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811Vol．8No．11201411ChineseJournalofEnvironmentalEngineeringNov.2014SBＲ11*2111．3000722．300221。SBＲ-PCＲ-DGGESBＲ。MBＲSBＲSBＲ、γ-。。16SrDNAPCＲ-DGGEX703A1673-9108201411-4813-07SuccessionofmicrobialcommunityinSBＲtreatingnatatoriumwastewaterunderdifferentsludgeloadingshocksZhaoShuanghong1SunBaosheng1YuFengqing2ZhangXiaoxue1QiGengshen11．SchoolofEnvironmentalScienceandTechnologyTianjinUniversityTianjin300072China2．TianjinCaringTechnologyDevelopmentCo．Ltd．Tianjin300221ChinaAbstractSludgeloadingcouldaffectthegrowthpatternofmicroorganismdirectlywhenthesludgeload-ingchangedmicrobialcommunitystructurewouldchangeobviouslyinashortperiodoftime．Inordertoinvesti-gatetheeffectsofsludgeloadingshockonmicrobialcommunityconstructureinSBＲactivatedsludgePCＲ-dena-turinggradientgelelectrophoresisPCＲ-DGGEwasusedtoanalyzethemicrobialcommunitystructureofaSBＲtreatingnatatoriumwastewaterunderthedifferentshocksofsludgeloading．TheresultsindicatedthatmicrobialcommunitystructurehadobvioussuccessioninSBＲtreatingnatatoriumwastewatercultivatedofMBＲunderthedifferentshocksofsludgeloading．Thediversityindexincreasedandtendedtostabilizewiththesludgeloadingshockincreasingbutthediversityindexdropedundertheexcessivesludgeloading．ThedominantbacteriainSBＲsludgewereunculturedbacteriumEnterobacteriaceaeTolumonassp．andGammaproteobacteriumbacteri-aect．Duetotheself-modulationofthemicroorganismstheactivatedsludgeformsthecommunitystructurewhichadaptstothedifferentsludgeloads．Keywordssludgeloadingshock16SrDNAPCＲ-DGGEactivatedsludgemicrobialcommunitystruc-ture2013－10－222013－11－291990—。E-mailzhaoshuanghong927@126．com*E-mailbaosheng_sun@sina．com。。、1。Ns-2。NS。8。。。PCＲ-DGGEDNAＲNA3456。PCＲ-DGGEMBＲSBＲSBＲ。。11．1SBＲ1112mm、106mm、500mm400mm4L。6∶4。。。MBＲ25d。1Fig．1SchematicdiagramoftheprocessCODBOD。、、。7。Ns0.2～0.5kgBOD5/kgMLSS·dNs0.05～0.15kgBOD5/kgMLSS·dNs0.03～0.08kgBOD5/kgMLSS·dNs1.5～3kgBOD5/kgMLSS·d871。2dPCＲ-DGGE。1Table1OperationconditionsMLSS-CODmg/LkgCOD/kgMLSS·dmg/L/hNS1NS22506～18571860～21390．02～0．030．10～0．2338～39287～2948844NS3Ns42254～29183106～45970．37～0．480．55～0．73481～497762～8536422NS5NS6NS75132～58732160～30472072～30891．08～1．381．45～1．841．91～2．561585～1681885～985885～985332111418411SBＲ1．2DNA1．2．110mL8mL3000×g4min。14min。DNA。1．2．2DNADNA-\\-9。1．3PCＲ16SrDNAV3F357-GC5'-CGCCCGC-CGCGCCCCGCGCCCGGCCCGCCGCCCCCGCCCCCCTACGGGAGGCAGCAG-3'Ｒ5185'-ATTACCGCG-GCTGCTGG-3'“GC”240bp10。PCＲPCＲ94℃5min2094℃1min65～55℃1min72℃1min0.5℃1094℃1min55℃1min72℃1min72℃8min11。50μL10～100ngDNA25μL2×TaqPCＲMasterMix1μL25μmol/LF357-GC1μL25μmol/LＲ51850μL。1．416SrDNADGGEC．B．S．SCIENTIFICDGGE-2001PCＲ1213。1．5DGGE1.5mL50μL60～100℃20min4℃DNA。5000r/min5min6μLPCＲ。PCＲ1.5%PCＲDNA。1．6DGGEBio-ＲadQuantityOne4．26DGGE。Shannon-Wiener。Cs。DGGE。HH=－∑si=1PilogPi=－∑si=1ni/Nlogni/N1Pi=ni/NniiN。Cs=2j/a+b2jABabAB。22．1DGGESBＲ7PCＲDGGE2。2SBＲ。Ns1、Ns2。Ns3、Ns4。Ns5、Ns6Ns7。2SBＲUnculturedbacteri-um14。15。SBＲ51848γ-Gammaproteobacterium、TolumonasEnterobacteriaceaebacterium。2NsPCＲDGGEFig．2DGGEprofileofbacteriaunderdifferentsludgeloadingshocksSBＲBanda、b、c、e。BandaEnterobacteriaceaebacterium16Enter-obactercloscae17。BandeTolumonas。216SrDNATable2SequencinganalysisofpartofmicrobialcommunityofdominantbacteriaNCBIaAB714459DHL-1399%bGU738449F1Q32TO03DCNL0100%cGU905013SINI91197%dJN125355FJ790139MBfＲ_Br-195%γ-S14-196%eJF808898Ｒ15-1898%fHQ609675AS-11100%gHQ482732Pohang_WWTP_December．2005_395994%hJQ480506Transfer_2C0698%。Bandd、hm。Banddγ-Gammaproteobacteri-umγ-Gammaproteobacterium。。14MBＲγ-。Kapley18γ-。proteobacterium。、。BanddAzoarcus19。BandfNs3、Ns4Ns5Ns。618411SBＲ。BandgNs1Ns4、Ns5、Ns6Ns7。2Ns3、Ns4、Ns5、Ns6Ns7NS。NsNs。2．2QuanlityoneDGGEShannonSBＲ3。3ShannonFig．3Shannon-wienerindexofsamplesundersludgeloadingshock3Ns1SBＲ0.57。0.89。。SBＲ。Ns6Ns7。2．3NS4Ns1Ns1。4Ns6、Ns7Ns150%SBＲ。Ns1SBＲ。4Fig．4SimilarityindexofdominantbacteriaundersludgeloadingshockUPGMA5。5。Ns6、Ns7Ns1、Ns2Ns3、Ns5。Ns6、Ns7。Ns1、Ns2、Ns3Ns580%、71848。5DGGEFig．5ClusteranalysisofbacterialDGGEundersludgeloadingshock31SBＲ。。SBＲ。2SBＲ。。733。3SBＲγ-GammaproteobacteriumEnterobacteriaceaeSBＲ。1．．2009361023-1028ChenYiSunBaoshengHuangXingetal．Studyofmi-crobialpopulationsuccessioninMBＲatdifferentstages．ChineseJournalofEnvironmentalEngineering2009361023-1028inChinese2．．20023．PCＲ-DGGE．2010304729-734MouJieSunBaoshengChenYi．MicrobialcommunitystructureinamembranebioreactordeterminedusingPCＲ-DGGE．ActaScientiaeCircumstantiae2010304729-734inChinese4．．2004106826-830LuLiqongXuYatongLiangJun．Perspectiveofmolecu-larbiologicaltechniquesappliedinmicrobiologicaldiversityandmicrobiologicalecology．ChineseJournalofApplied＆EnvironmentalBiology2004106826-830inChi-nese5NambaA．ShigenobuY．KobayashiM．etal．Anewprimerfor16SrDNAanalysisofmicrobialcommunitiesas-sociatedwithPorphyrayezoensis．FisheriesScience2010765873-8786SanzJ．L．KochlingT．MolecularbiologytechniquesusedinwastewatertreatmentAnoverview．ProcessBiochemis-try2007422119-1337FangH．H．P．ChuiH．K．MaximumCODloadingca-pacityinUASBreactorsat37℃．JournalofEnvironmentalEngineering19931191103-1198．．20059ＲosenbergerS．LaabsC．LesjeanB．etal．Impactofcolloidalandsolubleorganicmaterialonmembraneper-formanceinmembranebioreactorsformunicipalwaste