对硝基苯酚冲击对UASB反应器中污泥活性和微生物种群的影响梅琴

UASB＊梅　琴　高国庆　虞方伯　刘连成　沈　标＊＊(　　210095):(UpflowAnaerobicSludgeBlanket,UASB),(p-NP)。PCR-DGGE。,p-NPCOD,;,20mgL40mgLp-NP16d27d。p-NP,,p-NP。p-NPMethanosaetasp.Methanomicrobiasp.。Chloroflexisp.、Bacteroidesp.Anaerovibriosp.;Rheinheimerasp.20mgLp-NP,40mgLp-NP。Flavobacteriasp.p-NP,p-NP。:,,:Q939.9　　:A　　:0253-2654(2007)06-1123-06Effectofp-nitrophenolShockonSludgeActivityandMicrobialPopulationsandinUASBReactor＊MEIQin　GAOGuo-Qing　YUFang-Bo　LIULian-Cheng　SHENBiao＊＊(NanjingAgriculturalUniversity,EnvironmentMicrobiologyKeyandOpenLaboratoryofDepartmentofAgriculture,Nanjing　210095)Abstract:Effectofp-nitrophenolshockonmicrobialpopulationsandsludgeactivityinUASBreactorwasstudiedbyDGGE-PCRof16SrDNAfragmentsanddetectionofCODremovingandbiogasyield.Theresultsshowedthatp-nitrophenolseriouslyinhibitedthesludgeactivity,resultinginthedropofbiogasandCODremovingrate.The40mgLp-nitrophenolhadmoreinhibitionthan20mgLp-nitrophenol.Itwouldtake27and16daysrespectivelyforreactortorecoverafter40mgLand20mgLp-nitrophenolshock.Thediversityofeubacteriaandmethanogenswerealsoeffectedbythep-nitrophenolshock.Thevariationofeubacteriawasmorethanthatofmethanogensafterp-nitrophenolshock.ThedropofbiogaswasmainlyrelatedtothevariationofMethanosaetasp.andMethanomicrobiasp.afterp-nitrophenolshock.AmongtheeubacteriathepopulationofChloroflexisp.、Bacteroidesp.andAnaerovibriosp.decreasedgreatlyafterp-nitrophenolshock.Andmore,theRheinheimeraspdisappearedafter40mgLp-NPtreatment.ButtheFlavobacteriasp.appearedafterp-nitrophenolshock,whichwasprobablyrelatedtothedegradationofp-NP.Keywords:p-nitrophenolshock,Bacteriapopulationsdiversity,Sludgeactivity.　　＊(No.50308011)　＊＊　Tel:[phone],E-mail:[e-mail]:2007-04-04,:2007-06-01　　,,。,,,,,,。,,,,。、,,·1123·　200734(6)DOI:10.13344/j.microbiol.china.2007.06.028、、、、。,,[1,2]。,2-NP、4-NP、2,4-DNP“”,10ngL[3]。,p-NP(p-AP),500,p-APNH4-N,NO3-N,NO2-N。p-NPUASB,p-NPCOD、,。1　1.1　UASB,,20cm、50cm,2L,30℃。1.2　,1d,,8.2gL。1.3　,,COD∶N∶P=200∶5∶1。,Co、B、Cu、Ni、Fe、Zn、Mg、Mn,,pH7.2～8.0。1.4　SSVSS,COD[4],,3%[3]。p-NPp-AP[5,6]。1.5　1.5.1　:,,,50d,COD85%,,,COD1000mgL,(HRT)1d。1.5.2　p-NP:4,2p-NP20mgL,240mgLp-NP,2d。,420mgLp-NP(1),620mgLp-NP(2),8p-NP5mgL。,40mgLp-NP,2830,2,p-NP5mgL。p-NP。1.5.3　DNA:p-NP2d,()2mL,8000rmin5min-70℃。DNA[7]。1.5.4　PCR:PCR16SrDNAV3,333-GC-F(5′-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGG-GGGGTCCAGGCCCTACGGGG-3′)533R(5′-TTACCGCGGCTGCTGGCAC-3′)[8];16SrDNAV3P2(5′-ATTACCGCGGCTG-CTGG-3′)P3(5′-CGCCCGCCGCGCGCGGCGGGC-GGGGCGGGGCACGGGGGGCCTACGGGAGGCAGCAG-3′)[9],PCR[8]。“reconditioningPCR”PCRDNA[9]。1.5.5　[10]:PCR-DGGEBio-radDCodeSystem,8%,40%～60%(100%7molL40%),1×TAE(40mmolLTris,20mmolL,1mmolLNa2EDTA),200V10min,80V,60℃14h～15h。DGGEGelDoc2000(Bio-rad),QuantityOne。1.5.6　PCR:DGGEDNA,ddH2O3,5min.,30μLddH2O,50℃～60℃2h,,8000rmin5min。1μLDNAPCR,P2(GC)、P3,333F(GC)、533R,·1124·200734(6)2.0%,PMD19-T,E.coliDH5α,、。NCBI(http:),,,。2　2.1　p-NPCOD1,4,620mgLp-NP,COD,COD,50%,p-NP,COD,8p-NP5mgL,1,COD50%。,,COD、,24,16。2840mgLp-NP(3),COD448mgL,COD56.2%,,、。、p-NP5mgL,p-NP。304,COD700mgL,COD34%,p-NP10mgLd,,,25d。　　　1　p-NPCOD2.2　p-NP2p-NP。,,。p-NP。2　p-NP2.3　p-NPp-AP3,41,12hp-NP1.92mgL,p-AP16.98mgL,p-NPp-AP84.9%;2,12hp-NP1.34mgL,p-AP17.3mgL,p-NPp-AP86.5%;3,12hp-NP6.43mgL,48h0.37mgL,p-AP32.94mgL,82.35%;4,12hp-NP6.28mgL,48h0.93mgL,p-AP34.07mgL,82.35%。48h,p-AP,,p-NP、p-AP。DeliaTeresaSponza,p-NP,80%～90%p-NPp-AP,UASB[3].·1125·　200734(6)3　p-NPp-NP4　p-NPp-AP2.4　p-NP5,p-NP。DGGE9,A(Methanomicrobiasp.)B(Methanosaetasp.),,p-NP(20mgL),3、42A、B。p-NP(20mgL)。(40mgL),A、B。C(Methanogeniumsp.)F(Methanospirillumsp.),。CF,,,。D、G、HI,E,p-NP。G、H,,p-NP,G、Hp-NP。QuantityOneDGGE,5　16SrDNAV3DGGE:1;2;3～6p-NPNote:1.inoculatedsludge;2.sludgebeforeshock;3～6.sludgeafterfourp-NPshockDicecoefficient(Cs)DGGE,Cs[8]。DGGE1,6。1　PCR-DGGE(Cs)1234561100.0245.2100.0340.789.3100.0446.152.446.3100.0550.833.130.061.6100.0655.935.432.356.485.7100.06　DGGE　　6,6,44%,。20mgLp-NP,,,,23Cs89.3,·1126·200734(6),88%。,481%,p-NP。2316d,,45。40mgLp-NP20mgLp-NP,560.74。2.5　DGGEDNA2Ⅰ、Ⅱ、Ⅲ[11],Ⅱ,93%～100%。5Methanogeniumsp.、Methanospirillumsp.、Methanocorpusculumsp.Methanobacteriumsp.,Methanocorpusculumsp.。,,Methanomicrobiasp.Methanosaetasp.,Methanogeniumsp.Methanospirillumsp.。,Methanosaetasp.,。Methanosaetasp.40%,。p-NP,Methanosaetasp.,,Methanosaetasp.。Methanosaetasp.,。A,BMethanomicrobiasp.,,。p-NP,Methanosaetasp.Methanomicrobiasp.。,70%,,,,。2.6　p-NP7,,。、,342Ⅰ,Ⅱ,Ⅴ,Ⅲp-NP,,p-NP。,43,,p-NP。54,,p-NP。,Ⅳ。2　DGGEDNA%AMethanomicrobiasp.93EF489006BMethanomicrobiasp.93EF489007CMethanogeniumsp.98EF489008DMethanoplanussp.96EF489009EMethanosaetasp.100EF489010FMethanospirillumsp.96EF489011GMethanocorpusculumsp.99EF489012HMethanocorpusculumsp.99EF489013IMethanobacteriumsp.98EF4890147　16SrDNAV3DGGE:1;2;3～6p-NP3,8,23,58%,20mgLp-NP,,,43,。453　PCR-DGGE(Cs)1234561100.0248.3100.0345.557.8100.0447.651.754.7100.0532.057.852.347.5100.0642.531.540.630.524.6100.0·1127·　200734(6),40mgLp-NP,。,40mgLp-NP,6,。8　DGGE2.7　DGGEDNAⅠChloroflexi97%,Chloroflexi,,(FISH),Chloroflexi,[12],。7,p-NP,(Chloroflexi),,。Ⅱ(Anaerovibriosp.)100%,Anaerovibriosp.,,、,。p-NP,,,,,COD,COD38%。Ⅲ(Flavobacteriasp.)