我国南北区域城市污水处理系统内真菌群落的差异
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摘要
【背景】微生物是污水处理系统内污染物去除的主体,真菌作为其中不可或缺的一部分,在去除有毒化合物、提高生物转化等方面发挥着重要作用,应引起人们的关注。【目的】明确我国南北区域城市污水处理系统内真菌群落的多样性、组成差异及其影响因素。【方法】采集我国北方23个污水处理厂的90份活性污泥样品和南方37个污水处理厂的121份活性污泥样品,提取上述样本的总DNA,并应用Illumina MiSeq平台进行ITS高通量测序,结合多种数量生态学分析方法对数据进行分析。【结果】我国南北区域城市污水处理系统内真菌群落结构存在显著差异,真菌群落多样性南方显著高于北方;我国南北区域真菌群落的组成差异显著,南方群落以Sordariomycetes和Glomeromycetes为优势菌纲,以Ophiocordycep和Alternaria为优势菌属,北方群落以Tremellomycetes和Saccharomycetes为优势菌纲,以Trichosporon和Saccharomyces为优势菌属,其中Tremellomycetes和Sordariomycetes是常见的病原菌,对下游生物或者人类的健康具有潜在威胁,Trichosporon属的丝状真菌若大量异常增殖会引发污泥膨胀现象,影响系统的稳定运行;地理纬度、年平均气温、进水NH_4~+浓度、进水总氮浓度是导致我国南北区域真菌群落存在差异的重要影响因子。【结论】我国南北区域真菌群落多样性、组成存在显著差异,地理因素、所在城市的气候因素和进水污染物浓度对真菌群落结构影响显著。针对南北区域,应合理调控相应区域内的重要功能菌群,以调控污水处理厂的高效稳定运行,同时密切关注其中的条件致病菌和易引发污泥膨胀的菌群,建立面向种群的优化控制系统以管控风险。
[Background] Microorganisms are main agents to remove pollutants in activated sludge(AS) in wastewater treatment plants(WWTPs), among them, fungi should be paid attention to as they play important roles in removing toxic compounds and improving biological transformation, etc. [Objective]The major research will be focus on to characterize and compare the diversity, composition of fungal communities in WWTPs located in north and south China, determine the influence factors of fungal community structure. [Methods] Ninety AS samples from 23 WWTPs in north China and 121 AS samples from 37 WWTPs in south China were collected, respectively. After DNA extraction and amplification, high-throughput sequencing of ITS gene was carried out using Illumina MiSeq platform.Data were processed by a variety of quantitative ecological analysis methods. [Results] Fungal community structures in AS system between north and south China were significantly different.Diversity of fungal community in south China was higher than that of north China. The dominant classes and genera in fungal community were Sordariomycetes and Glomeromycetes classes, and Ophiocordycep and Alternaria genera in south China. While they were Tremellomycetes and Saccharomycetes classes, and Trichosporon and Saccharomyces genera in north China. Members of the classes Tremellomycetes and Sodariomycetes are opportunistic pathogens, which may be potential threats to downstream organisms or human health. Abnormal growth of members in Trichosporon genus will cause sludge bulking and affect the stable operation of WWTPs. Latitude, annual average air temperature, concentration of NH_4~+ and the total nitrogen in influent were identified as the important factors that influence fungal community structure in WWTPs. [Conclusion] Fungal communities between north and south China were significantly different in diversity and composition as a results of geographic factors, climate factors and concentration of pollutants in influent. Accordingly, toward the efficient and stable operation of the treatment plants, it is necessary to regulate the different important communities in south and north China. Furthermore, we should pay close attention to the opportunistic pathogenic fungi and some agents that easily cause sludge bulking. Establishment of population-oriented optimal control system is necessary to control the risks.
[Background] Microorganisms are main agents to remove pollutants in activated sludge(AS) in wastewater treatment plants(WWTPs), among them, fungi should be paid attention to as they play important roles in removing toxic compounds and improving biological transformation, etc. [Objective]The major research will be focus on to characterize and compare the diversity, composition of fungal communities in WWTPs located in north and south China, determine the influence factors of fungal community structure. [Methods] Ninety AS samples from 23 WWTPs in north China and 121 AS samples from 37 WWTPs in south China were collected, respectively. After DNA extraction and amplification, high-throughput sequencing of ITS gene was carried out using Illumina MiSeq platform.Data were processed by a variety of quantitative ecological analysis methods. [Results] Fungal community structures in AS system between north and south China were significantly different.Diversity of fungal community in south China was higher than that of north China. The dominant classes and genera in fungal community were Sordariomycetes and Glomeromycetes classes, and Ophiocordycep and Alternaria genera in south China. While they were Tremellomycetes and Saccharomycetes classes, and Trichosporon and Saccharomyces genera in north China. Members of the classes Tremellomycetes and Sodariomycetes are opportunistic pathogens, which may be potential threats to downstream organisms or human health. Abnormal growth of members in Trichosporon genus will cause sludge bulking and affect the stable operation of WWTPs. Latitude, annual average air temperature, concentration of NH_4~+ and the total nitrogen in influent were identified as the important factors that influence fungal community structure in WWTPs. [Conclusion] Fungal communities between north and south China were significantly different in diversity and composition as a results of geographic factors, climate factors and concentration of pollutants in influent. Accordingly, toward the efficient and stable operation of the treatment plants, it is necessary to regulate the different important communities in south and north China. Furthermore, we should pay close attention to the opportunistic pathogenic fungi and some agents that easily cause sludge bulking. Establishment of population-oriented optimal control system is necessary to control the risks.
引文
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[9]Xiao CQ,Wu XY,Liu TT,et al.Microbial community structure of activated sludge for biosolubilization of two different rock phosphates[J].Applied Biochemistry and Biotechnology,2017,182(2):742-754
[10]Wu B,Tian JQ,Bai CM,et al.The biogeography of fungal communities in wetland sediments along the Changjiang River and other sites in China[J].The ISME Journal,2013,7(7):1299-1309
[11]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
[12]Zhang T,Shao MF,Ye L.454 Pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants[J].The ISME Journal,2012,6(6):1137-1147
[13]Wang XH,Wen XH,Deng Y,et al.Distance-decay relationship for biological wastewater treatment plants[J].Applied and Environmental Microbiology,2016,82(16):4860-4866
[14]Zhang B,Wu LW,Wen XH.Potential source environments for microbial community in wastewater treatment plants(WWTPs)in China[J].Environmental Science,2019,DOI:10.13227/j.hjkx.201901147(in Chinese)张冰,吴林蔚,文湘华.全国城市污水处理厂中微生物群落的溯源分析[J].环境科学,2019,DOI:10.13227/j.hjkx.201901147
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[16]Wu LY,Wen CQ,Qin YJ,et al.Phasing amplicon sequencing on Illumina MiSeq for robust environmental microbial community analysis[J].BMC Microbiology,2015,15:125
[17]Caporaso JG,Lauber CL,Walters WA,et al.Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms[J].The ISME Journal,2012,6(8):1621-1624
[18]Guo X,Feng JJ,Shi Z,et al.Climate warming leads to divergent succession of grassland microbial communities[J].Nature Climate Change,2018,8(9):813-818
[19]Chao AN.Nonparametric estimation of the number of classes in a population[J].Scandinavian Journal of Statistics,1984,11(4):265-270
[20]Segata N,Izard J,Waldron L,et al.Metagenomic biomarker discovery and explanation[J].Genome Biology,2011,12(6):R60
[21]Feng K,Zhang ZJ,Cai WW,et al.Biodiversity and species competition regulate the resilience of microbial biofilm community[J].Molecular Ecology,2017,26(21):6170-6182
[22]Wang Y,Liu MK,Miao XC,et al.Effect of urban water supply system on fungi population and community structure[J].Microbiology China,2019,46(1):20-28(in Chinese)王钰,刘明坤,苗小草,等.城市供水系统对水中真菌数量和群落结构的影响[J].微生物学通报,2019,46(1):20-28
[23]Xia Y,Hu M,Wen XH,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(5):18509
[24]Wei ZY,Liu YY,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
[25]Buée M,Reich M,Murat C,et al.454 Pyrosequencing analyses of forest soils reveal an unexpectedly high fungal diversity[J].New Phytologist,2009,184(2):449-456
[26]Wubet T,Christ S,Sch?ning I,et al.Differences in soil fungal communities between european beech(Fagus sylvatica L.)dominated forests are related to soil and understory vegetation[J].PLoS One,2012,7(10):e47500
[27]Klaubauf S,Inselsbacher E,Zechmeister-Boltenstern S,et al.Molecular diversity of fungal communities in agricultural soils from Lower Austria[J].Fungal Diversity,2010,44(1):65-75
[28]Moll J,Hoppe B,K?nig S,et al.Spatial distribution of fungal communities in an arable soil[J].PLoS One,2016,11(2):e0148130
[29]Wang P,Xiao HY,Yuan RQ,et al.Fungal community in the estuarine sediment of Poyang Lake[J].Acta Scientiae Circumstantiae,2018,38(5):1949-1956(in Chinese)王鹏,肖汉玉,袁瑞强,等.鄱阳湖入湖河口沉积物真菌群落结构[J].环境科学学报,2018,38(5):1949-1956
[30]Wang L,Song ZW,Xu AL,et al.Phylogenetic diversity of airborne microbes in Qingdao downtown in autumn[J].Chinese Jouranl of Applied Ecology,2015,26(4):1121-1129(in Chinese)王琳,宋志文,徐爱玲,等.青岛市秋季空气微生物群落多样性[J].应用生态学报,2015,26(4):1121-1129
[31]Mothapo N,Chen HH,Cubeta MA,et al.Phylogenetic,taxonomic and functional diversity of fungal denitrifiers and associated N2O production efficacy[J].Soil Biology and Biochemistry,2015,83:160-175
[32]Findley K,Rodriguez-Carres M,Metin B,et al.Phylogeny and phenotypic characterization of pathogenic Cryptococcus species and closely related saprobic taxa in the Tremellales[J].Eukaryotic Cell,2009,8(3):353-361
[33]Heitman J.Microbial pathogens in the fungal kingdom[J].Fungal Biology Reviews,2011,25(1):48-60
[34]?ehulka J,KubátováA,Hubka V.Cephalotheca sulfurea(Ascomycota,Sordariomycetes),a new fungal pathogen of the farmed rainbow trout Oncorhynchus mykiss[J].Journal of Fish Disease,2016,39(12):1413-1419
[35]Landeka Dragi?evi?T,Zano?ki Hren M,Gmajni?M,et al.Biodegradation of olive mill wastewater by Trichosporon cutaneum and Geotrichum candidum[J].Arhiv za Higijenu Rada i Toksikologiju,2010,61(4):399-405
[36]Marrot B,Barrios-Martinez A,Moulin P,et al.Biodegradation of high phenol concentration by activated sludge in an immersed membrane bioreactor[J].Biochemical Engineering Journal,2006,30(2):174-183
[37]Wang JL,Chen C.Biosorption of heavy metals by Saccharomyces cerevisiae:a review[J].Biotechnology Advances,2006,24(5):427-451
[38]Ye JS,Yin H,Mai BX,et al.Biosorption of chromium from aqueous solution and electroplating wastewater using mixture of Candida lipolytica and dewatered sewage sludge[J].Bioresource Technology,2010,101(11):3893-3902
[39]Chesson P.Mechanisms of maintenance of species diversity[J].Annual Review of Ecology and Systematics,2000,31:343-366
[40]Hubbell SP.The Unified Neutral Theory of Biodiversity and Biogeography[M].Princeton:Princeton University Press,2001
[41]Wang XH,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
[42]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
[43]Hu M,Wang XH,Wen XH,et al.Microbial community structures in different wastewater treatment plants as revealed by 454-pyrosequencing analysis[J].Bioresource Technology,2012,117:72-79
[44]Xia Y,Wen XH,Zhang B,et al.Diversity and assembly patterns of activated sludge microbial communities:a review[J].Biotechnology Advances,2018,36(4):1038-1047
[45]Maza-Márquez P,Vilchez-Vargas R,Kerckhof FM,et al.Community structure,population dynamics and diversity of fungi in a full-scale membrane bioreactor(MBR)for urban wastewater treatment[J].Water Research,2016,105:507-519
[46]Bahram M,Hildebrand F,Forslund SK,et al.Structure and function of the global topsoil microbiome[J].Nature,2018,560(7717):233-237
[2]More TT,Yan S,Tyagi RD,et al.Potential use of filamentous fungi for wastewater sludge treatment[J].Bioresource Technology,2010,101(20):7691-7700
[3]Liu J,Li J,Tao YQ,et al.Analysis of bacterial,fungal and archaeal populations from a municipal wastewater treatment plant developing an innovative aerobic granular sludge process[J].World Journal of Microbiology&Biotechnology,2017,33(1):14
[4]Weber SD,Ludwig W,Schleifer KH,et al.Microbial composition and structure of aerobic granular sewage biofilms[J].Applied and Environmental Microbiology,2007,73(19):6233-6240
[5]Zhou YJ,Fu HY,Fan XF,et al.Isolation of filamentous fungi capable of enhancing sludge dewaterability and study of mechanisms responsible for the sludge dewaterability enhancement[J].Environmental Science,2015,36(2):612-618(in Chinese)周雨珺,付豪逸,范先锋,等.改善污泥脱水性能的丝状真菌的分离及其促进污泥脱水的机制初探[J].环境科学,2015,36(2):612-618
[6]Zheng SK,Sun JY,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
[7]Li H,Tan L,Ning SX,et al.Reactor performance and microbial community dynamics during aerobic degradation and detoxification of Acid Red B with activated sludge bioaugmented by a yeast Candida tropicalis TL-F1 in MBR[J].International Biodeterioration&Biodegradation,2015,104:149-156
[8]He MY,Tan L,Ning SX,et al.Performance of the biological aerated filter bioaugmented by a yeast Magnusiomyces ingens LH-F1 for treatment of Acid Red B and microbial community dynamics[J].World Journal of Microbiology&Biotechnology,2017,33(2):39
[9]Xiao CQ,Wu XY,Liu TT,et al.Microbial community structure of activated sludge for biosolubilization of two different rock phosphates[J].Applied Biochemistry and Biotechnology,2017,182(2):742-754
[10]Wu B,Tian JQ,Bai CM,et al.The biogeography of fungal communities in wetland sediments along the Changjiang River and other sites in China[J].The ISME Journal,2013,7(7):1299-1309
[11]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
[12]Zhang T,Shao MF,Ye L.454 Pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants[J].The ISME Journal,2012,6(6):1137-1147
[13]Wang XH,Wen XH,Deng Y,et al.Distance-decay relationship for biological wastewater treatment plants[J].Applied and Environmental Microbiology,2016,82(16):4860-4866
[14]Zhang B,Wu LW,Wen XH.Potential source environments for microbial community in wastewater treatment plants(WWTPs)in China[J].Environmental Science,2019,DOI:10.13227/j.hjkx.201901147(in Chinese)张冰,吴林蔚,文湘华.全国城市污水处理厂中微生物群落的溯源分析[J].环境科学,2019,DOI:10.13227/j.hjkx.201901147
[15]White TJ,Bruns T,Lee S,et al.Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[A]//Innis MA,Gelfand DH,Sninsky JJ,et al.PCR Protocols:A Guide to Methods and Applications[M].San Diego:Academic Press,1990:315-322
[16]Wu LY,Wen CQ,Qin YJ,et al.Phasing amplicon sequencing on Illumina MiSeq for robust environmental microbial community analysis[J].BMC Microbiology,2015,15:125
[17]Caporaso JG,Lauber CL,Walters WA,et al.Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms[J].The ISME Journal,2012,6(8):1621-1624
[18]Guo X,Feng JJ,Shi Z,et al.Climate warming leads to divergent succession of grassland microbial communities[J].Nature Climate Change,2018,8(9):813-818
[19]Chao AN.Nonparametric estimation of the number of classes in a population[J].Scandinavian Journal of Statistics,1984,11(4):265-270
[20]Segata N,Izard J,Waldron L,et al.Metagenomic biomarker discovery and explanation[J].Genome Biology,2011,12(6):R60
[21]Feng K,Zhang ZJ,Cai WW,et al.Biodiversity and species competition regulate the resilience of microbial biofilm community[J].Molecular Ecology,2017,26(21):6170-6182
[22]Wang Y,Liu MK,Miao XC,et al.Effect of urban water supply system on fungi population and community structure[J].Microbiology China,2019,46(1):20-28(in Chinese)王钰,刘明坤,苗小草,等.城市供水系统对水中真菌数量和群落结构的影响[J].微生物学通报,2019,46(1):20-28
[23]Xia Y,Hu M,Wen XH,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(5):18509
[24]Wei ZY,Liu YY,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
[25]Buée M,Reich M,Murat C,et al.454 Pyrosequencing analyses of forest soils reveal an unexpectedly high fungal diversity[J].New Phytologist,2009,184(2):449-456
[26]Wubet T,Christ S,Sch?ning I,et al.Differences in soil fungal communities between european beech(Fagus sylvatica L.)dominated forests are related to soil and understory vegetation[J].PLoS One,2012,7(10):e47500
[27]Klaubauf S,Inselsbacher E,Zechmeister-Boltenstern S,et al.Molecular diversity of fungal communities in agricultural soils from Lower Austria[J].Fungal Diversity,2010,44(1):65-75
[28]Moll J,Hoppe B,K?nig S,et al.Spatial distribution of fungal communities in an arable soil[J].PLoS One,2016,11(2):e0148130
[29]Wang P,Xiao HY,Yuan RQ,et al.Fungal community in the estuarine sediment of Poyang Lake[J].Acta Scientiae Circumstantiae,2018,38(5):1949-1956(in Chinese)王鹏,肖汉玉,袁瑞强,等.鄱阳湖入湖河口沉积物真菌群落结构[J].环境科学学报,2018,38(5):1949-1956
[30]Wang L,Song ZW,Xu AL,et al.Phylogenetic diversity of airborne microbes in Qingdao downtown in autumn[J].Chinese Jouranl of Applied Ecology,2015,26(4):1121-1129(in Chinese)王琳,宋志文,徐爱玲,等.青岛市秋季空气微生物群落多样性[J].应用生态学报,2015,26(4):1121-1129
[31]Mothapo N,Chen HH,Cubeta MA,et al.Phylogenetic,taxonomic and functional diversity of fungal denitrifiers and associated N2O production efficacy[J].Soil Biology and Biochemistry,2015,83:160-175
[32]Findley K,Rodriguez-Carres M,Metin B,et al.Phylogeny and phenotypic characterization of pathogenic Cryptococcus species and closely related saprobic taxa in the Tremellales[J].Eukaryotic Cell,2009,8(3):353-361
[33]Heitman J.Microbial pathogens in the fungal kingdom[J].Fungal Biology Reviews,2011,25(1):48-60
[34]?ehulka J,KubátováA,Hubka V.Cephalotheca sulfurea(Ascomycota,Sordariomycetes),a new fungal pathogen of the farmed rainbow trout Oncorhynchus mykiss[J].Journal of Fish Disease,2016,39(12):1413-1419
[35]Landeka Dragi?evi?T,Zano?ki Hren M,Gmajni?M,et al.Biodegradation of olive mill wastewater by Trichosporon cutaneum and Geotrichum candidum[J].Arhiv za Higijenu Rada i Toksikologiju,2010,61(4):399-405
[36]Marrot B,Barrios-Martinez A,Moulin P,et al.Biodegradation of high phenol concentration by activated sludge in an immersed membrane bioreactor[J].Biochemical Engineering Journal,2006,30(2):174-183
[37]Wang JL,Chen C.Biosorption of heavy metals by Saccharomyces cerevisiae:a review[J].Biotechnology Advances,2006,24(5):427-451
[38]Ye JS,Yin H,Mai BX,et al.Biosorption of chromium from aqueous solution and electroplating wastewater using mixture of Candida lipolytica and dewatered sewage sludge[J].Bioresource Technology,2010,101(11):3893-3902
[39]Chesson P.Mechanisms of maintenance of species diversity[J].Annual Review of Ecology and Systematics,2000,31:343-366
[40]Hubbell SP.The Unified Neutral Theory of Biodiversity and Biogeography[M].Princeton:Princeton University Press,2001
[41]Wang XH,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
[42]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
[43]Hu M,Wang XH,Wen XH,et al.Microbial community structures in different wastewater treatment plants as revealed by 454-pyrosequencing analysis[J].Bioresource Technology,2012,117:72-79
[44]Xia Y,Wen XH,Zhang B,et al.Diversity and assembly patterns of activated sludge microbial communities:a review[J].Biotechnology Advances,2018,36(4):1038-1047
[45]Maza-Márquez P,Vilchez-Vargas R,Kerckhof FM,et al.Community structure,population dynamics and diversity of fungi in a full-scale membrane bioreactor(MBR)for urban wastewater treatment[J].Water Research,2016,105:507-519
[46]Bahram M,Hildebrand F,Forslund SK,et al.Structure and function of the global topsoil microbiome[J].Nature,2018,560(7717):233-237