城市污水处理厂各工艺阶段内分泌干扰物活性变化规律研究马军

29120091　　　　　　ActaScientiaeCircumstantiaeVol.29,No.1Jan.,2009:(No.705013)SupportedbytheFoundationfortheMajorPrograminInnovationEngineeringfromtheMinistryofEducationofChina(No.705013):(1962—),,(),E-mail:[e-mail];＊()Biography:MAJun(1962—),male,professor(Ph.D.),E-mail:[e-mail];＊Correspondingauthor,,.2009.[J].,29(1):63-67MaJ,WenG,ShaoXL.2009.Investigationontheestrogenicityofdifferentprocessesinasewagetreatmentplant[J].ActaScientiaeCircumstantiae,29(1):63-67马军＊,文刚,邵晓玲哈尔滨工业大学,城市水资源开发利用(北方)国家工程研究中心,城市水资源与水环境国家重点实验室,哈尔滨150090:2008-04-07　　　:2008-12-02:-.,EC505.95×10-11mol·L-1,.,16.13～21.6517.10～22.85ng·L-1,、;A/O,3.37～6.76ng·L-1,、;58.1%～84.2%,75.2%,、.,,,.:;;;:0253-2468(2009)01-63-05　　　:X703　　　:AInvestigationontheestrogenicityofdifferentprocessesinasewagetreatmentplant　MAJun＊,WENGang,SHAOXiaolingNationalEngineeringResearchCenterofUrbanWaterResources,StateKeyLaboratoryofUrbanWaterResourcesandEnvironment,HarbinInstituteofTechnology,Harbin150090Received7April2008;　　　accepted2December2008Abstract:Asolidphaseextraction/yeasttwo-hybridsystemwasusedtoinvestigatetheestrogenicityofwastewaterindifferentseasonsfromeachtreatmentunitofasewagetreatmentplantlocatedinthenortheastpartofChina.Resultsshowedthatyeasttwo-hybridsystemhadarelativelyhighsensitivitywithEC50valueof5.95×10-11mol·L-1.Theestrogenicactivitiesinrawsewageandtheeffluentofprimarysedimentationwere16.13～21.65,17.10～22.85ng·L-1EstradiolEquivalents(EEQs),respectively,withthehighestEEQsinsummerandthelowestinwinter.Theestrogenicactivityintheeffluentofsecondarysedimentationwasreducedto3.37～6.76ng·L-1EEQsaftertheA/Oprocess,withthehigherlevelinspringandwinter,thelowerinsummerandautumn.Thetotalandaverageremovalratesofthewholetreatmentprocesswere58.1%～84.3%and75.2%,respectively,withthehighestremovalrateinsummer,thelowestinwinter.Additionally,itwasshownthatthesecondarybiotreatmentprocessplayedadominantroleintheremovalofestrogenicactivity,whereastheprimarytreatmentprocesshadverylimitedremovalefficiencysuchthattheestrogeniclevelevenincreasedcomparedwiththatinrawsewage.Keywords:solidphaseextraction;yeasttwo-hybridsystem;endocrinedisruptingchemicals;sewagetreatmentplant1　(Introduction)(EndocrineDisruptingChemicals,EDCs)、,、、,(Colbornetal.,1993),、、、.,ng·L-1EDCs,DOI:10.13671/j.hjkxxb.2009.01.029　　　　　　　　　　29、,,、、(Samiretal.,2006).,.,.EDCs,、EDCs.EDCs,EDCs,EDCs,,;,.,EDCs.()、,,1.-,、EDCs.1　Fig.1　FlowchartofasewagetreatmentplantlocatedinnortheastpartofChina2　(Materialsandmethods)2.1　主要试剂(17β-Estradiol,E2)、(DimethylSulphoxide,DMSO)β-D-(o-Nitrophenylβ-D-galactopyranoside,ONPG)Sigma,HPLCDikma,,(yeastnitrogenbasewithoutaminoacids)Difco,DO/-Leu/-HisClontech,,(18.2MΨ·cm)Milli-Q.2.2　样品采集与预处理2006820074、、、,4L、,20L.1μm(Whatman),pH2,.500℃2h.WatersC18,5mL5mLMilli-Q,(SupelcoVisiprepTMDL)10mL·min-1.10mL,/(80/20,)10mL2.2,,DMSO.125～2000.,/,,DMSO.,31000mL,.2.2.3　双杂交酵母测试方法.GRIP1,,,、,(Raoetal.,2004).(Maetal.,2007).,SD/-Trp/-Leu,30℃、130r·min-1,600nm(OD600)2～6,641:OD6000.75,0.995mL1.5mL,5μL,.200μL96,4,30℃、800r·min-14h.,OD600,150μL,120μL(21.51gNa2HPO4·12H2O、6.22gNaH2PO4·2H2O、0.75gKCl0.25gMgSO4·7H2O1L().3.33mL(0.1%)270μL(β-)20μL,30℃、1050r·min-110min,40μLONPG(13.3mmol·L-1,),30℃、800r·min-1,100μLNa2CO3(1mol·L-1),30℃、600r·min-110min,.200μL96,OD420.,5μLDMSO(),5μLE2DMSO,.(1)β-U:U=OD420-OD′420t×V×OD600(1):tONPG(min);V(mL);OD600、OD420600、420nm;OD′420420nm.2　17β--Fig.2　Dose-effectcurvefor17β-estradiolactivity3　(Results)3.1　17β-雌二醇剂量-活性效应关系DMSO,E2,β-,E2-,2,EC505.95×10-11mol·L-1,16.18ng·L-1.(Routledgeetal.,1996;Wuetal.,2004;Heetal.,2002)E2EC5010-10～3×10-9mol·L-1,EC50,.3　Fig.3　Profileofestrogenicactivityalongthewastewatertreatmentprocess3.2　污水处理厂各阶段内分泌干扰活性测定结果3,16.13～21.65ng·L-1(EEQs),19.14ng·L-1EEQs.Anders(2003)1.6～30ng·L-1EEQs,;Hashimoto(2007)83ng·L-1EEQs,.17.10～22.85ng·L-1EEQs,19.34ng·L-1EEQs.A/O,3.37～6.76ng·L-1EEQs,4.82ng·L-1EEQs,Anders(2003)(0.1～15ng·L-1),Hashimoto(2007)(23ng·L-1).58.1%～84.2%,75.2%.,,,,(7.7%±5.5%),Anders(2003)0.1ng·L-1EEQs.　3,.,3,.65　　　　　　　　　　29、5.54%、0.92%6.01%.,A/O,、、A/O69.2%、85.1%、82.0%60.5%,.3,,;,,.(84.2%),(58.1%).4　(Discussion)、3,.,,,;,(Hashimotoetal.,2007),,,(Desbrowetal,1998;Barontietal.,2000),,.A/O,(Johnsonetal.,2001).Ren(2007)(estrone,E1)、E2、17α-(ethinylestradiol,EE2),,.,,,.,,,A/O,,,,.Ren(2007)(4℃),EDCs,EDCs,,EDCs,.,EDCs,EDCs,.EDCs,EDCs、、、,(Byrns,2001).,(SRT).(4℃),EDCs,(45℃)EDCs(Renetal.,2007);SRT,EDCs(Hashimotoetal.,2007).,、EDCs,,EDCs;,,、、EDCs.5　(Conclusions)1),.2)16.13～21.65ng·L-1EEQs,19.14ng·L-1EEQs,,.3.37～6.76ng·L-1EEQs,4.82ng·L-1EEQs,,.3)58.1%～84.2%,,.4),;.:感谢中国科学院生态环境研究中心王子健老师和马梅老师提供菌种以及学习机会.:马军(1962—),男,博士生导师,长江学者特聘教授,第六届“中国青年科学家奖”获得者,主要研究方向为水的深度处理技术.E-mail:[e-mail].(References):AndersS,AllardSS,MatsE.2003.Removalofestrogenicityin661:Swedishmunicipalsewagetreatmentplants[J].WaterResearch,37(18):4433—4443BarontiC,CuriniR,D′AscenzoG,etal.2000.Monitoringnaturalandsyntheticestrogensatactivatedsludgesewagetreatmentplantsandinreceivingriverwater[J].EnvironmentalScienceandTechnology,34(24):5059—5066ByrnsG.2001.Thefateofxenobioticorganiccompoundsinwastewatertreatmentpla