高藻水中两虫检测的预处理方法研究魏魏于建伟张冬青李红岩安伟杨敏

“”魏魏，于建伟，张冬青，李红岩，安伟，杨敏(中国科学院生态环境研究中心环境水质学国家重点实验室，北京100085)针对密度梯度分离纯化法用于高藻水中贾第鞭毛虫和隐孢子虫(“两虫”)检测存在的问题，以铜绿微囊藻配制的模拟水源水为对象，探讨了氯氧化和加压两种预处理方法对于改善“两虫”回收率的效果。研究发现，预氯化处理虽然能有效破坏藻细胞，仍会对后续的分离纯化及镜检过程带来较大的干扰，而且藻细胞在沉淀过程中会裹带“两虫”，导致回收率下降;而通过加压处理破坏藻细胞气囊，可促进藻细胞的分离，使“两虫”回收率达到与免疫磁分离方法相当的效果，且操作简便，处理成本低，具有良好的应用前景。氯氧化;加压处理;高藻水;贾第鞭毛虫;隐孢子虫TU991A1000－4602201403－0039－05PretreatmentforDetectionofCryptosporidiumandGiardiainHighAlgae-ladenSourceWaterWEIWeiYUJian-weiZHANGDong-qingLIHong-yanANWeiYANGMinStateKeyLaboratoryofEnviromentalAquaticChemistryＲesearchCenterforEco-EnvironmentalSciencesChineseAcademyofSciencesBeijing100085ChinaAbstractAddressingtheproblemofdetectingCryptosporidiumandGiardiainhighalgae-ladensourcewaterusingthedensitygradientseparationmethodtheperformanceoftwopretreatmentmethodschlorinationandpressurizationinimprovingtherecoveryofparasitesinMicrocystisaeruginosa-containingwatersampleswascompared．Althoughpre-chlorinationcoulddestroythealgalcellswitharelativelylowdoseadverseeffectswereobservedinthesubsequentpurificationandmicroscopyprocess．InadditionCryptosporidiumandGiardiawerecarriedwithalgalcellsinthesedimentationprocesswhichdecreasedtherecoveryrate．Ontheotherhandpressurizationcoulddestroythealgalcellvesiclesandpromotesep-arationofthealgalcellsachievingarecoveryequivalenttothoseobtainedbyusingimmunomagneticsep-arationIMS．Duetoitseasyoperationandlowtreatmentcostpressurizationhasagoodapplicationprospect．Keywordschlorinationpressurizationhighalgae-ladensourcewaterCryptosporidiumGiardia2013yyxm0121277169“”《》GB5749—2006。GB/T5750．12—2006USEPA·93·第30卷第3期2014年2月中国给水排水CHINAWATEＲ＆WASTEWATEＲVol．30No．3Feb．2014815－Ｒ－05－002/1。—/—23。“”45。“”。、6。。0．5MPa94%7。“”。“”、—“”。1试验材料1.1、。DynalL10DynalMPC－1DynalMPC－S5L0～1MPa。1.270%～100%FDAmg/mLBG－11PBSpH7．2～7．4PBS+0．1%Tween80Percoll－1．10～1．15g/cm310×SLTMBufferABG/C0．45μm－30%、70%90%。1550mL。“”1×106。BG－1114h∶10h2926℃。400～650lx。2试验方法2.12.1.11L2L23mg/L1、2、5、1530min。10mL0．45μm0．01mol/L5mL。0、412h。2.1.21L0．50．8MPa10min10min5mL。2.2FDA530nm。500μL2μLFDA、7min8。683．5nmFL1515～545nmFL3＞650nmFL1FDAFL39。FL1FL310000Flowjo。2.3“”—2。40mL15mL4mL2mLPercoll－1050g、20℃10min“”CA·04·第30卷第3期中国给水排水．watergasheat．com。。10mLpH7．00±0．0510×SLTMBufferAB1mLG/C100μL18r/min、1h。DynalL10DynalMPC－11mL1×SL－BufferA1．5mLDynalMPC－S50μL、0．1mol/LHCl2。“”FITC、DAPIDIC。3结果与讨论3.1“”“”24200μL4800μLPBS“”10/μL。50μL51。56．5%2．1%。649541。1“”Tab．1ConfirmationofnumbersofCryptosporidiumandGiardia123456456326646636435175085985475433.23.2.1①1×108cells/L23mg/L、。1。15min99%30min。5mg/L30min。4h25%12h30%。1Fig．1Changeofresidualchlorineandalgalcellsurvival②“”35mg/L30min0、412hPercoll－、2。·14·．watergasheat．com魏魏，等:高藻水中“两虫”检测的预处理方法研究第30卷第3期2“”Fig．2EffectofchlorinationondetectionofCryptosporidiumandGiardiainalgae-containingwater21×107cells/L—“”65%≥5×107cells/L35%。“”。“”。1×108cells/L4h、66、18212h103、210。“”—。3.2.2①1×108cells/L0．8MPa10min10min5mL3。3a0．8MPaFSCForwardScatter、SSCSideScatter。10。、。0．8MPa10min、10min40%。3Fig．3Effectofpressurizationonalgalcells②“”5×107cells/L1×108cells/L10min、10min“”1×108cells/L4。4—“”Fig．4Effectofpressurization－sedimentationtreatmentondetectionofCryptosporidiumandGiardiainalgae-containingwater0．8MPa“”0．5MPa。4结论“”·24·第30卷第3期中国给水排水．watergasheat．com。0．8MPa、—“”。1HsuBMHuangCHsuYFetal．EvaluationoftwoconcentrationmethodsfordetectingGiardiaandCrypto-sporidiuminwaterJ．WaterＲes2001352419－424．2．/“”J．200925278－80．3．“”J．201329737－40．4ＲodgersMＲFlaniganDJJakubowskiW．Identifica-tionofalgaewhichinterferewiththedetectionofGiardiacystsandCryptosporidiumoocystsandamethodforalle-viatingthisinterferenceJ．ApplEnvironMicrobiol199561103759－3763．5．J．200319699－101．6．J．200521111323－1325．7BrookesJDGanfGGBurchMD．SeparationofformsofMicrocystisfromAnabaenainmixedpopulationsbytheapplicationofpressureJ．AustralianJournalofMarineandFreshwaterＲesearch1994455863－868．8LinTFChangDWLienSKetal．Effectofchlorina-tiononthecellintegrityoftwonoxiouscyanobacteriaandtheirreleasesofodorantsJ．JournalofWaterSupplyＲesearchandTechnology—Aqua2009588539－551．9FranklinNMStauberJLLimＲP．Developmentofflowcytometry-basedalgalbioassaysforassessingtoxici-tyofcopperinnaturalwatersJ．EnvironToxicolChem2001201160－170．10XiaoXHanZYChenYXetal．OptimizationofFDA－PImethodusingflowcytometrytomeasuremetabolicactivityofthecyanobacteriaMicrocystisaeruginosaJ．PhysicsandChemistryoftheEarth2011369/11424－429．1987－。[e-mail]2013－09－26·34·．watergasheat．com魏魏，等:高藻水中“两虫”检测的预处理方法研究第30卷第3期