好氧反硝化细菌的筛选鉴定及处理硝酸盐废水的研究苏俊峰

1,2,3,1＊,1,1,1(1.,　150090;2.,　710055;3.,　150080):,,5.f1、f2、f3、f5f7TN90.4%、91.2%、94.6%、95.6%97%,.16SrDNA,,f1、f3f5Pseudomonassp.,f2f4Paracoccussp..5.,98.16%.,,3.56mgL.COD87.24%,.:;;:X172;X703　:A　:0250-3301(2007)10-2332-04:2006-11-22;:2007-01-15:(50521140075);(QC05C05);(ZD200501):(1977～),,,,E-mail:[e-mail]＊,E-mail:[e-mail]tionofAerobicDenitrifiersandDisposetheWastewaterofNO-3-NSUJun-feng1,2,WANGJi-hua3,MAFang1,GAOShan-shan1,WEILi1,WANGChen1(1.SchoolofMunicipalandEnvironmentalEngineering,HarbinInstituteofTechnology,Harbin150090,China;2.SchoolofEnvironmentalandMunicipalEngineering,XianUniversityofArchitectureandTechnology,Xian710055,China;3.DepartmentofBiologyandEnvironment,HarbinNormalUniversity,Harbin150080,China)Abstract:Afterdomesticatingactivatedsludgetoenrichedaerobicdenitrifiers,5aerobicdenitrifierswereisolatedfromit.TheremovalratesofTNwere90.4%,91.2%,94.6%,95.6%and97%byf1,f2,f3,f5andf7,respectively.Accordingtothephysiologicalbiochemicalindex,fivestrainsofaerobicdenitrifiersweregenerallyidentifiedasPseudomonassp.,Paracoccussp.,respectively.Theestablishmentofcontinuousflowreactorsurveyedvariousindices.Andtheefficiencyofnitratenitrogenremovalcanreachupto98.16%,whenthereactorworkssmoothly.Thenitriteconcentrationofeffluentwasatalowlevelandthemaximumvaluewasnomorethan3.56mgL.Atthesametime,theefficiencyofCODremovalwas87.24%,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.,、、、、,,52810200710　　　　　　ENVIRONMENTALSCIENCEVol.28,No.10Oct.,2007DOI:10.13227/j.hjkx.2007.10.026,5.1　1.1　FM[14](gL):1.0;5.0;KNO31.0.(SG,gL):Na2HPO4·7H2O5.0;KH2PO41.0;MgSO4·7H2O0.1;5.5;KNO32.0;NaNO20.5;2mL;pH7～7.5.(gL):EDTA50.0;ZnSO42.2;CaCl25.5;MnCl2·4H2O5.06;FeSO4·7H2O5.0;(NH4)6Mo7O2·4H2O1.1;CuSO4·5H2O1.57;CoCl2·6H2O1.61;pH=7.0.1.2　1180mm,3L,SBR;,:※※※(0.3h)※(0.15h).12h,3d1～2h,.28d,,[15].1.3　10mL90mL,,.,,(SG)342.SG100mLSG,(),9,,:30℃,140rmin,40h.TOC-VCPN-TNM-1TN,TN.1.4　,KNO3、Na2HPO4、KH2PO4、MgSO4·7H2O、MnSO4、NaAC.1.1.5　1　Table1　WaterqualityofthesyntheticwastewaterCODNO-3-NNO-2-NTNpHmg·L-1149.57～356.7146.50～103.860.07～8.4754.38～114.466.8～7.51　Fig.1　Sketchofcontinuousflowreactor　　1,,12L,5,.、,,pHDOpH.,,3d.20～25℃.1.6　2.2　Table2　MainanalyticalitemsandanalyticalmethodsNO-2-NNO-3-NNH+4-NTN-NCOD1.7　DNA500μLPBS,12000r·min-1,10min,1,300μLDNA37℃30min,5min,.80μL10%SDS,65℃1h,233310:,12000r·min-1,10min,1.5mL.30μLNaAC10min,12000r·min-15min,1,400μL(24∶1).12000r·min-15min,400μL,,12000r·min-15min,200μL1.2.0.6,.-20℃,1h.12000r·min-120min,.6～8h.20μLTE.1.8　16SrDNA,16SrDNABLASTGenBank16SrDNA.,NCBIBlastnCluastx、PhyloDrawGenBank[16],.2　2.1　,SBR1,,42,TN90%5.f1、f2、f3、f5f7,90.4%、91.2%、94.6%、95.6%97%,.,TN30%,5.2.2　16SrRNABSF820BSR145120[9]DNA,3μL1%.,T-(),M13.BLASTGenBank16SrDNA.16SrDNA,f1、f3f5Pseudomonassp.99%,f2f4Paracoccussp.99%,,f1、f3f5Pseudomonasp.,f2f4Paracoccussp.,2.2　5NJFig.2　Phylogenictreeof5sampleandsimilarsequencesinGenBank,constructedbyneighbor-joiningmethod2.3　3.3,10d,,11～20d,,96.92mgL,,21～30d1.78mgL,98.16%.,3.56mgL,1.35mgL.4,TN10d;11～20d,TN,.21～30d,TN2.37mgL,TN1.09mgL,TN96.71%,,.3　NO-3-NFig.3　VariationofNO-3-Nintheeffluentofcontinuousflowreactor5,15dCOD2334　　　　　　284　TNFig.4　VariationofTNintheeffluentofcontinuousflowreactor,15dCOD,16～30dCOD87.24%.COD,,COD.5　CODFig.5　VariationofCODintheeffluentofcontinuousflowreactor3　(1)SBR1,5,f1、f2、f3、f5f7,90.4%、91.2%、94.6%、95.6%97%,.(2)16SrDNA,f1、f3f5Pseudomonasp.99%,f2f4Paracoccussp.99%.,f1、f3f5Paracoccussp.,f2f4Paracoccussp..(3)21～30d98.16%.,3.56mgL.21～30d,TN96.71%,,.16～30dCOD87.24%,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):1