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39420184ENVIＲONMENTALSCIENCEVol．39No．4Apr．2018YH01+YH02SBＲ112*11121．5106302．510630-Delftiasp．YH01+Acidovoraxsp．YH02SBＲ．NO－3-N、TNCOD12.1%、9.2%9.4%YH01+YH02PCAUPGMA4DelftiaAcidovoraxYH01+YH02SBＲ．-SBＲX172A0250-3301201804-1773-09DOI10．13227/j．hjkx．2017082532017-08-312017-09-17-U15012332016A0303120092015B0909030701991～E-mail913421208@qq．com*E-mailcao_g@163．comAerobicDenitrificationandMicrobialCommunityShiftinSBＲBioaugmentedwithStrainsYH01andYH02CHENHai-sheng1CAOGang12*ZHANGDi1HUANGZheng-zheng1MOCe-hui121．SchoolofEnvironmentJinanUniversityGuangzhou510630China2．GuangdongEngineeringCenterforEnvironmentContaminationControlandＲestorativeMaterialsGuangzhou510630ChinaAbstractTheenhancedaerobicdenitrificationcapabilityofthemixedstrainsYH01+YH02inutilizingpotassiumnitrateandthedynamicchangesinthemicrobialcommunitycomponentduringtheperiodofoperationwereevaluated．ThemicrobialcommunityindifferentstagesoftheSBＲwasanalyzedbyusinghigh-throughputsequencingtechnologyafterinoculationwithYH01+YH02．TheresultsshowedthattheNO－3-NTNandCODremovalefficienciesincreasedby12.1%9.2%and9.4%respectively．Therelativeabundancesofthemicrobesinthemicrobialcommunityincreasedatthegenuslevelandthediversityinthemicrobialcommunitydecreasedafterenhancement．PrincipalcomponentanalysisandUPGMAanalysisrevealedthattheperiodofSBＲoperationwasroughlydivinedintofourphases．TherelativeabundancesofDelftiaandAcidovoraxincreasedduringtheperiodofoperationandYH01+YH02exhibitedexcellentcompatibilitywiththeSBＲecosystemandplayedanimportantpartinaerobicdenitrification．Keywordsheterotrophicnitrification-aerobicdenitrificationmixedstrainsSBＲreactorhigh-throughputsequencingtechnologymicrobialcommunitycomponent-Ｒobertson11988-ThiosphaerapantotrophaBacillus2、Pseudomonas3、Alcaligenes4、Acinetobacter5-．-．-2～5．6～8OchrobactrumanthropiXH02、Bacillussp．K5AcinetobacterjuniiYB．-．9101139．12．SBＲ-YH01+YH02．PCAUPGMA．11.11.1.1-Delftiasp．YH01Acidovoraxsp．YH02．YH01、YH02500mLLB30℃、120r·min－124h．1∶1D6002.0．MLSS2500～3000mg·L－1SVI90～120mL·g－1．1.1.2KNO3COD950～1050mg·L－1NO－3-N190～210mg·L－1pH7.4～7.6．1L50mLK2HPO45.0g·L－1MgSO4·7H2O2.5g·L－1NaCl2.5g·L－1FeSO4·7H2O0.05g·L－1MnSO4·4H2O0.05g·L－1．1.2SBＲSBＲ3L．．SBＲ12h2h9h25min30min5min．3.5L·min－130℃．SBＲ3d．30%3d．TN、NH+4-N、NO－2-NCOD．1.327F5'-AGAGTTTGATCMTGGCTCAG-3'1492F5'-TTGGYTACCTTGTTACGACT-3'DNAPCＲ．．1.4．YH01+YH02．YH01+YH022d9DNA．TruSeqTMDNASamplePrepKitDNA．DNA16SrDNA338F5'-ACTCCTACGGGAGGCAGGAC-3'806Ｒ5'-GGACTACHVGGGTWTCTAAT-3'PCＲ．PCＲ10ng20μL4μLdNTPs25mmol·mL－12μLDNA100ng50mmol·mL－10.8μL．PCＲ95℃3min3095℃30s45℃30s72℃42s72℃10min．PCＲ2%AxyPrepDNAPCＲ．QuantiFluorTM-STPromegaPCＲ．PCＲIlluminaMiSeq．．QIIME97%OTUOTU0.001%13．GreengenesOTU．AceChao14ShannonSimpson15CanocoMothur．47714YH01+YH02SBＲ22.116SrDNAYH01、YH02DNAPCＲDNAGenBankKY419588、KY419589Blast．1YH01DelftiaYH02AcidovoraxProteobacteria．116SrDNAYH01、YH02Fig．1PhylogenetictreeofthestrainsYH01andYH02basedonthecompletesequenceofthe16SrDNAgene2.2YH01YH02-YH01、YH02YH01+YH02NO－3-N220mg·L－1、NO－2-N220mg·L－1．2a～2cNO－3-N24hNO－3-N、TN、COD．2a2bYH01、YH02、YH01+YH0224hNO－3-N88.7%、85.1%、95.3%TN67.2%、70.3%、87.2%YH01+YH02．2c24hYH01、YH02、YH01+YH02COD88.2%、89.4%、90.1%．NO－2-N2d～2f24hYH01、YH02YH01+YH02NO－2-N、TN、COD．24hYH01、YH02YH01+YH02NO－2-N88.2%89.0%90.1%TN85.3%、88.9%88.9%COD89.0%、90.1%91.2%．NO－2-NYH01、YH02YH01+YH02NO－3-N．Ochrobactrumsp．XH02+Pseudomonassp．XH0310Pseudomonassp．B2+Streptromycessp．A9+Fusariumsp．F39．16．YH01+YH02YH01、YH02YH01+YH02．2.3SBＲ3a．．10～31d．NO－3-N、TNCOD73.4%、64.5%80.7%．31d．31d．3b31～34d34～52dNO－3-N、TNCOD12.1%、9.2%9.4%．YH01+YH025771392YH01、YH02、YH01+YH02Fig．2CharacteristicsofdenitrificationbyYH01YH02andYH01+YH023Fig．3ChangesinNandCODconcentrationsintheperiodofstart-upandenhancement．17．1819．YH01+YH02YH01+YH02、．2.4SBＲAceChaoShannonSimpson4．ChaoAce40dAce417Chao420．ShannonSimpson3.710.0652d3.600.05．620218．YH01+YH02YH01+YH0267714YH01+YH02SBＲ4Fig．4Abundanceanddiversityindexes225Fig．5Changesintherelativeabundanceofmicroorganismsatthephylumlevel．．2.5592719Proteobacteria、Bacteroidetes、Planctomycetes777139Chloroflexi62.1%±4.1%、22.2%±8.0%、7%±3%3.3%±2.0%．ProteobacteriaOchrobactrumanthropiXH026Proteobacteria．Proteobacteria、ChloroflexiPlanctomycetesYH01+YH027.0%、3.4%3.1%．Proteobacteria、ChloroflexiPlanctomycetes23～25．Bacteroidetes31.0%14.6%YH01+YH02Bacteroidetes15.0%．6Fig．6HeatmapofthebacterialcommunityatthegenuslevelSBＲ30Heatmap6．．UnculturedSaprospiraceae、Thauera、Luteimonas3%Proteobacteria．UnculturedSaprospiraceae、ThaueraLuteimonasYH01+YH0252d7.1%、0.5%7.5%YH01+YH02．UnculturedSaprospiraceaeUnculturedSaprospiraceae．87714YH01+YH02SBＲDelftiaAcidovorax7．DelftiaAcidovorax0.09%0.1%52d2.2%3.5%．DelftiaAcidovorax10Delftiasp．YH01+Acidovoraxsp．YH02SBＲ10%YH01+YH02．Delftia、26Acidovorax27Delftia+Acidovorax．7DelftiaAcidovoraxFig．7ＲelativeabundancesofDelftiaandAcidovorax2.62.6.1PCAPCAPCAPCA2829．9OTUPCA831～34d37d31～34d37dYH01+YH02．40～46d3．40～46d49～52dSBＲ．SBＲ4YH01+YH02SBＲ．8PCAFig．8Principalcomponentanalysisofthebacterialcommunity2.6.23031．UPGMA20．9931～34d37d．40～46d．97713949～51d40～46d．31～34、37、40～4649～52d432．．9UPGMAFig．9ClusteranalysisofthebacterialcommunitybyUPG