本研究著重於三段式生物程序組合中硝化槽之功能評估及微生物生態探討，由於硝化菌的特性，生長緩慢，常成為氮去除的限制步驟，所以在本研究中嘗試以分子生物方法探討硝化菌群在處理含氮工業廢水之好氧系統中之特性，提供後續研究除氮程序設計之參考。
本研究接續2000~2003年以三段式流體化床生物組合程序(高溫厭氧脫氨/中溫無氧脫硝/中溫好氧硝化)，處理具丙烯腈(Polyacrylonitrile, PAN)人造纖維製程廢水，此廢水成分複雜且含有抑制性難生物分解之物質，包含有機污染物及高濃度氮化物(TKN/COD = 0.15~0.26)，並含有高濃度硫酸鹽(SO42- = 882~1762 mg/L)，且缺乏生物作用所需磷源(TP/COD = 3.3´10-5)，因此PAN廢水為難生物處理之高氮工業廢水。在三段式生物程序中，藉高溫厭氧微生物將PAN廢水中有機氮脫氨裂解，並將大分子有機碳化物初步降解成小分子有機酸類，由硝化槽將氨氮氧化成硝酸鹽，供給第二段無氧脫硝槽以高溫厭氧槽出流殘留有機碳做電子接受者，與硝化槽迴流水中硝酸鹽還原成氮氣，達成水中氮鹽去除之目的。
　　本研究接續2002年陳氏之研究，將好氧硝化流體化床從新啟動，初期以半批分次程序啟動，試圖增殖自營性硝化菌於反應槽中，結果顯示此期間氨氮均能完全轉化為硝酸鹽，硝化功能良好，但是在擔體粒狀活性碳(Granular activated carbon, GAC)上未能有效形成生物膜。再者於進流水中添加蔗糖，試圖以增殖有機碳氧化菌於GAC上，幫助硝化菌附著，結果仍未能有效促使生物膜生成。在三段式組合程序串聯操作時，好氧硝化槽之進流水質視前段處理效能而定，此期間分為兩試程，其一進流COD約為110 mg/L，氨氮為65 mg N/L，此期間COD約可去除25 %，氨氮之去除率受限於微生物量為50 %。另一試程進流COD約為290 mg/L，氨氮為30 mg N/L，此期間COD約可去除27 %，氨氮之去除率為96 %。可見好氧槽對有機物仍有處理效果，而氨氮之去除與負荷有關。以掃瞄式電子顯微鏡(Scanning electron microscope, SEM)作為菌相觀察可發現初期培養自營性硝化菌時，發現污泥不易附著GAC之特性，而添加蔗糖結果顯示有絲狀菌的形成，好氧反應槽啟動初期GAC上以桿菌為主，而三段式組合程序串聯操作後可發現桿菌及少許球菌，菌種趨向更多元化。
　　本研究中更進一步以分子生物方法探測好氧槽中菌群結構，以三段式程序組合操作時172天之懸浮污泥做16S rDNA基因選殖實驗，將71個不同菌落經DGGE篩選後可辨識出33個不同相對位置的條帶，將篩選到菌落出現比例較高者作定序分析，其中Nitrospira sp.佔16.9 %，為亞硝酸氧化菌之一屬；亦分析到Actinobacteria佔14.1 %，可能與絲狀菌形成相關；有三株屬於a-Proteobacteria，其中與Nitrobacter winogradskyi相似佔系統中5.6 %。
以專一性引子配合尾端修飾限制片段多型性 (Terminal restriction fragment length polymorphism, T-RFLP)分析各試程中硝化菌之分佈情形，發現在植種污泥及反應槽操作期間均可偵測到氨氧化菌及亞硝酸氧化菌的存在，在植種中存在的氨氧化菌以Nitrosospria為主，經氨氮馴養後轉變為以Nitrosomonas為主，而在整個反應槽操作期間均有Nitrobacter及Nitospria存在，由此可見生長環境影響菌群結構，以分子生物方式監測硝化菌比起傳統方式更能明確了解硝化菌特性，對未來控制程序條件提供了一些相關訊息。

　　This study evaluated performance and microbial ecology of nitrification in a three-satge fluidizaed bed bioprocess, thermophilic anaerobic/ anoxic denitrification/ aerobic nitrification, treating synthetic fiber manufacturing wastewater. The main component in the wastewater is polyacrylonitrile(PAN). The PAN wastewater contains not only organic pollutants but also high concentrtion of organic nitrogen(TKN/COD = 0.15~0.26) and sulfate(SO42- = 882~1762 mg/L), and lacked phosphorous source(TP/COD = 3.3´10-5). The industral wastewater was complex and inhibitive for biodegradation. Because nitrifying bacteria grow slowly and are easily inhibited, nitrification usually becomes a rate limiting step of nitrogen removal. This study used molecular biology techniques to investigate charaterestics of nitrifying bacteria in aerobic system treating industrial wastewater containing organic nitrogen.
　　In the three-stage process, thermophilic anaerobes degraded organic nitrogen to ammonia and converted longe-chain organic carbon to volatile acid. Ammonia oxidized to nitrate in the nitrification reactor. Nitrate and residual organic carbon from thermophilic anaerobic reactor provided denitrification reaction.
During the study, the aerobic nitrification was able to convert ammonia to nitrate efficiently, but biofilm formation on granular activated carbon(GAC) was ineffectively. Therefore, inducing organic carbon oxidizing bacteria adhered on GAC by adding sucrose in influent was employed to promote nitrifying bacteria biofilm formation. Results showed biofilm formation, howere wasn’t significant. In the three-stage process, the water guality of nitrification influent was determined by the first and second reactors. In one period, COD and ammonia concentration of nitrification influent were 110 mg/L and 65 mg N/L, and removal rate were 25 % and 50 %. In another period, COD and ammonia concentration of influent were 290 mg/L and 30 mg N/L, and removal rate were 27 % and 96 %. In conclusion, aerobic nitrification reactor was able to remove partial COD, but ammonia removal rate depedded ammonia loading.
　　According to the observation of scanning electron microscope(SEM), rod bacteria were dominant bacteria in the start-up period, and filamentous bacteria and cocci were observed at high organic volume loading.The cloning library was constructed for the sample taken at day 172 in three-strage process. Based on cloning-secguence analysis from 71 clones, Nitrospira sp., Nitrobacter winogradskyi, and Actinobacteria were 16.9 %, 5.6 %, and 14.1 %. In addition, the microbial diversity of ammonia oxidizing bacteria(AOB) and nitrite oxidizing bacteria(NOB) were monitored using terminal restriction fragment length polymorphism(T-RFLP). Nitrosomonas spp., Nitrobacter and Nitrospira sp. were dominant AOB and NOB in all operational periods.

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陳賢焜，鄭幸雄，(1994)，「生物膜化脫硝程序處理高濃度氨氮廢水之研究」，國立成功大學環境工程研究所博士論文。
陳文欽，鄭幸雄，(1997)，「固定生物流體化床處理高氮樹脂廢水之特性研究」，國立成功大學環境工程研究所博士論文。
陳彥男，鄭幸雄，(2002)，「三段式流體化床生物程序處理壓克力人造纖維製程廢水之程序研究」，國立成功大學環境工程研究所碩士論文。
陳莉容，李季眉，(2001)，「利用微生物將丙烯晴轉換成丙烯醯氨之研究」，中興大學環境工程系碩士論文。
陳文欽, 鄭幸雄, (1999)「樹脂製程之生物分解特性研究」, 第二十四屆廢水處理技術研討會論文集, pp. 15-20。
羅文益，鄭幸雄 (1995)「生物脫硝流體化床程序處理高氮壓克力製程廢水」，國立成功大學環境工程研究所碩士論文。
邱金窗，鄭幸雄 (1996)「脫硝污泥對高濃度有機氮工業廢水之生物分解特性研究」，國立成功大學環境工程研究所碩士論文。
周素圓，鄭幸雄，(1997)，「硝化脫硝程序中生物活性評估技術之研究」，國立成功大學環境工程研究所碩士論文。
蔡宗岳，鄭幸雄，(1998)，「兩段式生物脫硝及硝化組合程序去除ABS製程廢水中有機物與含氮化合物之研究」，國立成功大學環境工程研究所碩士論文。
林泓胤，鄭幸雄，(1999)，「三段式流體化床生物程序處理高氮工業廢水之程序研究」，國立成功大學環境工程研究所碩士論文。
鍾志宏，鄭幸雄，(2000)，「三段式生物程序組合處理ABS及PAN製程廢水之功能評估」，國立成功大學環境工程研究所碩士論文。
彭欽鑫，鄭幸雄，(2001)，「三段式流體化床三相生物程序處理人造纖維製程廢水之生物分解性研究」，國立成功大學環境工程研究所碩士論文。
吳坤龍，鄭幸雄，(2003)，「高溫厭氧菌分解PAN廢水之族群變化與功能評估」，國立成功大學環境工程研究所碩士論文。
莊蕙萍，鄭幸雄，(2004)，「三段式流體化床生物程序處理PAN廢水之程序功能評估與微生物族群動態變化之探討」，國立成功大學環境工程研究所碩士論文。
張淑惠，李季眉，(1996)，「氮系化合物分解微生物培養及篩選之研究」，國立中興大學環境工程所碩士論文。
王俊欽，李季眉，(1997)，「高氮廢水中丙烯腈分解菌及脫氮菌之研究」，國立中興大學環境工程所碩士論文。
鄭幸雄，陳佩足，許佩瑜(1998)，「氣泡呼吸儀批次試驗探討丙烯腈生物脫氨機制」，第二十三屆廢水處理技術研討會論文集，第248~255頁。
鄭幸雄，陳賢焜，林煜唐 (1995)， "Enhanced Biodegradation of Organic Nitrogenous compounds in Resin Manufacturing Wastewater by Anoxic Denitrification and Oxic Nitrification Process",第二十屆廢水處理技術研討會論文集, pp. 2-1~2-5。
鄭幸雄，李季眉，陳文欽，周素圓，王俊欽，蔡宗岳 (1997)，「丙烯晴氧化及脫硝分解功能評估」，第二十二屆廢水處理技術研討會論文集，448-455頁。
鄭幸雄，李季眉，陳文欽，周素圓，王俊欽，蔡宗岳 (1997)，「丙烯晴氧化及脫硝分解功能評估」，第二十二屆廢水處理技術研討會論文集,台中, pp. 448-455。
鄭幸雄, 鍾志宏, 彭欽鑫 (2000)「固定化生物程序處理高氮工業廢水之探討」, 第二十五屆廢水處理技術研討會論文集, pp.234-239。
鄭幸雄, 陳彥男, 吳坤龍, 莊蕙萍 (2002)「三段式流體化床生物程序處理壓克力纖維製程廢水之程序研究」, 第二十七屆廢水處理技術研討會論文集, w-a-o11。
鄭幸雄, 莊蕙萍, 楊雅斐, 陳彥男(2002)「高溫厭氧菌分解PAN廢水反應中氫氣對有機氮裂解之影響」, 第二十七屆廢水處理技術研討會論文集, w-a-o24。
鄭幸雄, 莊蕙萍, 陳彥男, 彭欽鑫(2002)「厭氧污泥、脫硝污泥及硝化污泥對PAN廢水之生物可分解性研究」, 第二十七屆廢水處理技術研討會論文集, w-a-o31。
鄭幸雄, 吳坤龍, 曾怡禎, 莊蕙萍(2003)「高溫厭氧流體化床處理PAN人造纖維廢水之微生物族群群態研究」, 第二十八屆廢水處理技術研討會論文集。
鄭幸雄, 莊蕙萍, 楊雅斐(2003)「三段式流體化床生物程序中脫硝流體化床處理壓克力製程廢水之研究」, 第二十八屆廢水處理技術研討會論文集。
鄭幸雄, 楊雅斐, 莊蕙萍(2003)「硝化污泥在不同pH與溫度下對氨氮降解之影響」, 第二十八屆廢水處理技術研討會論文集。