A/O工艺中污泥浓度对微生物群落结构的影响
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摘要
采用A/O工艺处理淀粉厂区废水,结合Miseq高通量测序技术研究了不同污泥浓度(MLSS)对A/O工艺脱氮效果及微生物群落结构的影响.结果显示,当污泥浓度为4066mg/L时(1号池),氨氮(NH_4~+-N)平均去除率高达90.2%;而污泥浓度为2985mg/L时(2号池),去除率仅为67.2%.两池体中优势菌门均为变形菌门、拟杆菌门、绿弯菌门和厚壁菌门,其中变形菌门和拟杆菌门平均丰度分别为40.65%、30.28%和25.25%、33.27%,其相对丰度比例差异较大.在功能基因层面,两池体中所有8个功能基因类别(KEGG,京都基因与基因组百科全书)排序相同,氮代谢相关基因中,硝化酶、反硝化酶、氨化酶的相关功能基因均在1号池中含量高.该工艺中污泥浓度对菌群结构影响显著,高污泥浓度有利于形成高效脱氮菌群结构.
The effect of different sludge concentration(MLSS) on denitrification efficiency and microbial community structure in A/O process for starch industry wastewater treatment were investigated. The 16 S r DNA Miseq high throughput sequencing technique was used for bacterial commnutiy analysis. The results showed that the average removal rate of ammonia nitrogen(NH_4~+-N) were respectively 90.2% and 67.2% when the concentration of sludge were 4066 mg/L and 2985 mg/L. The dominant bacteria were Proteobacteria, Bacteroidetes, Chloroflexi and Firmicutes in both of the pools; while the average abundance ratio of Proteobacteria and Bacteroides were 40.65% and 30.28% in the pool No. 1,25.25% and 33.27% in the pool No. 2 respectively. In the functional gene level, all of the eight categories(KEGG, Kyoto Encyclopedia of Genes and Genomes) had a same order. The content of nitrogen metabolism related genes, such as, nitrifying ferments, denitrifying enzymes and ammonifying enzymes genes were higher in the pool No. 1. In this study, the sludge concentration has a significant effect on the bacterial structure, and the high sludge concentration is beneficial to the formation of high efficiency denitrifying bacterial community.
The effect of different sludge concentration(MLSS) on denitrification efficiency and microbial community structure in A/O process for starch industry wastewater treatment were investigated. The 16 S r DNA Miseq high throughput sequencing technique was used for bacterial commnutiy analysis. The results showed that the average removal rate of ammonia nitrogen(NH_4~+-N) were respectively 90.2% and 67.2% when the concentration of sludge were 4066 mg/L and 2985 mg/L. The dominant bacteria were Proteobacteria, Bacteroidetes, Chloroflexi and Firmicutes in both of the pools; while the average abundance ratio of Proteobacteria and Bacteroides were 40.65% and 30.28% in the pool No. 1,25.25% and 33.27% in the pool No. 2 respectively. In the functional gene level, all of the eight categories(KEGG, Kyoto Encyclopedia of Genes and Genomes) had a same order. The content of nitrogen metabolism related genes, such as, nitrifying ferments, denitrifying enzymes and ammonifying enzymes genes were higher in the pool No. 1. In this study, the sludge concentration has a significant effect on the bacterial structure, and the high sludge concentration is beneficial to the formation of high efficiency denitrifying bacterial community.
引文
[1]龚灵潇.缺氧/好氧生物膜工艺处理低碳氮比生活污水的脱氮特性[D].北京:北京工业大学,2013.
[2]成官文,吴志超,黄翔峰,等.沸石强化A/O生物脱氮工艺氨氮去除机理研究[J].环境工程学报,2010,4(3):540-546.
[3]张鹏娟.A/O生物脱氮工艺影响因素及强化运行效果的试验研究[D].郑州:郑州大学,2013.
[4]徐峥勇.基于亚硝化、厌氧氨氧化与反硝化的脱氮耦合工艺及其控制策略研究[D].长沙:湖南大学,2011.
[5]赵继红,楼燕,余志晟,等.城市污水厂(A/O工艺)微生物群落结构及其动态变化[J].生态环境,2008,17(3):898-902.
[6]Wintzingerode F V,G?bel U B,Stackebrandt E.Determination of microbial diversity in environmental samples:Pitfalls of PCR-based r RNA analysis[J].FEMS Microbiology Reviews,1997,21(3):213-229.
[7]王绍祥,杨洲祥,孙真,等.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,2014,(3):196-203.
[8]孙豆豆.强化A/O工艺低温脱氮效能与菌群特性研究[D].哈尔滨:哈尔滨工业大学,2015.
[9]赵文莉.复合碳源填料深度反硝化脱氮特性研究[D].北京:北京工业大学,2015.
[10]户海燕,瞿思宜,张正哲,等.分子生物技术在厌氧氨氧化工艺研究中的应用[J].环境科学与技术,2016,(1):20-26+43.
[11]李鹏,毕学军,王军,等.常规和倒置A2/O工艺活性污泥微生物群落结构的比较[J].中国环境科学,2017,37(3):1137-1145.
[12]窦娜莎,王琳.不同温度下曝气生物滤池运行效能与微生物群落结构[J].环境工程学报,2016,10(6):2800-2806.
[13]李志静,孙宝盛,张笑雪,等.不同供氧策略和氨氮浓度下SBR中微生物群落的演变[J].环境工程学报,2017,11(1):359-365.
[14]Plósz B G,Jobbágy A,Jr G C.Factors influencing deterioration of denitrification by oxygen entering an anoxic reactor through the surface[J].Water Research,2003,37(4):853-863.
[15]邓仁健,张金松,曲志军.污泥浓度对双重后置反硝化工艺脱氮除磷的影响[J].环境科学研究,2014,27(7):797-803.
[16]辛海霞,季诚昌,张凡,等.MLSS对FBR处理高氨氮印染废水的影响[J].东华大学学报(自然科学版),2015,41(5):692-695.
[17]国家环境保护总局.水和废水监测分析方法[M].4版.北京:中国环境科学出版社,2002:258-281.
[18]Tanja M,Steven L.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2936.
[19]Patrick D S,Sarah L W,Thomas R,et al.Introducing mothur:Open-Source,Platform-Independent,Community-Supported Software for Describing and Comparing Microbial Communities[J].Applied&Environmental Microbiology,2009,75(23):7537-7541.
[20]Robert C Edgar.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[21]Langille M G,Zaneveld J,Caporaso J G,et al.Predictive functional profiling of microbial communities using 16S r RNA marker gene sequences[J].Nature Biotechnology,2013,31(9):814-821.
[22]封莉,张立秋,吕炳南.污泥浓度对膜生物反应器运行特性的影响研究[J].哈尔滨工业大学学报,2003,35(3):307-310.
[23]Kawasaki K,Maruoka S,Katagami R,et al.Effect of initial MLSS,on operation of submerged membrane activated sludge process[J].Desalination,2011,281(20):334-339.
[24]张恩华,戴幼芬,肖勇,等.还原Cr(Ⅵ)的混菌胞外聚合物和细菌群落结构分析[J].中国环境科学,2017,37(1):352-357.
[25]Chung C M,Cho K W,Kim Y J,et al.Enhanced biological nitrogen removal in MLE combined with post-denitrification process and EF clarifier[J].Bioprocess&Biosystems Engineering,2012,35(4):503-511.
[26]Yang X,Zhou N,Chen M,et al.Analysis of microbial community structure in MBR with different ammonia concentrations using fluorescence in situ hybridization[J].Journal of Southeast University:Nature Science Edition,2013,43(2):380-385.
[27]宋云龙,张金松,朱佳,等.基于高通量测序的微生物强化污泥减量工艺中微生物群落解析[J].中国环境科学,2016,36(7):2099-2107.
[28]黄菲,梅晓洁,王志伟,等.冬季低温下MBR与CAS工艺运行及微生物群落特征[J].环境科学,2014,35(3):1002-1008.
[29]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(10):72-79.
[30]Ye L.,Shao M.F.,Zhang T,et al.Analysis of the bacterial community in a laboratory-scale nitrification reactor and a wastewater treatment plant by 454-pyrosequencing[J].Water Research,2011,45(15):4390–4398.
[31]李卫华,孙英杰,刘子梁,等.序批式生物反应器填埋场脱氮微生物多样性分析[J].环境科学,2016,37(1):342-349.
[32]杨华,黄钧,赵永贵,等.陶厄氏菌Thauera sp.strain TN9的鉴定及特性[J].应用与环境生物学报,2013,19(2):318-323.
[33]陈亚平,叶宏,王娟,等.亚硝化单胞菌的分离鉴定及其降解特性的研究[J].环境科学与技术,2007,30(9):24-25.
[34]Ma Q,Qu Y Y,Shen W L,et al.Bacterial community compositions of coking wastewater treatment plants in steel industry revealed by Illumina high-throughput sequencing[J].Bioresource Technology,2015,179:436-443.
[35]李哿.活性污泥—生物膜复合系统脱氮除磷试验研究[D].济南:山东建筑大学,2011.
[36]Ning L,Jin H,Han Y,et al.Pathways in bacterial and archaeal communities dictated by ammonium stress in a high solid anaerobic digester with dewatered sludge[J].Bioresource Technology,2017,241:95-102.
[37]田美,刘汉湖,申欣,等.百乐克(BIOLAK)活性污泥宏基因组的生物多样性及功能分析[J].环境科学,2015,36(5):1739-1748.
[38]Yu K,Zhang T.Metagenomic and metatranscriptomic analysis of microbial community structure and gene expression of activated sludge[J].Plos One,2012,7(5):e38183.
[2]成官文,吴志超,黄翔峰,等.沸石强化A/O生物脱氮工艺氨氮去除机理研究[J].环境工程学报,2010,4(3):540-546.
[3]张鹏娟.A/O生物脱氮工艺影响因素及强化运行效果的试验研究[D].郑州:郑州大学,2013.
[4]徐峥勇.基于亚硝化、厌氧氨氧化与反硝化的脱氮耦合工艺及其控制策略研究[D].长沙:湖南大学,2011.
[5]赵继红,楼燕,余志晟,等.城市污水厂(A/O工艺)微生物群落结构及其动态变化[J].生态环境,2008,17(3):898-902.
[6]Wintzingerode F V,G?bel U B,Stackebrandt E.Determination of microbial diversity in environmental samples:Pitfalls of PCR-based r RNA analysis[J].FEMS Microbiology Reviews,1997,21(3):213-229.
[7]王绍祥,杨洲祥,孙真,等.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,2014,(3):196-203.
[8]孙豆豆.强化A/O工艺低温脱氮效能与菌群特性研究[D].哈尔滨:哈尔滨工业大学,2015.
[9]赵文莉.复合碳源填料深度反硝化脱氮特性研究[D].北京:北京工业大学,2015.
[10]户海燕,瞿思宜,张正哲,等.分子生物技术在厌氧氨氧化工艺研究中的应用[J].环境科学与技术,2016,(1):20-26+43.
[11]李鹏,毕学军,王军,等.常规和倒置A2/O工艺活性污泥微生物群落结构的比较[J].中国环境科学,2017,37(3):1137-1145.
[12]窦娜莎,王琳.不同温度下曝气生物滤池运行效能与微生物群落结构[J].环境工程学报,2016,10(6):2800-2806.
[13]李志静,孙宝盛,张笑雪,等.不同供氧策略和氨氮浓度下SBR中微生物群落的演变[J].环境工程学报,2017,11(1):359-365.
[14]Plósz B G,Jobbágy A,Jr G C.Factors influencing deterioration of denitrification by oxygen entering an anoxic reactor through the surface[J].Water Research,2003,37(4):853-863.
[15]邓仁健,张金松,曲志军.污泥浓度对双重后置反硝化工艺脱氮除磷的影响[J].环境科学研究,2014,27(7):797-803.
[16]辛海霞,季诚昌,张凡,等.MLSS对FBR处理高氨氮印染废水的影响[J].东华大学学报(自然科学版),2015,41(5):692-695.
[17]国家环境保护总局.水和废水监测分析方法[M].4版.北京:中国环境科学出版社,2002:258-281.
[18]Tanja M,Steven L.FLASH:fast length adjustment of short reads to improve genome assemblies[J].Bioinformatics,2011,27(21):2957-2936.
[19]Patrick D S,Sarah L W,Thomas R,et al.Introducing mothur:Open-Source,Platform-Independent,Community-Supported Software for Describing and Comparing Microbial Communities[J].Applied&Environmental Microbiology,2009,75(23):7537-7541.
[20]Robert C Edgar.UPARSE:highly accurate OTU sequences from microbial amplicon reads[J].Nature Methods,2013,10(10):996-998.
[21]Langille M G,Zaneveld J,Caporaso J G,et al.Predictive functional profiling of microbial communities using 16S r RNA marker gene sequences[J].Nature Biotechnology,2013,31(9):814-821.
[22]封莉,张立秋,吕炳南.污泥浓度对膜生物反应器运行特性的影响研究[J].哈尔滨工业大学学报,2003,35(3):307-310.
[23]Kawasaki K,Maruoka S,Katagami R,et al.Effect of initial MLSS,on operation of submerged membrane activated sludge process[J].Desalination,2011,281(20):334-339.
[24]张恩华,戴幼芬,肖勇,等.还原Cr(Ⅵ)的混菌胞外聚合物和细菌群落结构分析[J].中国环境科学,2017,37(1):352-357.
[25]Chung C M,Cho K W,Kim Y J,et al.Enhanced biological nitrogen removal in MLE combined with post-denitrification process and EF clarifier[J].Bioprocess&Biosystems Engineering,2012,35(4):503-511.
[26]Yang X,Zhou N,Chen M,et al.Analysis of microbial community structure in MBR with different ammonia concentrations using fluorescence in situ hybridization[J].Journal of Southeast University:Nature Science Edition,2013,43(2):380-385.
[27]宋云龙,张金松,朱佳,等.基于高通量测序的微生物强化污泥减量工艺中微生物群落解析[J].中国环境科学,2016,36(7):2099-2107.
[28]黄菲,梅晓洁,王志伟,等.冬季低温下MBR与CAS工艺运行及微生物群落特征[J].环境科学,2014,35(3):1002-1008.
[29]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(10):72-79.
[30]Ye L.,Shao M.F.,Zhang T,et al.Analysis of the bacterial community in a laboratory-scale nitrification reactor and a wastewater treatment plant by 454-pyrosequencing[J].Water Research,2011,45(15):4390–4398.
[31]李卫华,孙英杰,刘子梁,等.序批式生物反应器填埋场脱氮微生物多样性分析[J].环境科学,2016,37(1):342-349.
[32]杨华,黄钧,赵永贵,等.陶厄氏菌Thauera sp.strain TN9的鉴定及特性[J].应用与环境生物学报,2013,19(2):318-323.
[33]陈亚平,叶宏,王娟,等.亚硝化单胞菌的分离鉴定及其降解特性的研究[J].环境科学与技术,2007,30(9):24-25.
[34]Ma Q,Qu Y Y,Shen W L,et al.Bacterial community compositions of coking wastewater treatment plants in steel industry revealed by Illumina high-throughput sequencing[J].Bioresource Technology,2015,179:436-443.
[35]李哿.活性污泥—生物膜复合系统脱氮除磷试验研究[D].济南:山东建筑大学,2011.
[36]Ning L,Jin H,Han Y,et al.Pathways in bacterial and archaeal communities dictated by ammonium stress in a high solid anaerobic digester with dewatered sludge[J].Bioresource Technology,2017,241:95-102.
[37]田美,刘汉湖,申欣,等.百乐克(BIOLAK)活性污泥宏基因组的生物多样性及功能分析[J].环境科学,2015,36(5):1739-1748.
[38]Yu K,Zhang T.Metagenomic and metatranscriptomic analysis of microbial community structure and gene expression of activated sludge[J].Plos One,2012,7(5):e38183.