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432010 CHINACIVILENGINEERINGJOURNALVol.432010:,,:2010-09-29MBR李 军1 乔文燕1,2 王朝朝1 张雪松1 陈旭鸾1(1.,100124;2.,100086):、,SND。,MBR。,,DNA,PCR,(DGGE)。,,,。:;MBR;PCR-DGGE:X705 :A:1000-131X(2010)-0445-05AnalysisofmicrobialcommunitystructureandtheremovalofnitrogeninhybridMBRforlandfillleachatetreatmentLiJun1 QiaoWenyan1,2 WangZhaozhao1 ZhangXuesong1 ChenXuluan1(1.TheKeyLaboratoryofBeijingforWaterQualityScienceWaterEnvironmentRecoveryEngineering,BeijingUniversityofTechnology,Beijing100022,China;2.BeijingHaidianDistrictEnvironmentalSanitationScientificResearchInstitute,Beijing100086,China)Abstract:Theeffectsofdissolvedoxygen,temperatureonnitriteaccumulation,andthecontributionofSNDtototalnitrogenremovalwerestudied.TheresultsshowedthattheremovaleffectofHybridMBRProcessforlatelandfillleachatewasgood.Inthedifferentstages,thesampleswerecollectedrespectivelyfrommixedliquorandbiofilmofaMBR.DNAwasextractedfromthesixsamples,then16SrDNAfragmentswereamplifiedfromthetotalDNA,andthenwereusedfordenaturinggradientgelelectrophoresis(DGGE)analysis.Theresultsindicatedthatanoticeablechangetookplaceinmicrobialstructureininitialstages.Inthewholeoperation,bacterialcommunitystructuresarehigher.Keywords:landfillleachate;hybridmembranebioreactor(HMBR);PCR-DGGEE-mail:jglijun@bjut.edu.cn1 1.1 ,1,,COD,,。1.2 1,、、1 Table1 MainwaterqualityindexsduringexperimentCODcr(mg/L)(mg/L)pH(mg/L)BOD5/CODcr1986～34211000～22008.0～8.39000～142000.12～0.26DOI:10.15951/j.tmgcxb.2010.s2.003·446 · 2010,。24L,1∶3,。,。,,5cm,,。3(、)。,,,,0.2kg/(m3·d),,。3,1∶9、5∶5、,,6,1～6,-20℃。,(NO2--N/NOx--N)95%,。90%,COD50%。1 Fig.1 Thedevicesketchofreactor1.3 DOFApH,,。229℃、pH7.5FA。,MLSS4500mg/L、pH7.5、29℃。FA2.1mg/L、3.5mg/L、5.7mg/L7.8mg/L,DO0.5mg/L、1mg/L1.5mg/L。,2:2 FATable2 ConcentrationoftotalNH4+-NandFA(mg/L)50100150200FA1.42.84.25.7300500700900FA8.514.119.825.42 DOFAFig.2 EffectsofFAonaccumulationrateofnitriteindiffererntdissolvedoxygenconcentration2,DO0.5mg/L,FA,DO1mg/L1.5mg/L,FA。FA,FA5.7mg/L,DO1mg/L1.5mg/LDO0.5mg/L。FA5.7mg/L,DO,。,FA,DO,DO(0.5mg/L);FA,DO,FA(5.7mg/L),。DO0.5mg/L,312mg/L48mg/L18h;DO1mg/L,309mg/L51mg/L10h;DO1.5mg/L,301mg/L42mg/L9h。DO1mg/L1.5mg/L,DO0.5mg/L。1.4 ,DO1.0mg/L(:20℃、25℃、30℃)。,600mL,200mL,20℃、25℃、30℃,DO1.0mg/L1d,。,,, 43 ·MBR·447 ·。20℃,47%54%,,,30℃。3。3 Fig.3 Theremovalrateofammonianitrogenandthenitriteaccumulaterateindifferenttemperature1.5 COD4,COD。4 CODFig.4 ProcessvarialblecurveofCODandnitrogenduringsteadyoperation:2,1mg/L,30℃。,,2109mg/L870.7mg/L。,,30.4mg/L,8.6mg/L。,,,,,559.9mg/L,20.1mg/L,。、96%91%。,65%,,pH,,SND,,,,pH,。COD3420mg/L,,COD1781mg/L,,COD961mg/L,COD72%,,COD。1.6 DNAZhou[1]SDS。DNA10,PCR。。PCRPCR,PCR[2]。2 2.1 DNAPCRDNA1%,EB,,DNA23kb。PCR1%,PCR433bp,。2.2 PCRDGGE,15mlSYBR-Gold,UPV,。,,5。5 DGGEFig.5 DGGEprofileofthemicrobialcommunitiesinthereactor·448 · 20105,,4612、13,。,,,,,,。5,,,,,。,,。,;,;;,,;,。2.3 3,(95%)。,,(、、、Hyphomicrobiumsp.、Castellanielladenitrificanssp.Alcaligenessp.),,Hyphomicrobiumsp.,Castellanielladeni-trificanssp.Alcaligenessp.。,,。,[3](Co-mamonassp.)。,[4],(UnculturedAeromonassp.)。,。3 16SrDNATable3 Resultsof16SrDNAsequencesusingBLASTinGenBankaAM292279Nitrobactersp.Natopartial16SrRNAgene,strainNato96%bEF076127UnculturedChloroflexibacteriumcloneAD00716SribosomalRNAgene,partialsequence97%cAY856379Nitrosomonassp.OZK1116SribosomalRNAgene,partialsequence97%dFJ529954UnculturedbacteriumcloneMABRDTU116SribosomalRNAgene,partialsequence98%eAJ551095Hyphomicrobiumsp.wp14partial16SrRNAgene,isolatewp1499%fEU312975ProteobacteriumBF-416SribosomalRNAgene,partialsequence99%gAF169545Bacillussp.NRS-81016SribosomalRNAgene,partialsequence96%hEU179737PseudomonasputidastrainMG-Y216SribosomalRNAgene,partialsequence95%iDQ414434UnculturedNitrospirasp.clone1fromactivatedsludge16SribosomalRNAgene,partialsequence98%jAF338213NitrosococcusoceanistrainC-2716SribosomalRNAgene,partialsequence99%kFJ393072UnculturedAeromonassp.cloneMFC-B162-B0216SribosomalRNAgenegene,partialsequence98%lEU652562UnculturedbacteriumcloneC8S-13716SribosomalRNAgene,partialsequence100%mDQ640676UnculturedBacteroidetesbacteriumcloneSkagenf9316SribosomalRNAgene,partialsequence98%nU82826CastellanielladenitrificansstrainNKNTAU16SribosomalRNAgene,completesequence97%oAJ005450Alcaligenesdefragrans16SrRNAgene,strain65Phen98%pDQ822531BacteriumQM4616SribosomalRNAgene,partialsequence96%qEU107758Comamonassp.P3-316SribosomalRNAgene,partialsequence97%3 (1)FA,FA,DO。。20℃,47%54%, 43 ·MBR·449 ·,,30℃。(2),,,。(3),,。(4),,、,,。[1]ZhouJ,BrunsMA,TiedjeJM.DNArecoveryfromsoilsofdiversecomposition[J].AppliedMicrobiologyandBiotechnology,1996,62(2):316-322[2]BaeW,BaekSC,ChungJ,etal.Optimaloperationfactorsfornitriteaccumulationinbatchreactors[J].Biodegradation,2002,12:359-366[3]BalaSS,YanS,TyagiRD,etal.Isolationofextracellularbiopolymerproducingmicroorganismsfromwastewatersludgeforsludgesettlinganddewatering[C]//ProceedingsofWaterEnvironmentFederation,2006:473-489[4],,,.MBR[J].,2008,28(11):2192-2199(ZhangBin,SunBaosheng,JiMin,etal.Analysisandsuccessionofmicrobialcommunitystructureinamembranebioreactor[J].ActaScientiaeCircumstantiae,2008,