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研究生:王澤瑋
研究生(外文):Ze-Wei Wang
論文名稱:低氮負荷操作連續式Anammox-UASB在不同水力停留時間之生物除氮效能及菌相變動
論文名稱(外文):Treatment efficiency and microorganism community variation of a continuous flow Anammox-UASB system under different hydraulic retention time with low nitrogen loading
指導教授:洪俊雄洪俊雄引用關係
口試委員:張怡塘張育傑
口試日期:2016-07-20
學位類別:碩士
校院名稱:國立中興大學
系所名稱:環境工程學系所
學門:工程學門
學類:環境工程學類
論文種類:學術論文
論文出版年:2016
畢業學年度:104
語文別:中文
論文頁數:111
中文關鍵詞:厭氧氨氧化作用厭氧氨氧化微生物Anammox-UASB分子生物技術
外文關鍵詞:AnammoxAnammox microorganismsAnammox-UASBmolecular methods
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近年來學術界積極研究新穎的生物除氮程序厭氧氨氧化(Anammox)系統,目標在解決傳統生物處理氨氮廢水須經由硝化脫硝等複雜程序之困擾,此類系統具有高效能除氮能力,無須額外添加碳源,且節省成本等優點,已成功應用於處理高氨氮濃度之工業廢水及生活污水。另一方面,為因應更嚴格之放流水排放標準,必須開發出可以將較低的進流氨氮濃度處理至符合排放廢水標準的應用技術。

本研究目的為以實驗室自行操作之低進流氨氮濃度(約50 ppm)批次厭氧氨氧化反應槽為目標,使用之菌種為本團隊自台灣地區垃圾滲出水厭氧操作系統中所分離出之厭氧氨氧化微生物,將批次反應槽改裝為連續式Anammox-UASB反應槽,並進一步監測系統水質與微生物定性定量分析,藉此探討系統除氮效能、微生物變化之關係及顆粒污泥組成。

對照系統水質結果得知,在不同水力停留時間操作之下其除氮效能皆可趨於穩定,氨氮去除率達93%以上,其中最高除氮速率達0.090 kg-N/m3/d。系統從批次操作改建為連續流操作,內部厭氧氨氧化族群種類從Candidatus Brocadia sp. 40、Candidatus Kuenenia stuttgartiensis、Candidatus Brocadia sinica為主,轉變成Candidatus Kuenenia stuttgartiensis、Candidatus Brocadia sinica及Candidatus Anammoxoglobus propionicus為主,且Candidatus Anammoxoglobus propionicus亮帶訊號更為強烈,厭氧氨氧化微生物族群之基因表現量平均在1.0 x 108 copies/μg DNA。顆粒污泥部分,內部厭氧氨氧化族群以Candidatus Brocadia sp. 40、Candidatus Kuenenia stuttgartiensis、Candidatus Brocadia sinica及Candidatus Anammoxoglobus propionicus為主,厭氧氨氧化微生物族群之基因表現量為1.0 x 109 copies/μg DNA,NGS(Next Generation Sequencing)分析結果得知,顆粒污泥中厭氧氨氧化微生物占整體約15%,而綠色不含硫微生物(Chloroflexi)在顆粒污泥中明顯偏多,約占21%,推測綠色不含硫微生物可能有助於顆粒之形成。


Application of ammonia anaerobic oxidization systems on nitrogen removal has attracted countless research attentions in recent years. This technique differs from the traditional nitrification/denitrification process with higher nitrogen removal efficiency and saving money on aeration as well as no requirement of carbon source addition. Several application of Anammox systems have been successfully established on treatment high ammonia concentration wastewater. However, it is necessary to develop a better Anammox operating technology for low strength of ammonia containing wastewater to meet the effluent standard.

Therefore, the main goal of this proposal is to operate a long-term operating low-ammonia feeding (around 50 ppm) batch Anammox reactor established in our lab using Anammox microorganisms as seeding. These Anammox microorganisms were previously enriched from landfill leachate wastewater treatment system located in central Taiwan. In the first stage, modify the batch-operated reactor to continuous Anammox-UASB reactor. Then during the operation time, water quality, microbial community structure, interaction microorganisms and granular sludge were detected for monitoring and discussing the variation of nitrogen loading and nitrogen removal efficiency.

Results showed that removal efficiency of ammonium is more than 93% and the highest nitrogen removal rate is 0.090 kg-N/m3/d under different hydraulic retention time. The Anammox systems with different hydraulic retention time of the operation, the internal Anammox microorganisms changed from Candidatus Brocadia sp. 40, Candidatus Kuenenia stuttgartiensis, Candidatus Brocadia sinica mainly into Candidatus Kuenenia stuttgartiensis, Candidatus Brocadia sinica and Candidatus Anammoxoglobus propionicus mainly.On the other hand, Real-Time PCR analysis showed that the quantification of gene copies of Anammox microorganisms maintain at 108 copies/µg DNA. Granular sludge fraction, the internal Anammox microorganisms contained Candidatus Brocadia sp. 40, Candidatus Kuenenia stuttgartiensis, Candidatus Brocadia sinica and Candidatus Anammoxoglobus propionicus mainly. Real-Time PCR analysis showed that the quantification of gene copies of Anammox microorganisms maintain at 1.0 x 109 copies/µg DNA. NGS (Next Generation Sequencing) analysis showed that, in the granular sludge Anammox microorganisms accounted for about 15% of overall. Green non-sulfur bacteria (Chloroflexi) may contribute to the formation of granular sludge.


目錄 v
圖目錄 viii
表目錄 x
第一章 前言 1
第一節 研究緣起 1
第二節 研究目的 2
第二章 文獻回顧 3
第一節 氮循環 3
一、 生物固氮作用 4
二、 硝化作用 4
三、 脫硝作用 5
第二節 含氮污染物來源及其危害 6
第三節 含氮廢水處理程序 9
一、 好氧硝化/脫硝程序 9
二、 SHARON程序 9
三、 ANAMMOX程序 10
四、 結合SHARON+ANAMMOX程序 10
五、 CANON程序 10
六、 OLAND程序 11
七、 DEMON程序 11
八、 SNAD程序 11
第四節 厭氧氨氧化作用及厭氧氨氧化微生物 13
一、 厭氧氨氧化作用及其應用 13
二、 厭氧氨氧化微生物 16
三、 厭氧氨氧化微生物構造 19
四、 厭氧氨氧化微生物作用機制 21
五、 影響厭氧氨氧化微生物生長之因素 24
第五節 分子生物技術對厭氧氨氧化微生物研究之應用 26
一、 聚合酶鏈鎖反應 26
二、 變性梯度凝膠電泳法 28
三、 即時定量聚合酶鏈鎖反應(Real-time PCR) 28
四、 次世代定序分析(Next Generation Sequencing, NGS) 30
第六節 厭氧氨氧化微生物與其他微生物之共生關係 34
一、 厭氧氨氧化微生物與其他微生物之共生關係 34
二、 利用次世代定序分析探討共生關係之相關文獻 35
第七節 Anammox-UASB處理程序相關研究 37
一、 上流式厭氧污泥床(UASB) 37
二、 Anammox-UASB相關文獻 37
第八節 文獻閱讀心得及研究方向擬定 39
第三章 材料與方法 41
第一節 實驗架構 41
第二節 實驗設備 42
第三節 厭氧氨氧化生物反應槽 43
一、 薄膜生物反應槽設計(馴養期) 43
二、 連續式Anammox-UASB反應槽設計(實驗期) 44
第四節 厭氧氨氧化生物反應槽操作 46
一、 人工合成基質 46
二、 系統植種來源 46
三、 厭氧氨氧化微生物馴養 47
四、 連續式Anammox-UASB反應槽操作 47
第五節 水質分析方法 48
一、 氨氮 48
二、 亞硝酸鹽氮 48
三、 硝酸鹽氮 49
第六節 分子生物技術 50
一、 樣本前處理 50
二、 DNA萃取 50
三、 聚合酶鏈鎖反應(PCR) 51
四、 變性梯度凝膠電泳(DGGE) 54
五、 DNA序列分析 55
六、 即時定量聚合酶鏈鎖反應(Real-Time PCR) 55
七、 NGS定序 57
第四章 結果與討論 59
第一節 系統各階段操作及水質 60
一、 第一階段:批次馴養期 60
二、 第二階段:批次穩定期 60
三、 第三階段:HRT=48小時實驗期 61
四、 第四階段:HRT=36小時實驗期 61
五、 第五階段:HRT=24小時實驗期 62
第二節 系統各階段水質綜合討論 63
一、 系統各階段除氮效能 63
二、 氮化物反應劑量平衡 64
三、 系統各階段pH值及ORP值變動 65
第三節 系統各階段微生物族群分析 69
一、 厭氧氨氧化族群分析 69
二、 氨氧化族群分析 72
三、 亞硝酸鹽氧化族群分析 73
四、 脫硝族群分析 74
五、 真細菌族群分析 76
第四節 厭氧氨氧化微生物顆粒污泥分析 78
一、 污泥型態分析 78
二、 顆粒污泥微生物組成分析 79
第五節 綜合討論 87
第五章 結論與建議 89
第一節 結論 89
第二節 建議 91
第六章 參考文獻 93


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