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研究生:謝依蒨
研究生(外文):Yi-Chien Hsieh
論文名稱:壓力對蠟蚧輪枝菌孢子影響模式之探討
論文名稱(外文):Effect of pressure on germination rate of Verticillium lecanii spores.
指導教授:陳齊聖陳齊聖引用關係
指導教授(外文):Chee-Shan Chen
學位類別:碩士
校院名稱:朝陽科技大學
系所名稱:應用化學系碩士班
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:114
中文關鍵詞:蠟蚧輪枝菌固態發酵壓力孢子
外文關鍵詞:solid state fermentationverticillium lecaniisporepressure
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本研究探討蠟蚧輪枝菌固態發酵之培養基組成、接菌模式、溫度、濕度、壓力調控等操作條件,並進一步探討壓力對孢子之影響。系統除了使用壓力模式進行發酵期間內之調控外,在進氣部分也增加超音波製水霧器裝置,提供低溫過飽合水霧來維持培養基質之濕度,在兼顧發酵環境因子的同時,配合壓力最適操作條件下,達到高產孢量與孢子活性之維持。
實驗結果顯示同樣操作條件下,壓力為5psi、15psi、30psi (5次增/洩壓) 時發芽率分別為98%、77%、66%。在15psi加壓時間5min下比較不同加壓頻率(5次增/洩壓、10次增/洩壓),結果顯示發芽率分別為77%與67%,相同模式下30psi時則為66%與56%。改變壓力持續時間(5min/10min),以15psi觀察發芽率分別為67%與59%,30psi下為56%與47%。由實驗結果可知增加壓力頻率或加壓時間,其壓力操作條件越劇烈,孢子發芽率相對下降,且高壓力操作下變化顯著,但改以減緩增/洩壓(速率為0.2 psi/sec)模式於0-15psi操作觀察,孢子活性在24小時發芽率可達90%以上。將培養基組成改以米:米糠:稻殼以1:1:1比例混合,其接菌濃度1.22×107 spores/mL配合高接菌體積10 mL (1.58×106 spores / g dry solid)均勻噴灑接菌,最終單位產孢量可達1.72×1011 spores/g dry solid。
This research studied the solid state fermentation of Verticillium lecanii spore production for its potential application in the field of bio-insecticide. A fermentor using pressurized air for oxygen supply, heat and water balances was investigated. Operational parameters such as medium composition, inoculation strategy, pressure and humidity were studied. Results showed that using pressurized air system has the benefit of removal of heat and carbon dioxide generated from fermentation during the depressurizing stage. While moderate pressure helps in transferring oxygen into the solid state medium, the fermentation system tends to loose water during the pressurizing / depressurizing cycle. The addition of rice husk significantly improved mass transfer. The viability of spores produced was found to be seriously affected by the repeated pressurizing / depressurizing cycle at 30 psi (germination rate reduced to 66%). Time length of each pressurizing / depressurizing cycle also affected, to a lesser extent, the germination rate. The rate of pressure change, however, was found to be most important and should be controlled to be less than 0.2 psi/sec. Germination rate higher than 90% could be maintained at this rate of pressure change. Experiments also showed that water loss due to heat could be compensated by humidifying inlet air with mist generated by a supersonic vibration unit.
封面
中文摘要 Ⅰ
英文摘要 Ⅱ
誌謝 Ⅲ
圖目錄 Ⅳ
表目錄 Ⅶ
目錄 Ⅷ
第一章 緒論 1
第二章 文獻回顧 2
2.1 前言 2
2.2 常用農藥種類及其用途簡介 4
2.3 現階段台灣之蟲害管理技術 5
2.3.1農業防治 5
2.3.2物理防治 5
2.3.3化學防治 5
2.3.4 生物防治 6
2.3.5 費洛蒙、誘引劑誘殺防治 6
2.3.6 不孕性昆蟲防治 7
2.3.7 法規防治 7
2.4 生物性農藥與化學性農藥簡介 8
2.4.1 生物性農藥 8
2.4.2 化學農藥 9
2.5 生物性農藥概述 14
2.5.1 生物性農藥概分 14
2.5.2現今微生物之相關研究與分類 15
2.6 發酵量產 18
2.6.1 液態發酵(submerger fermenation) 19
2.6.2 固態發酵(soild state fermentation) 21
2.6.3 產程最適化之建立 26
2.7 菌種介紹 27
2.7.1 蠟蚧輪枝菌簡介 27
2.7.2 蠟蚧輪枝菌分類 28
2.7.3 蠟蚧輪枝菌的型態特徵 29
2.7.4 蠟蚧輪枝菌對昆蟲的致病作用與防治蟲害應用 34
2.7.5 環境因子對蠟蚧輪枝菌之影響 35
2.7.6 蠟蚧輪枝菌的生長與培養 38
2.7.7 菌種培養 39
第三章 儀器設備與實驗方法 40
3.1儀器設備 40
3.2 蠟蚧輪枝菌培養之實驗方法與流程 41
3.2.1 平面培養 41
3.2.2 基礎培養基搖瓶培養 43
3.2.3 太空包固態發酵培養 45
3.3 菌體生長型態觀察及孢子之計數 48
3.3.1 蠟蚧輪枝菌孢子計算公式 48
3.3.2 血球計數器計數(Hemocytometer) 48
3.3.3 數孢流程 49
3.4 孢子耗損程度判斷 51
3.4.1 發芽率(%)測定 51
3.4.2 methy blue染色 54
3.4.3 SEM觀察蠟蚧輪枝菌孢子型態 56
3.4.4 UVA280之測定 57
3.5 加壓供氧間歇除熱200公升固態發酵槽 59
3.5.1 發酵槽原理 60
3.5.2 發酵槽體設備簡介 61
第四章 模擬發酵之熱平衡實驗 62
4.1改善培養基組成與接菌模式探討 62
4.1.1 培養基通透性之改善對孢子產量之影響 63
4.1.2改變接菌濃度與體積對產孢之影響 67
4.2模擬發酵之熱平衡實驗 71
4.2.1 過飽和水氣對發酵時溫度變化之初步探討 72
4.2.2 超音波震盪霧化器 73
4.2.3 不同溫度下超音波製水霧量探討 74
4.3 水、空氣與熱之穿透實驗 77
4.3.1壓力對增壓速率與槽體容積之影響 77
4.3.2 導入超音波製霧之過飽和水氣對發酵時溫度變化之初步探討 78
第五章 壓力變化分析 83
5.1壓力操作模式對孢子發芽之影響 85
5.2 不同壓力條件對孢子發芽率之影響 88
5.2.1 不同壓力(0psi,15psi,30psi)下對孢子發芽率之影響 89
5.2.2 固定壓力(15 psi)下改變加壓頻率對發芽率之影響 90
5.2.3 固定壓力(30 psi)下改變加壓頻率對發芽率之影響 91
5.2.4 固定壓力(15 psi)下改變加壓時間對發芽率之影響 92
5.2.5固定壓力(30 psi)下改變加壓時間對發芽率之影響比較 93
5.2.6 改變增/洩壓操作條件對孢子發芽之影響 96
5.2.7 不同壓力配合緩慢洩壓操作對孢子發芽之影響 97
5.3 UV 蛋白質總量測定法驗證壓力對孢子影響之探討 98
第六章 結論與未來展望 104
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