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(SBBR),＊,,,,(, 410082):PCR、(DGGE),SBBR.,--,NH+4-N65%;2--.2,.,,NH+4-NNO-2-N,,.:SBBR;;;;:X703.1 :A :0250-3301(2007)10-2326-06:2006-12-18;:2007-03-19:(50478053);(863)(2003AA644010);(04JJ3004):(1981～),,,,E-mail:xuyaze@21cn.com＊,E-mail:yzh@hnu.cnApproachesofBiologicalNitrogenRemovalinaSingleSequencingBatchBiofilmReactorXUZheng-yong,YANGZhao-hui,ZENGGuang-ming,WANGRong-juan,XIAOYong,XUZhen(CollegeofEnvironmentalScienceandEngineering,HunanUniversity,Changsha410082,China)Abstract:TheconventionalmicroorganismtechniquesandthemoleculebiologicaltechniquessuchasPCRandDGGEwereutilizedtostudytheapproachesofbiologicalnitrogenremovalinasinglesequencingbatchbiofilmreactor(SBBR).Themainapproachofbiologicalnitrogenremoval,nolessthan65%ofthetotalNH+4-Nwasremovedinthisapproach,wascomposedofpartialnitrification,anaerobicammoniumoxidationanddenitrification.Thesecondapproachincludedtwainprocessessuchaspartialnitrificationanddenitrification,andthethirdonewasconventionalnitrogenremovalprocess(nitrificationanddenitrification).Allapproachesaccomplishedsimultaneouslyorasynchronously.Whentheapproachoccurredinthesimultaneousway,itperformedasnitrogen-loss.Theaccomplisherofasynchronouswaydependedonthemicrobe'sdifferentactivitybetweenaeratedphaseandanaerobicphase.First,ammoniawasconvertedtonitriteintheaeratedphasewhenthenitritebacteriaperformedhighactivity,andthenanaerobicphasewasoperatedinturnwhenthedenitrifyingbacteriaandtheanammoxbacteriaeliminatedthenitrogenformgases.Keywords:singleSBBR;approaches;partialnitrification;anaerobicammoniumoxidation;phylogeneticanalysis (sequencingbatchbiofilmreactor,SBBR),Gonzale1990[1].(SBR),,.(),1[2,3].[4～8],(SBBR),,,();,、,.SBBR-[8],2a.,,,,.2810200710 ENVIRONMENTALSCIENCEVol.28,No.10Oct.,2007DOI:10.13227/j.hjkx.2007.10.0251 1.1 ,COD580～1500mg·L-1,BOD5150～420mg·L-1,NH+4-N440～590mg·L-1,NO-3-NNO-2-N,pH7.8～8.2.30℃NH4ClNH+4-N450mg·L-1,300mg·(L·d)-1,pH7.8.1.2 SBBR1,3L,,1∶3.,32～34℃.PLC、,1.2～1.4mg·L-1.500mL0.01mol·L-1().1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14.1 SBBRFig.1 SchematicchartoftheSBBRrector1.3 SBBRSBBR12h,4:①:;②:2,(O)2h,(A)1h,.,SBBR1.2～1.4mg·L-1;③:1;④:,0.5.2 SBBRFig.2 OperatingscheduleofSBBRduringreactionphase1.4 (1) NH+4-N、NO-3-NNO-2-N(ET99730,Lovibond,);DO(HI9143,Hanna,);pHpH(PenpHmeter,Lida,).(2),0.3g,8,S1～S8,-20℃;DNAK-CTAB[9].DNAGC-341F907R[10](GC-341F:5′-CCTACGGGAGGCAGCAG-3′;907R:5′-CCGTCAATTCCTTTGAGTTT-3′)PCR.PCR1%,QIAquikGelExtractionKit(Qiagen,Germany),60μLMilli-Q.30μLPCRDCodeSystem(Bio-Rad,USA),30%～60%,6%,140V,15h,55℃[11].,0.5μg·mL-1,GelDoc2000(BioRad,USA).DGGE,60μLMilli-Q24h,20μLPCR,DGGE,(Sangon,).GenBankBLAST,DNAStar5.0.2 2.1 、,、,232710:(SBBR),、.SBBR、.101NH+4-N、NO-3-NNO-2-N,3.NH+4-N,.NO-2-N,;,,[12,13],NO-2-N.NO-3-NNO-2-N,.3 Fig.3 CyclevariationofnitrogenconcentrationinSBBR2.2 (1).NH+4+OH-NH3+H2O(1) (2)[14]:ρFA=17ρ(NH+4-N×10pH)14(KbKw+10pH)(2),Kb,Kb=10-9.24(20℃);Kw,Kw=0.69×10-14(20℃);KbKw=e[6.344(273+T)].(1)(2),pH,,.,pH,(1),,,pH4,,pH,,pH.,pH.4 SBBRpHFig.4 CyclevariationofpHinSBBR,,,,,,.2.3 pHpH,pH,5.,pH,pH,1,pH0.11.,4:5.7、5.4、3.44.6,,0.5 pHFig.5 CyclevariationofpHinabsorptionliquidanddissipationcapacityineachphases2.4 NH+4-NNO-2-N[15],NO-2-NNH+4-N1.3[16,17].,35,NO-2-NNH+4-N2328 28,,1.1 1)mg·L-1Table1 Variationcharacteristicofnitrogenconcentrationineachphasemg·L-111223344NH+4-N-39.4-14.3-45.7-18.2-40.5-16.8-29.4-11NO-2-N22.4-20.729.7-29.127.2-27.615.4-16.6NO-3-N2.9-2.72.9-3.13.3-3.12.7-2.65.705.403.404.602)-8.4—-7.7—-6.6—-6.7—NO-2-NNH+4-N—1.45—1.60—1.64—1.511);2) 1,,,NH+4-N,NH+4-N.0.5%～2.0%,[18],,,[19];,1.2～1.4mg·L-1,,[20,21],NO-2-NNO-3-N.,NO-2-NNH+4-N,.,.,1molNH+4-N0.22molNO-3-N,3NO-3-N,.NO-3-N,[22].2.5 ,,.PCR-DGGES1～S881616SrDNA(6),(Sangon,),BLASTGenBank,DNAStar5.07.6 DGGEFig.6 DGGEresultsofbiofilmsamples7 Fig.7 Phylogenetictreebasedonthesequencesofbacteriainthebiofilm232910:(SBBR):CandidatusBrocadiaanammoxidans(5),Nitrosomonaseutropha(2)、Nitrosomonassp.Nm143(7)、Nitrosomonassp.Is425(8),Nitrobacterwinogradskyi(6),Hyphomicrobium(13)、Bacillusclausii(16).3 ,、、,(3)～(7).NH+4+1.5O2※NO-2+2H++H2O(3)NO-2+0.5O2※NO-3(4)NH+4+1.31NO-2※1.045N2+0.22NO-3+0.09e-(5)NO-2+4H++3e-※0.5N2+2H2O(6)NO-3+6H++6e-※0.5N2+3H2O(7) ,NH+4-N(1),NH+4-N,NH+4-NNO-2-NNO-3-N.,SBBRNO-3-N;(5),,4NO-3-N:0.8、0.73、0.630.64mg·L-1.NO-3-N.,NH+4-N().1,NO-2-NNH+4-N,NO-2-N.,NO-3-N,NO-3-NNO-2-N,,NO-3-NNO-2-N[23].,,NO-3-N,NO-2-N.,,.,1.2～1.4mg·L-1,3.2～4.5mg·L-1.2[24].,.,,,.,,、;,.8,、.SBBR3:--(Ⅰ),-(Ⅱ),-(Ⅲ).① ② ③ ④8 Fig.8 Schematicchartofnitrogenbio-conversionSBBR,NH+4-N,3.32,,,.,NO-3-NNO-2-N,3NH+4-N,2330 2813.7%.,NH+4-NⅠ,ⅠNO-2-N,NO-2-NⅡ.2ⅠⅡNH+4-NNO-2-N.,Ⅰ65%,Ⅱ7%.2 ⅠⅡ1)mg·L-1Table2 NitrogenconsumedbyapproachⅠandⅡmg·L-11234ⅠNH+4-N14.3018.2016.8011.0060.30NO-2-N18.7323.8422.0114.4178.99ⅡNO-2-N1.975.265.592.1915.011)ⅠNO-2-N3.1(5)4 (1)SBBR,--,65%.(2),.(3)PCR-DGGESBBR,DNA、SBBR.SBBR.:[1]GonzalesS,WildererPA.Phosphateremovalinabiofilmreactor[J].WaterScienceandTechnology,1990,23(7-9):1405～1416.[2],,,.[J].,2002,22(3):320～323.[3],,,.[J].,2004,25():74～77.[4]SartiA,PozziE,ChinaliaFA,etal.Theperformanceofananaerobicsequencingbatchbiofil