冬季低温下MBR与CAS工艺运行及微生物群落特征黄菲

35320143ENVIＲONMENTALSCIENCEVol．35No．3Mar．2014MBＲCAS*200092MBＲCAS454．MBＲＲ1Ｒ2CASＲ385.2%、56.1%、58.8%NH+4-N99.7%、99.7%、59.7%Ｒ2、Ｒ1、Ｒ3Ｒ3、Ｒ1、Ｒ2MBＲ45497%Ｒ2＞Ｒ3＞Ｒ1Ｒ2＞Ｒ1＞Ｒ3MBＲCASＲ1、Ｒ2、Ｒ3Nitrospira1.22%、1.64%、0.15%Zoogloea、Thauera、ComamonadaceaeComamonas5.8%、4.52%、15.21%、、TNMBＲ、．454X703.1A0250-3301201403-1002-07DOI10．13227/j．hjkx．2014．03．0262013-07-242013-10-22112312004028632012AA0634031989～[e-mail]*[e-mail]rformanceandMicrobialCommunityStructuresinMBＲsandCASProcessesatLowTemperatureHUANGFeiMEIXiao-jieWANGZhi-weiWUZhi-chaoStateKeyLaboratoryofPollutionControlandＲesourceＲeuseSchoolofEnvironmentalScienceandEngineeringTongjiUniversityShanghai200092ChinaAbstractInthispapertheperformanceofmembranebioreactorsMBＲandconventionalactivatedsludgeCASprocessesatlowtemperaturewasinvestigatedbyanalyzingtheireffluentqualityandmicrobialviability．454high-throughputpyrosequencingwasalsoappliedtostudythemicrobialcommunitystructures．TheresultsofthreesystemstwoMBＲsＲ1withhighsludgeconcentrationＲ2withlowsludgeconcentrationandoneCASＲ3showedthattheaverageremovalrateofNH+4-Nwas99.7%99.7%and59.7%respectivelyandtheaverageremovalrateofTNwas85.2%56.1%and58.8%respectively．Ｒ2showedthehighestspecificammoniumuptakerateSAUＲfollowedbyＲ1andＲ3Ｒ1showedthehighestspecificnitrateuptakerateSNUＲfollowedbyＲ2andＲ3.ItcouldbeconcludedthatMBＲswithhighsludgeconcentrationhadabetterperformanceofnitrogenremovalunderlowtemperatureoperation．454high-throughputpyrosequencinganalysisrevealedthatthemicrobialrichnesswasＲ2＞Ｒ3＞Ｒ1andthemicrobialdiversitywasＲ2＞Ｒ1＞Ｒ3at97%sequenceidentity．ThemicrobialstructureandbacterialabundancewerequitedifferentbetweenMBＲandCASsystems．ThedominantnitrifierinthisresearchwasNitrospiraandthetotalrelativeabundanceofnitrifiersinＲ1Ｒ2Ｒ3was1.22%1.64%and0.15%respectively．TheZoogloeaThaueraComamonadaceaeandComamonasmightbethedominantdenitrifersinthisstudyandthetotalrelativeabundanceofdenitrifierinＲ1Ｒ2andＲ3was5.8%4.52%and15.21%respectively．MBＲ'scharacteristicsoflongsolidretentiontimehighsludgeconcentrationandlowtotalnitrogenloadingwellsupportedtheaccumulationofnitrifiersanddenitrifiersandimprovedtheperformanceofbiologicalnitrogenremovalatlowtemperature．Keywordslowtemperaturemembranebioreactorconventionalactivitedsludgeprocess454high-throughputpyrosequencingmicrobialcommunitystructuresnitrogenremoval“”10%1．＜12℃、．membranebioreactorMBＲconventionalactivatedsludgeCAS、、、．MBＲ3MBＲCASHＲTSＲT．“”MBＲ2．3MBＲCAS4MBＲCASMBＲ、MBＲ．MBＲ．DGGE、、5～7．454DNA、、8910．454MBＲ．11.12012-12-01～2013-02-205.9℃±2.7℃3Ｒ1、Ｒ2、Ｒ3Ｒ1、Ｒ2AO-MBＲ．0.2μmＲ3A2/O．3463±167mg·L－147.05±11.62mg·L－135.95±9.21mg·L－1．31．11Table1OperatingconditionsofＲ1Ｒ2Ｒ3Ｒ1Ｒ2Ｒ3AO-MBＲAO-MBＲAAOHＲT/h132112～13SＲT/d100308～10MLSS/g·L－114.89±0.647.18±0.303.08±1.51MLVSS/g·L－110.97±0.475.45±0.272.34±1.16TN/VSS/kg·kg·d－10.009±0.0020.012±0.0020.041±0.0141MLSSMLVSSn=101.21.2.1COD、TN、NH+4-N、NO－3-NNO－2-N、MLSS、MLVSS．11．1.2.21213SAUＲ1LNH+4-N25mg·L－1NaHCO3pH7～8．DO5～6mg·L－120～30min3h．NH+4-NNH+4-NAUＲ．MLVSSSAUＲ．SNUＲCODNO－3-N300mg·L－125mg·L－1．5min．20～30min3h．NO－2-N+NO－3-NNO－2-N+NO－3-NNUＲ．MLVSSSNUＲ．1.2.3DNAPCＲＲ1、Ｒ2、Ｒ31．E．Z．N．ASoilDNAOMEGABiotek300135DNADNA14．DNA1%．PCＲ16SrＲNAV1-V3“5'454A、B-3'”AB．16SrＲNA533Ｒ5'-TTACCGCGGCTGCTGGCAC-3'27F5'-AGAGTTTGATCCTGGCTCAG-3'．PCＲTransGenAP221-02TransStartFastPuDNAPolymeraseTansGen20μL5×FastPfuBuffer4μL2.5mmol·L－1dNTPs2μL5μmol·L－1533Ｒ0.8μL5μmol·L－127F0.8μLFastPuPolymerase0.4μLDNA10ng．GeneAmp9700ABIPCＲ95℃2min2595℃30s55℃30s72℃30s72℃5min．AxyPreDNAAXYGENPCＲTris-HCl2%．PCＲQuantiFlourTM-STPromega．454454GSFLXTitaniuＲoche454．．①②150bp、Ns、2、6．OTU．OTU150bpSILVA106．OTUMOTHUＲhttp//．mothur．org/wiki/Main_Page97%95%．OTU．MOTHUＲChaohttp//．mothur．org/wiki/ChaoShannonhttp//．mothur．org/wiki/ShannonGood'scoveragehttp//．mothur．org/wiki/Coverage．400bpSILVA106OTU80%．22.11Ｒ1、Ｒ2、Ｒ3Fig．1EffluentqualityofＲ1Ｒ2Ｒ3Ｒ1、Ｒ2、Ｒ3TN、NH+4-N、NO－2-N、NO－3-N1．Ｒ1、Ｒ2、Ｒ3TN85.24%、56.05%、58.75%NH+4-N99.7%、99.7%、59.7%．MBＲＲ1、Ｒ2Ｒ3TNNH+4-NMBＲＲ1Ｒ2NH+4-NNH+4-N0.2mg·L－1MBＲＲ1Ｒ2TNNO－3-NＲ1Ｒ1TNTN．3Ｒ1A、、MBＲ．SAUＲSNUＲ．23SAUＲＲ2＞Ｒ1＞Ｒ3MBＲ．40013MBＲCASMBＲ0.0737kg·kg·d－1、0.0715kg·kg·d－1MBＲ0.0347kg·kg·d－10.1110kg·kg·d－115A2/O13℃．Ｒ3＞Ｒ1＞Ｒ233Ｒ1、Ｒ2、Ｒ30.7800、0.2825、0.2742kg·kg·d－1MBＲ、．3SNUＲ0.0625、0.0588、0.1225kg·kg·d－1、．2Ｒ1、Ｒ2、Ｒ3Table2MicrobialviabilityofＲ1Ｒ2Ｒ3Ｒ1Ｒ2Ｒ3SAUＲNH+4-N/VSS/kg·kg·d－10.05920.06730.0347SNUＲNO－3-N/VSS/kg·kg·d－10.07110.05180.1172SAUＲNH+4-N/VSS/kg·kg·d－10.07370.07150.1110SNUＲNO－3-N/VSS/kg·kg·d－10.06250.05880.1225MBＲMBＲ、、MBＲ．2.2316SrＲNAＲ1、Ｒ2、Ｒ39334、8687、9505473bp．SILVA10697%2610、2595、2281OTUＲ1、Ｒ2、Ｒ326000OTUＲ1、Ｒ2、Ｒ3Ｒ1、Ｒ2、Ｒ3．3Alpha-diversity．ChaoOTUOTU97%ChaoＲ2＞Ｒ3＞Ｒ1ChaoOTU．Shannon2Ｒ1、Ｒ2、Ｒ3Fig．2ＲarefactioncurveofsampleＲ1Ｒ2Ｒ33Ｒ1、Ｒ2、Ｒ3Table3ＲichnessanddiversityestimatorsofthebacteriainsampleＲ1Ｒ2Ｒ3OTUChaoShannonＲ1261074376.680.73Ｒ2259589596.840.69Ｒ3228178736.420.73Ｒ2、Ｒ1、Ｒ3．97%OTU3Ｒ1、Ｒ2、Ｒ319、17、183ProteobacteriaBacteroidetes．Ｒ1、Ｒ2、Ｒ3266、253、140500135MBＲ．MBＲＲ1、Ｒ2A2/OＲ3Ｒ1、Ｒ2Acidobacteria、Chloroflexi、Nitrospirae、PlanctomycetesＲ31%．Nitrospirae16Proteobacteria、Firmicutes17．．4Ｒ1、Ｒ2、Ｒ3、Fig．4PhylogeneticanalysisofsampleＲ1