CANON在SBAF中的快速启动及其微生物特征刘竹寒

38120171ENVIＲONMENTALSCIENCEVol．38No．1Jan．2017CANONSBAF1221*221．5106322．511458CANON．SBAFCANON．30±2℃、DOⅠ0.3～0.5mg·L－1Ⅱ～Ⅳ0.1～0.2mg·L－148dCANONNH+4-NTN99.9%86.5%．16SrDNA．ProteobacteriaPlanctomycetes26.6%17.8%β-ProteobacteriaNitrosomonasBrocadiaeCandidatusbrocadia．SBAFCANON、、．16SrDNAX703.1A0250-3301201701-0253-07DOI10．13227/j．hjkx．2016070852016-07-132016-08-212015100101991992～[e-mail]*[e-mail]-upofSBAFSystemAssistedCANONProcessandtheMicrobialAnalysisLIUZhu-han1YUEXiu2YUGuang-ping2JINLa-hua1*TANGJia-li2JIShi-ming21．SchoolofEnvironmentJinanUniversityGuangzhou510632China2．ShenyangInstituteofAutomationinGuangzhouChineseAcademyofSciencesGuangzhou511458ChinaAbstractLongperiodstart-upisoneofthemainrestrainingfactorsofthesingle-stagecompletelyautotrophicnitrogenremovalovernitriteCANONprocess．Thisstudyinvestigatedthefaststart-upoftheCANONprocessinitiatedbyasubmergedbiologicalaeratedfilterSBAFmethod．WithconventionalactivatedsludgefromthesecondarysedimentationtankofmunicipalwastewatertreatmentplantsastheseedsludgetheCANONprocesswassuccessfullystartedupaftertheacclimationofsludgemicroorganismsfor48daysundertheexperimentalconditionsof30±2℃organiccarbonfreeandcontrolleddissolvedoxygenstageⅠ0.3-0.5mg·L－1stageⅡ-Ⅳ0.1-0.2mg·L－1withthemaximumremovalratesofammonianitrogenandtotalnitrogenachievedat99.9%and86.5%respectively．Thepopulationstructurecharacteristicsofmicroorganismsinthesystemwerestudiedusinghigh-throughputsequencingof16SrDNAamplicon．TheresultsdemonstratedthatthetwodominantmicrobialstrainsinthesystemwereProteobacteriaandPlanctomycetesaccountingfor26.6%and17.8%respectively．ThemajorcontributorsofnitrogenremovalwereNitrosomonasinβ-ProteobacteriaandCandidatusbrocadiainBrocadiae．ThroughtheaboveexperimentsitwasrevealedthattheinvestigatedSBAFbasedCANONpossesseshadtheadvantagesoffaststart-upefficientbiologicalnitrogenremovalandstableoperationprocess．KeywordsSBAFCANONammonianitrogen16SrDNAsequencingmicrobialpopulationstructure2090completelyautotrophicnitrogenremovalovernitriteCANON62.5%、100%50%、12．ammoniaoxidizingbacteriaAOBanaerobicammoniumoxidationbacteriaAnAOB．AOBAnAOB34CANON56．AOBAnAOB．38CANON7．denaturinggradientgelelectrophoresisDGGE8．、Illumina16SrDNA、、、9．fluorescenceinsituhybridizationFISH、polymerasechainreactionPCＲ、DGGE10、、CANON．submergedbiologicalaeratedfilterSBAF、48dCANON．16SrDNA．11.1111cm38cm3.6L3L．2.5cm30±2℃．．O2dissolvedoxygenDO．2cm2.5cm．63r·min－1125mL·h－1hydraulicretentiontimeHＲT24h．1Fig．1SchematicdiagramoftheSBAFusedinthisstudy1.2．3dMLSS14.267g·L－1MLVSS8.361g·L－12L．1.3NH4Cl0.380g·L－1KH2PO40.025g·L－1MgSO40.010g·L－1CaCl20.020g·L－1NaHCO31.000g·L－10.35mL·L－1．FeCl3·6H2O3.515g·L－1MnCl2·4H2O0.359g·L－1CuSO4·5H2O0.075g·L－1ZnSO4·7H2O0.300g·L－1CoCl2·6H2O0.375g·L－1．1．1Table1KeywaterqualityoftheinfluentNH+4-Nmg·L－182.5～113.5NO－3-Nmg·L－12.0～2.3NO－2-Nmg·L－10.00pH—8.04～8.28CODmg·L－10～730±2℃SBAFN272h．．4521CANONSBAF16SrDNA．1.4NH+4-NNO－2-NN-1--NO－3-N11TN=NH+4-N+NO－2-N+NO－3-NCODpHDOpHPJBJ-260JPB-607A．1.5Illumina16SrDNA1DNAAgaroseGelElectrophoresisND-1000Nanodrop．2TruSeqCustomAmpliconSamplePrepKitPCＲ、PCＲAgilent2200TapeStationQubit2.0．3①poolingpooling2.1nmol·L－1②pooling2.1nmol·L－1PhixControl19∶1③2.0mol·L－1NaOH0.2mol·L－1NaOH④②③1∶15min⑤HT1④100420μL．4PairEndFlowCellMiSeq2500PairEnd2×100．5．cleandataPaired-endreadsTagsUniqueTagsUniqueTagsOTUoperationaltaxonomicunitsOTU、Alpha．22SBAF4Ⅰ1～16d、Ⅱ17～33d、CANONⅢ34～48dCANONⅣ49～76d48dtotalnitrogenremovalrateTNＲ74.3%386.5%CANON．2CANONFig．2VariationsofnitrogenconcentrationinCANONreactor3CANONFig．3VariationsofTNinCANONreactor2.1CANONNO－2-NNH+4-NAnAOBNO－2-N12CANON．AOBnitriteoxidizingbacteriaNOBNO－2-N．AOBNOBDOAOBNOB552380.6mg·L－12.2mg·L－113Hellinga141525℃AOBNOB．30±2℃IDO0.3～0.5mg·L－1．41dNO－2-N2.2mg·L－1NH+4-N34%2NO－3-N0.3mg·L－132.7%5Ⅲ～ⅣFig．5VariationsofnitrogenconcentrationineffluentstageⅢ-Ⅳ16．2dNO－2-NNH+4-N8d9d16dNO－2-N53mg·L－1．ⅡDO0.1～0.2mg·L－1AOBNO－2-N17～33dNO－2-N44～52mg·L－1．2.2CANON534～48dNO－2-NNH+4-NNO－2-N49mg·L－18mg·L－1NH+4-N27.5mg·L－10.5mg·L－1NH+4-NammonianitrogenremovalrateAＲＲAＲＲ88.5%99.5%TNＲ25.1%83.27%．NO－3-N5.1mg·L－112.9mg·L－1AnAOBNO－2-NNH+4-NN2NO－3-NNO－2-NNH+4-N4Ⅰ～ⅡFig．4EffectofnitriteaccumulationstageⅠ-ⅡNO－3-N17．TN16SrDNAγ-ProteobacteriaPseudomonas0.006%．CANON48dSBAFCANON．CANON34d12dAnAOB≥1010～1011·mL－11819．40d20N2CANON．49d6521CANONSBAFAＲＲ93.3%99.9%TNＲ76.1%86.5%68%．CANON1121ΔNO－3-N/ΔTNCANON0.127CANONNOB．ΔNO－3-N/ΔTN＞0.127NOBNO－2-NNO－3-NΔNO－3-NΔTNNO－2-NΔNO－3-N/ΔTN＜0.127ΔTN．6ΔNO－3-N/ΔTNNO－3-N＜NO－3-NΔNO－3-NΔNO－3-N/ΔTNCANONΔNO－3-N/ΔTN0.127．2.316SrDNA2.3.1CANONIllumina6CANONΔNO－3-N/ΔTNFig．6VariationsofΔNO－3-N/ΔTNinCANONreactorMiSeq16SrDNAOTU、Alpha2．Alphachao1、Shannon、Simpson．．2799%．2Alpha1Table2Alphadiversityofdifferentmicrobialsampleschao1ShannonSimpsonOTUTags/%15021.028.090.97430112416299.0625105.008.300.97449911492899.03112．chao1ShannonSimpsonOTUOTUShannonSimpson2.3.216SrDNA．5%401%11810%AOBAnAOBProteobacteriaPlanctomycetes26.6%17.8%CANON．Chloroflexi9.3%Chlorobi7.7%Gemmatimonadetes6.5%、Bacteroidetes4.7%、Acidobacteria4.2%、Actinobacteria2.3%．CANONAOBAnAOBAOBProteobact