Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/5444
標題: 不同型式無氧氨氧化系統之菌相結構分析
Microbial community studies from different design of anaerobic ammonia oxidation reactors.
作者: 邱榆鈞
Chiu, Yu-chun
關鍵字: 無氧氨氧化程序
Anaerobic ammonia oxidation reaction
部分硝化
無氧氨氧化細菌
氨氧化菌
分子生物技術
生物擔體
上流式厭氧污泥系統
Partial nitrification
Anaerobic ammonia oxidation bacteria
Ammonia oxidization bacteria
Molecular biotechnology
biological
UASB system
出版社: 環境工程學系所
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摘要: 無氧氨氧化反應(ANAMMOX,Anaerobic Ammonia Oxidation)是氮循環中較不明顯的分支,參與該反應之自營性無氧氨氧化菌之特性是能於無氧環境中將氨氮與亞硝酸根直接反應生成氮氣,其化學方程式為NH4+ + NO2- → N2 + 2H2O,與傳統之硝化、脫硝反應不同,這個反應對於氧氣濃度較低的環境中,具有維持氮循環平衡的重要功能。 參與此反應的微生物-無氧氨氧化細菌,生長速度較為緩慢,以傳統微生物方法進行培養十分耗時,利用較新穎之16S rDNA比對來進行分析,將會是鑑定無氧氨氧化菌較適合的方法。本研究以工研院所設計之不同類型無氧氨氧化反應槽,包括進流基質為氨氮與亞硝酸鹽之UASB系統和添加BioNET泡棉擔體之UASB系統以及進流基質為氨氮之添加BioNET泡棉擔體之Partial nitrification/Anammpx系統,操作在進流基質濃度逐漸提升之下探討其微生物族群結變動情形,利用PCR-DGGE和FISH方法探討不同無氧氨氧化系統下微生物菌群結構變化和除氮效能間的關係。 利用專一性Anammox引子組(PLA46 & Amx368-B)分析不同型態無氧氨氧化系統之Anammox族群,其結果顯示在DGGE圖譜上可得到多個不同位置的亮帶,而將其定序經由比對後發現這些序列與資料庫中已知菌種之相似度約為93-99%之間。由微生物組成來看,添加BioNET擔體之系統中菌相結構較繁多,但是存在於系統中之Anammox菌群的數量比例偏低,推測在添加BioNET擔體之UASB系統中,可能會有許多與無氧氨氧化族群競爭氮源之非無氧氨氧化族群增加於系統中,使整體菌相結構較為複雜,更使得在相同操作條件下除氮效能較UASB系統略差,但是系統整體之除氮效能仍然維持在80%左右。另一添加BioNET擔體之Partial nitrification/Anammpx系統中,其微生物多樣性較為複雜,尤其是其他微生物種類(如硝化菌群或異營性微生物等)相較於UASB系統多,推測可能是無氧氨氧化菌群與非無氧氨氧化菌群彼此間共同競爭基質,造成系統除氮效能不佳(N-removal rate大約30~70%)。 在總菌相結構上,存在於系統中之不屬於Anammox微生物包含氨氧化菌、Nitrospira sp.、異營性微生物以及未被定義之菌種(Uncultured bacterium)等。各菌群間互相競爭基質的結果明顯地影響厭氧脫硝效果,這些菌群間的相互作用值得進一步探討。
The anaerobic ammonia oxidation(ANAMMOX) reaction is a minor pathway in the nitrogen cycle. The anaerobic ammonia oxidation bacteria could directly oxidized ammonium and nitrite to nitrogen gas under anoxic conditions by the following equation NH4+ + NO2- → N2 + 2H2O. Anammox reaction, which is different from the traditional nitrification and denitrification reaction, is important for the balance of the nitrogen cycle under lower oxygen concentration environment. Microorganisms which participate in this reaction grow extremely slow. For this reason, using novel 16S rDNA based techniques to determine the phylogenetic identities of the dominant members of the microbial community is necessary. In this research, the microbial community structures of different type anaerobic ammonia oxidation reactors (developed by ITRI) including UASB system、UASB with BioNET cotton system and partial nitrification/Anommox system with BioNET cotton operated under various influent concentrations were studies using PCR-DGGE and FISH techniques. The relationship between the microbial community structures and nitrogen removal efficiency were compared. Experimental results showed that Anammox primer(PLA46 & Amx368-B) could successfully distinguish different Anammox species in all tested systems. Sequences collected from DGGE (denaturing gradient gel electrophoresis) gel have sequence identities ranging from 93-99% comparing to the 16S rDNA sequence in NCBI database. BioNET biological system has a complex microbial community but its Anammox cell count is low. There is a non-Anammox bacteria population which could compete nitrogen source with Anammox bacteria existed in the UASB with BioNET system and resulted a more complex microbial community. Nitrogen removal efficiencies for these two systems maintained at around 80% despite the difference in microorganism composition. Among these three systems, the Partial nitrification/Anammpx with BioNET system seems to have the most complex bacteria structures. Therefore, non-anammox bacteria could significantly compete substrate with Anammox bacteria and results a lower nitrogen removing efficiency (30~70%) in the Partial nitrification/Anammpx with BioNET system. As for the total microbial community analysis, non-Anammox bacterium include ammonia oxidization bacteria、Nitrospira sp., heterotrophic bacteria and unidentified bacterium co-existed with Anammox community in these tested systems. The competition between all these microorganisms significantly affected the denitrification efficiency. Further studies on exploring these particular bacterial interactions are needed.
URI: http://hdl.handle.net/11455/5444
其他識別: U0005-2107200816263400
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2107200816263400
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