复合嗜盐微生物处理高含盐废水的研究杨波

300192GB－4、GB－5、GB－12、16SrＲNAHalobacillushalophilus、Marinobactersp．Bacillusmethylotrophicus。33∶2∶148hCOD81．85%。25℃～40℃、pH6．0～9．0、NaCl6%～12%COD70%。X703A1673－6850201511－0033－07ＲesearchonHypersalineWastewaterTreatmentbyCompositeHalophilicMicroorganismYANGBoHAOJian－anZHANGXiao－qingZHANGAi－junJIANGTian－xiangWANGJingZHANGYu－shanTheInstituteofSeawaterDesalinationandMulti－purposeUtilizationSOATianjin300192ChinaAbstractThreehalophilicmicroorganismwasnamedGB－4GB－5andGB－12whichwereisolatedfromasoilsamplethatwascollectedfromTianjinportChina．ThesethreehalophilicbacteriawereidentifiedasHalobacillushalophilusMarinobactersp．andBacillusmethylotrophicusbasedonthemorphologicalcharacteristicsphysiologicalandbiochemicalcharacteristicsandphylo-geneticanalysisof16SrＲNAgenesequences．TheCODremovalratecanreach81．85%whenthreehalophilicbacteriastrainswith3∶2∶1wereculturedinhypersalinewastewaterforroutinelifefor48hwhichwassignificantlybetterthanthesinglestrainofhypersalinewastewater．Complexmicroorganismcangrowwellwhenculturedinhypersalinewastewaterunderthecultureconditionof25～40℃pH6．0～9．0NaClconcentrationwithinthescopeof6%～12%theCODremovalratewerealsokeptabove70%．KeywordshypersalinewastewaterhalophilicbacteriacomplexmicroorganismsCOD201[e-mail]3．5%TDSTotalDissolvedSolid1。、2、、、3。Na+、Ca2+、Cl－、SO2－44。、。334411201511JournalofSaltandChemicalIndustryDOI:10.16570/j.cnki.issn1673-6850.2015.11.011。、、5。、、6。。、、、7。。、“”。、。11．11．1．1。1．1．21250mg/L128mg/LKH2PO472mg/LK2HPO430mg/LNaCl12%pH7．5。。1．1．32216E5g/L1g/L0．01g/L1LpH7．6121℃20min。Gibbons5．0g/L12．0g/L6．0g/L3．0g/LKCl3．0g/LMgSO4·7H2O18．0g/LCaCl20．2g/LFeSO4·7H2O1mg/LNaCl70．0g/L1．5g/L1LpH7．5121℃20min。1．21．2．1、、10g90mL10－11mL10－19mL10－2。。2216E37℃48h。3～5。1．2．2Gibbons37℃、160r/min5000r/min5min1%100mL24h、48h72h。。8。1．2．3、、。《》9《》10。API。1．2．416SrＲNACTABDNA16SrＲNAfD15'－AGAGTTTGATCCTGGCTCAG－3'rp25'－ACG-GCTACCTTGTTACGACTT－3'11PCＲ95℃5min94℃1min56℃1min72℃2min3072℃10min。。BLAST16SrＲNAGeneBank16SrＲNA。DNAMAN。4344112015111．2．5GB－4、GB－5GB－12。1。1Tab．1ThedesigntableofstrainproportioningexperimentGB－4GB－5GB－12111121223133421252236231731383219332100mL250mL2mL137℃、160r/min48hCOD。1．2．6pH4．0、5．0、6．0、7．0、8．0、9．0、10．0、11．0pH15℃、20℃、25℃、30℃、35℃、40℃、45℃NaCl0%、3%、6%、9%、12%、15%。COD。1%30℃、160r/min48hOD600COD。22．124h～48h、6。62。2．224h、48h、72hCOD1。2Tab．2ThemorphologicalcharacteristicsofhalophilicmicroorganismsGB－2GB－3GB－4GB－5GB－8GB－12COD去除率/%微生物菌株100806040200-20-40GB-2GB-3GB-4GB-5GB-8GB-1224h48h72h1CODFig．1ＲemovaleffectofCODinhighsaltwastewaterbysaltmicrobialstrains1GB－4、GB－5、GB－8、GB－1272hCOD60%。GB－4、GB－5GB－1224hCOD45%。2．32GB－40．78μm×0．50μmGB－51．25μm×0．54μmGB－121．85μm×0．52μm。2Fig．2AFMphotosofhalophilicmicroorganismstrainsGB－4GB－5GB－12Vitek253。3～5。3GB－4GPTab．3MonitoringresultsonthestrainGB－4againstvarionsenzgmeactivityofGram－positivebacterialidentificationcardGP－α－+－C－－6．5%NaCl－D－－β－D－－D－－1－+D－－β－++－B－D－－α－+D－－－－－－－D－－－B－O/129－L－－D－－－β－－D－+－α－－L－－D－－－－2－+N－－β－－－L－－D－－β－－－“－”“+”4GB－5GPTab．4MonitoringresultsonthestrainGB－5againstvariousenzymeactivityofGram－positivebacterialidentificationcardGP－－－β－－－－β－－N－－β－－－L－－α－－L－－－－D－－－－β－－+－H2S－+－β－N－－D－－－－－－D－－D－－COUＲMAＲATE－γ－－D－－β－－－－0/129－β－－－－－－D－－5－－－L－－D－－+ELLMAN－D－－α－+L－－“－”“+”5GB－12GPTab．5MonitoringresultsonthestrainGB－12againstvariousenzymeactivityofGram－positivebacterialidentificationcardGPβ－+－－L－++α－+L－－ELLMAN+D－－+－D－－D－++α－－－－+D－－β－++D－－+D－－－α－－D－+6．5%NaCl+6344112015115Continue－D－－++N－－D－－+N－－β－D－－++－－+L－－B－CYCLODEXTＲINE+β－+B+D－－β－－+B－“－”“+”“－”“+”Vitek26h～8h3～5。Vitek2。Vitek。vitek2Vitek。2．416SrＲNA16SrＲNABLASTN3。16SrＲNAfD1rP2。0.05GB-5Marinobactersp.TB140（EU308448）Marinobacteradhaerensstrain_HP15（NR115177）MarinobacterflavimariisolateD1-1M（AB617558）Unculturedgammaproteobacterium（AM408940）Acinetobacterlwoffiistrain262XY6（KF818633）BacillussubtilisstrainYA-3（KM497438）BacillusvallismortisstrainNBIF-001（KP994551）BacillusmethylotrophicustrainPY3（KC790266）GB-12HalobacillusalkaliphilusstrainZSTB206（JQ068929）Halobacillushalophilusstrain3（NR075035）GB-4UnculturedFirmicutesbacteriumcloneKir21gryB10.a6（HM480231）EscherichiacolistrainATCC35469（KP941759.1）3Fig．3Thephylogenetictreeofhalophilicmicroorganismstrains16SrＲNA97%12。、。、、。、GB－4、GB－5GB－12Halobacillushalophilus、Mari-nobactersp．Bacillusmeth-ylotrophicus。2．51%100mL30℃、160r/min24h48hCOD。4。0h24h48hCOD/（mg·L-1）菌种组合400350300250200150100500123456789CK4Fig．4Strainsofcompositetestresults4。48hCOD824h48hCOD72．63%81．85%。COD。73。2．62．6．1COD。COD5。OD600COD去除率温度/℃OD6001.61.41.21.00.80.60.40.201009080706050403020100COD去除率/%152025303540455CODFig．5EffectoftemperatureongrowthofhalophilicbacteriaandremovalofCOD5CODCOD20℃～45℃。25℃～40℃OD600COD343．23mg/LCOD65%。COD。2．6．2pHCODpH。pHpH。pHpH。pH。6pH。pH6．0～9．0OD6001．2COD69%。pH5．010．0。pH值OD600COD去除率OD6004.01.81.61.41.21.00.80.60.40.201009080706050403020100COD去除率/%5.06.07.08.09.010.011.06pHCODFig．6EffectofpHongrowthofhalophilicbacteriaandremovalofCOD2．6．3COD。。NaCl7。OD600NaCl浓度/%01.81.61.41.21.00.80.60.40.201009080706050403020100COD去除率/%OD600COD去除率36912157NaClCODFig．7EffectofNaC