微氧折流反应器中产甲烷菌的分布与群落结构分析
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摘要
为了研究微氧条件下折流反应器处理生活污水的微生物学机理,针对产甲烷菌的MCR基因采用实时荧光定量PCR和构建克隆文库的方法,对反应器中的关键功能微生物产甲烷菌的分布特点与群落结构变化进行了分析。结果表明,微氧条件下折流反应器的环境条件可以保证产甲烷菌的增长,不同格室的产甲烷菌丰度均要高于接种污泥(1 720 copies/ng),其中经过微氧曝气处理的1#(2 864 copies/ng)和2#(2 282 copies/ng)格室产甲烷菌丰度低于厌氧状态的3#(3 508copies/ng)格室;微氧处理后折流反应器产甲烷菌的群落结构发生了明显的变化,接种污泥的产甲烷菌多属于Methanomicrobiales目的 Methanoregulaceae属(6/9),而保持微氧状态的1#格室中产甲烷菌多隶属于Methanomicrobiales目(6/10)的其他类群,2#格室的产甲烷菌则多隶属于Methanobacteriales目(5/10),保持厌氧状态的3#格室的产甲烷菌主要属于Methanomicrobiales目(3/10)和Methanosarcinales目(3/10);产甲烷菌MCR基因的多样性指数表明微氧曝气的引入降低了活性污泥中产甲烷菌的生物多样性。
In order to study the microbial mechanism of the baffled reactor treating domestic wastewater under microaerobic condition,the distribution and the community structure of methanogen which is the key functional microorganism in the reactor were analyzed by real-time quantitative PCR and clone library. The results showed that the environmental conditions of the baffled reactor under microaerobic operation would ensure the growth of methanogens. The abundance of methanogens in different cells were higher than those in the inoculation sludge( 1 720 copies /ng). The abundance of methanogens of the cell 1#( 2 864 copies/ng) and 2#( 2 282 copies /ng) which were treated by micro-oxygen aeration were lower than that of the cell 3#( 3 508 copies/ng) in the anaerobic state. There was a significant change of the community structure of methanogens in the baffled reactor under micoaerobic condition. The most of methanogens of the inoculation sludge fell into the family of Methanoregulaceae( 6 /9) associated with the order of Methanomicrobiales,while the methanogens of the cell 1#were mostly affiliated with the other families within the order of Methanomicrobiales( 6 /10). The methanogens of the cell 2#were mostly affiliated with the order of Methanobacteriales( 5 /10),and the methanogens of the cell 3#maintained in anaerobic state mainly belonged to the order of Methanomicrobiales( 3 /10) and Methanosarcinales( 3/10). The diversity indices based on the sequences of MCR gene showed that the introducing of micro-oxygen aeration reduced the biodiversity of methanogen in the activated sludge.
In order to study the microbial mechanism of the baffled reactor treating domestic wastewater under microaerobic condition,the distribution and the community structure of methanogen which is the key functional microorganism in the reactor were analyzed by real-time quantitative PCR and clone library. The results showed that the environmental conditions of the baffled reactor under microaerobic operation would ensure the growth of methanogens. The abundance of methanogens in different cells were higher than those in the inoculation sludge( 1 720 copies /ng). The abundance of methanogens of the cell 1#( 2 864 copies/ng) and 2#( 2 282 copies /ng) which were treated by micro-oxygen aeration were lower than that of the cell 3#( 3 508 copies/ng) in the anaerobic state. There was a significant change of the community structure of methanogens in the baffled reactor under micoaerobic condition. The most of methanogens of the inoculation sludge fell into the family of Methanoregulaceae( 6 /9) associated with the order of Methanomicrobiales,while the methanogens of the cell 1#were mostly affiliated with the other families within the order of Methanomicrobiales( 6 /10). The methanogens of the cell 2#were mostly affiliated with the order of Methanobacteriales( 5 /10),and the methanogens of the cell 3#maintained in anaerobic state mainly belonged to the order of Methanomicrobiales( 3 /10) and Methanosarcinales( 3/10). The diversity indices based on the sequences of MCR gene showed that the introducing of micro-oxygen aeration reduced the biodiversity of methanogen in the activated sludge.
引文
[1]Barber W.P.,Stuckey D.C.The use of the anaerobic baffled reactor(ABR)for wastewatertreatment:A review.Water Research,1999,33(7):1559-1578
[2]童昶,沈耀良,赵丹,等.厌氧反应器技术的发展及ABR反应器的工艺特点.江苏环境科技,2001,14(4):9-11Tong Chang,Shen Yaoliang,Zhao Dan,et al.Development of anaerobic reactor technology and process characteristics of ABR reactor.Jiangsu Environmental Science and Technology,2001,14(4):9-11(in Chinese)
[3]董春娟,吕炳南,陈志强.微氧条件下厌氧颗粒污泥和消化污泥特性研究.南京理工大学学报,2005,29(2):216-222Dong Chunjuan,LüBingnan,Chen Zhiqiang.Characteristic of anaerobic granular sludge and digestion sludge under microaerobic conditions.Journal of Nanjing University of Science and Technology,2005,29(2):216-222(in Chinese)
[4]董春娟,吕炳南.EGSB反应器在微氧条件下的运行特性.环境科学学报,2006,26(2):220-224Dong Chunjuan,LüBingnan.Performance characteristic of the EGSB reactor in the microaerobic conditions.Acta Scientiae Circumstantiae,2006,26(2):220-224(in Chinese)
[5]游胜,周兴求,牛晓君,等.IC反应器在微氧条件下的运行特性研究.中国给水排水,2009,25(9):16-19You Sheng,Zhou Xingqiu,Niu Xiaojun,et al.Performance characteristic of IC reactor under micro-aerobic condition.China Water&Wastewater,2009,25(9):16-19(in Chinese)
[6]蓝惠霞,邱献欢,隋冰冰,等.降解五氯酚(PCP)的微氧颗粒污泥的形成机理.环境科学学报,2009,29(2):273-278Lan Huixia,Qiu Xianhuan,Sui Bingbing,et al.The granulation mechanism of micro-aerobic granules for PCP degradation.Acta Scientiae Circumstantiae,2009,29(2):273-278(in Chinese)
[7]董春娟,吕炳南.厌氧颗粒污泥的性能研究.环境工程学报,2007,6(1):102-108Dong Chunjuan,LüBingnna.Characteristic research of anaerobic granules.Chinese Journal of Environmental Engineering,2007,6(1):102-108(in Chinese)
[8]Zhou W.,Imai T.,Ukita M.,et al.Effect of limited aeration on the anaerobic treatment of evaporator condensate from a sulfite pulp mill.Chemosphere,2007,66(5):924-929
[9]胡颖华,孙丰霞,高廷耀,等.活性污泥法污水厂剩余污泥微氧消化的实验研究.中国沼气,2005,23(1):3-6Hu Yinghua,Sun Fengxia,Gao Tingyao,et al.The research of excess sludge digestion in low oxygen concentration.China Biogas,2005,23(1):3-6(in Chinese)
[10]李刚,周兴求,伍健东.微氧条件下厌氧折流板反应器运行特性研究.环境工程学报,2009,3(7):1263-1267Li Gang,Zhou Xingqiu,Wu Jiandong.Study on performance characteristic of anaerobic baffled reactor under microaerobic conditions.Chinese Journal of Environmental Engineering,2009,3(7):1263-1267(in Chinese)
[11]Skiadas I.V.,GavalaG H.N.,Lyberatos.Modelling of the periodic anaerobic baffled reactor(PABR)based on the retaining factor concept.Water Research,2000,34(15):3725-3736
[12]Hales B.A.,Edwards C.,Ritchie D.A.,et al.Isolation and identification of methanogen-specific DNA from blanket bog peat by PCR amplification and sequence analysis.Applied and Environmental Microbiology,1996,62(2):668-675
[13]Hallam S.J.,Girguis P.R.,Preston C.M.,et al.Identification of methyl coenzyme M reductase A(mcrA)genes associated with methane-oxidizing archaea.Applied and Environmental Microbiology,2003,69(9):5483-5491
[14]Schloss P.D.,Handelsman J.Introducing DOTUR,a computer program for defining operational taxonomic units and estimating species richness.Applied and Environmental Microbiology,2005,71(3):1501-1506
[15]胡林林,王建龙,文湘华,等.低溶解氧条件下生物脱氮研究中的新现象.应用与环境生物学报,2003,9(4):444-447Hu Linlin,Wang Jianlong,Wen Xianghua,et al.Improvements on biological nitrogen removal under low dissolved oxygen condition.Chinese Journal of Appled and Environ.Biology,2003,9(4):444-447(in Chinese)
[16]Alvarez-Cohen L.,McCarty P.L.Two-stage dispersedgrowth treatment of halogenated aliphatic compounds by cometabolism.Environmental Science&Technology,1991,25(80):1387-1393
[17]丁维新,蔡祖聪.植物在CH4产生、氧化和排放中的作用.应用生态学报,2003,14(8):1379-1384Ding Weixin,Cai Zucong.Effect of plants on methane production,oxidation and emission.Chinese Journal of Applied Ecology,2003,14(8):1379-1384(in Chinese)
[2]童昶,沈耀良,赵丹,等.厌氧反应器技术的发展及ABR反应器的工艺特点.江苏环境科技,2001,14(4):9-11Tong Chang,Shen Yaoliang,Zhao Dan,et al.Development of anaerobic reactor technology and process characteristics of ABR reactor.Jiangsu Environmental Science and Technology,2001,14(4):9-11(in Chinese)
[3]董春娟,吕炳南,陈志强.微氧条件下厌氧颗粒污泥和消化污泥特性研究.南京理工大学学报,2005,29(2):216-222Dong Chunjuan,LüBingnan,Chen Zhiqiang.Characteristic of anaerobic granular sludge and digestion sludge under microaerobic conditions.Journal of Nanjing University of Science and Technology,2005,29(2):216-222(in Chinese)
[4]董春娟,吕炳南.EGSB反应器在微氧条件下的运行特性.环境科学学报,2006,26(2):220-224Dong Chunjuan,LüBingnan.Performance characteristic of the EGSB reactor in the microaerobic conditions.Acta Scientiae Circumstantiae,2006,26(2):220-224(in Chinese)
[5]游胜,周兴求,牛晓君,等.IC反应器在微氧条件下的运行特性研究.中国给水排水,2009,25(9):16-19You Sheng,Zhou Xingqiu,Niu Xiaojun,et al.Performance characteristic of IC reactor under micro-aerobic condition.China Water&Wastewater,2009,25(9):16-19(in Chinese)
[6]蓝惠霞,邱献欢,隋冰冰,等.降解五氯酚(PCP)的微氧颗粒污泥的形成机理.环境科学学报,2009,29(2):273-278Lan Huixia,Qiu Xianhuan,Sui Bingbing,et al.The granulation mechanism of micro-aerobic granules for PCP degradation.Acta Scientiae Circumstantiae,2009,29(2):273-278(in Chinese)
[7]董春娟,吕炳南.厌氧颗粒污泥的性能研究.环境工程学报,2007,6(1):102-108Dong Chunjuan,LüBingnna.Characteristic research of anaerobic granules.Chinese Journal of Environmental Engineering,2007,6(1):102-108(in Chinese)
[8]Zhou W.,Imai T.,Ukita M.,et al.Effect of limited aeration on the anaerobic treatment of evaporator condensate from a sulfite pulp mill.Chemosphere,2007,66(5):924-929
[9]胡颖华,孙丰霞,高廷耀,等.活性污泥法污水厂剩余污泥微氧消化的实验研究.中国沼气,2005,23(1):3-6Hu Yinghua,Sun Fengxia,Gao Tingyao,et al.The research of excess sludge digestion in low oxygen concentration.China Biogas,2005,23(1):3-6(in Chinese)
[10]李刚,周兴求,伍健东.微氧条件下厌氧折流板反应器运行特性研究.环境工程学报,2009,3(7):1263-1267Li Gang,Zhou Xingqiu,Wu Jiandong.Study on performance characteristic of anaerobic baffled reactor under microaerobic conditions.Chinese Journal of Environmental Engineering,2009,3(7):1263-1267(in Chinese)
[11]Skiadas I.V.,GavalaG H.N.,Lyberatos.Modelling of the periodic anaerobic baffled reactor(PABR)based on the retaining factor concept.Water Research,2000,34(15):3725-3736
[12]Hales B.A.,Edwards C.,Ritchie D.A.,et al.Isolation and identification of methanogen-specific DNA from blanket bog peat by PCR amplification and sequence analysis.Applied and Environmental Microbiology,1996,62(2):668-675
[13]Hallam S.J.,Girguis P.R.,Preston C.M.,et al.Identification of methyl coenzyme M reductase A(mcrA)genes associated with methane-oxidizing archaea.Applied and Environmental Microbiology,2003,69(9):5483-5491
[14]Schloss P.D.,Handelsman J.Introducing DOTUR,a computer program for defining operational taxonomic units and estimating species richness.Applied and Environmental Microbiology,2005,71(3):1501-1506
[15]胡林林,王建龙,文湘华,等.低溶解氧条件下生物脱氮研究中的新现象.应用与环境生物学报,2003,9(4):444-447Hu Linlin,Wang Jianlong,Wen Xianghua,et al.Improvements on biological nitrogen removal under low dissolved oxygen condition.Chinese Journal of Appled and Environ.Biology,2003,9(4):444-447(in Chinese)
[16]Alvarez-Cohen L.,McCarty P.L.Two-stage dispersedgrowth treatment of halogenated aliphatic compounds by cometabolism.Environmental Science&Technology,1991,25(80):1387-1393
[17]丁维新,蔡祖聪.植物在CH4产生、氧化和排放中的作用.应用生态学报,2003,14(8):1379-1384Ding Weixin,Cai Zucong.Effect of plants on methane production,oxidation and emission.Chinese Journal of Applied Ecology,2003,14(8):1379-1384(in Chinese)