化学强化除磷污水处理厂聚磷菌的分离筛选

IsolationandidentificationofphosphorusaccumulatingbacteriafromwastewatertreatmentplantbychemicalphosphorusremovalZHANGDan-dan1SUNYong-li1,2LIPeng-feng2LIJia-ju1QINGSong-yan11.SchoolofEnvironmentalScienceandSafetyEngineering,TianjinUniversityofTechnology,Tianjin300384,China;2.NorthChinaMunicipalEngineeringDesignandResearchInstituteCo.Ltd.,Tianjin300074,ChinaAbstractInrecentyears,itwasobservedthattheabilityofreleasingandaccumulatingphosphorusofsludgehasbeendeclinedorevendisappearedduringphosphorusremovedbychemicalprecipitationinwastewatertreatmentplant.Tosolvethisproblem,PAOswasseparatedfromlongrunningperiodsewagetreatmentwithinvertedA/A/Oprocessbyphosphorusremovalthroughchemicalprecipitation.PAOsdominantspeciesisolatedfromlong-termferrousexposurewatercanprovidepotentialstrainsourceforEBPRinwastewatertreatmentplantthroughchemicalphosphorusremoval.Inthispaper,basedondilution-platemethod,Blue-andwhite-coloredscreening,AlbertandPHBstainingmethod,3strainsPAOswereisolated.BetterabilityofabsorptionofphosphorusandphosphorusreleaseofP2andP19werefoundaccordingtoexperiments.Compared16SRNAoftwostrainswithGenbank,P2wasRaoultellaplanticolaandP19wasKlebsiellapneumoniae.Keywordsphosphorusaccumulatingbacteria;isolationandidentification;chemicalphosphorusremoval;Raoultellaplanti－cola;Klebsiellapneumonia1122111.3003842.300074A/A/O.poly-PPHB.P2P19.16SRNAGenbankP2Raoultellaplanticola98%P19Klebsiellapneumoniae98%.；；；；X-173Adoi10.3969/j.issn.1673-095X.2016.005.0132015-11-27.2014ZX07305001-003.1988—.1978—[e-mail].A[1].BOD[2]...[3]Fe2+FeSO4·7H2O15mg·L-1.[4]Fe3+Fe3+Fe3+1673-095X201605-0060-05JOURNALOFTIANJINUNIVERSITYOFTECHNOLOGY325201610Vol.32No.5Oct.2016201610Fe3+.nFeIII∶nP＞1∶1FeIIIpH5.50[5]...[6]SBR、1-β-PHB.[7]1.[8]、.[9]70.81%.[10]2/28℃80%...A/A/O..11.1A/A/O.65mg·L-1.1.2YG[11]MOPS[12][13][14].YG12h.1.3130℃12h.10-2、10-3、10-40.1mLYG30℃2～3d.YG.2.MOPS30℃2～3d..MOPSX-Pi5-bromo-4-chloro-3-indolyl-phosphate.3.PHB[15-17]poly-PAlbert[15-17].1.430℃12h4000r·min-15min=[ATP—BTP/ATP]×100%[18]1ATPBTP.1.5[19]752JSM-7500F2h30℃600nmOD600..1.6TaqDNAPolymeraseDNADNA16SrDNABSF85′-AGAGTTGATCCTGGCTCAG-3′BSR15415′-AAGGAGGTGATCCAGCCGCA-3′PCR16SrDNA.Genbank16SrDNABLASTMEGA5.0Neighbor-joining.61··325b1PHBapoly-PbFig.1ResultsofPHBstainingandpoly-Pstaining22.1YG1.1.[3-520].2.2YG.MOPS30℃3d.38P1P2…P38.PHB[21].、.2.33812hPHB3812hAlbert.138PHBAlbertpoly-PP2、P10、P19.PHB.P2、P10、P1912h15.61%、12.01%、18.02%.ATP.P10.P2、P10、P195mg·L-110mg·L-1.2.290%20mg·L-150%..20mg·L-1P1941.28%P1036.17%.P19.P10P2P19.P2P19.P2P192.2h.[22]..2~6h6h.21YGTab.1CountresultsofYGculturemediumCFU/mL10-210-310-4342±5077±1018±53.4×105376±5163±62±23.6×105a2Tab.2PhosphorusremovalefficiencyofPAOsunderdifferentconcentration/mg·L-1P2/%P10/%P19/%598.9790.3593.142036.1725.4441.2862··2016103P2aP19bFig.3PhylogenetictreeofP2andP190.01RaoultellaomithinolyticaATCC31898EnterobacteriaceaebacteriumLC007898GammaProteobacterriumAY982873KlebsiellaornithinolyticaY19662P2RaoultellaplanticolaKC456530KlebsiellaplanticolaY17663AlcaligenesfaecalisKF475853AlcaligenesfaecalisKF712271a100487159AlcaligenesfaecalisKF475853AlcaligenesfaecalisKF712271P19KlebsiellatrevisaniistrainATCC33558TKlebsiellapneumoniaeKM096433EnterobacteriaceaebacteriumLC007898RaoultellaornithinolyticastrainATCC31898RaoultellaplanticolastrainATCC33531b100668380670.02.NissimSwissa[27]..31.2poly-PPHBP2P1916SRNAGenbankRaoultellaplanticolaKlebsiellapneumoniae.3.P2P19P1920mg·L-141.28%.［1］，，，.A［J］.，2009（9）：.P2P19DNAPCR.Blast16SrDNAGenBbank16SrDNAP2Raoultellaplanticola98%P19Klebsiellapneumoniae98%.GenBankP2P1997%16rDNAMEGA5.0Neighbor-joining3Maxi－mumCompositeLikelihood.[23-24].[25-26]2Fig.2GrowthcurveoftwoPAOsstrains9.69.49.29.08.88.68.48.28.07.87.6logCFU/mLP2P1918161412108642020/h63··32532-36.［2］，，，.［J］.，2009，19：114-116.［3］，，，.BAF［J］.，2015（10）：4236-4243.［4］，，，.Fe3+［J］.，2015（1）：125-133.［5］，，.［J］.：，2015（2）：82-88.［6］，，，.1-β-［J］.：，2014（2）：202-206.［7］，，，.Alcaligenessp.ED-12［J］.，2014（3）：678-687.［8］，，，.［J］.，2014（12）：149-151.［9］，，，.、［J］.，2015（8）：986-991.［10］，，，.［J］.，2015（3）：1027-1032.［11］.［D］.：，2011.［12］.，ppk［D］.：，2005.［13］.、［D］.：，2009.［14］.PAOs［D］.：，2011.［15］，，，.GM1［J］.，2005，42（4）：635-641.［16］.［D］.：，2013.［17］.［D］.：，2010.［18］.PAOX［D］.：，2008.［19］.［D］.：，2007.［20］.［D］.：，2012.［21］MorohoshiT，YamashitaT，KatoJ，etal.Amethodforsc-reeningpolyphosphate-accumulatingmutantswhichremovephosphateefficientlyfromsyntheticwastewater［J］.JournalofBioscienceandBioengineering，2003，95（6）：637-640.［22］.、［D］.：，2009.［23］，，，.［J］.：，2014（4）：257-260.［24］.［D］.：，2013.［25］.［D］.：，2011.［26］，，，.N-4：，CN102391961A［P］.2012-03-28.［27］SwissaN，NitzanY，LangzamY，etal.Atrazinebiodegrada-tionbyamonocultureofRaoultellaplanticolaisolatedfromaherbicideswastewatertreatmentfacility［J］.InternationalBiodeteriorationandBiodegradation，2014，92：6-11.64··