城市污水处理系统真核微生物群落特性与地域性差异
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摘要
城市污水处理厂是控制水污染、保证城市与人类可持续发展的重要设施.真核微生物作为活性污泥系统中不可或缺的一部分,在指示活性污泥性质、预测出水水质、强化净化效果、保障系统稳定运行过程中发挥着重要作用.本研究以来自北京、深圳和无锡的14个城市污水处理厂的61份样品为研究对象,采用18S r DNA高通量测序技术并结合多种生态学及统计学相关分析方法,探讨了活性污泥系统内真核微生物群落特性与地域性差异.结果表明,不同空间尺度下,活性污泥系统内真核微生物群落优势种群组成相似,在Devision水平上,群落优势种群主要由真菌、原生动物(纤毛亚门)和后生动物组成,三者相对丰度之和高达86. 22%～89. 40%.不同城市污水处理厂活性污泥系统内真核微生物多样性存在差异,无锡群落的多样性(丰富度和香农-威纳多样性指数)最高,北京的最低,且3个城市的活性污泥系统内真核微生物群落结构存在显著的地域性差异. partial mantel test和多元回归矩阵分析结果表明,曝气池混合液温度和出水总氮浓度与活性污泥系统内真核微生物群落结构的相关性较强.研究结果深化了对污水处理系统中真核微生物的认识.
Wastewater treatment plants( WWTPs) are important facilities to control water pollution and ensure the sustainable development of cities and humans. As an indispensable part of the activated sludge( AS) system,eukaryotic microbes play important roles in indicating the properties of AS,predicting the quality of the effluent,enhancing the purification effect,and ensuring a stable performance of the system in WWTPs. In this study,61 AS samples from 14 full-scale WWTPs of Beijing,Shenzhen,and Wuxi were collected. Characteristics and regional heterogeneity of eukaryotic microbial community were elucidated via high-throughput sequencing( HTS) of the 18 S r DNA and multi ecological and statistical methods. Results showed that eukaryotic microbial communities in different scales shared similar main members,which were mainly composed of fungi,ciliophora,and metazoa in division level with their total relative abundance up to 86. 22%-89. 40%. Diversity of eukaryotic microbial community in WWTPs of different cites varied. Richness and Shannon Wiener index of eukaryotic microbial communities in the AS system of Wuxi were the highest,while that of Beijing was the lowest. Diversity of eukaryotic microbes from HTS in this study was higher than that of conventional methods,but lower than the diversity of bacteria in AS systems. Regional heterogeneity of eukaryotic microbial community structure was uncovered by nonmetric multidimensional scaling based on Bray-Curtis distance and dissimilarity analysis. Results of partial mantel test and multiple regression matrix analysis showed that the eukaryotic microbial community was significantly correlated with the temperature of the aeration tank mixture and the total nitrogen concentration of the effluent of the AS system. These results help deepen the understanding of eukaryotic microbes in WWTPs.
Wastewater treatment plants( WWTPs) are important facilities to control water pollution and ensure the sustainable development of cities and humans. As an indispensable part of the activated sludge( AS) system,eukaryotic microbes play important roles in indicating the properties of AS,predicting the quality of the effluent,enhancing the purification effect,and ensuring a stable performance of the system in WWTPs. In this study,61 AS samples from 14 full-scale WWTPs of Beijing,Shenzhen,and Wuxi were collected. Characteristics and regional heterogeneity of eukaryotic microbial community were elucidated via high-throughput sequencing( HTS) of the 18 S r DNA and multi ecological and statistical methods. Results showed that eukaryotic microbial communities in different scales shared similar main members,which were mainly composed of fungi,ciliophora,and metazoa in division level with their total relative abundance up to 86. 22%-89. 40%. Diversity of eukaryotic microbial community in WWTPs of different cites varied. Richness and Shannon Wiener index of eukaryotic microbial communities in the AS system of Wuxi were the highest,while that of Beijing was the lowest. Diversity of eukaryotic microbes from HTS in this study was higher than that of conventional methods,but lower than the diversity of bacteria in AS systems. Regional heterogeneity of eukaryotic microbial community structure was uncovered by nonmetric multidimensional scaling based on Bray-Curtis distance and dissimilarity analysis. Results of partial mantel test and multiple regression matrix analysis showed that the eukaryotic microbial community was significantly correlated with the temperature of the aeration tank mixture and the total nitrogen concentration of the effluent of the AS system. These results help deepen the understanding of eukaryotic microbes in WWTPs.
引文
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[30]Mc Gill B J,Etienne R S,Gray J S,et al.Species abundance distributions:moving beyond single prediction theories to integration within an ecological framework[J].Ecology Letters,2007,10(10):995-1015.
[31]Nemergut D R,Schmidt S K,Fukami T,et al.Patterns and processes of microbial community assembly[J].Microbiology and Molecular Biology Reviews,2013,77(3):342-356.
[32]Matsunaga K,Kubota K,Harada H.Molecular diversity of eukaryotes in municipal wastewater treatment processes as revealed by 18S rRNA gene analysis[J].Microbes and Environments,2014,29(4):401-407.
[33]Matsubayashi M,Shimada Y,Li Y Y,et al.Phylogenetic diversity and in situ detection of eukaryotes in anaerobic sludge digesters[J].PLo S One,2017,12(3):e0172888.
[34]Santoferrara L F,Grattepanche J D,Katz L A,et al.Pyrosequencing for assessing diversity of eukaryotic microbes:analysis of data on marine planktonic ciliates and comparison with traditional methods[J].Environmental Microbiology,2014,16(9):2752-2763.
[35]Kunin V,Engelbrektson A,Ochman H,et al.Wrinkles in the rare biosphere:pyrosequencing errors can lead to artificial inflation of diversity estimates[J].Environmental Microbiology,2010,12(1):118-123.
[36]Xia Y,Hu M,Wen X H,et al.Diversity and interactions of microbial functional genes under differing environmental conditions:insights from a membrane bioreactor and an oxidation ditch[J].Scientific Reports,2016,6:18509.
[37]Brte J,Logares R,Berney C,et al.Freshwater perkinsea and marine-freshwater colonizations revealed by pyrosequencing and phylogeny of environmental r DNA[J].The ISME Journal,2010,4(9):1144-1153.
[38]Chesson P.Mechanisms of maintenance of species diversity[J].Annual Review of Ecology and Systematics,2000,31:343-366.
[39]Hubbell S P.The unified neutral theory of biodiversity and biogeography[M].Princeton:Princeton University Press,2001.
[40]Wang X H,Hu M,Xia Y,et al.Pyrosequencing analysis of bacterial diversity in 14 wastewater treatment systems in China[J].Applied and Environmental Microbiology,2012,78(19):7042-7047.
[41]Liu J,Yang M,Qi R,et al.Comparative study of protozoan communities in full-scale MWTPs in Beijing related to treatment processes[J].Water Research,2008,42(8-9):1907-1918.
[42]Hu M,Wang X H,Wen X H,et al.Microbial community structures in different wastewater treatment plants as revealed by454-pyrosequencing analysis[J].Bioresource Technology,2012,117:72-79.
[43]Lepère C,Boucher D,Jardillier L,et al.Succession and regulation factors of small eukaryote community composition in a lacustrine ecosystem(Lake Pavin)[J].Applied and Environmental Microbiology,2006,72(4):2971-2981.
[44]Wei Z Y,Liu Y Y,Feng K,et al.The divergence between fungal and bacterial communities in seasonal and spatial variations of wastewater treatment plants[J].Science of the Total Environment,2018,628-629:969-978.
[45]Tyagi V K,Subramaniyan S,Kazmi A A,et al.Microbial community in conventional and extended aeration activated sludge plants in India[J].Ecological Indicators,2008,8(5):550-554.
[46]齐嵘,刘娟,杨敏,等.无机废水硝化过程中原生动物群落结构特征及其变化规律[J].环境科学学报,2013,33(1):53-57.Qi R,Liu J,Yang M,et al.Study on protozoan community structures during the inorganic wastewater nitrification process[J].Acta Scientiae Circumstantiae,2013,33(1):53-57.
[47]Bahram M,Hildebrand F,Forslund S K,et al.Structure and function of the global topsoil microbiome[J].Nature,2018,560(7717):233-237.
[48]Telford R J,Vandvik V,Birks H J B.Dispersal limitations matter for microbial morphospecies[J].Science,2006,312(5776):1015.
[49]Whitaker R J,Grogan D W,Taylor J W.Geographic barriers isolate endemic populations of hyperthermophilic archaea[J].Science,2003,301(5635):976-978.
[50]Cho J C,Tiedje J M.Biogeography and degree of endemicity of fluorescent Pseudomonas strains in soil[J].Applied and Environmental Microbiology,2000,66(12):5448-5456.
[51]Borcard D,Legendre P.Environmental control and spatial structure in ecological communities:an example using oribatid mites(Acari,Oribatei)[J].Environmental and Ecological Statistics,1994,1(1):37-61.
[52]Vuono D C,Benecke J,Henkel J,et al.Disturbance and temporal partitioning of the activated sludge metacommunity[J].The ISME Journal,2015,9(2):425-435.
[2]Madoni P.A sludge biotic index(SBI)for the evaluation of the biological performance of activated sludge plants based on the microfauna analysis[J].Water Research,1994,28(1):67-75.
[3]Madoni P.Protozoa in wastewater treatment processes:a minireview[J].Italian Journal of Zoology,2011,78(1):3-11.
[4]SalvadóH,Gracia M P.Determination of organic loading rate of activated sludge plants based on protozoan analysis[J].Water Research,1993,27(5):891-895.
[5]Hashimoto A,Kunikane S,Hirata T.Prevalence of Cryptosporidium oocysts and Giardia cysts in the drinking water supply in Japan[J].Water Research,2002,36(3):519-526.
[6]Rittmann B E,Mc Carty P L.环境生物技术原理与应用[M].文湘华,王建龙,译.北京:清华大学出版社,2012.
[7]Zheng S K,Sun J Y,Han H.Effect of dissolved oxygen changes on activated sludge fungal bulking during lab-scale treatment of acidic industrial wastewater[J].Environmental Science&Technology,2011,45(20):8928-8934.
[8]Eschenhagen M,Schuppler M,Roske I.Molecular characterization of the microbial community structure in two activated sludge systems for the advanced treatment of domestic effluents[J].Water Research,2003,37(13):3224-3232.
[9]Karayanni H,Christaki U,Van Wambeke F,et al.Evaluation of double formalin-Lugol's fixation in assessing number and biomass of ciliates:an example of estimations at mesoscale in NEAtlantic[J].Journal of Microbiological Methods,2004,56(3):349-358.
[10]郑祥,刘俊新.膜生物反应器系统中原生动物的群落特征[J].环境科学,2009,30(9):2635-2640.Zheng X,Liu J X.Protozoan community diversity in membrane bioreactor[J].Environmental Science,2009,30(9):2635-2640.
[11]Meng Z C,Xu K D,Dai R H,et al.Ciliate community structure,diversity and trophic role in offshore sediments from the Yellow Sea[J].European Journal of Protistology,2012,48(1):73-84.
[12]代仁海.黄东海底栖纤毛虫多样性及微型底栖生物群落结构特点[D].青岛:中国科学院研究生院(海洋研究所),2012.
[13]Moll J,Hoppe B,K9nig S,et al.Spatial distribution of fungal communities in an arable soil[J].PLo S One,2016,11(2):e0148130.
[14]Zhou J Z,Deng Y,Shen L N,et al.Temperature mediates continental-scale diversity of microbes in forest soils[J].Nature Communications,2016,7:12083.
[15]Wang Z,Xu G J,Xu H L.Determiningβ-diversity of protozoa for bioassessment in coastal ecosystems using community-based dispersions[J].Ecological Indicators,2017,72:47-52.
[16]Fuhrman J A,Steele J A,Hewson I,et al.A latitudinal diversity gradient in planktonic marine bacteria[J].Proceedings of the National Academy of Sciences of the United States of America,2008,105(22):7774-7778.
[17]Shi S J,Nuccio E,Herman D J,et al.Successional trajectories of rhizosphere bacterial communities over consecutive seasons[J].m Bio,2015,6(4):e00746-15.
[18]Wang Y,Sheng H F,He Y,et al.Comparison of the levels of bacterial diversity in freshwater,intertidal wetland,and marine sediments by using millions of Illumina tags[J].Applied and Environmental Microbiology,2012,78(23):8264-8271.
[19]Nagarkar M,Countway P D,Yoo Y D,et al.Temporal dynamics of eukaryotic microbial diversity at a coastal Pacific site[J].The ISME Journal,2018,12(9):2278-2291.
[20]Xia Y,Wen X H,Zhang B,et al.Diversity and assembly patterns of activated sludge microbial communities:a review[J].Biotechnology Advances,2018,36(4):1038-1047.
[21]Saunders A M,Albertsen M,Vollertsen J,et al.The activated sludge ecosystem contains a core community of abundant organisms[J].The ISME Journal,2016,10(1):11-20.
[22]Zhang T,Shao M F,Ye L.454 Pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants[J].The ISME Journal,2012,6(6):1137-1147.
[23]Schmidt P A,Bálint M,Greshake B,et al.Illumina metabarcoding of a soil fungal community[J].Soil Biology and Biochemistry,2013,65:128-132.
[24]Wang X H,Wen X H,Deng Y,et al.Distance-decay relationship for biological wastewater treatment plants[J].Applied and Environmental Microbiology,2016,82(16):4860-4866.
[25]Stoeck T,Bass D,Nebel M,et al.Multiple marker parallel tag environmental DNA sequencing reveals a highly complex eukaryotic community in marine anoxic water[J].Molecular Ecology,2010,19(S1):21-31.
[26]Feng K,Zhang Z J,Cai W W,et al.Biodiversity and species competition regulate the resilience of microbial biofilm community[J].Molecular Ecology,2017,26(21):6170-6182.
[27]Caporaso J G,Lauber C L,Walters W A,et al.Ultra-highthroughput microbial community analysis on the Illumina Hi Seq and Mi Seq platforms[J].The ISME Journal,2012,6(8):1621-1624.
[28]Li Y,He J Z,He Z L,et al.Phylogenetic and functional gene structure shifts of the oral microbiomes in periodontitis patients[J].The ISME Journal,2014,8(9):1879-1891.
[29]Legendre P,Lapointe F J,Casgrain P.Modeling brain evolution from behavior:a permutational regression approach[J].Evolution,1994,48(5):1487-1499.
[30]Mc Gill B J,Etienne R S,Gray J S,et al.Species abundance distributions:moving beyond single prediction theories to integration within an ecological framework[J].Ecology Letters,2007,10(10):995-1015.
[31]Nemergut D R,Schmidt S K,Fukami T,et al.Patterns and processes of microbial community assembly[J].Microbiology and Molecular Biology Reviews,2013,77(3):342-356.
[32]Matsunaga K,Kubota K,Harada H.Molecular diversity of eukaryotes in municipal wastewater treatment processes as revealed by 18S rRNA gene analysis[J].Microbes and Environments,2014,29(4):401-407.
[33]Matsubayashi M,Shimada Y,Li Y Y,et al.Phylogenetic diversity and in situ detection of eukaryotes in anaerobic sludge digesters[J].PLo S One,2017,12(3):e0172888.
[34]Santoferrara L F,Grattepanche J D,Katz L A,et al.Pyrosequencing for assessing diversity of eukaryotic microbes:analysis of data on marine planktonic ciliates and comparison with traditional methods[J].Environmental Microbiology,2014,16(9):2752-2763.
[35]Kunin V,Engelbrektson A,Ochman H,et al.Wrinkles in the rare biosphere:pyrosequencing errors can lead to artificial inflation of diversity estimates[J].Environmental Microbiology,2010,12(1):118-123.
[36]Xia Y,Hu M,Wen X H,et al.Diversity and interactions of microbial functional genes under differing environmental conditions:insights from a membrane bioreactor and an oxidation ditch[J].Scientific Reports,2016,6:18509.
[37]Brte J,Logares R,Berney C,et al.Freshwater perkinsea and marine-freshwater colonizations revealed by pyrosequencing and phylogeny of environmental r DNA[J].The ISME Journal,2010,4(9):1144-1153.
[38]Chesson P.Mechanisms of maintenance of species diversity[J].Annual Review of Ecology and Systematics,2000,31:343-366.
[39]Hubbell S P.The unified neutral theory of biodiversity and biogeography[M].Princeton:Princeton University Press,2001.
[40]Wang X H,Hu M,Xia Y,et al.Pyrosequencing analysis of bacterial diversity in 14 wastewater treatment systems in China[J].Applied and Environmental Microbiology,2012,78(19):7042-7047.
[41]Liu J,Yang M,Qi R,et al.Comparative study of protozoan communities in full-scale MWTPs in Beijing related to treatment processes[J].Water Research,2008,42(8-9):1907-1918.
[42]Hu M,Wang X H,Wen X H,et al.Microbial community structures in different wastewater treatment plants as revealed by454-pyrosequencing analysis[J].Bioresource Technology,2012,117:72-79.
[43]Lepère C,Boucher D,Jardillier L,et al.Succession and regulation factors of small eukaryote community composition in a lacustrine ecosystem(Lake Pavin)[J].Applied and Environmental Microbiology,2006,72(4):2971-2981.
[44]Wei Z Y,Liu Y Y,Feng K,et al.The divergence between fungal and bacterial communities in seasonal and spatial variations of wastewater treatment plants[J].Science of the Total Environment,2018,628-629:969-978.
[45]Tyagi V K,Subramaniyan S,Kazmi A A,et al.Microbial community in conventional and extended aeration activated sludge plants in India[J].Ecological Indicators,2008,8(5):550-554.
[46]齐嵘,刘娟,杨敏,等.无机废水硝化过程中原生动物群落结构特征及其变化规律[J].环境科学学报,2013,33(1):53-57.Qi R,Liu J,Yang M,et al.Study on protozoan community structures during the inorganic wastewater nitrification process[J].Acta Scientiae Circumstantiae,2013,33(1):53-57.
[47]Bahram M,Hildebrand F,Forslund S K,et al.Structure and function of the global topsoil microbiome[J].Nature,2018,560(7717):233-237.
[48]Telford R J,Vandvik V,Birks H J B.Dispersal limitations matter for microbial morphospecies[J].Science,2006,312(5776):1015.
[49]Whitaker R J,Grogan D W,Taylor J W.Geographic barriers isolate endemic populations of hyperthermophilic archaea[J].Science,2003,301(5635):976-978.
[50]Cho J C,Tiedje J M.Biogeography and degree of endemicity of fluorescent Pseudomonas strains in soil[J].Applied and Environmental Microbiology,2000,66(12):5448-5456.
[51]Borcard D,Legendre P.Environmental control and spatial structure in ecological communities:an example using oribatid mites(Acari,Oribatei)[J].Environmental and Ecological Statistics,1994,1(1):37-61.
[52]Vuono D C,Benecke J,Henkel J,et al.Disturbance and temporal partitioning of the activated sludge metacommunity[J].The ISME Journal,2015,9(2):425-435.