Effects of Pretreatment on Anaerobic Digestion of Biological Sludges

本論文目的是探討前處理對污泥消化之影響。前處理包含加入微生物菌群、增溫、凍融及調整酸鹼值等。微生物菌群處理是加入梭狀芽孢桿菌群(Clostridium spp.)。實驗中，運用代謝物分析與微生物社群分析(PCR-DGGE)結果來追蹤厭氧消化的代謝現象及菌群之變化。高性能液相層析儀(HPLC)所檢測到的代謝物主要以揮發酸(Volatile Fatty Acids, VFA)。微生物菌群監測目標主要以產酸及產甲烷菌為主，如:Clostridium spp.、Methanobacterium spp.。研究結果發現，污泥加入梭狀芽孢桿菌群培養液後，可大幅加速污泥厭氧消化及增加甲烷產量。然而先經過增溫、凍融及調整酸鹼值處理的污泥再加入梭狀芽孢桿菌培養液的樣品其產氣結果並未明顯優於單純加入梭狀芽孢桿菌群培養液的處理方式。The aim of this thesis is intended to investigate the influence of pretreatment on sludge digestion. Microbial inoculums, sterilization, frozen/thawed, acidification, and alkalization are used to pretreat the sludge. The microbial inoculum is clostridium inoculums. In our experiment, we used the HPLC and microbial community analysis (PCR-DGGE) to monitor the digestion pathway and the microbial population change. The major metabolite detected by HPLC are volatile fatty acids (VFA) and lactic acid. The main tracked bacteria in microbial community analysis are acetogens and methanogens, like Clostridium spp.、Methanobacterium spp.. The results show that the pretreatment of Microbial inoculums can substantially increases digestion rate and methane product. However, the sludge treated by adding Clostridium inoculums combined with other pretreatmentis aren't clearly better than only by Clostridium inoculums.目錄 頁次 中文摘要............................................. I 英文摘要............................................. II 目錄................................................. III 圖目錄............................................... V 表目錄............................................... VII 第一章 前言........................................ 1 第二章 文獻回顧.................................... 3 2-1厭氧消化.......................................... 3 2-1-1水解(hydrolysis) ............................... 4 2-1-2酸化(acidogenesis) ............................. 4 2-1-3甲烷化(methanogenesis) ......................... 7 2-2甲烷生成菌群(methanogenesis) ..................... 7 2-3運用分子生物技術觀察微生物族群變化................ 13 2-3-1 聚合酵素鏈鎖反應(PCR)的原理與應用.............. 14 2-3-2變性梯度凝膠電泳................................ 16 第三章 實驗材料與方法.............................. 20 3-1 污泥來源與菌種來源............................... 20 3-1-1 污泥採樣處..................................... 20 3-1-2 菌種........................................... 20 3-2 污泥前處理方法................................... 20 3-3 血清瓶厭氧批次消化實驗........................... 22 3-4檢測項目.......................................... 23 3-5 分子生物技術分析PCR&DGGE......................... 26 3-5-1 DNA萃取........................................ 27 3-5-2 PCR之操作條件.................................. 27 3-5-3變性梯度凝膠電泳DGGE............................ 31 第四章 結果與討論.................................. 34 4-1各種前處理對污泥樣品的影響........................ 34 4-2 微生物菌群整理................................... 37 4-3微生物監測結果與代謝物關係討論.................... 40 4-3-1原始污泥之代謝物變化與微生物結果討論........ 42 4-3-2接種污泥之代謝物變化與微生物結果討論........ 43 4-3-3 酸化污泥之代謝物變化與微生物結果討論....... 47 4-3-4 鹼化污泥之代謝物變化與微生物結果討論....... 50 4-3-5 凍融污泥之代謝物變化與微生物結果討論....... 52 4-3-6 滅菌污泥之代謝物變化與微生物結果討論....... 55 第五章 結論........................................ 65 第六章 參考文獻.................................... 6