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研究生:阮氏秋妝
研究生(外文):Nguyen Thi Thu Trang
論文名稱:白蝦攝食高粱酒糟之離體與體外消化率
論文名稱(外文):The in vitro and ex vivo digestibility of sorghum distillery residue fed to white shrimp (Litopenaeus vannamei)
指導教授:孫寶年孫寶年引用關係
指導教授(外文):Bonnie Sun Pan
學位類別:碩士
校院名稱:國立臺灣海洋大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:英文
論文頁數:57
中文關鍵詞:高粱酒糟消化率白蝦
外文關鍵詞:Sorghum distillery residuedigestibilityshrimp
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高粱酒糟(Sorghum distillery residue, SDR)為高粱酒之副產物,其粗蛋白含量為14% 。因此評估SDR應用於蝦類商用飼料,取代蛋白源之可行性,並減少高粱酒糟廢棄物。模擬SDR於白蝦體內之消化,故建立SDR之體外消化試驗,探討主要消化蛋白酶包含總蛋白酶 (total proteinase)、胰蛋白酶 (trypsin)、胰凝乳蛋白酶 (chymotrypsin) 及羧基胜肽酶 (carboxypeptidase) A 與 B。
白蝦中腸腺 (midgut gland) 蛋白酶活性於 60℃、 pH 2.0與 pH 7.0有最佳活性,胰蛋白酶及胰凝乳蛋白酶的活性於 pH 8.0最佳,羧基胜肽酶 A 及羧基胜肽酶 B於 pH 3.0有最佳活性。
體外(in vitro)消化試驗使用白蝦中腸腺粗酵素液,以兩階段進行消化:首先於 pH8.0、60℃ 消化 30 分鐘,接著於 pH3.0、60℃ 消化 30 分鐘。白蝦以各飼料飼養 8 週(in vivo)後,取其中腸腺進行體外消化試驗條件同上所述,即為離體試驗 (ex vivo)。
N-100 大豆粉 (粗蛋白含量49.22%) 和魚粉 (粗蛋白含量47.33%) 與白蝦中腸腺粗酵素液反應,模擬體外試驗,兩者之消化率無顯著差異(P > 0.05),但顯著較高於SDR 或添加聚乙二醇(polyethylene glycol, PEG)之SDR。SDR添加0.5% PEG 之蛋白消化率(27.03%) 顯著高於未添加PEG之SDR (12.31%)。

Sorghum distillery residue (SDR), a by-product from spirit fementation, contains 14% crude protein. Therefore, it is worthy to evaluate the potential of utilizing SDR as an ingredient in commercial shrimp feed to reduce the cost of protein source used in feed production and solve the waste disposal problem caused by the SDR. To develop an in vitro digestibility test of SDR simulating the in vivo digestion in white shrimp, activities of the major digestive proteases including total proteinase, trypsin, chymotrypsin and carboxypeptidase A & B were studied.
The proteinase activity from midgut gland of white shrimp (Litopenaeus vannamei) was found maximum at 60℃ with two optimal pHs of 2.0 and 7.0. The maximal trypsin and chymotrypsin activities were detected at pH 8.0. Carboxypeptidase A and B showed the highest activities at pH 3.0.
In vitro test was performed using enzymatic extracts from midgut gland of white shrimp to digest in two steps, first at pH 8.0, 60℃ for 30 min followed by digestion at pH 3.0, 60℃ for 30 min. For ex vivo digestibility, midgut glands of shrimp cultured for 8 weeks in in vivo test were taken out to do in vitro digestibility as described above.
There was no significant (P > 0.05) difference in in vitro digestibility between N-100 soybean meal (49.22% crude protein) and fish meal (47.33% crude protein), but significantly (P < 0.05) higher than SDR with or without polyethylene glycol. SDR treated with 0.5% PEG resulted in higher protein digestibility (27.03%) compared to SDR without PEG (12.31%).
In vitro and ex vivo digestibility of two kinds of iso-protein feeds, Feed A containing 14% N-100 soybean meal and 6% porcine and fish meal, and Feed B containing 14% porcine and fish meal and 6% N-100 soybean meal, showed that soybean meal can replace animal protein up to 14% in feed formulation for white shrimp. There is a difference in in vitro and ex vivo digestibilty of feeds added with SDR and without SDR. In the in vitro test, no significant (P > 0.05) difference in protein digestibility between feeds added with 20% SDR and without SDR for white shrimp. However, shrimp fed diets without SDR had higher specific growth rate and protein efficiency ratio than shrimp fed diets containing 20% SDR in the ex vivo test.

TABLE OF CONTENTS
摘要 I
ABSTRACT III
TABLE OF CONTENTS V
LIST OF FIGURES VIII
ABBREVIATIONS XI
1. INTRODUCTION 1
2. OBJECTIVES 3
3. LITERATURE REVIEW 4
3.1 PROTEOLYTIC ENZYME ACTIVITY OF SHRIMP 4
3.2 IN VITRO AND IN VIVO DIGESTIBILITY 6
3.3 SUBSTITUTION OF ANIMAL PROTEIN SOURCES BY PLANT PROTEIN SOURCES CONTAINING ANTI-NUTRITIONAL FACTORS 7
4. EXPERIMENTAL DESIGN 11
4.1 ENZYME ACTIVITY DETERMINATION 11
4.2 IN VITRO DIGESTIBILITY 12
4.3 IN VIVO DIGESTIBILITY 13
5. MATERIALS AND METHODS 14
5.1 PROXIMATE ANALYSIS 14
5.1.1 Crude protein content 14
5.1.2 Moisture and ash content 14
5.1.3 Crude lipid content 15
5.1.4 Analysis of gross energy 16
5.2 PREPARATION OF CRUDE ENZYME EXTRACT 17
5.3 TOTAL SOLUBLE PROTEIN DETERMINATION 18
5.4 DETERMINATION OF OPTIMAL PH AND TEMPERATURE OF PROTEOLYTIC DIGESTIVE ENZYMES 18
5.4.1 Total proteinase 18
5.4.2 Trypsin 20
5.4.3 Chymotrypsin 20
5.4.4 Carboxypeptidase A & B 21
5.5 PREPARATION OF FEEDS 21
5.6 IN VITRO DIGESTIBILITY TEST 24
5.7 ANALYSIS OF PHYSIOLOGICAL INDICES OF WHITE SHRIMP 25
5.7.1 Feeding trials 25
5.7.2 Specific growth rate, feed conversion ratio, protein efficiency ratio 25
5.7.3 Condition factor (K) (Abowei 2010). 26
5.7.4 Digestive somatic index (DSI) (Hidalgo et al., 1999). 26
5.8 STATISTICAL ANALYSIS 27
6. RESULTS AND DISCUSSIONS 28
6.1 OPTIMAL ACTIVITY OF PROTEOLYTIC DIGESTIVE ENZYMES OF WHITE SHRIMP 28
6.1.1 Total protease 28
6.1.2 Trypsin 31
6.1.3 Chymotrypsin 32
6.1.4 Carboxypeptidase A & B 33
6.2 IN VITRO DIGESTIBILITY OF FEEDS 35
6.3 THE CORRELATION OF IN VIVO DIGESTIBILITY WITH SGR, PER, FCR, K, DSI AND IN VITRO DIGESTIBILITY 43
7. CONCLUSIONS 50
8. REFERENCES 52

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