OLAND生物脱氮系统运行及其硝化菌群的分子生物学检测张丹

　2003,9(5):530～533　　ChinJApplEnvironBiol=ISSN1006-687X　2003-10-25　OLAND＊张丹　徐慧　刘耀平　张颖　陈冠雄＊＊　vanCleemputOswald1　VerstraeteWilly2(　　110016)(1;2　　9000)　　-(oxygen-limitedautotrophicnitrificationanddenitrificationsystem,OLAND)、COD.,,,0.1～0.3mgL-1,〔1(1.2±0.2)〕,,.,,、,,-Nitrosomonassp.,Nitrobactersp..4222　-;;;CLC　X172PERFORMANCEANDMOLECULARMONITORINGOFNITRIFYINGBACTERIAINNITRIFICATIONSTAGEOFOLANDSYSTEM＊ZHANGDan＊＊,XUHui,LIUYaoping,ZHANGYing,CHENGuanxiong＊＊,vanCleemputOswald1&VerstraeteWilly2(KeyLaboratoryofTerrestrialEcologicalProcess,InstituteofAppliedEcology,ChineseAcademyofSciences,Shenyang　110016,China)(1LaboratoryofAppliedPhysicalChemistry,2LaboratoryofMicrobialEcologyandTechnology,FacultyofAgriculturalandAppliedBiologicalSciences,GhentUniversity,Gent　CoupureLink653,B-9000,Belgium)Abstract　Atwo-stageoxygen-limitedautotrophicnitrificationanddenitrification(OLAND)systemwasappliedforthetreatmentofsludgedigesterliquorwithahighammoniumconcentrationandalowchemicaloxygendemand(COD).Asteadyratioofammoniumandnitrite〔1(1.2±0.2)〕fromtheeffluentinthefirststage(nitritationstage)wasattainedbycontrol-lingthedissolvedoxygenintherangeof0.1～0.3mgL-1,andtheretentionofbiomasswasachievedinthesystemwiththeimmersedpolyethersulfonemembrane.Performanceofnitritationstagewasfocusedinthisstudy,andamoleculartechnolo-gy—fluorescenceInsituhybridization(FISH)wasappliedtoanalyzetheshiftofmicrobialcommunityinthisstage.Itisshowedthattheamountofammoniumoxidizerswasstableandnitriteoxidizersdecreasedalittlebitwiththedropofdissolvedoxygen.Atthesametime,itwasfoundthatNitrosomonaswasthedominantbacteriaofammoniumoxidizerswhichoxidizeammoniumtonitrite,Nitrobacteristhedominantofnitriteoxidizerswhichoxidizenitritetonitrate.Fig4,Tab2,Ref22Keywords　oxygen-limitedautotrophicnitrificationanddenitrification(OLAND);sludgedigesterliquor;fluorescenceInsitu　　　　　hybridization(FISH);nitrogenremovalCLC　X172　　,,[1,2].,,H4+NO-2NO-2NO-3.,,Nitrosomonas,NitrsospiraNitrosococcus;NitrobacterNitrospira.:2002-12-12　　:2003-02-18＊-(BIL00/03)　SupportedbythefoundationofCina-Belgiuminternationalcooperation(No.BIL00/03)＊＊Correspondingauthor　(E-mail:[e-mail]),,NO-3NO-2[3].,-,:H4+※NO-2※N2,:NH+4※NO-2※NO-3※N2.25%,40%,、COD[4].、COD:、,,().,,,-(:,,:(completelyautotrophicnitrogenremovalovernitrite,CANON)、-(deammonifi-cation)、-(oxygen-limitedautotrophicnitrificationanddenitrificationsystem,OLAND)、SHARON-ANAMMOX(singlereactorsystemforhighrateammoniaremovalovernitrite-anaerobicam-moniumoxidationSHARON-ANAM-MOX)[6～9].-,NH+4,62.5%,,,H+4,62.5%,,,NH+4NO-2,.,,.SHARON,[10].、[11～13],.,[14].,-,,[15],62.5%,.[16],.1　1.1　2L,1.5L,,,pH,1.1　Fig1　Configurationofmembrane-assistedbioreactor(MBR)1.;2.;3.;4.;5.;6.;7.;8.pH;9.1.Temperaturesensor;2.Influent;3.Acidaddition;4.Baseaddition;5.Dissolvedoxygenprobe;6.Stirrer;7.Airsparger;8.pHprobe;9.Ef-fluentwithunder-pressurepumpconnectedtoimmersedhollowfibremi-crofiltrationmembranes.1.2　Avecom(ammoniumbindinginoculumliquid,ABIL).,.:ρ(NH+4-N)=1000mgL-1(NH4HCO3)、ρ(PO3-4-P)=20mgL-1(KH2PO4)、2mLL-1.,-,:ρ(NH+4-N)=(1028±90)mgL-1、ρ(COD)=(550±50)mgL-1、ρ(BOD)=(190±10)mgL-1.1.3　、、pH、pH,.NH+4-N、NO-2-N,NO-3-NCOD[16].1.4　1.4.1　　　10mL,(-SYTO9),,.1.4.2　　　OLAND,4%,,100%,-20℃.1.4.3　　　:NSO-190[17](β-,β-);NSM156[17](Nitrosomonassp.,Nitrosomonassp.);NIT-3[18](Nitrobactersp.,Nitrobactersp.),Ntspa0662[19](Nitrospirasp.,Nitrospirasp.),(-Cy3,-Cy5,-FLUOS).1.4.4　[20]　　2～10mL,45℃,,50%,80%100%3min,.,(,),9mL1mL(Cy3Cy520ngmL-1,Fluos50ngmL-1),.46℃1.5～3h.,(),48℃.,,,48℃10min,,,.1.4.5　　　,CCD,PC,Microimage4.0.531　5:OLAND　　2　2.1　2.1.1　　　,,1.1　Tab1　CompositionofsludgedigestorliquorfromAntwerpwastewatertreatmentplantpHρ(TAN＊)/mgL-1ρ(COD)/mgL-1ρ(NO2-N)/mgL-1ρ(NO3-N)/mgL-18.095564400.7＊TAN:totalammoniumnitrogen,ρ(N)=10mgL-1[21].2.1.2　　　(MBR1)(MBR2),、.2.,1600mL/min15mL/min,40,MBR1[1(1.2±0.2)][22],[14],,.3.2　Tab2　ParametersofnitrificationstageofOLANDsystemV/Lt(HRT)/dt(SRT)/dθ/℃pHDissolvedoxygenρ/mgL-11.51∞257.900.12　Fig2　Performanceofnitrificationstageoftwostagessuspendedsystem3　Fig3　MicrobialcommunityofnitrificationstageintheearlyandlatestageA.ABIL-;B.〔1(1.2±0.2)〕A.ABIL-inoculumsludge;B.Steadystagesludge　　2,,,,,d5,,,d24,,,d42,,.2.2　2.2.1　　　,,.3,,,,,,,,.,,.2.2.2　　　(d1,d13d42),,(NSO-190,NSM156,NIT-3Ntspa0662).(),Microimagedigitalanalysissoftware,(4).4　Fig4　Estimationofshiftsofmicrobialcommunityduringdifferentperiodssincenitrificationstarted-up　　(4),,,70%(),Nitrosomonassp.,.Nitrobactersp.Nitrospirasp.10%～12%0.5%～1%.,,,,,ρ(NO-3-N)=0.7mgL-1,,,.Nitrobactersp.,,,.,(Nitro-somonas),80%,532　　　　　　　　　　ChinJApplEnvironBiol　　　　　　　　　　　　　　　　　9,,Nitrobacter,Nitrospira,.3　-(OLAND)0.1～0.3mgL-1,[1(1.2±0.2)],,,90%.,:,70%,,13%10.5%,,Nitrosomonassp.Nitrobactersp.,,,.References1　KuaiL,VerstraeteW.Ammoniumremovalbytheoxygenlimitedau-totrophicnitrificationdenitrification(OLAND)system.Appl&EnvironMicrobiol,1998,64(11):4500～45062　ZhouSQ(),ZhouJL().Theadvancesininvestigationofnewtechnologiesonbiologicalnitrogenremoval.Tech&EquipEnvironPollutCont(),2000,1(6):11～193　FochtDD,VerstraeteW.Biochemicalecologyofnitrificationanddenitri