利用生物技術大量產製瘤胃細菌纖維分解酵素複合體

本試驗以瘤胃中之纖維分解菌F.succinogenes S85 為纖維素分解酵素及纖維素結合蛋白質（CBP）來源，並利用R.. albus 7 之XynC 基因產物為聚木糖分解酵素（xylanase）來源，測試添加纖維素結合蛋白質後之酵素活性表現。XynC 基因之產物 約為62 kDa，且具有高活性之聚木糖分解酵素活性。來自F.. succinogenes S85 之纖維素分解酵素經離子交換法及膠體過濾法純化後可得約120 及65 kDa 兩個主要蛋白質，且經活性染色確定其酵素活性。CBP利用含10％纖維二糖之磷酸鹽緩衝液配合1％CHAPS 進行回收。XynC 基因產物之聚木糖分解酵素與純化後之纖維素分解酵素，在加入CBP 後可有效提升其酵素作用 之特異性4 至10 倍，且發現CBP 可作用於不同菌株而來之分解酵素，提升酵素之效力。Attempts were made to examine enzymes activity after addition cellulose binding protein (CBP). Cellulase and CBP were purified from rumen cellulytic bacterium F.. succinogenes S85, and the gene encoding xylanase C (XynC) of rumen cellulytic bacterium R. albus 7 was be the source of xylanase. The gene encoded xylanase was 62 kDa, and detected high xylanase activity. Cellulase from F.. succinogenes S85, after passing ion exchange and gel filtration columns, present two protein peaks with cellulase activity, and the molecular weight was approximately 120 and 65 kDa, respectively. CBP was eluted with sodium phosphate buffer containing 10% cellobiose and 1% CHAPS. Enzymes activity were increasing from four to ten times after addition of CBP. The result showed that CBP can effect on both cellulytic and hemicellulytic enzyme from different bacteria species and improve their enzyme activity

纖維分解酵素於青貯製作應用方法之改善

本試驗採用兩種不同成熟度狼尾草與玉米，分別進行(1)對照組不添加任何物質;(2)添加乳酸菌(Lactobacillus plantarium);(3)同時添加乳酸菌以及纖維素與半纖維素分解酵素;(4)同時添加乳酸菌、纖維素與半纖維素分解酵素以及α-L-arabinofuranosidase 與feruloyl esterase 之青貯製作試驗。處理組2 乳酸菌的添加與對照組相比較，可以確保顯著較低的青貯pH 值與顯著較高的青貯評分，以及顯著較高的乾物質試管消化率。使用纖維分解酵素的處理組3 與4，顯著降低乾物質試管消化率。對兩種成熟度玉米之青貯，處理組2 乳酸菌或纖維分解酵素的使用與對照組相比較，沒有改善任何青貯的品質。泌乳羊飼養試驗顯示較老熟狼尾草四種處理之青貯，所得乾物質採食量(1837, 1743, 1817, 1834 g/day)、4%乳脂修正乳量(1826, 1804, 1549, 1724 g/day)與中洗纖維表面消化率(77, 79, 76,77%)並無顯著之差異。較幼嫩狼尾草的青貯亦是有相同結果。較老熟玉米四種處理之青貯泌乳羊飼養試驗，所得乾物質採食量(1142, 1039, 1096, 1103 g/day)、4%乳脂修正乳量(1744, 1501, 1665, 1706 g/day)與中洗纖維表面消化率(56, 56, 51,41%)並無顯著之差異。較幼嫩玉米的青貯，所得乾物質採食量(1175, 1294, 1187,1328 g/day)與4%乳脂修正乳量(1665, 1340, 1811, 1779 g/day)亦無顯著的處理組間之差異。整體而言，乳酸菌的添加可以改善狼尾草青貯的品質，但是無助益於玉米青貯。本試驗所採用纖維分解酵素的添加，並無法進一步改善青貯的纖維消化率。The present study used Napier grass and corn of two maturities to test the silage making by (1)control without additive, (2)added with Lactobacillus planetarium, (3)added with Lactobacillus planetarium in combination with cellulose and xylanase, (4)same as treatment 3 with addition of α-L-arabinofuranosidase and feruloyl esterase. Compared to the control, treatment 2 resulted in significantly lower silage pH and higher silage score along with higher in vitro dry matter digestibility (IVDMD). Usage of fibrolytic enzymes in treatments 3 and 4 resulted in significantly lower IVDMD than treatment 2. For corn silage making, usage of lactic acid bacterial inoculant or fibrolytic enzymes did not resulted in better silage quality than the control. Dairy goat lactation studies showed no difference of DM intake (1837, 1743, 1817, 1834 g/day), 4%FCM (1826, 1804, 1549, 1724 g/day) or apparent NDF digestibility(77, 79, 76, 77%) among treatment groups for matured Napier grass silages. Dairy goats fed less matured Napier grass silages had similar response as those fed matured Napier grass silages. For the matured corn silage, dairy goat lactation study showed no significant difference of DM intake (1142, 1039, 1096, 1103 g/day), 4%FCM (1744, 1501, 1665, 1706 g/day) and apparent NDF digestibility (56, 56, 51, 41%). For less matured corn silage, lactation study showed no significant difference of DM intake (1175, 1294, 1187, 1328 g/day) and 4%FCM (1665, 1340, 1811, 1779 g/day). In conclusion, lactic acid bacterial inoculant could improve the silage quality of Napier grass, but had no benefit toward corn silage quality. Application of fibrolytic enzymes in addition to lactic acid bacterial inoculant could not further improve fiber digestibility for both Napier grass and corn silages

延長光照處理對泌乳山羊之影響

延長光照為提升泌乳牛產能之重要方法;然而，此方法於泌乳羊產能影響並不清楚。本研究即企圖以延長光照提升泌乳山羊之產能。試驗動物為11 頭Alpine 經產羊及3 頭初產羊，依產次、分娩日期及體型平均分配於長日照組 (LP)與自然日照組(NP)。長日照組光照長度為16 小時 (0400 – 2000)，光照強度設 定大於 600 lux。實驗期為22 週 (2005 年二月至八月)。兩組皆採 NRC 建議泌乳山羊營養需要量配製之相同日糧配方。每日擠乳2 次，並於每週末二日記錄乳量並採集乳樣進行乳組成分析。結果顯示延長光照處理對於乳糖、乳脂率及乳生菌數皆未有影響 (P > 0.05)。長日照組原始乳量則於泌乳前期 (第3、6 及7 週) 有提升之趨勢 (3.37 vs. 2.34 kg / d;3.54vs. 2.24 kg / d;3.61 vs. 2.54 kg /d;P < 0.10)，4% 乳脂校正乳亦是相似結果。延長光照平均增加4% 乳脂校正乳乳量達30.2% (P = 0.208)，並可增加採食量 41.4% (P <0.0001)。The long daily photoperiod was proved to improve dairy cattle production; however, its effect on dairy goat was unclear. In this study, we intended to enhance dairy goat production by long daily photoperiod. Fourteen Alpine dairy goats (11 multiparous, 3 primiparous) were evenly assigned to long daily photoperiod (LP, 16 h of light and 8 h darkness/d) or natural daily photoperiod (NP) according to parity, kidding date, and body condition. Light were turn on at 0400 till 2000 for the LP group, and light intensity was set more than 600 lux at goats’ eyes level. The experimental period was 22 weeks after kidding (from Feb 2005 to Aug 2005). Same diet was offered to each group according to the NRC lactating dairy goat recommendation. Goats were milked twice daily, and milk were weighed and sampled on last two days each week. Results showed that long daily photoperiod had no effect on lactose and milk fat concentrations and raw milk total bacterial number (P > 0.05). Milk yield on weeks 3, 6, and 7 tended to be increased by long daily photoperiod (3.37 vs. 2.34 kg / d; 3.54 vs. 2.24 kg / d; 3.61 vs. 2.54 kg / d; P < 0.10). The 4% fat corrected milk (4%FCM) had similar results as milk yield. The long daily photoperiod increased 4%FCM and feed intake by 30.2% (P = 0.208) and 41.4% (P < 0.0001) on average