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1,2,3,1*,1,1,1(11,150090;21,710055;31,150080):,,5.f1f2f3f5f7TN9014%9112%9416%9516%97%,.16SrDNA,,f1f3f5Pseudomonassp1,f2f4Paracoccussp..5.,98116%.,,3156mgPL.COD87124%,.:;;:X172;X703:A:0250-3301(2007)10-2332-04:2006-11-22;:2007-01-15:(50521140075);(QC05C05);(ZD200501):(1977~),,,,E-mail:sjf1977518@sina.com.cn*,E-mail:mafang@hit.edu.cnIsolationandIdentificationofAerobicDenitrifiersandDisposetheWastewaterofNO-3-NSUJun-feng1,2,WANGJ-ihua3,MAFang1,GAOShan-shan1,WEILi1,WANGChen1(11SchoolofMunicipalandEnvironmentalEngineering,HarbinInstituteofTechnology,Harbin150090,China;21SchoolofEnvironmentalandMunicipalEngineering,XipanUniversityofArchitectureandTechnology,Xipan710055,China;31DepartmentofBiologyandEnvironment,HarbinNormalUniversity,Harbin150080,China)Abstract:Afterdomesticatingactivatedsludgetoenrichedaerobicdenitrifiers,5aerobicdenitrifierswereisolatedfromit.TheremovalratesofTNwere9014%,9112%,9416%,9516%and97%byf1,f2,f3,f5andf7,respectively1Accordingtothephysiologicalbiochemicalindex,fivestrainsofaerobicdenitrifiersweregenerallyidentifiedasPseudomonassp.,Paracoccussp.,respectively.Theestablishmentofcontinuousflowreactorsurveyedvariousindices.Andtheefficiencyofnitratenitrogenremovalcanreachupto98116%,whenthereactorworkssmoothly.Thenitriteconcentrationofeffluentwasatalowlevelandthemaximumvaluewasnomorethan3156mgPL.Atthesametime,theefficiencyofCODremovalwas87124%,whichdemonstratesthecoinstantaneousremovingofbothorganicandnitratenitrogen.Keywords:aerobicdenitrifiers;nitratewastewater;aerobicdenitrification,[1].,.2080,30%[2].[3~5],,-.-:(Alcaligenesfaecalis)[6](Pseudomonas)[7~9](Klebsiella)[7](Ochrobactrum)[10](Rhodococcus)[11](Zoogloea)[7][(Thiosphaerapantotropha),(Paracoccusdenitrificans)[12]]..,,..Gupta[13]Thiosphaerapantotropha.Kshirsagar[14]2.,,,52810200710ENVIRONMENTALSCIENCEVol.28,No.10Oct.,2007,5.1111FM[14](gPL):110;510;KNO3110.(SG,gPL):Na2HPO4#7H2O510;KH2PO4110;MgSO4#7H2O011;515;KNO3210;NaNO2015;2mL;pH7~715.(gPL):EDTA5010;ZnSO4212;CaCl2515;MnCl2#4H2O5106;FeSO4#7H2O510;(NH4)6Mo7O2#4H2O111;CuSO4#5H2O1157;CoCl2#6H2O1161;pH=710.1121180mm,3L,SBR;,:yyy(013h)y(0115h).12h,3d1~2h,.28d,,[15].11310mL90mL,,.,,(SG)342.SG100mLSG,(),9,,:30e,140rPmin,40h.TOC-VCPN-TNM-1TN,TN.114,KNO3Na2HPO4KH2PO4MgSO4#7H2OMnSO4NaAC.1.1151Table1WaterqualityofthesyntheticwastewaterCODNO-3-NNO-2-NTNpHPmg#L-1149157~35617146150~1031860107~814754138~114146618~7151Fig.1Sketchofcontinuousflowreactor1,,12L,5,.,,pHDOpH.,,3d.20~25e.1162.2Table2MainanalyticalitemsandanalyticalmethodsNO-2-NNO-3-NNH+4-NTN-NCOD117DNA500LLPBS,12000r#min-1,10min,1,300LLDNA37e30min,5min,.80LL10%SDS,65e1h,233310:,12000r#min-1,10min,115mL.30LLNaAC10min,12000r#min-15min,1,400LLP(24B1).12000r#min-15min,400LL,,12000r#min-15min,200LL1.2.016,.-20e,1h.12000r#min-120min,.6~8h.20LLTE.11816SrDNA,16SrDNABLASTGenBank16SrDNA.,NCBIBlastnCluastxPhyloDrawGenBank[16],.2211,SBR1,,42,TN90%5.f1f2f3f5f7,9014%9112%9416%9516%97%,.,TN30%,5.21216SrRNABSF8P20BSR1451P20[9]DNA,3LL1%.,T-(),M13.BLASTGenBank16SrDNA.16SrDNA,f1f3f5Pseudomonassp.99%,f2f4Paracoccussp.99%,,f1f3f5Pseudomonasp.,f2f4Paracoccussp.,2.25NJFig.2Phylogenictreeof5sampleandsimilarsequencesinGenBank,constructedbyneighbor-joiningmethod2133.3,10d,,11~20d,,96192mgPL,,21~30d1178mgPL,98116%.,3156mgPL,1135mgPL.4,TN10d;11~20d,TN,.21~30d,TN2137mgPL,TN1109mgPL,TN96171%,,.3NO-3-NFig.3VariationofNO-3-Nintheeffluentofcontinuousflowreactor5,15dCOD2334284TNFig.4VariationofTNintheeffluentofcontinuousflowreactor,15dCOD,16~30dCOD87124%.COD,,COD.5CODFig.5VariationofCODintheeffluentofcontinuousflowreactor3(1)SBR1,5,f1f2f3f5f7,9014%9112%9416%9516%97%,.(2)16SrDNA,f1f3f5Pseudomonasp.99%,f2f4Paracoccussp.99%.,f1f3f5Paracoccussp.,f2f4Paracoccussp..(3)21~30d98116%.,3156mgPL.21~30d,TN96171%,,.16~30dCOD87124%,COD,.:[1],,,.[J].,2005,11(2):226~228.[2]PatureauD,ZumsteinE,DelgenesJP,etal.Aerobicdenitrificationisolationfromdiversenaturalandmanagedecosystems[J].MicrobEcol,2000,39:145~152.[3]DingAZ,FuJM,ShengGY.Evidenceofaerobicdenitrification[J].ChinScieBull,2000,45(3):2779~2782.[4]XieSG,ZhangXJ,WangZS.Integratedstudyonbiochemicalmechanisminbiofiltersystem[J].ActaSciCircumst,2002,22(5):557~561.[5]RobertsonLA,KuenenJG.Aerobicdenitrification:acontroversyrevived[J].ArchMicrobiol,1984,13(9):351~354.[6]OttesS,SckalkJ,KuenenJG.HydeoxylamineoxidationandsubsequentnitrousoxideproductionbytheheterotrophicammoniaoxidizerAlcaligenesfaecalis[J].ApplMicrobiolBiotechnol,1999,51(2):255~261.[7]TakayaN,MariaAntoninaBCS,YasushiS,etal.Aerobicdenitrificationbacteriathatproducelowlevelsofnitrousoxide[J].ApplEnvirMicrobio,2003,69(6):3152~3157.[8],,,.[J].,2006,8:28~30.[9],,,.[J].,2006,38(4):575~577.[10]PaiSL,ChongNM,ChenCH.Potentialapplicationsofaerobicdenitrifiyingbacteriaasbio-agentsinwastewatertreatment[J].BioresourceTechnology,1999,68(2):179~185.[11],,.[J].,2004,25:75~78.[12]LukowT,DiekmannH.AerobicdenitrificationbyanewlyisolatedheterotrophicbacteriumstrainTL1[J].BioTechnolLett,1997,11(19):1157~1159.[13]GuptaAB,GuptaSK.SimultaneouscarbonandnitrogenremovalinamixedcultureaerobicRBCbiofilm[J].W