不同时期排泥对培养SBBR生物膜的影响
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摘要
序批式生物膜反应器(sequencing biofilm batch reactor, SBBR)是应用广泛的污水处理方法。为探究不同时期排泥对SBBR污染物去除效果与微生物群落结构的影响,本研究设置了挂膜初期、中期和后期进行排泥的反应器处理生活污水,同时结合16S rDNA高通量测序技术对微生物群落结构进行分析,并采用机器学习(machine learning, ML)的方法,在传统的微生物优势种分析基础上,对测序数据进行深度挖掘,寻找造成组间差异的关键物种。水质测定结果显示,COD去除效果在不同时期排泥的SBBR间没有明显差异,出水COD均低于30mg/L。挂膜中期排泥的SBBR的NH_3-N去除率先达到稳定且高于前期和后期排泥的系统。高通量测序结果显示,各SBBR中微生物优势种均以降解有机物的物种为主。挂膜中期排泥的SBBR中,ML筛选得到的NH_3-N去除相关物种(Hydrogenophaga、Gemmata和Nitrospira)与差异关键物种丰度更高,微生物群落结构稳定性更强,可从微生物层面解释分析SBBR污染物去除效果的差异。
Sequencing biofilm batch reactor(SBBR) is applied to sewage treatment. In order to investigate the effects of different sludge drainage stages on the pollutant removal and microbial community structure, three reactors were installed to treat domestic sewage. Activated sludge was removedin reactors at the initial, middle, and late culturing stages, respectively. Meanwhile, the microbialcommunity structure and traditional microbial dominant species were analyzed by 16 S rDNA high-throughput sequencing technology. Moreover, machine learning(ML) also played a role in excavating thesequencing data in depth and finding key species which led to the differences among groups. The waterquality analysis results showed that there was no significant difference in COD removal efficiency amongreactors with different sludge drainage stages, and the COD of effluent was all lower than 30mg/L. TheNH_3-N removal efficiency of SBBR in sludge discharge in the middle stage was the first to reach a stablelevel and higher than that in the early and late stage of sludge discharge system. The results of high-throughput sequencing showed that the dominant microbial species in SBBR were mainly those that withhigh organic-pollutants-degrading capability. In SBBR, the NH_3-N removal related species (Hydrogenophaga,Gemmata and Nitrospira)screened by ML had higher abundance and stronger stability of microbial community structure,which could explain the difference of pollutant removal efficiency of SBBR from the microbial level.
Sequencing biofilm batch reactor(SBBR) is applied to sewage treatment. In order to investigate the effects of different sludge drainage stages on the pollutant removal and microbial community structure, three reactors were installed to treat domestic sewage. Activated sludge was removedin reactors at the initial, middle, and late culturing stages, respectively. Meanwhile, the microbialcommunity structure and traditional microbial dominant species were analyzed by 16 S rDNA high-throughput sequencing technology. Moreover, machine learning(ML) also played a role in excavating thesequencing data in depth and finding key species which led to the differences among groups. The waterquality analysis results showed that there was no significant difference in COD removal efficiency amongreactors with different sludge drainage stages, and the COD of effluent was all lower than 30mg/L. TheNH_3-N removal efficiency of SBBR in sludge discharge in the middle stage was the first to reach a stablelevel and higher than that in the early and late stage of sludge discharge system. The results of high-throughput sequencing showed that the dominant microbial species in SBBR were mainly those that withhigh organic-pollutants-degrading capability. In SBBR, the NH_3-N removal related species (Hydrogenophaga,Gemmata and Nitrospira)screened by ML had higher abundance and stronger stability of microbial community structure,which could explain the difference of pollutant removal efficiency of SBBR from the microbial level.
引文
[1]郑桂林. SBBR同步硝化反硝化的影响因素对溶解氧在生物膜中扩散的影响研究[D].广州:广州大学, 2017.ZHEN G L. Study on the effect of SBBR simultaneous nitrification anddenitrification on the distribution of dissolved oxygen in biofilm[D].Guangzhou:Guangzhou University, 2017.
[2]杨涛,郝学凯,陈宝玉,等. Al3+对序批式生物膜反应器(SBBR)中污泥脱氢酶活性(DHA)和胞外聚合物(EPS)的影响[J].环境科学学报, 2018, 38(4):1453-1459.YANG T, HAO X K, CHEN B Y, et al. Effects of Al3+ondehydrogenase activity(DHA)and extracellular polymeric substances(EPS)of activated sludge in a sequencing batch biofilm reactor(SBBR)[J]. Acta Scientiae Circumstantiae, 2018, 38(4):1453-1459.
[3]刘舒巍,张从轩,杨兴桐,等. SBBR与人工湿地组合工艺脱氮除磷[J].环境工程学报, 2017, 11(8):4527-4534.LIU S W, ZHANG C X, YANG X T, et al. Biological phosphorus andnitrogen removal by combined process of sequencing biofilm batchreactor and constructed wetland[J]. Chinese Journal of EnvironmentalEngineering, 2017, 11(8):4527-4534.
[4]王玉,姚倩,彭党聪,等.不同C/N值下SBBR中生物膜硝化特性研究[J].中国给水排水, 2016, 32(5):23-27.WANG Y, YAO Q, PENG D C, et al. Nitrification performance ofbiofilm in SBBR with different C/N ratios[J]. China Water&Wastewater, 2016, 32(5):23-27.
[5]郑桂林,张朝升,曹勇锋,等. DO在SBBR生物膜中传递及对同步硝化反硝化的影响[J].中国给水排水, 2016, 32(3):22-26.ZHENG G L, ZHANG C S, CAO Y F, et al. Transfer of DO in biofilmand effect of DO on simultaneous nitrification and denitrification inSBBR[J]. China Water&Wastewater, 2016, 32(3):22-26.
[6]秦宇,方芳,郭劲松. pH值对SBBR自养脱氮系统效能及功能菌数量的影响[J].中国给水排水, 2012, 28(17):36-39.QIN Y, FANG F, GUO J S. Effect of pH on SBBR autotrophic nitrogenremoval process[J]. China Water&Wastewater, 2012, 28(17):36-39.
[7]周戈,赵小晶,梁红,等.不同温度对SBBR同步短程硝化反硝化处理炼油催化剂废水的影响[J].环境工程学报, 2015, 9(6):2869-2874.ZHOU G, ZHAO X J, LIANG H, et al. Effects of different temperatureson treatment of catalyst wastewater with simultaneous shortcutnitrification-denitrification in a SBBR[J]. Chinese Journal ofEnvironmental Engineering, 2015, 9(6):2869-2874.
[8]罗晓,郑向阳,赵丛丛,等. A/O工艺中污泥浓度对微生物群落结构的影响[J].中国环境科学, 2018, 38(1):275-283.LUO X, ZHENG X Y, ZHAO C C, et al. Effects of sludgeconcentration on microbial community structure in A/O process[J]. ChinaEnvironmental Science, 2018, 38(1):275-283.
[9] XU Y, WANG C, HOU J, et al. Long term effects of cerium dioxidenanoparticles on the nitrogen removal micro-environment andcommunity dynamics of a sequencing batch biofilm reactor[J].Bioresource Technology, 2017, 245:573-580.
[10] WANG J f, DING L L, LI K, et al. Estimation of spatial distribution ofquorum sensing signaling in sequencing batch biofilm reactor(SBBR)biofilms[J]. Science of the Total Environment, 2018, 612:405-414.
[11] WANG J Y, RONG H W, ZHANG C S. Evaluation of the impact ofdissolved oxygen concentration on biofilm microbial community insequencing batch biofilm reactor[J]. Journal of Bioscience andBioengineering, 2018, 125(5):532-542.
[12] MIAO L, ZHANG Q, WANG S Y, et al. Characterization of EPScompositions and microbial community in an anammox SBBR systemtreating landfill leachate[J]. Bioresource Technology, 2018, 249:108-116
[13] GUNNEMANN S. Machine learning meets databases[J]. DatenbankSpektrum, 2017, 17:77-83.
[14] BECK D, FOSTER J A. Machine learning techniques accuratelyclassify microbial communities by bacterial vaginosis characteristics[J]. Plos One, 2014, 9(2):e87830.
[15] YATSUNENKO T, REY F E, MANARY M J, et al. Human gutmicrobiome viewed across age and geography[J]. Nature, 2012, 486(7402):222-227.
[16]樊明颢.不同温度对SBBR脱氮能力和菌群结构的影响研究[D].雅安:四川农业大学, 2012.FAN M H. Effects of temperature on denitrification capability andmicrobial community structure in SBBR[D]. Ya’an:SichuanAgricultural University, 2012.
[17]安卫星,高娜,夏明,等.动胶菌属系统分类、生理特征及其在活性污泥中的作用[J].应用与环境生物学报, 2016, 22(6):1167-1174.AN W X, GAO N, XIA M, et al. Physiological characteristics andsystematic classification of the Zoogloea species and their role in the activated sludge[J]. Chinese Journal of Applied&EnvironmentalBiology, 2016, 22(6):1167-1174.
[18]范丽莎.复极性三维电极耦合曝气生物滤池处理城市污水的试验研究[D].济南:济南大学, 2015.FAN L S. The research on the treatment of city sewage by complexthree-dimensional electrode coupled biological aerated filter[D].Jinan:University of Jinan, 2015.
[19]侯洁.生物炭对潜流人工湿地生物脱氮影响机理研究[D].重庆:西南大学, 2017.HOU J. Influences of biochar on biological nitrogen removal insubsurface flow constructed wetland[D]. Chongqing:SouthwestUniversity, 2017.
[20]杨浩,张国珍,杨晓妮,等. 16S rRNA高通量测序研究集雨窖水中微生物群落结构及多样性[J].环境科学, 2017, 38(4):1704-1716.YANG H, ZHANG G Z, YANG X N, et al. Microbial communitystructure and diversity in cellar water by 16S r RNA high-throughputsequencing[J]. Environmental Science, 2017, 38(4):1704-1716.
[21]张洪勋,郝春博,白志辉.嗜酸菌研究进展[J].微生物学杂志, 2006(2):68-72.ZHANG H X, HAO C B, BAI Z H. Advance in acidophile[J]. Journalof Microbiology, 2006(2):68-72.
[22]陈瑜,刘婉薇,李良芳,等.功能性消化不良患者唾液菌群的研究[J].重庆医学, 2017, 46(13):1789-1791, 1796.CHEN Y, LIU W W, LI L F, et al. Analysis on saliva microbiome inpatients with functional dyspepsia[J]. Chongqing Medicine, 2017, 46(13):1789-1791, 1796.
[23]韦云萍,赵云,张金龙,等.嗜糖假单胞菌麦芽四糖淀粉酶的枯草芽孢杆菌表达及酶学性质分析[J].食品与发酵工业, 2014, 40(5):70-76.WEI Y P, ZHAO Y, ZHANG J L, et al. Properties of the enzymeexpressed by the optimized Pseudomonas accharophila maltotetraohydrolase gene(mta)in Bacillus subtilis[J]. Food andFermentation Industries, 2014, 40(5):70-76.
[24]陈接锋,许旭萍,李惠珍.球衣菌属的研究概况[J].环境科学与技术, 2002(6):43-46, 49.CHEN J F, XU X P, LI H Z. Research overview of Schizophyllum[J].Environmental Science and Technology, 2002(6):43-46, 49.
[25] BRONWYN D H, RAYMOND L C, WENDY M, et al. Using the Ginicoefficient with BIOLOG substrate utilisation datato provide analternative quantitative measure for comparing bacterial soilcommunities[J]. Journal of Microbiological Methods, 1997, 30(1):91-101.
[26]李青,成小英.不同填料生物反应器中脱氮微生物群落比较分析[J].安全与环境学报, 2017, 17(6):2360-2365.LI Q, CHENG X Y. Discriminative analysis of denitrifying microbialcommunities in bioreactors with different materials as biofilm carriers[J]. Journal of Safety and Environment, 2017, 17(6):2360-2365.
[27]王国宏.再论生物多样性与生态系统的稳定性[J].生物多样性,2002, 10(1):126-134.WANG G H. Further thoughts on diversity and stability in ecosystems[J]. Biodiversity Science, 2002, 10(1):126-134.
[28]沈烽,黄睿,王司辰,等.不同OTUs划分阈值对湖泊浮游细菌α多样性的影响[J].化学与生物工程, 2016, 33(2):59-60.SHEN F, HUANG R, WANG S C, et al. Effects of different OTUscutoff values onα-diversity of Bacterioplankton in lake[J]. Chemistry&Bioengineering, 2016, 33(2):59-60.
[2]杨涛,郝学凯,陈宝玉,等. Al3+对序批式生物膜反应器(SBBR)中污泥脱氢酶活性(DHA)和胞外聚合物(EPS)的影响[J].环境科学学报, 2018, 38(4):1453-1459.YANG T, HAO X K, CHEN B Y, et al. Effects of Al3+ondehydrogenase activity(DHA)and extracellular polymeric substances(EPS)of activated sludge in a sequencing batch biofilm reactor(SBBR)[J]. Acta Scientiae Circumstantiae, 2018, 38(4):1453-1459.
[3]刘舒巍,张从轩,杨兴桐,等. SBBR与人工湿地组合工艺脱氮除磷[J].环境工程学报, 2017, 11(8):4527-4534.LIU S W, ZHANG C X, YANG X T, et al. Biological phosphorus andnitrogen removal by combined process of sequencing biofilm batchreactor and constructed wetland[J]. Chinese Journal of EnvironmentalEngineering, 2017, 11(8):4527-4534.
[4]王玉,姚倩,彭党聪,等.不同C/N值下SBBR中生物膜硝化特性研究[J].中国给水排水, 2016, 32(5):23-27.WANG Y, YAO Q, PENG D C, et al. Nitrification performance ofbiofilm in SBBR with different C/N ratios[J]. China Water&Wastewater, 2016, 32(5):23-27.
[5]郑桂林,张朝升,曹勇锋,等. DO在SBBR生物膜中传递及对同步硝化反硝化的影响[J].中国给水排水, 2016, 32(3):22-26.ZHENG G L, ZHANG C S, CAO Y F, et al. Transfer of DO in biofilmand effect of DO on simultaneous nitrification and denitrification inSBBR[J]. China Water&Wastewater, 2016, 32(3):22-26.
[6]秦宇,方芳,郭劲松. pH值对SBBR自养脱氮系统效能及功能菌数量的影响[J].中国给水排水, 2012, 28(17):36-39.QIN Y, FANG F, GUO J S. Effect of pH on SBBR autotrophic nitrogenremoval process[J]. China Water&Wastewater, 2012, 28(17):36-39.
[7]周戈,赵小晶,梁红,等.不同温度对SBBR同步短程硝化反硝化处理炼油催化剂废水的影响[J].环境工程学报, 2015, 9(6):2869-2874.ZHOU G, ZHAO X J, LIANG H, et al. Effects of different temperatureson treatment of catalyst wastewater with simultaneous shortcutnitrification-denitrification in a SBBR[J]. Chinese Journal ofEnvironmental Engineering, 2015, 9(6):2869-2874.
[8]罗晓,郑向阳,赵丛丛,等. A/O工艺中污泥浓度对微生物群落结构的影响[J].中国环境科学, 2018, 38(1):275-283.LUO X, ZHENG X Y, ZHAO C C, et al. Effects of sludgeconcentration on microbial community structure in A/O process[J]. ChinaEnvironmental Science, 2018, 38(1):275-283.
[9] XU Y, WANG C, HOU J, et al. Long term effects of cerium dioxidenanoparticles on the nitrogen removal micro-environment andcommunity dynamics of a sequencing batch biofilm reactor[J].Bioresource Technology, 2017, 245:573-580.
[10] WANG J f, DING L L, LI K, et al. Estimation of spatial distribution ofquorum sensing signaling in sequencing batch biofilm reactor(SBBR)biofilms[J]. Science of the Total Environment, 2018, 612:405-414.
[11] WANG J Y, RONG H W, ZHANG C S. Evaluation of the impact ofdissolved oxygen concentration on biofilm microbial community insequencing batch biofilm reactor[J]. Journal of Bioscience andBioengineering, 2018, 125(5):532-542.
[12] MIAO L, ZHANG Q, WANG S Y, et al. Characterization of EPScompositions and microbial community in an anammox SBBR systemtreating landfill leachate[J]. Bioresource Technology, 2018, 249:108-116
[13] GUNNEMANN S. Machine learning meets databases[J]. DatenbankSpektrum, 2017, 17:77-83.
[14] BECK D, FOSTER J A. Machine learning techniques accuratelyclassify microbial communities by bacterial vaginosis characteristics[J]. Plos One, 2014, 9(2):e87830.
[15] YATSUNENKO T, REY F E, MANARY M J, et al. Human gutmicrobiome viewed across age and geography[J]. Nature, 2012, 486(7402):222-227.
[16]樊明颢.不同温度对SBBR脱氮能力和菌群结构的影响研究[D].雅安:四川农业大学, 2012.FAN M H. Effects of temperature on denitrification capability andmicrobial community structure in SBBR[D]. Ya’an:SichuanAgricultural University, 2012.
[17]安卫星,高娜,夏明,等.动胶菌属系统分类、生理特征及其在活性污泥中的作用[J].应用与环境生物学报, 2016, 22(6):1167-1174.AN W X, GAO N, XIA M, et al. Physiological characteristics andsystematic classification of the Zoogloea species and their role in the activated sludge[J]. Chinese Journal of Applied&EnvironmentalBiology, 2016, 22(6):1167-1174.
[18]范丽莎.复极性三维电极耦合曝气生物滤池处理城市污水的试验研究[D].济南:济南大学, 2015.FAN L S. The research on the treatment of city sewage by complexthree-dimensional electrode coupled biological aerated filter[D].Jinan:University of Jinan, 2015.
[19]侯洁.生物炭对潜流人工湿地生物脱氮影响机理研究[D].重庆:西南大学, 2017.HOU J. Influences of biochar on biological nitrogen removal insubsurface flow constructed wetland[D]. Chongqing:SouthwestUniversity, 2017.
[20]杨浩,张国珍,杨晓妮,等. 16S rRNA高通量测序研究集雨窖水中微生物群落结构及多样性[J].环境科学, 2017, 38(4):1704-1716.YANG H, ZHANG G Z, YANG X N, et al. Microbial communitystructure and diversity in cellar water by 16S r RNA high-throughputsequencing[J]. Environmental Science, 2017, 38(4):1704-1716.
[21]张洪勋,郝春博,白志辉.嗜酸菌研究进展[J].微生物学杂志, 2006(2):68-72.ZHANG H X, HAO C B, BAI Z H. Advance in acidophile[J]. Journalof Microbiology, 2006(2):68-72.
[22]陈瑜,刘婉薇,李良芳,等.功能性消化不良患者唾液菌群的研究[J].重庆医学, 2017, 46(13):1789-1791, 1796.CHEN Y, LIU W W, LI L F, et al. Analysis on saliva microbiome inpatients with functional dyspepsia[J]. Chongqing Medicine, 2017, 46(13):1789-1791, 1796.
[23]韦云萍,赵云,张金龙,等.嗜糖假单胞菌麦芽四糖淀粉酶的枯草芽孢杆菌表达及酶学性质分析[J].食品与发酵工业, 2014, 40(5):70-76.WEI Y P, ZHAO Y, ZHANG J L, et al. Properties of the enzymeexpressed by the optimized Pseudomonas accharophila maltotetraohydrolase gene(mta)in Bacillus subtilis[J]. Food andFermentation Industries, 2014, 40(5):70-76.
[24]陈接锋,许旭萍,李惠珍.球衣菌属的研究概况[J].环境科学与技术, 2002(6):43-46, 49.CHEN J F, XU X P, LI H Z. Research overview of Schizophyllum[J].Environmental Science and Technology, 2002(6):43-46, 49.
[25] BRONWYN D H, RAYMOND L C, WENDY M, et al. Using the Ginicoefficient with BIOLOG substrate utilisation datato provide analternative quantitative measure for comparing bacterial soilcommunities[J]. Journal of Microbiological Methods, 1997, 30(1):91-101.
[26]李青,成小英.不同填料生物反应器中脱氮微生物群落比较分析[J].安全与环境学报, 2017, 17(6):2360-2365.LI Q, CHENG X Y. Discriminative analysis of denitrifying microbialcommunities in bioreactors with different materials as biofilm carriers[J]. Journal of Safety and Environment, 2017, 17(6):2360-2365.
[27]王国宏.再论生物多样性与生态系统的稳定性[J].生物多样性,2002, 10(1):126-134.WANG G H. Further thoughts on diversity and stability in ecosystems[J]. Biodiversity Science, 2002, 10(1):126-134.
[28]沈烽,黄睿,王司辰,等.不同OTUs划分阈值对湖泊浮游细菌α多样性的影响[J].化学与生物工程, 2016, 33(2):59-60.SHEN F, HUANG R, WANG S C, et al. Effects of different OTUscutoff values onα-diversity of Bacterioplankton in lake[J]. Chemistry&Bioengineering, 2016, 33(2):59-60.