好氧颗粒污泥和活性污泥细菌种群结构对五氯酚污染的响应研究

26920069ActaScientiaeCircumstantiaeVol.26,No.9Sep.,2006:(No.BK2005402)SupportedbytheNaturalScienceFoundationofJiangsuProvince(No.BK2005402):(1974),,;3(),E2mail:[e-mail]:LIUHe(1974),male,Ph.D.;3Correspondingauthor,E2mail:[e-mail],,,.2006.[J].,26(9):1445-1450LiuH,LiGW,YUNJ,etal.2006.Theresponseofbacterialcommunitywithinaerobicgranularsludgeandactivatedsludgetopentachlorophenolcontamination[J].ActaScientiaeCircumstantiae,26(9):1445-14501,3,1,1,1,21.,2140362.,214036:2005212212:2006207215:2006207227:(T2RFLP)(PCP).,PCP30mgL-1,COD38%77%,1020mgL-1PCP,56%15%.,PCP20mgL-1,13%58%,1020mgL-1PCP,86%40%,PCPCOD,.PCP,;PCP30mgL-1,,TRFs2614,,.:;;;;T2RFLP:025322468(2006)0921445206:X172:ATheresponseofbacterialcommunitywithinaerobicgranularsludgeandactivatedsludgetopentachlorophenolcontaminationLIUHe1,3,LIGuangwei1,YUNJiao1,CHENJian1,21.LaboratoryofEnvironmentalBiotechnology,SchoolofBiotechnology,SouthernYangtzeUniversity,Wuxi2140362.KeyLaboratoryofIndustrialBiotechnology,MinistryofEducation,SouthernYangtzeUniversity,Wuxi214036Received12December2005;receivedinrevisedform15July2006;accepted27July2006Abstract:Theperformanceofaerobicgranularsludgeforthetreatmentofwastewaterwasstudied.TheresponseofthebacterialcommunitytodifferentconcentrationsofPentachlorophenol(PCP)wasmonitoredbyterminalrestrictionfragmentlengthpolymorphism(T2RFLP)technique.WhenPCPconcentrationwas30mgL-1,CODremovalratesoftheaerobicgranularsludgeandactivatedsludgewere38%and77%,whichdecreasedby56%and15%,respectively.Inaddition,theammoniaremovalratesoftheaerobicgranularsludgeandactivatedsludgewere13%and58%,whichdecreasedby86%and40%,respectively.ThesedataindicatedthattheimpactofPCPontheaerobicgranularsludgewasstrongerthanthatofactivatedsludgeandtheinfluenceonremovalofammonianitrogenwasstrongerthanthatofCODremoval.TheresultsoftheT2RFLPanalysisshowedthatthechangeofbacterialcommunitywithintheaerobicsludgewasmoreapparentthanthatofactivatedsludge.WhenPCPwas30mgL-1,thenumberofTerminalRestrictionFragments(TRFs)decreasedfrom26to14inaerobicgranularsludgebutmaintainedstableinactivatedsludge.Itissuggestedthatthebacterialspeciesnumberwithintheaerobicgranularsludgedecreasedbutthatinactivatedsludgekeptstable.Thechangetrendofthebacterialdiversitywasconsistentwiththeperformanceofthewastewatertreatment.Keywords:Aerobicgranularsludge,activatedsludge,bacterialcommunity,Pentachlorophenol,T2RFLP1(Introduction),,,.26(,2000;,2002).,,,(Yuanetal.,2002)..(Tohetal.,2003)(2001;Yangetal.,2003),(,2003),,,.,,,,.,16SrDNA(Forneyetal.,2004),PCR2DGGE(,2005).,.SBR,(T2RFLP)(PCP),.2(Materialsandmethods)2.1SBR,,84mm,260mm,1500mL,900mL.(,2003),.,MLSS56gL-1,2.0mgL-1,CODCr700800mgL-1,4050mgL-1,pH7.0,(,2003).4h,10min,3.5h,20min.PCP0.02molL-1NaOH,1gL-1,.2.2pHpHS23,DO,COD,NH+42N,MLSS(AmericanPublicHealthAssociation,1998).2.3DNA200mgDNA.50mg,(Fastprep,ThermoBIO101)(4.5ms-1,30s,4),DNA(3SDNAIsolationKitV2.1,)DNA,.DNA018%,SYBRGreenDNA.2.416SrDNAPCR16SrDNA.8227F(52agagtttgatcmtggctcag23)(Marshetal.,2000),1378R(52cggtgtgtacaaggcccgggaacg23)(Heueretal.,1997),56-(FAM)..50LPCR:Taq5L,MgCl22.5mmolL-1,dNTPs(each)0.2mmolL-1,0.4mmolL-1,40ng,Taq(TaKaRa)5,ddH2O31L.:9510min,Taq,951min,6030sec.,721.5min,30,7210min.PCRPCR(TaKaRa),.2.5T2RFLPPCRAluI(TaKaRa),20L,,AluI8,10Lbuffer2L,DNA200ng;372h,6510min.T2RFLP.2.6TRFsRDP(:analyses.html)T2RFLP,TRFLPTRFs,.Sorensens(Eileretal.,2004)Dxy(Dxy=2Nxy/(Nx+Ny),NxNyTRFs,NxyTRFs.3(Results)3.1PCPCOD120d,COD90%,93%,.SBRPCP,PCP,1.1,PCP1020mgL-1,COD,PCP30mgL-1,COD38%77%,56%15%,.,PCP1020mgL-1,,PCP30mgL-1,13%58%,86%40%.,PCP30mgL-1,PCP;,,PCPCOD,PCP.1PCP(a.COD,b.)Fig.1TheeffectofdifferentPCPonperformancesofaerobicgranularsludgeandactivatedsludge(a.CODremoval,b.Ammoniaremoval)3.2T2RFLP2ABI370016SrDNAPCRAluI.(TRFs),.PCP.3.3PCP2,1%TRFs.,1%TRFs90%,TRFs.32TRFs1%PCP.,PCP,TRFs2614,,.COD,,COD.3.4PCP,TRFs(1%),2PCP(4).7441262PCPT2RFLP(A.,B.)Fig.2T2RFLPprofilesofaerobicgranularsludgeandactivatedsludgeatdifferentPCPconcentration.(A.Profileofgranularsludge,B.Profileofactivatedsludge)3PCPTRFsFig.3ChangesofTRFsnumberwithinaerobicgranularsludgeandactivatedsludgeatdifferentPCPconcentration,.,61bp101bp122bp230bp242bpPCP10mgL-1;222bp429bp10mgL-1PCP,PCP20mgL-1;146bp190bp256bp271bp292bp440bp20mgL-1PCP,30mgL-1PCP.PCP30mgL-1,TRFs82bp175bp198bp229bp,TRFs69bp57bp71bpPCP.,PCP30mgL-1,TRFs272bp262bp240bp.84419:4PCPTRFs(A.,B.)Fig.4TheconstitutionvarietyofTRFswithinaerobicgranularsludgeandactivatedsludgeunderdifferentPCPconcentration(A.aerobicgranularsludge,B.activatedsludge)3.5PCP2PCP,1.1PCPTable1ThesimilaritycomparisonofdifferentPCPconcentrationsofaerobicgranularsludgeandactivatedsludgePCP0mgL-110mgL-120mgL-130mgL-10mgL-110mgL-120mgL-130mgL-10mgL-1-0.5820.6530.4820.8470.7660.77910mgL-10.7950.5590.7580.71420mgL-10.6000.8391,PCP102030mgL-1PCP,48.2%65.3%,76.6%84.7%,,PCP.4(Discussion),PCP30mgL-12,,COD.PCP,PCP.,,,.,,,.,,.(MLSS20gL-1),,2,,PCP,.PCP,(T2RFLP).,PCP,.COD,COD.,T2RFLP,PCP,944126,.5(Conclusions)1)PCP30mgL-1,COD1020mgL-156%15%,86%40%,PCPCOD,.2)PCP,PCP30mgL-1,TRFs2614,,COD