生物膜脱氮滤池的微生物群落结构特性
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摘要
为探究生物膜脱氮滤池脱氮效能差异的微生物因素,设置组1(包括接种污泥、稳定期滤料、反冲洗后滤料微生物变化)和组2(包括反冲洗前滤料、反冲洗后不同时间段滤料微生物变化)两组实验,通过高通量测序,研究不同阶段微生物群落结构、丰度和多样性的分布情况.结果表明:接种污泥与滤料表面微生物群落结构和多样性在门水平下差异不大,但在属水平下差异显著,而相对丰度在两种分类水平下始终差异显著.反冲洗后0～4 h滤料表面微生物的OTU数目、Shannon指数、Chao1指数变化,在滤池30 cm处先升后降,而在60 cm处曲折上升,也即二者的微生物丰度和多样性变化趋势.反冲洗前后Proteobacteria始终占主导地位,相对丰度为79%～90%,Proteobacteria中Betaproteobacteria占主导优势,反冲洗后Betaproteobacteria数量相对减少,后逐渐恢复.总反硝化优势菌属相对丰度在反冲洗1 h后30 cm处由78%左右下降到70%左右,60 cm处由68%左右下降到64%左右,此时出水总氮达到最高值7.7 mg·L~(-1),之后总反硝化优势菌属及出水TN浓度逐渐恢复至正常水平,这种消长变化表明滤池脱氮效果与总反硝化优势菌属相对丰度密切相关,滤池运行状态的改变使得反硝化优势菌属的群落结构差异显著.
In order to investigate the effect of microorganisms on nitrogen removal efficiency of a denitrification biofilm filter, two experiments were conducted, i.e., experiment 1(inoculated sludge, filter media in a stable period and filter media after backwashing) and experiment 2(the filter media before backwashing and after backwashing at different periods). High-throughput sequencing was applied to analyze the distribution of microbial community structure, abundance and diversity at different stages. There were no differences of the microbial community structure and diversity of the inoculated sludge and filter media at the phylum level. However, a significant difference at the genus level was observed. Meanwhile, the relative abundance was always significant at the two classification levels. In the first four hours after backwashing, the OTU number, Shannon index and Chao1 index of the microbes on the surface of the filter media were increased first and then decreased at 30 cm. In addition, it ascended with an upward tendency at 60 cm. Proteobacteria was dominant before and after backwashing with a relative abundance of 79%～90%. Betaproteobacteria was dominant in Proteobacteria. Its number decreased after backwashing and then recovered gradually. In the first 1 h after backwashing, the relative abundance of the total dominant denitrifiers decreased from 78% to 70% at 30 cm, from 68% to 64% at 60 cm. At the same time, the total nitrogen of the effluent reached to the highest value of 7.7 mg·L~(-1). The total dominant denitrifiers and the total nitrogen of the effluent were gradually recovered to its normal level. The results of this study demonstrated that the nitrogen removal efficiency was closely related to the relative abundance of the total dominant denitrifiers. The difference of the community structure of the dominant denitrifiers was due to changes of the operational state of the filter.
In order to investigate the effect of microorganisms on nitrogen removal efficiency of a denitrification biofilm filter, two experiments were conducted, i.e., experiment 1(inoculated sludge, filter media in a stable period and filter media after backwashing) and experiment 2(the filter media before backwashing and after backwashing at different periods). High-throughput sequencing was applied to analyze the distribution of microbial community structure, abundance and diversity at different stages. There were no differences of the microbial community structure and diversity of the inoculated sludge and filter media at the phylum level. However, a significant difference at the genus level was observed. Meanwhile, the relative abundance was always significant at the two classification levels. In the first four hours after backwashing, the OTU number, Shannon index and Chao1 index of the microbes on the surface of the filter media were increased first and then decreased at 30 cm. In addition, it ascended with an upward tendency at 60 cm. Proteobacteria was dominant before and after backwashing with a relative abundance of 79%～90%. Betaproteobacteria was dominant in Proteobacteria. Its number decreased after backwashing and then recovered gradually. In the first 1 h after backwashing, the relative abundance of the total dominant denitrifiers decreased from 78% to 70% at 30 cm, from 68% to 64% at 60 cm. At the same time, the total nitrogen of the effluent reached to the highest value of 7.7 mg·L~(-1). The total dominant denitrifiers and the total nitrogen of the effluent were gradually recovered to its normal level. The results of this study demonstrated that the nitrogen removal efficiency was closely related to the relative abundance of the total dominant denitrifiers. The difference of the community structure of the dominant denitrifiers was due to changes of the operational state of the filter.
引文
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Thomsen T R, Kong Y H, Nielsen P H. 2007. Ecophysiology of Abundant Denitrifying Bacteria in Activated Sludge[J]. Fems Microbiology Ecology, 60(3): 370-382
Vanden Hoven R N, Santini J M. 2004. Arsenite Oxidation By the Heterotroph Hydrogenophaga Sp. Str. Nt-14: the Arsenite Oxidase and Its Physiological Electron Acceptor[J]. Biochimica Et Biophysica Acta: Bioenergetics, 1656(2-3): 148-155
王绍祥,杨洲祥,孙真,等. 2014. 高通量测序技术在水环境微生物群落多样性中的应用[J]. 化学通报, 77(3): 196-203
Xu X L, Liu G H, Wang Y Y, et al. 2018. Analysis of Key Microbial Community During the Start-up of Anaerobic Ammonium Oxidation Process with Paddy Soil as Inoculated Sludge[J]. Journal of Environmental Sciences, 64: 317-327
窦娜莎. 2013. 曝气生物滤池处理城市污水的效能与微生物特性研究[D]. 青岛:中国海洋大学
窦娜莎,王琳. 2011. 16S rDNA克隆文库法分析Biostyr曝气生物滤池处理城市污水的细菌多样性研究[J]. 环境科学学报, 31(10): 2117-2124
Feng Y, Li X, Song T, et al. 2017. Effect of Backwashing on the Microbial Community Structure and Composition of a Three Dimensional Particle Electrode Coupled with Biological Aerated Filter Reactor (tde-baf)[J]. Ecological Engineering, 101: 21-27
Fischer S G, Lerman L S. 1979. Length-independent Separation of Dna Restriction Fragments in Two-dimensional Gel Electrophoresis[J]. Cell, 16(1): 191-200
Figuerola E L M, Erijman L. 2007. Bacterial Taxa Abundance Pattern in an Industrial Wastewater Treatment System Determined By the Full Rrna Cycle Approach[J]. Environmental Microbiology, 9(7): 1780-1789
高建锋,杨碧印,赵建树,等. 2016. 反硝化生物滤池用于再生水脱氮效能及动力学研究[J]. 环境工程学报, 10(1): 199-204
刘水,李伟光,郜玉楠,等. 2009. 生物因子控制反冲洗对BEAC滤池优势菌稳定性影响研究[J]. 环境科学, 30(12): 3573-3578
路莹. 2013. 浅层地下水系统石油类污染物的生物降解机制研究[D]. 吉林:吉林大学
Mao Y P, Xia Y, Zhang T. 2013. Characterization of Thauera-dominated Hydrogen-oxidizing Autotrophic Denitrifying Microbial Communities By Using High-throughput Sequencing[J]. Bioresource Technology, 128(1): 703-710
Shen Z Q, Zhou Y X, Hu J, et al. 2013. Denitrification Performance and Microbial Diversity in a Packed-bed Bioreactor Using Biodegradable Polymer as Carbon Source and Biofilm Support[J]. Journal of Hazardous Materials, 250(Supplement C): 431-438
Thomsen T R, Kong Y H, Nielsen P H. 2007. Ecophysiology of Abundant Denitrifying Bacteria in Activated Sludge[J]. Fems Microbiology Ecology, 60(3): 370-382
Vanden Hoven R N, Santini J M. 2004. Arsenite Oxidation By the Heterotroph Hydrogenophaga Sp. Str. Nt-14: the Arsenite Oxidase and Its Physiological Electron Acceptor[J]. Biochimica Et Biophysica Acta: Bioenergetics, 1656(2-3): 148-155
王绍祥,杨洲祥,孙真,等. 2014. 高通量测序技术在水环境微生物群落多样性中的应用[J]. 化学通报, 77(3): 196-203
Xu X L, Liu G H, Wang Y Y, et al. 2018. Analysis of Key Microbial Community During the Start-up of Anaerobic Ammonium Oxidation Process with Paddy Soil as Inoculated Sludge[J]. Journal of Environmental Sciences, 64: 317-327