低温条件下城市污水厂污泥膨胀的生物学成因陆鑫

107Vol．10No．720167ChineseJournalofEnvironmentalEngineeringJul．201611*22111．2100232．450006。16SrＲNA。414HaliscomenobacterTrichococcus。BLAST3ＲichococcusFlocculiformis、CandidatusHaliscomenobactercalcifugiens、NostocoidalimicolaⅠ。X172A1673-9108201607-3925-06DOI10．12030/j．cjee．201502026Biologicalcausesofsludgebulkinginmunicipalwastewatertreatmentplantunderlow-temperatureconditionLUXin1LIUBo1*TANYunfei2WANGXiaoling2WANGDepeng1DINGXinchun11．SchooloftheEnvironmentNanjingUniversityNanjing210023China2．ZhengzhouSewagePurificationCompanyZhengzhou450006ChinaAbstractThisstudyusedopticalmicroscopyandscanningelectronmicroscopySEMtoobservemicrobi-almorphologyinbulkingsludge．Largeamountsoffilamentsbetweensludgeflocswereassociatedwithsludgebul-king．Toclassifythemicrobialcausesofsludgebulkinga16SrDNAgenelibrarywasconstructedandatotalof4phylaand14generaofbacteriawereidentified．AtthegeneralevelHaliscomenobacterandTrichococcuswerethemainbulkingbacteriadetected．BLASTresultsidentifiedthreebacterialspecieswithhighsimilaritytoTrichococ-cusflocculiformisCandidatusHaliscomenobactercalcifugiensandNostocoidalimicolaIasrelevantforsludgebul-kinginwastewatertreatmentplants．Keywordsactivatedsludgefilamentousbulkingclonelibrarycommunitystructure2012ZX07204-001-0012015－02－032015－04－151990—。[e-mail]*[e-mail]。1。22。90%。3。GUO414NostocoidalimicolaIII、Mycobacteriumfortuitum、Type1863Microthrixparvicella。STOＲM5、DO、、pH。。10。。16SrＲNA。11．1100mL20131114．3℃。1∶1－20℃GB18918-2002ASV3073%SVI242mL·g－1。1Fig．1Annualchangeofwatertemperatureinsewagetreatmentsystem1．2SEMNikon-EclipseTi-US-3400N。1．3DNA16SrＲNAPCＲ4000r·min－110min200mgDNA。FastDNADNA。DNANanoDropND-20001%DNA。16SrＲNA27f5'-AGAGTTT-GATCMTGGCTCAG-3'1492r5'-CGGYTACCTT-GTTACGACTT-3'DNAPCＲ。PCＲ50μL10×PCＲbufferMg2+free2．5μLMgCl225mmol·L－11．5μLdNTP5．0mmol·L－12．0μLTaqDNA5U·μL－10．2μL50μmol·L－11．0μL50μmol·L－11．0μLcDNA2．0μL50μL。PCＲ95℃5min95℃1min55℃1min72℃1．5min3572℃10min。1．416SrＲNAPCＲTakarapGM-TAmpX-GalIPTG。M13。1．5DNA16SrＲNA50。http//．ncbi．nlm．nih．gov/tools/vecscreen/。DNAman97%OTUoperationaltaxonomicunitOTUＲDP＞80%。OTUBLASTNCBIMEGA6neighbor-joininganalysis。22．14002。。SEM10003。、、。629372400Fig．2Microbialmorphologyofactivatedsludgeunderopticalmicroscope×40031000Fig．3MicrobialmorphologyofactivatedsludgeunderSEM×1000。。SEZGIN6。7。23。。2．216SrＲNA50DNAmanOTU97%1OTU40OTU。ＲDPphylum4a4Proteobacteria、Bacteroidetes、FirmicutesVer-rucomicrobia56%、22%、6%6%。10%4Fig．4Microbialcommunitystructureofactivatedsludge。Pro-teobacteria4classα-Proteobacteria4%、β-Proteobacteria38%、γ-Proteobacteria10%Δ-Proteobacteria4%。Pro-teobacteriaβ-Proteobacteria8。Proteobacteria、9。。38。MicrothrixparvicellaActinobacteriaProteobacteria10。Actinobacteria。genus。4b14Haliscomenobacter16%、Azospira10%、Dok-donella6%、Trichococcus4%、Aquabacterium2%、Desulfomicrobium2%、Dongia2%、Georgfuchsia2%、Inhella2%、Lactococcus2%、729310Luteolibacter2%、Ｒamlibacter2%、Terrimonas2%Tolumonas2%。44%10%。11。。。HaliscomenobacterTrichococcus122%。Haliscom-enobacter。13Haliscomenobacter≤10℃。HaliscomenobacterZoogloeaHaliscomenobacter。。2．3BLAST7NCBI88%～99%90%36%97%。。Nitrosomonassp．2OTU2AOB14。UnculturedDenitratisomasp．2OTU2、Dechloromonassp．1OTU1、Ｒhodanobactersp．1OTU1Pseudo-monassp．1OTU112。Ｒhodocy-clussp．2OTU615。。1OTU-23、24、28Tri-chococcusflocculiformis、CandidatusHaliscomenobactercalcifugiens、NostocoidalimicolaⅠTrichoccusHaliscomenobacterＲDP。Trichococcusflocculiformis4℃16。。CandidatusHaliscomenobactercalcifugiens16SrＲNAHaliscomenobact-erhydrossis17。。NostocoidalimicolaⅠMicrothrixparvicella7。3。。。OTU5OTU7BLASTOTU11Proteobacteria。。3。HaliscomenobacterTrichococcusHaliscomenobacter。。3Trichococcusflocculiformis、CandidatusHaliscomenobactercalcifugiens、NostocoidalimicolaⅠ。。82937116SrＲNATable116SrＲNAgeneclonelibraryofmicroorganismfrombulkingsludgeOTUGenebank/%NCBI211AlphaproteobacteriumP-2096AM411930α-Proteobacteria11UnculturedDenitratisomasp．96KF956508β-Proteobacteria51GeorgfuchsiatoluolicastrainG5G694NＲ_115995β-Proteobacteria91Nitrosomonassp．Nm8696AY123798β-Proteobacteria101UnculturedAzospirasp．91FJ393129β-Proteobacteria111Dechloromonassp．JJ95AY032611β-Proteobacteria121Ｒamlibactersp．P-896AM411936β-Proteobacteria131UnculturedAquabacteriumsp．98KM263172β-Proteobacteria151Nitrosomonassp．Is79A390AJ621026β-Proteobacteria161UnculturedＲhodocyclussp．91JF808914β-Proteobacteria201UnculturedDenitratisomasp．93JF808868β-Proteobacteria221Denitratisomaoestradiolicum98KF810120β-Proteobacteria251Inhellainkyongensis96NＲ_114244β-Proteobacteria385UnculturedＲhodocyclussp．97JF808916β-Proteobacteria141Ｒhodanobactersp．7B-39391KF441674γ-Proteobacteria181UnculturedgammaproteobacteriumcloneAS2192EU283356γ-Proteobacteria191Tolumonasauensis95NＲ_074805γ-Proteobacteria331Pseudomonassp．90HQ860327γ-Proteobacteria373UnculturedDokdonellasp．95JN679143γ-Proteobacteria341Desulfomicrobiumhypogeium96NＲ_114508Δ-Proteobacteria41UnculturedSaprospiraceaebacterium97EU177763Bacteroidetes241CandidatusHaliscomenobactercalcifugiens96AJ786328Bacteroidetes311UnculturedSphingobacterialesbacterium90JX114399Bacteroidetes393UnculturedSaprospirace