不同酵母菌组合处理含油废水的效能及酵母菌群落结构研究吕文洲

,,(,　315211):10,SBR,PCR-DGGE.,O2、G1、W1,3、;O4、G2,;,G1,O2W1.:;;PCR-DGGE;;:X172;X703.1　:A　:0250-3301(2008)09-2488-05:2007-12-10;:2008-03-27:(Y507701);(20070970);(2006B100071);(2007A610057):(1974～),,,,,E-mail:[e-mail]unityStructureofOil-ContainingWastewaterTreatmentSystemInoculatedbyDifferentYeastStrainsComplexLWen-zhou,LIUYing,ZHUJian-lin(DepartmentofEnvironmentalEngineering,CollegeofArchitecturalCivilEngineeringandEnvironment,NingboUniversity,Ningbo315211,China)Abstract:Tenyeaststainsweregroupedandappliedinpilot-scalesequencingbatchreactorstotreatoil-containingwastewater.TheefficiencyandstabilityofdifferentreactorswerediscussedandyeastcommunitystructurewasinvestigatedbyPCR-DGGEmethod.Theresultsshow:thegroupconsistingofO2,G1andW1ismarkedlysuperiortoothersinefficiencyandstabilityrespects;thegroupabsenceofthese3stainsfailstoformasystemwithhighefficiencyandgoodstability;O4andG2strainsleadtoturbidsupernatantfluidandareeliminatedfromsystemstepbystep;thedistributionofyeastcellsinsettlementsludgevarieswithdifferentstains.Whenaerationisstopped,G1depositsintolowerlayerbutO2orW1distributesevenly.Keywords:yeaststrainscomplex;oil-containingwastewater;PCR-DGGE;efficiency;communitystructure　　Myuzer[1]1993DGGE,.,PCR-DGGE[2～7].,[8,9]、[10～13]、[14][15],、.,,.2000Cocolin[16]26SrDNAD1D2PCR-DGGE,[17].Prakitchaiwattana[18]Nielsen[19]PCR-DGGE.,.10,,,.,3.,10,33,PCR-DGGE,,、.29920089　　　　　　ENVIRONMENTALSCIENCEVol.29,No.9Sep.,2008DOI:10.13227/j.hjkx.2008.09.0421　1.1　:5;32、、.COD,80%.1.1　Table1　YeaststrainsandcorrespondingcodesinexperimentsO2CandidatropicalisG1CandidalipolyticaO3CanidaboidiniiG2CandidaintermediaO3iRhodotorulaglutinisG3CandidapseudolambicaO4CandidautilisW1CandidahalophilaO5TrichosporoncutaneumW2Trichosporonasteroids1.2　10,T1:G1、G2、O3i、W2、O2、O3、O47;T2:G1、G2、O3i、W2、W1、G3、O57.PCR-DGGE,G1、O2W1().3,.A:O3、O4、W2、O3i、G2(T1G1、O2);B:G3、O5、W2、O3i、G2(T2G1、W1);C:T1;D:G1、O2、W1.1.3　42LSBR,,.,YPD24h,,.22h2h,.pH5.5.11dMLSS、SVI、SS、pH、CODCOD.1.4　1.4.1　COD:CTL-12COD;CODCOD,;CODCODCOD,COD.pH:HM-14pH;SS:.[20].1.4.2　PCR-DGGE(1)　11dSBR.,;,D.200～500μL,8000rm,,,-21℃.(2)DNA　Harju[21]DNA,,PCR-DGGEDNA.-80℃5min,95℃2min,1.DNA(i-Mupid@J,)DNA.DNAmarker()λDNAHindⅢ.(3)PCR　:PCR:NL-1(5′-GCATATCAATAAGCGGAGGAAAAG-3′)NL-4(5′-GGTCCGTGTTTCAAGACGG-3′).PCR:NL1-GC(5′-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCATATCAATAAGCGGAGGAAAAG-3′)LS2(5′-ATTCCCAAACAACTCGACTC-3′)[18]..:①:50μL.0.2mLPCR:10×PCR5μL;10nmolDNTP4μL;10pmol1.5μL;DNA1μL,Taq2.5U,50μL;②PCR:95℃5min;3695℃45s55℃45s72℃50s;72℃10min;③PCR:2PCR,.PCR600bp;300bp.DNAmarker()100bpDNAladdermarker.24899:(4)DGGE　:(DcodeTM,Bio-Rad,);(UNIVERSALHOOD-S.N.75S02564,Bio-Rad,).:DGGE8%(37.5∶1,),30%～50%(100%7molL40%).:PCR50μL6×loadingbuffer10μL,,30μL.:60℃、140V4～6h,,1%EB20min,2,5min,.1.5　DGGEPCR1μLPCR,DGGE.DGGE,22,.2　2.1　A、B、C、D4SBR.111d4MLSSSVI.,CDMLSS10gL16gL,SVI60mLg,;A、BMLSS4gL,,,SVI.C,AT1O2G1,BT2G1W1,O2、G1G1、W1.CD,DMLSSC60%,,C,3,.2.2　SSpH211dpHSS.A、B,SS1700mgL1100mgL,SS,SS.CSS380mgL,C1　11dMLSSSVIFig.1　MLSSandSVIindifferentstrainscomplexafter11dcultivation,DSS12mgL,D,.,pHA～D,,pH,.,G1、O2、W1SS,.2　11dpHSSFig.2　EffluentpHandSSindifferentstrainscomplexafter11dcultivation2.3　COD3CODCOD.COD.,CCOD,D.C、D,CG1、O2,.A、BCODC、D,A、B,G1、O2、W1.COD2h,2490　　　　　　29,DCOD,,SS.CODD,,.3　11dCODCODFig.3　FilteredandsuspendedeffluentCODindifferentstrainscomplexafter11dcultivation2.4　10DGGE,DGGE,10DNAPCR-DGGE,4.,10,O2W1、O3G3、O4O5,(ALL)7.DGGE,..4　10DGGEFig.4　DGGEimageof10yeaststrains'referencesystem2.5　,2h,A、B,PCR-DGGE,56.DGGE,,.;.,,.5,D,3,A、BG2,W2、O4O5;CO4G2,O4G2.,G2、O4、O5、W2,.A～D:A～D;M:,5　DGGEFig.5　DGGEimageofyeastcommunitystructureofsupernatantfluidindifferentSBR6.A、BG2,,ABG2,G2,;G2.A、B,SVI,.6D1、D2DDGGE,O2W1;G1,,,G1,,,O2W1.24919:D1、D2:D6　DGGEFig.6　DGGEimageofyeastcommunitystructureofsettlementsludgeindifferentSBR1MLSS,,11dAB,G2,,,,MLSS.C,O2G1、G2,O4,G2O4(5),G2O4C.G2O4,、PCR-DGGE.G2O4,.3　(1)O2、G1、W1,3、.(2)O4、G2,.(3),G1,O2W1.:[1]　MuyzerG,EllenCDW,AndreGU.Profilingofcomplexmicrobialpopulationsbydenaturinggradientgelelectrophoresisanalysisofpolymerasechainreactiongenescodingfor16SrRNA[J].ApplEnvironMicrobiol,1993,59:695-700.[2]　KreuzingerN,FarnleitnerA,WandlG,etal.Molecularbiologicalmethods(DGGE)asatooltoinvestigatenitrificationinhibitioninwastewatertreatment[J].WaterSciTechnol,2003,47(11):165-172.[3]　LaParaTM,NakatsuCH,PanteaLM,etal.Stabilityofthebacterialcommunitiessupportedbyaseven-stagebiologicalprocesstreatingpharmaceuticalwastewaterasrevealedbyPCR-DGGE[J].WaterRes,2002,36(3):638-646.[4]　NodaN,YoshieS,MiyanoT,etal.PCR-DGGEanalysisofdenitrifyingbacteriainametallurgicwastewatertreatmentprocess[J].WaterSciTechnol,2002,46(1-2):333-336.[5]　TamK,YangCH,MatsumotoMR,etal.ComparisonofPCR-DGGEandselectiveplatingmethodsformonitoringthedynamicsofamixedculturepopulationinsyntheticbrewerywastewater[J].BiotechnolProg,2005,21(3):712-719.[6]　YoshieS,NodaN,MiyanoT,etal.CharacterizationofmicrobialcommunityinnitrogenremovalprocessofmetallurgicwastewaterbyPCR-DGGE[J].WaterSciTechnol,2002,46(11-12):93-98.[7]　DavidM,Stamper,MarianneWalch,JacobsRN.Bacterialpopul