採樣 26
七. 免疫指標分析 27
八. 實驗飼料與全魚一般化學成份分析 30
九. 消化率測定 32
十. 胺基酸分析 34
十一. 強制灌食與血液游離胺基酸濃度分析 36
十二. 統計分析 37
結果 45
討論 72
結論 81
參考文獻 82
附錄 103
附錄表一、免疫分析用藥品配製 103
附錄表二、點帶石斑魚餵食不同飼料4週再分別灌食不同飼料後之血清游離甲硫胺酸濃度隨時間的變化 104
附錄表三、點帶石斑魚餵食不同飼料4週再分別灌食不同飼料後之血清游離甲硫胺酸/游離必需胺基酸(%)隨時間的變化 105
附錄表四、實驗飼料胺基酸濃度指標 106

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