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第 21 筆
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論文名稱: | 藍點鸚嘴魚幼魚與成魚蛋白酶活性的分佈
| Distribution of protease activities in gut of juvenile and adult parrotfish (Scarridae)
研究生: | 盧恩得 En-Te Lu
指導教授: | 孫寶年 Bonnie Sun Pan
| 陳麗淑 Li-Shu Chen
學位類別: | 碩士
學校名稱: | 國立台灣海洋大學
系所名稱: | 食品科學系
畢業學年度: | 91
語文別: | 英文
關鍵字: | 腸道 enteropeptidase
| 蛋白酶 protease
| 鸚哥魚 parrotifish
| 鸚嘴魚 trypsin
全文點閱次數: | 2

全文影像連結:

電子檔1

論文頁數: | 72
摘要: | 鸚嘴魚為珊瑚礁的優勢魚種，經濟價值高，目前尚未以人工繁養殖。在台灣所有之鸚嘴魚中以藍點鸚嘴魚量最多，其成魚為雜食性魚類，本研究探討鸚嘴魚由幼魚發育至成魚（IP型）期間，其腸道型態與各腸段比例、腸道組織表面積及消化道蛋白酶活性之分佈與變化，以瞭解成長中其食性變化，供未來繁養殖之參考。
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| 藍點鸚嘴魚之腸道，依其肌肉組織形態分為食道（Ⅰ）、小腸囊（Ⅱ）、前腸（Ⅲ）、中腸前段（Ⅳ）、中腸後段（Ⅴ）、後腸（Ⅵ）等六段，腸道第Ⅳ、Ⅴ段之醣酶及蛋白酶活性高於其他腸段，故選擇此二段作進一步研究。剛捕獲成魚腸道呈非淨空狀態時，第Ⅴ段呈粉紅色，但經蓄養後呈色褪去。以共軛焦點顯微鏡觀察呈粉紅色之中腸後段，藻紅蛋白分佈在黏膜層，其他腸段則無。
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| 由幼魚發育至成魚期，以alcian blue periodic acid-Schiff（AB-PAS）染色法觀察第IV段與第Ⅴ段腸道，成魚及幼魚黏液細胞主要分佈在腸道黏膜層，以第V段腸道分佈較多；成魚第V段腸道總黏膜表面積比第IV段多18.7 %，幼魚腸道第V段比第IV段多12.5%；第V段腸道之皺襞較深，黏膜係數（腸道黏膜表面積/每克魚體重）平均為第IV段之1.18倍。以偶氮基（Azo）染料法觀察幼魚與成魚腸黏膜層胺基胜肽酶活性，主要分佈於腸道第V段。內切酶以酶原形式存在，經活化後之胰蛋白酶活性約為胰凝乳蛋白酶之2倍，第V段為蛋白質消化吸收之主要腸段，由幼魚發育至成魚，食性與腸道形態與各段長度比例已趨固定。
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| 為瞭解鸚嘴魚對飼料蛋白質之消化作用，以成魚之第IV段與第V段腸道粗酵素液模擬鸚嘴魚體內消化作用，生魚肉被腸道粗酵素液分解之速率最快，其次依序為紅魚粉、白魚粉與黃豆粉，鸚嘴成魚具有雜食性的消化能力，可作為未來選擇飼料配方之參考。
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| Parrotfish is a dominant fish species living in coral reefs with very high economic value. It has not yet been cultivated artificially. Scarus ghobban has the largest population among all parrotfish in Taiwan. The adult fish are omnivorous. The objective of this study was to understand the morphology and protease distribution in the intestine and the ontogenesis from juvenile fish into initial phase (IP) of adult fish.
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| The digestive tract of parrotfish are divided into 6 segments: oesophagus (I), intestine bulbs (II), anterior intestine (III), anterior of middle intestine (IV), posterior of middle intestine (V), posterior intestine (VI) according to their muscular structural patterns. Activities of amylase and protease were found higher in segment IV and V than the other segments. Therefore, this study was focused on these two segments.
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| Five adult parrotfish instantly caught, and dissected, the segment V appeared pinkish in color, which disappeased after being cultured in tank.  The pink segment observed with confocal microscope showed fluorescence of phycobiliprotein distributed throughout the mucosal layer, while the other segments did not have this fluorescent pigmentation.
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| Observation of segment IV and V using histochemical stainings, alcian blue periodic acid-Schiff (AB-PAS), Goblet cells distributed mainly at mucosal layer being more concentrated in segment V of both adult and juvenile fish. For adult fish, the mucosal area of segment V was 18.7% more than that of the segment IV. For juvenile fish, the former was 12.5% more than the segment IV. Mucosal folds were deeper in segment V than segment IV. The intestine mucosal coefficient (intestine mucosal area (cm)/ per gram body weight) of the segment V averaged 18% more than that of the segment IV.
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| Aminopeptidase activities of both juvenile and adult fish distributed mainly in the mucosal layer in segment V of intestine detected after azo dye staining. Endopeptidase existed in the form of zymogen. After activation using endopeptidase, trypsin activity were almost twice of the chymotrypsin activity. Segment V was found the main segment for protein digestion and absorption in intestine.
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| An in vitro digestion test was developed to measure feed protein digestibility simulating the digesting process in adult parrotfish. Raw fish was the most readily digeslible diet, followed by brown fish meal, white fish meal and soybean meal. The results obtained by this study may be applicable to feed formulation for culturing parrotfish in the future.
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論文目次: | 中文摘要    I
| 英文摘要    Ⅲ
|
| 壹、前言    1
|
| 貳、文獻整理    4
| 一、魚類之食性    4
| 二、消化道型態及酵素隨成長發育之變化    5
| （一）消化道型態之變化    5
| （二）消化酵素之變化    7
| 三、鸚嘴魚的生活史及分類    9
| 四、鸚嘴魚的消化系統    11
| 五、鸚嘴魚之攝食及食性    12
| 六、酵素之測定方法    13
| （一）生化性測定法    13
| （二）組織化學測定方法    14
| 七、體外消化模擬試驗    16
|
| 參、材料與方法    18
| 一、藍點鸚嘴魚    18
| 二、生物指標之測定    18
| 三、實驗架構    21
| 四、腸道組織觀察    23
| （一）腸道黏液細胞染色    23
| （二）腸黏膜表面積與腸黏膜係數之計算    23
| （三）冷凍切片    24
| （四）粉紅色腸道之觀察    24
| （五）腸道組織leucine aminopeptidase活性染色    25
| 五、蛋白酶活性測定    25
| （一）腸道粗酵素液之製備    25
| （二）胰蛋白酶 (trypsin) 活性    26
| （三）胰凝乳蛋白酶 (chymotrypsin) 活性    26
| （四）胰蛋白酶原 (trypsinogen) 之活化    27
| （五）胰凝乳蛋白酶原 (chymotrypsinogen) 之活化    27
| （六）白胺酸胜肽酶（Leucine aminopeptidase）
| 及其最適pH值    28
| 六、蛋白質定量    28
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| 七、體外消化率之測定    29
| 八、統計分析    29
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| 肆、結果與討論    30
| 一、鸚嘴魚幼魚與成魚之生物指標之比較    30
| 二、幼魚與成魚腸道第IV段與第V段之差異    37
| （一）型態    37
| （二）第V段腸道粉紅色素之分佈    39
| （三）黏液細胞之分佈    42
| （四）腸道絨毛之表面積    46
| （五）胺基胜肽酶之分佈    48
| 三、成魚腸道中之蛋白酶與酶原    51
| （一）胰蛋白酶與酶原    51
| （二）胰凝乳蛋白酶與酶原    51
| 四、飼料蛋白質之體外消化率    54
|
| 伍、結論    58
| 陸、參考文獻    60
|
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2009 年 11 月 22 日 23:45