CANNED工艺细菌种群结构变化的FISH研究

23920109ResearchofEnvironmentalSciencesVol.23No.9Sep.2010CANNEDFISH12121*31.1000122.4101283.730030FISH/CANNEDFA.ρFA0.5mg/LAOBNOB0.9%2.8%ρFA12.4～48.6mg/LAOBNOB33.6%～47.4%1.0%～2.9%.ρFA0.5～12.4mg/L、7.2%～15.3%7.2%～10.3%9.1%～14.3%ρFA48.6mg/L334.5%44.2%60.8%ρFAρFA3SEM.FISHX703.1A1001－6929201009－1185－07StudyofBacterialCommunityChangesinCANNEDProcessbyFISHMethodZHANGWei-hua12WANGHai-yan1ZENGQing-ru2ZHOUYue-xi1GEJian-tuan31.ResearchCenterfortheWaterPollutionControlTechnologyChineseResearchAcademyofEnvironmentalSciencesBeijing100012China2.CollegeofResourcesandEnvironmentHunanAgriculturalUniversityChangsha410128China3.EnvironmentalSciencesDesignResearchAcademyofGansuProvinceLanzhou700030ChinaAbstractScanningelectronmicroscopeSEMandfluorescenceinsituhybridizationFISHwereusedtostudythebacterialmorphologyandcommunityofthecompleteautotrophicnitrite-nitrificationandelectrochemicalbio-denitrificationcombinedprocessCANNEDwhichwasoperatedforammoniumwastewatertreatmentatdifferentinfluentconcentrationsoffreeammoniaFA.WhenthemassconcentrationofFAwas0.5mg/Linthenitrite-nitrificationparttheAOBandNOBwere0.9%and2.8%ofthebacteriarespectively.Itcouldachieveshort-termnitrificationwhenthemassconcentrationofFAwas12.4-48.6mg/L.AOBandNOBwere33.6%-47.4%and1.0%-2.9%ofbacteriarespectivelyundertheabovementionedconditions.Intheelectrochemicalbio-denitrificationpartthethiobacillusdenitrificansanammoxbacteriaandeubacteriawere7.2%-15.3%7.2%-10.3%and9.1%-14.3%oftotalbacteriarespectivelywhentheinfluentFAwas0.5-12.4mg/L.WhenthemassconcentrationofFAincreasedto48.6mg/Ltheabovethreekindsofbacteriawere34.5%44.2%and60.8%oftotalbacteriarespectivelywhichindicatedthatthegrowthofthebacteriawasacceleratedwiththeconcentrationofFAincreasing.TheabovementionedresultswereinaccordancewiththeSEMresults.Keywordsnitrite-nitrificationelectrochemicalbio-denitrificationfreeammoniaFISHSE[e-mail].*[e-mail]、1-3.SHARON－ANAMMOXOLANDOxygenLimitedAutotrophicNitrificationDenitrificationCANONCompletelyAutotrophicNitrogenremovalOverNitriteinonesinglereactor4.23、、C/N、HRT1d、22%5.209098%.、－200%6-8./SulfurLimestoneAutotrophicDenitrificationSLADWANG90.66kg/m3·d10.11.12/CompleteAutotrophicNitrite-NitrificationandElectrochemicalBio-DenitrificationCombinedProcessCANNEDMBRρ1000mg/L95%12.MBR、.FISH208013.rRNA14DNADNA.PCRFISH15-17.、18.SEMρFAρFA.11.1.MBR〔1a〕485mm300mm450mm43L40L.ρDO0.2～0.6mg/L19..1b.670mm×400mm×284mm106L2.5cm1.1—2—3—4—5—6—7—8—9—PLC10—DO1Fig.1Experimentalequipments68119CANNEDFISH1Table1Theparametersofthecathodeandanode///cm2/mm400mm×276mm×10mm400mm×276mm×10mm101124288251∶1.40LDH1720A－6.1.2MBRpHNaHCO3MBRρFA.MBRρDO0.2～0.6mg/L32～35℃HRT24h.MBRρMLSS2200mg/LρMLSS1200mg/L.MBR.MBRρ200mg/LρFA〔ρFA〕0.5mg/LMBRM1E1ρFA12.4mg/LMBRM2E2MBRρ700mg/LρFA48.6mg/LMBRM3E3.23.2Table2TheoperationparametersandresultsoftheMBRHRT/hρDO/mg/LpHρFA/mg/Lρ/mg/Lρ/mg/L/%ρ/mg/L/%ρ/mg/L/%M1210.548.027.880.5216.148.8846.73258.63M2240.168.178.3012.4214.829.260136.3632.798M3230.368.258.3648.6765.7294.562415.4540.0100.3Table3Theoperationparametersandresultsoftheelectrochemicalbiorealtor/V/ApHρ/mg/L/%/%E10.80.047.6079.46722E22.00.357.847.06146E32.41.738.5441.8261200mL30min3000r/min5minFISHSEM20.1.3ρρICS－1000DionexISC－1000ρ21752ρMLSSpHpHS－25pHρDOYSIDO200.1.4FISHFISH4%46℃1～3h48℃20%Tris－HCl20min.FISH4100mg/L.7811234FISH22-26Table4ThemessageforFISHprobes5′－3′w/%NSO190β－CGATCCCCTGCTTTTCTCC190～20820NIT3CCTGTGCTCCATGCTCCG1035～104840Thi－dTGCTGGTACCGTCATTAGCACACGATG446～47235Amx820AAAACCCCTCTACTTAGTGCCC820～84140EUB338ⅡGCAGCCACCCGTAGGTGGT338～3552022.1ρFA2.HRT24hρDO0.2～0.6mg/L30～35℃78～136ρ200mg/L137～170ρ700mg/L60%～85%.2MBRFig.2TheoperationresultsoftheMBRforthenitrite-nitrification2MBR78～90ρFA0.5～2.0mg/L83.8%5%61%～77%ρMBR91～136ρFA8.1～12.5mg/L63.5%34%～85%98%0%137～170ρFA48.6～62.0mg/L54.2%50%100%.ρFA50%.ρFA8.1～62.0mg/LAOB.〔ρFA40～70mg/L〕27.2.2FA3FA.15～20℃.378～900.16AρFA0.5～1.6mg/Lρρ48.86.7mg/L、70.0%21.5%91～1360.42AρFA8.1～12.4mg/Lρρ23.6～141.66～183mg/L、60%～97%10.6%～64.3%ρρ.137～1701.61AρFA48.6～62.0mg/Lρρ294.5～312.5278.4～415.4mg/L、2%～7%61%.ρFA.2.3FISHEUB33888119CANNEDFISH3Fig.3TheoperationresultsoftheelectrochemicalbiorealtorDAPI28..AOBNSO190NOBNIT3Thi－dAmx820EUB338Ⅱ.2.3.1FISHM1M2M3E1E2E3615～20FISH4～9.Image－ProPlus5.4M1DAPINSO190Fig.4ThefluorescenceimagesofDAPIandNSO190probeforM15M2DAPINSO190Fig.5ThefluorescenceimagesofDAPIandNSO190probeforM26E2DAPIThi－dFig.6ThefluorescenceimagesofDAPIandThi-dprobeforE27E3DAPIThi－dFig.7ThefluorescenceimagesofDAPIandThi-dprobeforE38E2DAPIAMX820Fig.8ThefluorescenceimagesofDAPIandAMX820probeforE25ρFA0.5mg/LM1AOBNOB0.9%2.8%ρFA12.4mg/LM2AOBNOB9811239E3DAPIAMX820Fig.9ThefluorescenceimagesofDAPIandAMX820probeforE333.6%2.9%ρFA48.6mg/LM3AOBNOB47.4%1.0%.ρFA10～50mg/LAOBNOB.5CANNEDFISHTable5TheresultsofFISHforsludgesamplesinCANNEDprocess/%NSO190NIT3Thi－dAmx820EUB338ⅡM10.92.838.918.525.1M233.62.90048.3M347.41.04.43.566.0E1——15.310.314.3E2——7.27.29.1E3——34.544.260.8ρFA0.5mg/L38.9%ρFA12.4～48.6mg/L33.6%～47.4%.ρFA0.5～12.4mg/LE1E2FISH、7.2%～15.3%