31320103 ENVIRONMENTALSCIENCEVol.31,No.3Mar.,2010UASB1,1,2,1*,1,1(1., 050018;2., 100862):,(fluorescenceinsituhybridization,FISH)UASB.,(),(methanogens)、(Methanobacteriales)(Methanosarcinales),.;;(25.50±8.63)%~(48.67±8.87)%;1.0~2.0mm,(47.08±8.26)%(48.67±8.87)%..1.311~1.562g/(g·d),COD,,.:;;;;:X172 :A :0250-3301(2010)03-0725-06:2009-05-11;:2009-06-08:(50878072);(E2008000694,E2009000709):(1976~),,,,,E-mail:liuchun02@mails.tsinghua.edu.cn*,E-mail:yangjingliang@hebust.edu.cnAnalysisofMethanogenicCommunityofAnaerobicGranularSludgeinaFull-scaleUASBTreatingAvermectinWastewaterLIUChun1,MAJun-ke1,WUGen2,YANGJing-liang1,GUOJian-bo1,LIZai-xing1(1.SchoolofEnvironmentalScienceandEngineering,HebeiUniversityofScienceandTechnology,Shijiazhuang050018,China;2.BasicResearchServiceCenter,MinistryofScienceandTechnology,Beijing100862,China)Abstract:Methanogensisconsideredtobeimportantfunctionalmicrobialpopulationinanaerobicgranularsludge.Fluorescenceinsituhybridizationwasusedtoanalyzemethanogensofanaerobicgranularsludgeinafull-scaleUASBtreatingavermectinwastewater.Theresultsindicatedthatthedistributionformsofmethanogens,MethanobacterialesandMethanosarcinaleswereidenticalonthesurfaceandinnerfaceofgranularsludgesamplesindifferentformationstages(withdifferentdiameters),althoughtherelativeabundancesofthesemethanogensweredifferent.Therelativeabundancesofmethanogensontheinnerfacewerelargerthantheseonthesurfaceofgranularsludgesamples.TherelativeabundancesofMethanobacterialeswerelargerthantheseofMethanosarcinales.Therelativeabundancesofmethanogensinallgranularsludgesamplesrangedfrom(25.50±8.63)%to(48.67±8.87)%.Themaximumrelativeabundancesofmethanogenswereobtainedinmaturegranularsludgewithdiameterof1.0-2.0mm,(47.08±8.26)%onthesurfaceand(48.67±8.87)%ontheinnerface,respectively.Theavermectinresidueinwastewatershowedpossibleinhibitioneffectonmethanogens.Themaximumspecificmethanogenicactivitiesofgranularsludgesamplesrangedfrom1.311to1.562g/(g·d),varyingassameasCODremovalratiosandrelativeabundancesofmethanogens,implyingthestrongcorrelationofmethanogenswithbioactivityofgranularsludge.Keywords:anaerobicgranularsludge;methanogens;fluorescenceinsituhybridization(FISH);relativeabundance;avermectinwastewater ,,[1~4].,、.,(FISH),[5,6].[7~9]FISH,,,.Montero[10]FISH..,DOI:10.13227/j.hjkx.2010.03.017 31[11,12][13].,,,.Chen[14]、、;Liu[15]PCPPCP.,.,.,,.UASB,FISH,.1 1.1 UASB(500m3),VSS/SS0.941.UASB0.6mm、0.6~1.0mm、1.0~2.0mm、2.0mm4,14.8%、5.3%、60.3%19.6%,VSS/SS0.905、0.940、0.9570.945.1.0~2.0mm,;0.6~1.0mm;2.0mm;0.6mm.1.2 FISH1.2.1 4:UNIV1392、MPB1、MEB859MSSH859,,1.(Takara).1.2.2 250μL1.5mL,34%.4℃1h,10000r/min5min,,1×PBS3,,50%、80%100%,3min.1μL0.1nmol/μL9μL(0.9mol/LNaCl,0.01%SDS,0.02mol/LTris-HCl,,pH7.2),,(UVP,HB-1000,),46℃5h.1 FISHTable1 ProbesusedinFISH(5′-3′)UNIV1392ACGGGCGGTGTGTAC5′FITC[8]MPB1CATGCACCACCTCTCAGC5′FITC[12]MEB859GGACTTAACAGCTTCCCT5′FITC(5′)[16]MSSH859CTCACCCATACCTCACTCGGG5′FITC[16] 46℃(0.01%SDS,0.02mol/LTris-HCl,NaCl,pH7.2),46℃ddH2O3,.:,.1.2.3 (Motic,BA200,),30FISH(Moticam2206).MoticFluo1.0,,,(1)[17].(%)=×100%(1)1.3 :(VSS)4,35℃,.,7263:UASB:1%、1%、0.25%、0.25%、0.05%NH4Cl、0.02%K2PO40.005%MgCl2(),pH7.2,COD4000~5500mg/L[18],1.6~2.2kg/(m3·d).,K,(2)A[19].A=24×K×fV×S(2),A,g/(g·d);K,L/h;f,g/L,2.574;V,L;S,g/L.COD:8hCOD,.2 2.1 2.1.1 3(Methanogens)、(Methanobacteriales)(Methanosarcinales),.,,.11.0~2.0mm、.;,[1(d)].24..,,.,,3.,,,1.0~2.0mm,2.0mm.,0.6mm、0.6~1.0mm、1.0~2.0mm2.0mm4(25.50±8.63)%、(39.48(a);(b);(c);(d)()()1 1.0~2.0mmFig.1 Distributionsofdifferentmethanogensonthesurfaceofsludgesamplewithdiameterof1.0-2.0mm(a)0.6mm;(b)0.6~1.0mm;(c)1.0~2.0mm;(d)2.0mm2 Fig.2 DistributionofMethanogensonthesurfaceofsludgesampleswithdifferentdiameters±9.05)%、(47.08±8.26)%(43.69±9.26)%.,,;1.0~2.0mm,;,.,,.1.0~2.0mm,(43.68±7.98)%(35.31±6.24)%,727 313 Fig.3 Relativeabundancesofdifferentmethanogensonthesurfaceofgranularsludge(47.08±8.26)%..,1.0~2.0mm1.23,,.2.1.2 1.0~2.0mm2.0mm,FISH,,.41.0~2.0mm.,,5.,.1.0~2.0mm2.0mm,1.0~2.0mm2.0mm(48.67±8.87)%、(45.01±8.97)%;,,1.0~2.0mm(44.94±8.78)%、(37.87±6.58)%,1.18,2.0mm.,,1.0~2.0mm(37.87±6.58)%、(35.31±6.24)%.Jupraputtasri[12]FISH,(50.7±6.9)%;Liu[8](Slot-blothybridization)rRNA,rRNA62.4%..,40mg/L[20],..(a);(b)4 1.0~2.0mmFig.4 DistributionofMethanosarcinalesontheinnerfaceandsurfaceofsludgesamplewithdiameterof1.0-2.0mm5 Fig.5 Relativeabundancesofdifferentmethanogensontheinnerfaceofgranularsludge2.2 ,,.4COD,,2.,COD,,1.0~2.0mm,1.562g/(g·d),COD7283:UASB88.64%,2.0mm..,,.,1.0~2.0mm1.231.18,,.,.2 CODTable2 CODremovalratioandmaximumspecificmethanogenicactivityofsludgesampleswithdifferentdiameters/mm0.60.6~1.01.0~2.02.0/g·(g·d)-11.3111.3941.5621.467COD/%76.3181.2188.6485.813 (1)(),.(2)():(25.50±8.63)%~(48.67±8.87)%;1.0~2.0mm;;..(3)1.311~1.562g/(g·d),COD,,.:[1] ChangYJ,NishioN,NagaiS.CharacteristicsofgranularmethanogenicsludgegrownonphenolsyntheticmediumandmethanogenicfermentationofphenolicwastewaterinaUASBReactor[J].JFermentBioeng,1995,79(4):348-353.[2] FukuzakiS,NishioN,NagaiS.Highrateperformanceandcharacterizationofgranularmetha