請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/5681
標題: 除氮系統中與無氧氨氧化菌共存之微生物組成分析
Microbial Community Analysis of the Microorganisms Coexist with the Anammox Bacteria in the Biological Nitrogen Removal System
作者: 張婷婷
Chang, Ting-Ting
關鍵字: Anaerobic ammonia oxidation reaction
無氧氨氧化程序
Anaerobic ammonia oxidation bacteria
Molecular biotechnology
無氧氨氧化細菌
分子生物技術
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)為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特性為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,與傳統硝化、脫硝反應不同,此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。本研究以工研院所設計之不同型式無氧氨氧化反應槽為研究對象,包括進流基質為氨氮與亞硝酸鹽之上流式厭氧污泥化系統(UASB)系統及進流基質為氨氮添加BioNET泡棉擔體之Partial nitrification/Anammox系統,利用聚合酶鏈鎖反應-變性梯度凝膠電泳(PCR-DGGE)、Cloning和螢光原位雜合(FISH)等分子生物技術探討不同型式無氧氨氧化系統之微生物菌群結構變化和除氮效能間的關係,此外亦探討與無氧氨氧化菌共存的微生物組成。 由微生物族群DGGE圖譜可發現,在UASB系統中確實存在著無氧氨氧化菌Candidatus Brocadia fulgida,同時系統中也存在著氨氧化菌ammonia-oxidizing Betaproteobacteria,推測氨氧化菌的作用在將氨氮轉換為亞硝酸鹽氮,並與無氧氨氧化菌Candidatus Brocadia fulgida共同將反應槽內之氨氮轉換為氮氣。此外,系統中亦存在有亞硝酸鹽氧化菌Nitrobacter sp.,且約占總菌數的5%左右,系統中亦存在有脫硝菌Steroidobacter denitrificans及梭狀芽孢桿菌Clostridium celatum、酸化菌Syntrophus sp.、Butyrivibrio fibrisolvens、硫酸還原菌Desulfobulbus sp.等。推測這些與生物除氮無直接關係的微生物扮演的角色可能為與有機質或蛋白質分解有關,進而釋放二氧化碳,提供給無氧氨氧化菌Candidatus Brocadia fulgida等自營菌所需之碳源,或可以消耗系統中的溶氧,提供無氧氨氧化菌適合生長的厭氧環境。 而添加BioNET泡棉擔體之Partial nitrification / Anammox系統則測定不到無氧氨氧化菌,污泥樣本中存在有氨氧化菌、亞硝酸鹽氧化菌Nitrospira sp.與Nitrobacter sp.、酸桿菌Bacterium Ellin、脫硝菌Defluvicoccus vanus、亞硝酸鹽氧化菌Nitrospira sp.、Uncultured bacterium、紫色不含硫菌Rubrivivax gelatinosus、革蘭氏陰性桿菌Aquimonas sp.與尚未定序的菌種等。 無氧氨氧化菌Candidatus Brocadia fulgida是目前為止發現唯一會自發螢光的無氧氨氧化菌,由於現今對於無氧氨氧化菌Candidatus Brocadia fulgida相關的結構與特性尚未釐清,且與其共生的微生物族群研究甚少,因此與無氧氨氧化菌Candidatus Brocadia fulgida共生的菌群間相互作用值得更進一步探討。
The anaerobic ammonia oxidation (ANAMMOX) reaction is one of side pathways 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. In this research, the microbial community structures of different type anaerobic ammonia oxidation reactors(developed by ITRI) including upflow anaerobic sludge bed (UASB) system and partial nitrification/Anammox system with BioNET cotton that operated under various influent concentrations were studied using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE)、Cloning and fluorescence in situ hybridization (FISH) techniques. Not only the relationship between the microbial community structures and nitrogen removal efficiency were compared, the possible roles of microorganisms co-existed with the anammox bacteria community were studied as well. Gene sequences obtained from DGGE patterns showed that anaerobic ammonia oxidation bacteria “Candidatus Brocadia fulgida” existed in the UASB system. Application of FISH quantification on both ammonia and nitrite oxidizing bacteria in the UASB indicated the existence of ammonia oxidizing β-proteobacteria(10%) and nitrite oxidizing bacteria Nitrobacter sp.(5%). We hypothesize that ammonia oxidizing bacteria converted ammonium to nitrite then being used by anammox bacteria to ensure total nitrogen removal. Results from DGGE experiments revealed that heterotrophic bacteria including Steroidobacter denitrificans、Clostridium celatum、Syntrophus sp.、Butyrivibrio fibrisolvens and Desulfobulbus sp. existed in the UASB system. We hypothesize that heterotrophic bacteria might be involved in the degradation of complex compounds in the UASB system to reduce the oxygen concentration and release carbon dioxide for aiding the survival of anammox bacteria. Ammonia oxidizing bacteria、nitrite oxidizing bacteria、Bacterium Ellin、Defluvicoccus vanus、Rubrivivax gelatinosus、Aquimonas sp. and certain uncultured bacterium existed in the Partial nitrification/Anammox with BioNET system. However, no anammox bacteria were detected. Candidatus Brocadia fulgida exhibited the common characteristics of anammox bacteria and its the biofilm aggregates could show strong autofluorescence. It is the only known anammox species exhibiting this feature. Detailed characteristics of this anammox bacterium remains unknown, so does research on the microbial community co-existed with Candidatus Brocadia fulgida. Further studies on Candidatus Brocadia fulgida as well as its interactions with the other microorganisms are needed.
URI: http://hdl.handle.net/11455/5681
其他識別: U0005-1507200923275500
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1507200923275500
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