3DBER-S反硝化脱氮性能及其菌群特征
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摘要
针对污水处理厂尾水TN去除问题,采用16S r DNA克隆文库法,探究了3DBER-S(三维电极生物膜耦合硫自养脱氮工艺)的强化脱氮机制及其菌群特征.结果表明,I(电流)和HRT(水力停留时间)对3DEBR-S中氢自养和硫自养反硝化作用所占比例的影响较大,但对脱氮效率影响不显著.当进水C/N〔ρ(CODCr)/ρ(TN)〕为1、ρ(NO3--N)为35 mg/L、I为300 m A、HRT为4 h时,NO3--N和TN去除率可分别稳定在80%和74%以上.16S r DNA克隆文库结果显示,反应器中β变形菌纲为优势菌群,占47.89%〔以OUT(操作单元)计〕.在β变形菌纲中,与具有反硝化功能的陶厄氏菌属(Thauera)相似的细菌所占比例最大,为52.94%;与可分别利用硫和氢为电子供体进行反硝化脱氮的硫杆菌属(Thiobacillus)和食酸菌属(Acidovorax)相似的细菌分别占17.65%和14.71%.3DBER-S中存在异养联合氢自养和硫自养反硝化协同去除硝酸盐氮的作用,可为反硝化脱氮提供充足的电子供体,节约了有机碳源消耗,并保证了稳定高效的脱氮效果.
To strengthen the removal TN for wastewater treatment plant(WWTP) effluent,the performance and bacterial community of a coupling 3-dimensional biofilm-electrode with sulfur autotrophic denitrification technology(3DBER-S) was investigated under different operating condition experiments and 16 S r DNA clone library.It was found that the influences of electric current and hydraulic retention time(HRT) were obvious on the proportion of hydrogen and sulfur autotrophic denitrification process in 3DBER-S,but there was no significant impact on the denitrification performance.When CODCr/TN(C/N) was 1 and NO3--N concentration was 35 mg/L in the influent,NO3--N and TN removal rates were above 80% and 74% respectively under conditions of electric current of 300 m A and HRT of4 h.The analysis of 16 S r DNA clone library showed that the dominant populations were β-Proteobacteria(47.89%) in the biofilm of3DBER-S.52.94% of the β-Proteobacteria consisted of a bacteria similar to Thauera that could perform denitrification.Moreover,the bacteria similar to Thiobacillus and Acidovorax accounted for 17.65% and 14.71% respectively of the β-Proteobacteria,which can use sulfur and hydrogen as an electron donor for denitrification respectively.There is a mutual complementation among heterotrophic,hydrogen and sulfur autotrophic denitrification processes,which supply 3DBER-S with enough electron donors.Then the stable and highperformance nitrogen removal can be maintained under different operating conditions,even if the carbon source of the influent is insufficient.
To strengthen the removal TN for wastewater treatment plant(WWTP) effluent,the performance and bacterial community of a coupling 3-dimensional biofilm-electrode with sulfur autotrophic denitrification technology(3DBER-S) was investigated under different operating condition experiments and 16 S r DNA clone library.It was found that the influences of electric current and hydraulic retention time(HRT) were obvious on the proportion of hydrogen and sulfur autotrophic denitrification process in 3DBER-S,but there was no significant impact on the denitrification performance.When CODCr/TN(C/N) was 1 and NO3--N concentration was 35 mg/L in the influent,NO3--N and TN removal rates were above 80% and 74% respectively under conditions of electric current of 300 m A and HRT of4 h.The analysis of 16 S r DNA clone library showed that the dominant populations were β-Proteobacteria(47.89%) in the biofilm of3DBER-S.52.94% of the β-Proteobacteria consisted of a bacteria similar to Thauera that could perform denitrification.Moreover,the bacteria similar to Thiobacillus and Acidovorax accounted for 17.65% and 14.71% respectively of the β-Proteobacteria,which can use sulfur and hydrogen as an electron donor for denitrification respectively.There is a mutual complementation among heterotrophic,hydrogen and sulfur autotrophic denitrification processes,which supply 3DBER-S with enough electron donors.Then the stable and highperformance nitrogen removal can be maintained under different operating conditions,even if the carbon source of the influent is insufficient.
引文
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[18]OSAKA T,YOSHIE S,TSUNEDA S,et al.Identification of acetate or methanol assimilating bacteria under nitrate-reducing conditions by stable-isotope probing[J].Microbial Ecology,2006,52(2):253-266.
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[23]MAO Yanping,XIA Yu,ZHANG Tong.Characterization of Thauera-dominated hydrogen-oxidizing autotrophic denitrifying microbial communities by using high-throughput sequencing[J].Bioresource Technology,2013,128:703-707.
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[26]VASILIADOU I A,PAVLOU S,VAYENAS D V.Dynamics of a chemostat with three competitive hydrogen oxidizing denitrifying microbial populations and their efficiency for denitrification[J].Ecological Modelling,2009,220:1169-1180.
[27]NALCACI O O,BOKE N,OVEZ B.Potential of the bacterial strain Acidovorax avenae subsp.avenae LMG 17238 and macro algae Gracilaria verrucosa for denitrification[J].Desalination,2011,274:44-53.
[28]陶天申,杨瑞馥,东秀珠.原核生物系统学[M].北京:化学工业出版社,2007:295-296.
[29]鲍立宁,洪桂云,黄显怀.电极生物膜脱氮工艺中反硝化菌相分析[J].安徽建筑工业学院学报(自然科学版),2004,12(5):1-4.BAO Lining,HONG Guiyun,HUANG Xianhuai.Analysis of denitriying bacteria in biofilm-electrode process[J].Journal of Anhui Institute of Architecture&Industry(Science and Technology),2004,12(5):1-4.
[30]姜磊,徐成斌,马溪平,等.1株好氧反硝化菌的分离鉴定和反硝化特性研究[J].环境科学与技术,2013,36(3):12-15.JIANG Lei,XUN Chengbin,MA Xiping,et al.Identification and denitrification characteristics of an aerobic denitirfier[J].Environmental Science&Technology(China),2013,36(3):12-15.
[31]LABRENZ M,GROTE J,MAMMITZSCH K,et al.Sulfurimonas gotlandica sp.nov.,a chemoautotrophic and psychrotolerant epsilonproteobacterium isolated from a pelagic redoxcline,and an emended description of the genus Sulfurimonas[J].International Journal of Systematic and Evolutionary Microbiology,2013,63:4141-4148.
[32]田春杰,陈家宽,钟扬.微生物系统发育多样性及其保护生物学意义[J].应用生态学报,2003,14(4):509-612.TIAN Chunjie,CHEN Jiankuan,ZHONG Yang.Phylogenetic diversity of microbes and its perspectives in conservation biology[J].Chinese Journal of Applied Ecology,2003,14(4):509-612.
[2]官策,郁达伟,郑祥,等.我国人工湿地在城市污水处理厂尾水脱氮除磷中的研究与应用进展[J].农业环境科学学报,2012,31(12):2309-2320.GUAN Ce,YU Dawei,ZHENG Xiang,et al.Removing nitrogen and phosphorus of effluent from wastewater treatment plants by constructed wetlands in China:an overview[J].Journal of AgroEnvironment Science,2012,31(12):2309-2320.
[3]罗国芝,鲁璐,杜军,等.循环水养殖用水中反硝化碳源研究现状[J].渔业现代化,2011,38(3):11-16.LUO Guozhi,LU Lu,DU Jun,et al.Current status of the research about the carbon source of the heterotrophic denitrification of recirculating aquaculture system[J].Fishery Modernization,2011,38(3):11-16.
[4]ZHAO Yingxin,FENG Chuanping,WANG Qinghong,et al.Nitrate removal from groundwater by cooperating heterotrophic with autotrophic denitrification in a biofilm-electrode reactor[J].Journal of Hazardous Materials,2011,192:1033-1039.
[5]GHAFARI S,HASAN M,AROUA M K.Bio-electrochemical removal of nitrate from water and wastewater:a review[J].Bioresource Technology,2008,99:3965-3974.
[6]ZHOU Minghua,FU Wenjing,GU Heyan,et al.Nitrate removal from groundwater by a novel three-dimensional electrode biofilm reactor[J].Electrochimica Acta,2007,52:6052-6059.
[7]SUN Yimin,NEMATI M.Evaluation of sulfur-based autotrophic denitrification and denitritation for biological removal of nitrate and nitrite from contaminated waters[J].Bioresource Technology,2012,114:207-216.
[8]QAMBRANI N A,OH S E.Influence of reactive media composition and chemical oxygen demand as methanol on autotrophic sulfur denitrification[J].Journal of Microbiology and Biotechnology,2012,22(8):1155-1160.
[9]WANG Dongjin,LIU Huijuan,QU Jiuhui,et al.Using the combined bioelectrochemical and sulfur autotrophic denitrification system for groundwater denitrification[J].Bioresource Technology,2009,100:142-148.
[10]李素梅,郝瑞霞,孟成成.三维电极生物膜反应器低温启动试验研究[J].中国给水排水,2013,29(5):101-105.LI Sumei,HAO Ruixia,MENG Chengcheng.Start-up of threedimensional electrode biofilm reactor at low temperature[J].China Water&Wastewater,2013,29(5):101-105.
[11]李建婷,纪树兰,刘志培,等.16S r DNA克隆文库方法分析好氧颗粒污泥细菌组成[J].环境科学研究,2009,22(10):1218-1223.LI Jianting,JI Shulan,LIU Zhipei,et al.Analysis of bacterial composition of aerobic granular sludge with 16S r DNA clone library[J].Research of Environmental Sciences,2009,22(10):1218-1223.
[12]PARK H I,KIM D K,CHOI Y J,et al.Nitrate reduction using an electrode as direct electron donor in a biofilm-electrode reactor[J].Process Biochemistry,2005,40:3383-3388.
[13]庞朝晖,刘迎云,刘文碧.温度和电流对电极生物膜反硝化效果的影响[J].南华大学学报:自然科学版,2007,21(2):60-63.PANG Zhaohui,LIU Yingyun,LIU Wenbi.The effect of temperature and current on ration biofilm-electrode denitrification[J].Journal of University of South China:Science and Technology,2007,21(2):60-63.
[14]GHAFARI S,HASAN M,AROUA M K.Nitrate remediation in a novel upflow bio-electrochemical reactor(UBER)using palm shell activated carbon as cathode material[J].Electrochimica Acta,2009,54:4164-4171.
[15]胡传侠,杨昌柱,杨群,等.固定化三维电极-生物膜法去除污水中硝酸盐氮[J].环境科学与技术,2008,31(2):83-87.HU Chuanxia,YANG Changzhu,YANG Qun,et al.Study on removal of nitrate from by immobilized three-dimension electrodebiofilm reactor[J].Environmental Science&Technology(China),2008,31(2):83-87.
[16]BATCHELOR B,LAWRENCE A W.Autotrophic denitrification using elemental sulfur[J].Journal of the Water Pollution Control Federation,1978,50(8):1986-2001.
[17]XIA Siqing,LI Jixiang,WANG Rongchang,et al.Tracking composition and dynamics of nitrification and denitrification microbial community in a biofilm reactor by PCR-DGGE and combining fish with flow cytometry[J].Biochemical Engineering Journal,2010,49(3):370-378.
[18]OSAKA T,YOSHIE S,TSUNEDA S,et al.Identification of acetate or methanol assimilating bacteria under nitrate-reducing conditions by stable-isotope probing[J].Microbial Ecology,2006,52(2):253-266.
[19]DANIEL L M C,POZZI E,FORESTI E,et al.Removal of ammonium via simultaneous nitrification-denitrification nitriteshortcut in a single packed-bed batch reactor[J].Bioresource Technology,2009,100(3):1100-1107.
[20]ETCHEBEHERE C,ERRAZQUIN M I,DABERT P,et al.Community analysis of a denitrifying reactor treating landfill leachate[J].FEMS Microbiology Ecology,2002,40(2):97-106.
[21]OSAKA T,SHIROTANI K,YOSHIE S,et al.Effects of carbon source on denitrification efficiency and microbial community structure in a saline wastewater treatment process[J].Water Research,2008,42(14):3709-3718.
[22]毛跃建.废水处理系统中重要功能类群Thauera属种群结构与功能的研究[D].上海:上海交通大学,2009.
[23]MAO Yanping,XIA Yu,ZHANG Tong.Characterization of Thauera-dominated hydrogen-oxidizing autotrophic denitrifying microbial communities by using high-throughput sequencing[J].Bioresource Technology,2013,128:703-707.
[24]HAO Ruixia,LI Sumei,LI Jingbing,et al.Denitrification of simulated municipal wastewater treatment plant effluent using a three-dimensional biofilm-electrode reactor:operating performance and bacterial community[J].Bioresource Technology,2013,143:178-186.
[25]KELLY D P,WOOD A P.Confirmation of Thiobacillus denitrificans as a species of the genus Thiobacillus in the beta-subclass of the Proteobacteria,with strain NCIMB 9548 as the type strain[J].International Journal of Systematic and Evolutionary Microbiology,2000,50:547-550.
[26]VASILIADOU I A,PAVLOU S,VAYENAS D V.Dynamics of a chemostat with three competitive hydrogen oxidizing denitrifying microbial populations and their efficiency for denitrification[J].Ecological Modelling,2009,220:1169-1180.
[27]NALCACI O O,BOKE N,OVEZ B.Potential of the bacterial strain Acidovorax avenae subsp.avenae LMG 17238 and macro algae Gracilaria verrucosa for denitrification[J].Desalination,2011,274:44-53.
[28]陶天申,杨瑞馥,东秀珠.原核生物系统学[M].北京:化学工业出版社,2007:295-296.
[29]鲍立宁,洪桂云,黄显怀.电极生物膜脱氮工艺中反硝化菌相分析[J].安徽建筑工业学院学报(自然科学版),2004,12(5):1-4.BAO Lining,HONG Guiyun,HUANG Xianhuai.Analysis of denitriying bacteria in biofilm-electrode process[J].Journal of Anhui Institute of Architecture&Industry(Science and Technology),2004,12(5):1-4.
[30]姜磊,徐成斌,马溪平,等.1株好氧反硝化菌的分离鉴定和反硝化特性研究[J].环境科学与技术,2013,36(3):12-15.JIANG Lei,XUN Chengbin,MA Xiping,et al.Identification and denitrification characteristics of an aerobic denitirfier[J].Environmental Science&Technology(China),2013,36(3):12-15.
[31]LABRENZ M,GROTE J,MAMMITZSCH K,et al.Sulfurimonas gotlandica sp.nov.,a chemoautotrophic and psychrotolerant epsilonproteobacterium isolated from a pelagic redoxcline,and an emended description of the genus Sulfurimonas[J].International Journal of Systematic and Evolutionary Microbiology,2013,63:4141-4148.
[32]田春杰,陈家宽,钟扬.微生物系统发育多样性及其保护生物学意义[J].应用生态学报,2003,14(4):509-612.TIAN Chunjie,CHEN Jiankuan,ZHONG Yang.Phylogenetic diversity of microbes and its perspectives in conservation biology[J].Chinese Journal of Applied Ecology,2003,14(4):509-612.