摘要
反硝化厌氧甲烷氧化(Denitrifying anaerobic methane oxidation，DAMO)耦联了甲烷氧化和硝酸盐/亚硝酸盐的还原过程，具有脱氮和温室气体减排双重作用，对全球碳氮循环研究，温室气体减排和废水脱氮新技术开发等方面具有重要作用。但是DAMO微生物生长缓慢，脱氮速率低，对其长期培养特性认识不足等问题限制了该过程走向工程应用。本论文通过长期对比运行DAMO微生物反应器，进一步认识SBR中硝酸盐型和亚硝酸型DAMO过程速率和微生物学特性的差异，调查膜生物反应器(Membrane bioreactor, MBR)中的DAMO过程和运行模式对SBR 中DAMO过程强化效应。主要取得了以下结论：
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①在相同条件下分别以硝酸盐和亚硝酸盐在SBR中长期运行DAMO，前者微生物优势菌为DAMO古菌和DAMO细菌，后者的微生物优势菌为DAMO细菌；DAMO古菌混合体系达到稳定比DAMO细菌晚150 d ，速率上升期比细菌长70 d；DAMO细菌最大脱氮速率达到80.9 mg-N/(L·d)，DAMO古菌为32.6 mg-N/(L·d) ，DAMO古菌最终脱氮速率为DAMO菌的40 %左右；反应器中DAMO古菌拷贝数比DAMO细菌低出一个量级。以硝酸盐为电子受体的DAMO体系的微生物富集速率和运行效率的提升仍需进一步研究。
②DAMO古菌和DAMO细菌混合体系可以长期稳定共存，DAMO古菌可能是混合体系脱氮活性的限制性因素。DAMO古菌和DAMO细菌混合体系速率达到稳定后，培养800 d脱氮活性未下降，反应体系中没有亚硝酸盐的积累。
③MBR能够稳定培养DAMO细菌，具有一定优势。低速波动时间相较原SBR 时间缩短一半，快速提升达到最终稳定的时间较短，动力学参数NO2--N半饱和常数Ks为0.62 mg/L，远小于已报道值12.6 mg/L。MBR运行一年未发生严重膜污染。
④排泥对SBR中亚硝酸盐型DAMO过程的脱氮性能提升和功能微生物群落优化有明显效果：与对照组相比，调控组的脱氮速率明显增加，从调控前的17.00 mg-N/(L·d)提升到到70.61 mg-N/(L·d)，显示出工程应用潜力；调控组相对比活性提高显著，对比对照组最高提升5.88倍。调控后，实验组DAMO细菌由1.17×108 copies/g d，增长到4.02×108 copies/g d，提高了3.44倍。DAMO细菌所占比例由38.3 %上升到67.7%，纯度提高了1.77倍。minimoto
关键词：反硝化厌氧甲烷氧化，序批式反应器，长期运行，膜生物反应器，排泥
ABSTRACT
Denitrifying anaerobic methane oxidation (DAMO) couples methane oxidation and nitrate/nitrite reduction process and has the dual functions of nitrogen removal and greenhou gas emission reduction. It plays an important role in the rearch of the global carbon and nitrogen cycle, the reduction of greenhou gas, and the development of new technologies for the nitrogen removal from wastewater. However, the problems that it is difficult for DAMO to grow, the rate of nitrogen rem
oval is too low to meet actual needs, and the lack of knowledge of long-term culture performance limits its application to engineering. So we conducted a long-term comparison of the cultivation of DAMO microbial reactors in order to further understand the differences in the process rate and microbiological characteristics of nitrate and nitrite type DAMO in SBR. And a membrane bioreactor (MBR) was designed to investigate the DAMO performance. Furthermore we studied the enhancement effect of running mode in SBR. The main conclusions are as follows:
①Under the same conditions, nitrate and nitrite were ud as electron acceptor of DAMO in the long-term operation of SBR. As a result a The reactor containing only the substrate of nitrate cultures the mixture system of DAMO archaea and DAMO bacteria, and the dominant microorganism cultured in the reactor containing nitrite substrate is the DAMO bacteria. Morover, the DAMO archaeal mixed system reached stability 150 days later than the DAMO bacteria, and the time of rate increa was 70 days longer than that of the bacteria. As for the performance of nitrogen removal, the maximum nitrogen removal rate of DAMO bacteria reached 80.9 mg-N/(L·d), and that of DAMO was 32.6 mg-N/(L·d). The final nitrogen removal rate of DAMO archaea system was only about 40% of that of DAMO bacteria and the DAMO bacterial copy number is one order of magnitude higher than the DAMO archaea. Further studies on the microbial enrichment rate and operational efficiency of DAMO system with nitrate as electron acceptor still need further study.
②In the long-term culture, DAMO archaea and DAMO bacteria can stably coexist, and DAMO archaea may be the limiting factor in the nitrogen removal activity of the mixed system. After the rate of the DAMO bacteria mixture system of DAMO was stable, the nitrogen removal activity did not decrea during the 800 d and there was no accumulation of nitrite in the reaction system.
重庆大学硕士学位论文
③MBR can stably cultivate DAMO bacteria but also has certain advantages. The low-speed fluctuation time is reduced by half compared with the original SBR time, and the time for rapid improvement to achieve final stabilization is short. The kinetic parameter of NO2--N half saturation constant Ks is 0.62 mg/L, far less than the reported value of 12.6 mg/L. Further more, no vere membrane fouling occurs in the MBR operation for one year.
④Sludge removal has obvious effect on nitrogen removal performance and functional microbial community optimization in nitrite type DAMO process in SBR. Compared with the control group, the denitrification rate of the control group is significantly incread, from 17.00 mg-N/(L·d) before the control to 70.61 mg-N/(L·d), showing the application potential of the process. And the contrast activity of the control group is significantly improved. The highest level of the control group was 5.88
七年级数学补习times higher than that in the control group. After adjustment, he copies of DAMO bacteria incread from 1.17×108 copies/g d to 4.02×108 copies/g d, increasing by 3.44 times. But also, the proportion of DAMO bacteria incread from 38.3% to 67.7%, and the purity incread by 1.77 times.
Key words：Denitrification anaerobic methane oxidation, SBR,Long-term contrast culture, MBR, Sludge
IV
目录
目录
甜心辣舞中文摘要.......................................................................................................................................... I 英文摘要....................................................................................................................................... III 1 绪论 . (1)
1.1 问题的提出及研究意义 (1)
1.1.1问题的提出 (1)
interchange1.1.2 研究的意义 (2)
1.2 国内外研究现状 (2)
1.2.1 DAMO微生物的富集及其影响因素 (2)
1.2.2 DAMO微生物的生理特性与代谢机理 (6)
1.2.3 DAMO过程的工程化应用潜力 (13)
1.3研究内容与方案 (17)
qv1.3.1 主要研究内容 (17)
1.3.2 技术路线 (17)
1.3.3 难点与创新点 (18)重庆科技学院是几本
2 SBR中长期运行DAMO过程的比较研究 (21)
2.1 材料与方法 (21)
2.1.1 接种物与培养基质 (21)
2.1.2 反应器运行条件 (22)
2.1.3 分析指标与测试方法 (24)
pirate bay2.2 结果 (27)
2.2.1 基质转化速率 (27)
2.2.2 功能微生物种群结构 (29)
2.3 讨论 (30)
2.3.1长期运行DAMO古菌更难培养 (30)
2.3.2 DAMO古菌系统能够长期稳定 (31)
2.3.3 换水出现短期活性提升 (31)
2.4 本章小结 (31)
3 MBR中运行DAMO过程实验研究 (33)
3.1 材料与方法 (34)
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3.1.1 MBR系统构建 (34)
3.1.2 接种与运行方案 (36)
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重庆大学硕士学位论文
3.1.3 MBR中NO2-降解动力学实验方法 (38)
3.1.4 分析测试与计算方法 (39)
3.2 结果 (39)
3.2.1 基质去除速率 (39)
3.2.2 基质去除动力学 (41)
3.2.3 膜通量变化 (41)
3.3 讨论 (42)
3.3.1 MBR培养DAMO细菌优势 (42)
3.3.2 MBR培养DAMO细菌动力学 (44)
3.3.3 MBR中基质更新影响 (44)
3.4 本章小结 (45)
4 SBR反应器中DAMO过程的强化 (47)
4.1 材料与方法 (47)
4.1.1 实验材料 (47)
4.1.2 运行模式优化方案 (47)
镁光灯4.1.3 分子生物学测试 (48)
4.2 结果与讨论 (50)
4.2.1 微生物群落结构优化 (50)
4.2.2 脱氮速率性能强化 (52)
4.4 本章小结 (56)
5 结论与展望 (57)
5.1 主要结论 (57)
5.2 工作展望 (57)
致谢 (59)
参考文献 (61)
附录 (73)
A. 作者攻读硕士学位期间的论文 (73)
B. 作者攻读硕士期间参研的课题 (73)
VI
