IC反应器处理啤酒废水的效能及其微生物群落动态分析朱文秀

33820128ENVIRONMENTALSCIENCEVol．33No．8Aug．2012IC*21412235℃IC．ICCOD20kg·m3·d－1COD85%VSS210mL·g·d－1DGGEDGGEDGGEF420DGGEF420Methanosaetasp．MethanosarcinamazeiMethanosaetasp．F42020kg·m3·d－1F420VSS0.16μmol·g－1Methanosaetasp．UPGAMAShannon．ICShannonPCR-DGGEF420X797A0250-3301201208-2715-082011-10-202011-12-068632009AA0330041986～[e-mail]*[e-mail]lCommunityDynamicCharacteristicsofanInternalCirculationReactorTreatingBreweryWastewaterZHUWen-xiuHUANGZhen-xingRENHong-yanRUANWen-quanSchoolofEnvironmentandCivilEngineeringJiangnanUniversityWuxi214122ChinaAbstractAlab-scaleinternalcirculationreactorICfedbyartificialbrewerywastewaterwasoperatedwithincreasingvolumetricloadingrateunder35℃continuously．Thereactorperformanceandtherelationshipbetweenmicrobialcommunitystructureandbioactivityintheanaerobicsludgewereinvestigatedduringtheoperation．TheCODremovalefficiencywasabove85%furthermorethemaximumvolumetricloadingrateVLRandthemaximumspecificmethanogenicactivitySMAofthereactorcouldbeupto20kg·m3·d－1and210mL·g·d－1respectively．TheresultsfromthedehydrogenaseandthebacteriaDGGEexperimentsdemonstratedthatthedehydrogenasevariationtendencywaspositivelycorrelatedtototallightintensityofthewholebacteriaDGGEbandsforeachsample．ThetotallightintensityofthewholebacteriaDGGEbandscanuseasareferentialindexforbiomassliveweightinanaerobicsystem．MoreoverthecoenzymeF420contentrelatedtotherelativeabundanceofMethanosaetabasedoncoenzymeF420andarchaebacteriaDGGEanalysis．AsthevolumetricloadingrateincreasedMethanosaetabecamesignificantlydominantwhichwasaccompaniedbythecoenzymeF420contentincreasing．Thecontentcouldbeupto0.16μmol·g－1meanwhilethesuperiorityofMethanosaetabecamesignificantlyobviousUPGAMAanalysisandShannonindexalsoconfirmedthedynamicchangesofmicrobialcommunitystructureduringtheoperation．KeywordsinternalcirculationreactorICShannonindexPCR-DGGEdehydrogenasecoenzymeF4202080．．1t10～20m31．ASBR2、UASB3、AFB4、、．IC、、．DOI:10.13227/j.hjkx.2012.08.02033．、．PCR-DGGE、、5、、、6．IC．PCR-DGGEF420．11.1．2000mg·L－1FeCl3·4H2O80μg·L－1NiCl2·7H2O2μg·L－1CuCl2·2H2O1.2μg·L－1ZnCl22μg·L－1MnCl2·4H2O20μg·L－1CoCl2·2H2O80μg·L－1NH46Mo7O2·2H2O3.6μg·L－1．pHNaHCO3pH7.0．1.286%mVSS/mTSS0.760.9～2mm10L1/2．1.31IC180mm10mm20L．．．35±1℃．1ICFig．1DiagramofICreactor1.41.4.1COD、SS、VSSpH7F42089TTC10．1.4.2PCR-DGGEMO-BIOPowerSoilDNAIsolationKitDNAPCRPCR16SrDNAV3．Muzyer11F341R534PARC109FPRUN518R12～14．DGGEF341PARC109F5'GC20．PCR25μLFermentasPCRMasterMix2×PCR．PCR94℃4min94℃1min65℃1min0.5℃72℃1min2094℃1min55℃1min72℃1min1072℃6minPCR94℃4min94℃45s58.5℃45s0.5℃72℃45s2194℃45s48.5℃45s72℃45s1072℃6min．PCR1%．DGGEDcode61728ICBio-Rad8%35%～55%30%～60%．1×TAE80V60℃12hSYBRGreenIDGGEQuantityOne．1.4.3DGGEGCPCR．5．GenBankBlast．1.4.4DGGEShannonH'H'DGGE．Shannon15H'=－Σni/Nlgni/NniN．22.12.1.1COD106d．．、COD2000mg·L－1COD3kg·m3·d－116h．．3kg·m3·d－18、10、15、18、20kg·m3·d－1COD2000mg·L－1COD20kg·m3·d－12.4h．COD、COD2CODCODCOD85%．15kg·m3·d－120kg·m3·d－1COD．22kg·m3·d－1COD20kg·m3·d－110dCOD80%15dCOD85%．2CODCODFig．2Influent/effluentCODconcentrationandchangesofremovalrate2.1.23Fig．3Specificmethanogenicactivityatdifferentvolumetricloadingrates25mL100mLSMA．3VSS56mL·g·d－1210mL·g·d－12.75．7172333kg·m3·d－1．COD．2.2DGGE16．．、QuantityOneDGGE．43kg·m3·d－1TF/SS5.3mg·g·h－14.1mg·g·h－112.7mg·g·h－1．DGGE．DGGE16SrDNAV3DGGE．4DGGEFig．4RelationbetweendehydrogenaseandintensityofDGGEbandforbacteriaatdifferentvolumetricloadingrates2.3F420F4208Delafontaine17F420．5F420F420．2、3、41F420．20kg·m3·d－1F420VSS0.16μmol·g－11．F420．5F420Fig．5RelationbetweenF420andarchaealcommunitystructureatdifferentvolumetricloadingrates2.42.4.1PCR-DGGE、DGGEDGGEQuantity81728ICOneUPGAMA6DGGE．R1122R322121、2、3、4、5、6、7、8、9、13．R2．141461561514．20kg·m3·d－1．UPGAMA6．81、2、3、45、6、87．20kg·m3·d－1．R1R2～R73、8、10、15、18、20kg·m3·d－1R822kg·m3·d－16DGGEFig．6DGGEfingerprintofbacteriaunderdifferentvolumetricloadingrates2.4.2PCR-DGGE、DGGEDGGEQuantityOneUPGAMA7．52、3、42215．14、Roest18．7DGGEFig．7DGGEfingerprintofarchaeaunderdifferentvolumetricloadingratesQantityOneDGGE1R2～R871.4%、69.1%、80.5%、47.7%、44.6%、25.9%、22.9%．R3R180.5%R822.9%3、8kg·m3·d－170%22.9%IC．9172331Table1DicecoefficientcomparingthesimilaritiesofPCR-DGGEfingerprints123456781100.071.469.180.547.744.625.922.9271.4100.075.685.659.846.934.136.5369.175.6100.079.653.449.027.231.4480.585.679.6100.055.151.226.432.1547.759.853.455.1100.050.935.354.1644.646.949.051.250.9100.052.547.1725.934.127.226.435.352.5100.052.6822.936.531.432.154.147.152.6100.0ShannonH'8．19、20．3kg·m3·d－1Shannon15kg·m3·d－1Shannon．2.56723．8ShannonFig．8Shannonindexofgranularsludgethroughoutoperationalprocess2Table2PhylogeneticrelationshipofdifferentclonesequencesofbacteriaGenBank%Streptomycesclavuligerus98%、Vibriosp．90%、Flavobacteriumpsychrophilum98%、Serratiasp．98%、Clostridiumbeijerinckii98%、UnculturedVeillonellaceaebacterium98%、Unculturedbacterium98%、Unculturedbacterium98%、UnculturedLactococcussp．100%、UnculturedClostridialesbacterium98%1、2、3、4、5、6、7、8、9、13UnculturedLactococcussp．100%14UnculturedVeillonellaceaebacterium98%、Lactococcuslactissubsp．98%6、153Table3Phylog