甲烷厌氧氧化反硝化微生物的富集

甲烷厌氧氧化反硝化微生物的富集(论文17000字)
摘要
目前，硝酸盐污染是全球水体恶化的重要原因之一，已严重影响到人们的健康与生活，亟待解决。甲烷作为温室气体之一，可引起严重的温室效应。甲烷厌氧氧化反硝化（denitrifying anaerobic methane oxidation,DAMO）即在厌氧环境下，同步实现甲烷厌氧氧化和反硝化反应，在处理污水脱氮和控制温室气体排放工艺中具有广阔的应用前景。本文以骆马湖、佛手湖、东海中水样为原始接种源，人工模拟硝酸盐污染水体，构建中空纤维生物膜和厌氧血清瓶富集反应器，培养富集甲烷氧化反硝化菌群，测定水样中硝酸盐氮、亚硝酸盐氮浓度，确定甲烷厌氧氧化反硝化效果。实验结果表明，骆马湖富集的甲烷厌氧氧化反硝化微生物的反硝化效果最好，硝酸氮去除率达到80.75%，中空纤维膜生物膜反应器可缩短微生物富集时间，但活性依旧较低。总结了实验过程中存在的问题，提出微生物测序是下一步研究方向，并对以后关于DAMO的发展方向做出了展望，如优化反应器设计、对功能微生物影响因素进行研究，采用电化学方法加快反硝化速率等。
关键词：甲烷厌氧氧化反硝化微生物DAMO硝酸盐氮富集

Enrichment of denitrifying methane anaerobic oxidation microorganisms
Abstract
At present, nitrate pollution is an important cause of the deterioration of global water bodies, which has seriously affected people's health and life and needs to be resolved. As one of the greenhouse gases, methane can cause a serious greenhouse effect. Denitrifying anaerobic methane oxidation (DAMO) is a simultaneous realization of methane anaerobic oxidation and NO3-/NO2-denitrification in an anaerobic environment.It has broad application prospects in the process of treating wastewater denitrification and controlling greenhouse gas emissions. In this paper, the water samples from Luoma Lake, Fossil Lake and East China Sea in Zhoushan were used as research objects to artificially simulate nitrate-contaminated water bodies. Hollow fiber biofilm and anaerobic serum bottle reactors were constructed for different inoculum and culture medium substrates to culture and enrich methane oxidation. The denitrifying bacteria group measures the concentration of nitrate nitrogen and nitrite nitrogen in the water sample to determine the anaerobic oxidation and denitrification effect of methane. The data showed that the anaerobic oxidation and denitrification effect of methane in Loma Lake was the best, and the nitrogen nitrate removal rate reached 80.75%. The hollow fiber membrane biofilm reactor can shorten the microbial enrichment time, but the activity is still low. The problems in the experiment process are summarized. It is proposed that the PCR amplification and sequencing of microbial genes is the next research direction, and the future development direction of DAMO is prospected. For example, the design of the reactor is optimized, the research on the factors affecting the functional microorganisms and use the electrochemical method.
Key Words:Methane ; anaerobic oxidation denitrification ; microorganism ; DAMO ; nitrate nitrogen ; enrichment

目录
摘要    - I -
Abstract    - II -
第一章绪论    - 1 -
1.1研究背景    - 1 -
1.1.1地表地下水污染问题    - 1 -
1.1.2硝酸盐污染危害    - 1 -
1.1.3国内外研究现状    - 1 -
1.1.4生物反硝化    - 2 -
1.1.5甲烷氧化反硝化    - 2 -
1.1.6 好氧甲烷反硝化机理    - 4 -
1.1.7 厌氧甲烷反硝化机理    - 4 -
1.2课题研究内容    - 5 -
1.3研究目的、意义及技术路线    - 6 -
1.3.1研究目的    - 6 -
1.3.2研究意义    - 6 -
1.3.3技术路线    - 6 -
第二章实验材料与方法    - 8 -
2.1实验装置    - 8 -
2.1.1中空纤维生物膜反应器    - 8 -
2.1.2厌氧血清瓶反应器    - 10 -
2.2水样采集    - 11 -
2.2.1骆马湖    - 11 -
2.2.2佛手湖    - 12 -
2.2.3东海    - 12 -
2.3微生物富集方法与水样检测方法    - 13 -
2.3.1微生物富集方法    - 13 -
2.3.2水样检测方法    - 15 -
2.3.3DAMO效果评定方案    - 18 -
第三章实验结果与讨论    - 19 -
3.1骆马湖水样中甲烷厌氧氧化反硝化微生物的富集    - 19 -
3.1.1硝酸根浓度变化规律    - 19 -
3.1.2亚硝酸根浓度变化规律    - 20 -
3.1.2 eqp含量变化规律    - 21 -
3.2佛手湖水样中甲烷厌氧氧化反硝化微生物的富集    - 22 -
3.2.1硝酸根浓度变化规律    - 22 -
3.2.2亚硝酸根浓度变化规律    - 23 -
3.2.2 eqp含量变化规律    - 23 -
3.3东海水样中甲烷厌氧氧化反硝化微生物的富集    - 24 -
3.3.1硝酸根浓度变化规律    - 24 -
3.3.2亚硝酸根浓度变化规律    - 27 -
3.3.3 eqp含量变化规律    - 29 -
3.4本章小结    - 30 -
第四章结论与展望    - 32 -
4.1结论与存在问题    - 32 -
4.1.1结论    - 32 -
4.1.2存在问题    - 33 -
4.2展望    - 33 -
参考文献    - 35 -
致谢    - 39 -