本研究為探討厭氧氨氧化(Anammox)作用發生於人工濕地之可行性，參考污水廠之自營性脫氮程序(CANON)，設計實驗模槽及操作條件。因CANON程序為單槽中完成部分硝化及Anammox兩種作用，因此選用垂直流人工濕地(VFCW)做為實驗模槽，利用其深度及濕地停留時間(RT)長之特性，藉此達到微好氧及厭氧分層共存之環境。此外，因CANON中兩種參與之作用皆由自營菌所促使，為避免彼此競爭而導致自營菌生長受限，因此進流水中不添加有機碳源，以抑制異營菌的作用及生長。
VFCW中填充礫石做為微生物附著之媒介，並藉此阻礙氧氣傳輸至深層水域。以含氨氮之進流水進行RT控制在5天條件下之批次實驗。實驗初始係以附著性微生物(礫石上之生物膜)為主要參與作用之微生物，處理含氨氮之校園生活污水，但在RT控制在5天之情況下，仍難以達到厭氧環境，且處理效果不佳。因此第二階段實驗除了利用原有之礫石表面附著性微生物外，再添加懸浮性微生物(厭氧污泥)於模槽底部，並改以人工廢水做為實驗進流水。實驗結果顯示，保留部分懸浮性微生物於VFCW中，可提高模槽中附著性微生物之富集。試程操作24小時內，在硝化作用旺盛，促使溶氧不但可達部分硝化進行之條件，也可能已有Anammox作用。而在RT控制為5天時，總氮去除率最高可達97.5%。進一步以不同操作條件的對照實驗，排除可能出現於VFCW系統中之其他脫氮途徑。同時，PCR實驗顯示實驗模槽中確實有Anammox菌種存在。另外，實驗結果亦顯示此系統在種植水生植物之情形下，明顯有助於整體總氮之去除率。

In this study, the possibility of driving Anammox process in constructed wetland systems is investigated. The experimental designs are based on completely autotrophic nitrogen removal over nitrogen removal over nitrite (CANON), which can couple Anammox process with partial nitrification in one single unit. To reach the purpose, in this study a vertical-flow constructed wetland (VFCW) system with deep depth and high RT was used. The VFCW system was installed with gravels,on which biofilm was formed and attached, and fill with some anaerobic sludge for suspension growth in the system. In order to inhibit the effects of heterotrophic denitrification, low levels of chemical oxygen demand (10.14-11.59 mg/L) were controled in the inflow of the systems.
The experimental results show that the process of nitrification is very significant in the first day of test run, while the concentrations of DO were decreased down to below 1 mg/L, and the concentrations of NH4+-N were decreased with increasing NOx(NO2--N+NO3--N). Comparing with the amounts of NH4+-N decreased, the amounts of NO2--N increased was far less than it, while the same situation was shown until the fifth day. The percentage of TN removed were reached to 97.5% (PS) in the fifth day, which was the end of test run. Though the test of Polymerase Chain Reaction (PCR),it was helpful to know the Anammox bacteria did exist in the VFCW system, which might occur from the first day to the fifth day. It was also shown that the plants in VFCW system could enhance the removal of TN. That was because the plants could absorb NO3--N, which was produced by those processes like nitrification and Anammox.

摘要 I
ABSTRACT III
目錄 IV
圖目錄 VI
表目錄 IX
第一章 前言 1
1.1研究動機 1
1.2研究目的 3
第二章 文獻回顧 4
2.1 濕地介紹 4
2.1.1 濕地定義 4
2.1.2 濕地功能 5
2.1.3 人工濕地介紹 5
2.1.4 人工濕地種類 7
2.1.5 人工濕地處理污染物之機制 12
2.2 生物脫氮機制 15
2.2.1 傳統生物脫氮原理 15
2.2.2 新式生物脫氮技術 18
2.3 案例介紹 30
2.3.1 Anammox於生物膜反應槽之應用 30
2.3.2 Anammox於人工濕地之應用 32
第三章 材料與方法 34
3.1系統設計 34
3.2模槽設計及濾料特性 34
3.3濕地系統植物選擇 34
3.4模槽植種及馴養 35
3.5樣品採集及分析方法 36
3.6 實驗設計及流程 40
第四章 結果與討論 44
4.1第一階段實驗 44
4.2 第二階段實驗 54
4.2.1修正設計後之實驗 54
4.2.2高濃度氨氮試驗 62
4.2.3 24小時連續監測試驗 68
4.3 第三階段實驗 73
4.3.1 其他影響去除率之因素 73
4.4 微生物實驗 81
4.4.1 模槽總菌落數分析 81
4.4.2 Anammox菌種確認 81
4.5綜合討論 83
第五章 結論與建議 86
5.1結論 86
5.2建議 86
第六章 參考文獻 87
附錄 照片 97

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Green, M.B., Haberl, R. (Eds.), Constructed wetlands for wastewater treatment
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Huihui Chen, Sitong Liu, Fenglin Yang, Yuan Xue, Tao Wang., 2009. The development of simultaneous partial nitrification, ANAMMOX and denitrification (SNAD) process in a single reactor for nitrogen removal. Bioresource Technology 100, 1548–1554.
Hwang, S., K. Jang, H. Jang, J. Song, W. Bae, 2006. Factors afftcting nitrous oxide production: a comparison of biological nitrogen removal processes with partial and complete nitrification. Biodegradation 17, 19–29.
Isaka, K., T. Sumino, S. Tsuneda., 2007. High nitrogen removal performance at moderately low temperature utilizing anaerobic ammonium oxidation reactions, J. Biosci. Bioeng. 103 (5), 486–490.
Isaka, K. Y. Date, Y. Kimura, T. Sumino, S. Tsuneda, 2008. Nitrogen removal performance using anaerobic ammonium oxidation at low temperatures, FEMS Microbiol. Lett. 282 (1), 32–38.
Itokawa, H., K. Hanaki, T. Matsuo, 2001. Nitrous oxide production in high-loading biological nitrogen removal process under low COD/N ratio condition. Water Res 35(3), 657-64.
Jan Vymazal, 2007. Removal of nutrients in various types of constructed wetlands. Science of the Total Environment 380, 48-65.
Jaroszynski, L.W., N. Cicek, R. Sparling, J.A. Oleszkiewicz., 2011. Importance of the operating pH in maintaining the stability of anoxic ammonium oxidation (Anammox) activity in moving bed biofilm reactors, Bioresour. Technol. 102 (14), 7051–7056.
Jetten, M. S. M., S. Logemann, G. Muyzer, L. A. Robertson, S. deVries, M. C. M. vanLoosdrecht and J. G. Kuenen,1997. Novel principles in the microbial conversion of nitrogen compounds. Antonie Van Leeuwenhoek International. Journal of General and Molecular Microbiology 71(1-2), 75-93.
Jetten, M. S. M., M. Wagner, J. Fuerst, M. van Loosdrecht,G. Kuenen, and M. Strous, 2001. Microbiology and application of the anaerobic ammonium oxidation (‘anammox’) process. Current Opinion in Biotechnology 12(3), 283-288.
Jetten, M.S.M., O. Sliekers, M. Kuypers, T. Dalsgaard, L. van Niftrik, I. Cirpus, K. van de Pas-Schoonen, G. Lavik, B. Thamdrup, D. Le Paslier et al., 2003. Anaerobic ammonium oxidation by marine and freshwater planctomycete-like bacteria. Applied Microbiology and Biotechnology 63(2), 107-114.
Kadlec, R. H., Knight, R. L.,1996. Treatment Wetlands. Lewis Publishers, Michigan.
Kalyuzhnyia, S., M. Gladchenkoa, A. Mulderb, Bram Versprillec., 2006. DEAMOX—New biological nitrogen removal process based on anaerobic ammonia oxidation coupled to sulphide-driven conversion of nitrate into nitrite. Water Res. 40, 3637– 3645.
Kartal, B., J. Rattray, L.A. van Niftrik, J. van de Vossenberg, M.C. Schmid, R.I. Webb, et al., 2007. Candidatus ‘Anammoxoglobus propionicus’ a new propionate oxidizing species of anaerobic ammonium oxidizing bacteria. Syst. Appl. Microbiol. 30, 39–49.
Kartal, B., M. Koleva, R. Arsov, W. van der Star, M.S.M. Jetten, M. Strous., 2006. Adaptation of a freshwater Anammox population to high salinity wastewater. J. Biotechnology 126 (4), 546–553.
Keisuke Hanaki, Chalermraj Wantawin and Shinichiro Ohgaki., 1990.Nitrification at low levels of dissolved oxygen with and without organic loading in a suspended-growth reactor. Water Res. 24,297-302.
Körner, H., Zumft WG., 1989. Expression of denitrification enzymes in response to the dissolved oxygen level and respiratory substrate in continuous culture of Pseudomonas stutzeri. Appl Environ Microbiol. 55(7), 1670-6.
Kuai, L., and W. Verstraete, 1998. Ammonium removal by the oxygen-limited autotrophic nitrification-denitrification system. Applied and Environmental Microbiology 64(11), 4500-4506.
Kuypers, M. M. M., A. O. Sliekers, G. Lavik, M. Schmid, B. B. Jørgensen, J. G. Kuenen, J. S. Sinninghe Damste, M. Strous and M. S. M. Jetten., 2003. Anaerobic ammonium oxidation by anammox bacteria in the black sea.Nature 422, 608.
Lackner, S., C. Lindenblatt, H. Horn., 2012. ‘Swinging ORP’ as operation strategy for stable reject water treatment by nitritation–Anammox in sequencing batch reactors, Chem. Eng. J. 180, 190–196.
Liao Dexiang, LI Xiaoming, YANG Qi, ZENG Guangming, GUO Liang, YUE Xiu., 2008. Effect of inorganic carbon on anaerobic ammonium oxidation enriched in sequencing batch reactor, J. Environ. Sci 20 (8), 940–944.
Lindsay MR, Webb RI, Strous M, Jetten MS, Butler MK, Forde RJ, Fuerst JA., 2001. Cell compartmentalisation in planctomycetes: novel types of structural organisation for the bacterial cell. Arch Mi crobiol 175(6),413-29.
Ling Wang, Tian Li, 2011. Anaerobic ammonium oxidation in constructed wetlands with bio-contact oxidation as pretreatment. Ecological Engineering 37, 1225–1230.
Mark Poth and Dennis D. Focht, 1985. 15N Kinetic Analysis of N2O Production byNitrosomonas europaea: an Examination of Nitrifier Denitrification. Appl Environ Microbiol. 49(5), 1134–1141.
Morris Michael C., 1999. The effects of substrate particle size, depth and vegetation on ammonia removal in a vertical flow constructed wetland. In: J. Vymazal (Ed.) Nutrient Cycling and Retention in Natural and Constructed Wetlands. Backhuys Publishers, Leiden, The Netherlands, 31-40.
Mulder, A., A. A. Van de graaf, L. A. Robertson and J. G. Kuenen., 1995. Anaerobic ammonium oxidation discovered in a denitrifying fluidized-bed reactor. Fems Microbiology Ecology 16(3), 177-183.
Platzer, C., 2000. Development of Reed Bed System - a Europea Perspective. In Proceeding of 7th international conference on wetland system for water pollution control.11-16 Nov. Floria.
Quan, Z.X., S.K. Rhee, J.E. Zuo, Y. Yang, J.W. Bae, J.R. Park, et al. 2008. Diversity of ammonium-oxidizing bacteria in a granular sludge anaerobic ammonium-oxidizing (anammox) reactor. Environ Microbiol 10, 3130–3139.
Ren-Cun Jin, Guang-Feng Yang, Jin-Jin Yu, Ping Zheng., 2012. The inhibition of the Anammox process: A review. Chemical Engineering Journal 197, 67–79.
Ritchie GA, Nicholas DJ., 1972. Identification of the sources of nitrous oxide produced by oxidative and reductive processes in Nitrosomonas europaea. Biochem J 126(5), 1181-91.
Rittmann, B. E., Perry L. McCarty., 2001. Environmental Biotechnology: Principles and Applications.
Schmid, M., K. Walsh, R. Webb, W.I.C. Rijpstra, K. van de Pas-Schoonen, , M.J. Verbruggen, et al. 2003. Candidatus ‘Scalindua brodae’, sp. nov., Candidatus ‘Scalindua wagneri’, sp. nov., two new species of anaerobic ammonium oxidizing bacteria. Syst Appl Microbiol 26, 529–538.
Schmid, M., U. Twachtmann, M. Klein, M.Strous, S. Juretschko, M. Jetten, et al., 2000. Molecular evidence for genus level diversity of bacteria capable of catalyzing anaerobic ammonium oxidation. Syst Appl Microbiol 23, 93–106.
Schmidt, I, Bock E., 1997. Anaerobic ammonia oxidation with nitrogen dioxide by Nitrosomonas eutropha. Arch Microbiol 167(2-3),106-11.
Show SP, Fredine CG. 1956. Wetlands of the United States: Their Extent, and Their Value for Waterfowl and Other Wildlife. Circular 39, U.S. Fish and Wildlife Service, U.S. Department of Interior, Washington, D.C., 67.
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