Applicability of Non-conventional Protein Feedstuff

本研究旨在以體外模擬消化試驗探討非傳統性蛋白質原料及添加外源蛋白之消化情形，並與肉雞活體試驗相對照。 體外模擬消化試驗採3×3複因子處理，以大豆粕（soybean meal；SBM）、葵花籽粕（sunflower meal；SFM）及菜籽粕（canola meal；CM）為蛋白質來源，並分別配合一完全不添加外源酵素之對照組及添加等力價（0.05 unit / g feed）之鳳梨酵素（bromelain；750 units / g solid）或微生物來源之蛋白（microbial protease；45 units / g solid）進行試驗。經體外模擬消化試驗後三種原料之蛋白質、胺基酸消化率及水溶性蛋白質含量皆以葵花籽粕較其餘二者為佳（P<0.05），添加外源蛋白皆可明顯改善胺基酸消化率。體外消化後分子量大於150 kDa以上之水溶性蛋白質的比例，大豆粕高於葵花籽粕與菜籽粕；而分子量小於1 kDa之比例，大豆粕顯著低於其餘兩者，此結果與SDS - PAGE之蛋白分佈結果一致。 可利用率試驗採3×3複因子處理，以三種蛋白質原料配製成等蛋白之半純化飼糧及配合與體外模擬消化試驗相同之三種酵素處理。90隻14日齡及45隻35日齡愛拔益加種公雞各分配至45個試驗單位並逢機分為9處理組進行之。結果顯示，三週齡及六週齡雞隻之蛋白質、胺基酸可利用率及採食後1小時所測得之血漿游離胺基酸含量均以菜籽粕及葵花籽粕較大豆粕為佳（P < 0.05），添加微生物來源之蛋白具顯著改善之效果（P < 0.05），但添加鳳梨酵素則否。 生長試驗採4×2複因子處理，以704隻愛拔益加種雛雞逢機分為32組，處理為葵花籽粕取代飼糧中大豆粕蛋白質之0、5、10、20 ％，及有無添加微生物來源之蛋白。結果顯示，於等能量等蛋白條件下，葵花籽粕取代大豆粕及酵素處理，未顯著影響肉雞生長期之生長性狀（P > 0.05），但於肉雞肥育期及全期，葵花籽粕取代量增加，採食量與體增重均隨之增加（P < 0.05），飼料效率則無顯著影響，添加酵素顯著改善其生長性狀。 以體外模擬消化試驗、肉雞可利用率試驗顯示葵花籽粕之蛋白質為三種蛋白質原料中最易被消化酵素水解之蛋白質，然蛋白質原料於肉雞飼糧之應用效果，需綜合飼糧中所有影響因子，故除葵花籽粕之蛋白質及胺基酸含量不足外，另需考慮葵花籽粕之之飼糧纖維對生長所造成之影響，本試驗以葵花籽粕取代20 ％大豆粕蛋白質對肉雞生長並無不良影響。Applicability of Non-conventional Protein Feedstuff in Broiler Diets The purpose of this study was to evaluate the changes in a non-conventional protein feedstuff with exogenous protease supplement using an in-vitro digestive trial contrasted to an in-vivo broiler trial. Soybean meal (SBM), sunflower meal (SFM) and canola meal (CM) were used as the protein feed source, with or without the same activity of bromelain (750 units / solids) or microbial protease (45 units / solids) administered in 9 treatments for a 3 × 3 factorial experiment. The results showed that protein and amino acid digestibility and the percentage of water-soluble protein of SFM after in vitro digestion was significantly higher than the other trial diets. There was a significant affect on amino acid digestibility form the exogenous proteases supplement. The water soluble protein distribution showed that the percentage of molecular weight greater than 150 kDa in the SBM was higher than the other trial proteins. The molecular weight less than 1 kDa in SFM was significantly lower than the other. The SDS - PAGE polypeptide distribution showed the same soluble protein trend. Ninety, fourteen day old and 45, thirty-five day old Arber Acer broilers used for the in vivo availability trial were randomly divided into 9 groups for a 3 × 3 factorial experiment. The experimental diet was formulated into an isonitrogenic semipurified diet with the same protein and exogenous proteases supplement used in the in—vitro trial. The result showed that the SFM protein and amino acid availability was higher than SBM and CM in the growth and finishing periods. The plasma total free amino acid concentration taken in the first hour after a meal had same trend as the availability data (P < 0.05). Compared with the control, the addition of microbial protease had a significant effect on improving protein and amino acid availability and total free amino acids in the plasma. The bromelain supplement showed no improvement on protein and amino acid availability. Seven hundred and four Arber Acer chicks used for the growth trial were randomly divided into 32 pens for a 4 × 2 factorial experiment. The treatment involved four levels of SFM protein substitute of SBM protein (0, 5, 10 and 20 %) in isoenergenic and isonitrogenic diets, with or without microbial protease supplement. Results showed that there were no significant differences in feed intake, efficiency and weight gain during the growing period between all treatments. During the finishing period and for the entire trial period, feed intake and weight gain increased with increasing levels of SFM in the diet. Addition of microbial protease to the diet had a significant effect on improving growth performance. In-vitro digestive trial and in-vivo availability trial showed SFM protein is hydrolysed by digestive enzymes more easily than the other protein source. We must think upon all effect to realize the applicability of SFM. So, besides the amount of crude protein and amino acid is not enough to broilers. High dirtary fiber content in SFM may affect broilers' performance. There was no significant effect on feed efficiency when a SFM protein substitute of 20 % SBM protein was used in the growth trial.目 錄 壹、 中 文 摘 要7 貳、 前 言9 參、 文 獻 檢 討12 一、飼糧蛋白質於單胃動物體內之消化12 （一）胃內之消化及消化後蛋白質之分佈12 （二）腸道內之消化及消化後蛋白質之分佈12 （三）刷狀緣上之消化13 （四）細胞內之消化14 二、飼糧蛋白質於動物體內之吸收14 （一）吸收方式14 （二）血漿內游離胺基酸之變化15 三、飼糧蛋白質於動物體內之排出16 （一）內源性氮耗損16 1.飼糧中影響內源性氮的因子16 （1）乾物質採食量16 （2）飼糧纖維17 （3）抗營養因子17 2.內源性氮耗損之組成18 3.飼糧蛋白質與內源性氮之關係18 （二）後腸發酵之菌體蛋白質19 （三）排泄物中之尿酸態氮20 四、影響飼糧蛋白質利用之因子20 （一）胺基酸之組成及平衡20 （二）加工處理及方式21 五、非傳統性蛋白質原料之一般特性22 （一）葵花籽粕之一般特性22 （二）菜籽粕之一般特性24 六、體外模擬消化試驗及活體試驗之比較26 肆、 材 料 與 方 法28 一、材 料28 （一）飼料蛋白質原料28 （二）蛋白28 二、方 法29 Ⅰ.利用in vitro模擬消化試驗探討非傳統性蛋白質原料及蛋白應用之效果29 （一）兩段式in vitro模擬消化試驗29 （二）飼料原料之近似分析及蛋白質消化率之計算31 （三）in vitro模擬消化試驗後之胺基酸消化率31 （四）in vitro模擬消化試驗後之蛋白分佈32 （五）in vitro模擬消化試驗後水溶性蛋白質之含量及計算33 （六）in vitro模擬消化試驗後水溶性蛋白質分子量之分佈33 Ⅱ.以動物可利用率試驗探討非傳統性蛋白質原料及蛋白添加之效果34 （一）試驗動物34 （二）試驗設計及方法34 （三）分析方法37 1.飼料及排泄物之分析與利用率測定37 2.血漿游離胺基酸之測定37 Ⅲ.肉雞飼糧應用非傳統性蛋白質飼料（葵花籽粕）及補充蛋白之效果探討39 （一）試驗動物39 （二）試驗設計及方法39 三、統計分析43 （一）分析方法43 （二）試驗模式43 伍、 結 果 與 討 論44 Ⅰ.利用IN VITRO模擬消化試驗探討非傳統性蛋白質原料及蛋白應用之效果44 一、原料之組成分及胺基酸組成44 二、in vitro模擬消化試驗後之蛋白質及胺基酸消化率49 三、in vitro模擬消化試驗後之蛋白分佈54 四、in vitro模擬消化試驗後之水溶性蛋白質含量57 五、in vitro模擬消化試驗後之水溶性蛋白質分子量分佈59 Ⅱ.以動物消化試驗探討非傳統性蛋白質原料及蛋白應用之效果62 一、蛋白質可利用率62 二、胺基酸可利用率65 三、血漿游離胺基酸變化69 Ⅲ.肉雞飼糧應用非傳統性蛋白質原料（葵花籽粕）及補充蛋白之效果探討73 陸、 結 論80 柒、 參 考 文 獻81 捌、 英 文 摘 要96 玖、小 傳9