北京市售酸奶中益生菌的分离鉴定及耐药性检测

55239520109JOURNALOFHYGIENERESEARCHVol.39No.5Sep.20101000-8020201005-0552-03··123100021、6、、、、。6。MRS、36±1℃、31523715。52261238.7%1×106cfu/mlg1961.3%14。。AIsolationandidentificationofprobioticsfromyoghourtinBeijingDONGYinpingLIFengqinCUIShenghuiYUHongxiaInstituteofNutritionandFoodSafetyChineseCenterforDiseaseControlandPreventionBeijing100021ChinaAbstractObjectiveTocarryoutisolationandtheviableplatecountofprobioticsfromyoghourt.Toinvestigatetheantimicrobialsusceptibilityofstrainstopenicillingentamicinampicillinvancomycinclindamycinanderythromycin.MethodsProbioticswererecoveredandcountedfromyoghourtbyplatecountmethod.TherecoveredisolateswerecharacterizedbyGram＇sstainbiochemicaltestantimicrobialsusceptibilitytest.ResultsMRSmediumand36±1℃weretheoptimumconditionofculture.Intotal52probioticisolateswererecoveredfrom31yoghourtproducts1238.7%productsshowedalowerprobioticquantitythan1×106cfu/mlofthenationalstantardand1961.3%productswerehigherthan1×106cfu/ml.Gram＇sstainshowedthat37isolatesweregrampositivebacilliand15isolateswerepositivecoccus.These52isolatesweregroupedintosixspeciesbybiochemicalmethodincludingL.bulgaricusS.thermophilusL.acidophilusL.caseiL.delbrueckiiandL.fermentum.All52isolatesweresusceptibletopenicillinandampicillinoneisolatewasresistanttobothclindamycinanderythromycinfourisolateswereresistanttovancomycin.ConclusionMostofyoghourtproductscontainedsufficientviableprobioticsasrequiredbythenationalstandard.Asmallquantityofisolateswereresistanttosomeantimicrobialsandmulti－drugresistantisolateswerealsofound.Keywordsyoghourtprobioticsiso[e-mail]3。1989FoodandDrugAdministrationFDAAmericanAssociationofFeedControlOfficialsAAFCO41。20021881、211、82136、8。199661。、。。、5.5532。HUMMEL、、TOSI、、、、、、、、、3-5。。、、。11.11.1.1Lactobacilliuscaseisubspcasei1.2435、Lactobacilliusdelbrueckiisubspbulgaricus1.2161、Lactobacilliusplantanum1.2158、Lactobacilliusacidophilus1.2686、Bifidobacteriumadolescentis1.2190。E.faecalisATCC29212、E.coliATCC25922。1.1.2penicillinPEN、gentamicinGEN、ampicillinAMP、vancomycinVAN、clindamycinCLIN、erythromycinERY。1.1.320079～2008331431、、、2、34。1.1.4MRSBectonDickinson、MHBectonDickinson、、BHI。1.1.5NaH2PO4、Na2HPO4、NaCl、、API50CHLBiomerieux。1.1.6phosphatebuffersolutionPBSNaH2PO45.56g、Na2HPO423g、NaCl87.68g1000ml1000mlpH7.4121℃15min1∶10。1.1.7MIR-2624℃～50℃±1℃、MSL-3705、SANYOGENboxJar2.5LREF96127Biomerieux、AVANTI30BECKMAN、phoenixSpeBectonDickinson、METTLER-AE240-、VisionBiomed。1.21.2.1MRS、MH36±1℃48h。MRSMH。MRS、36±1℃。1.2.21.2.2.125mlg225mlPBS1∶101ml9mlPBS1∶1001∶106。1∶104、1∶1051∶1061mlMRSPBS。36±1℃46～48h。20min。1.2.2.2、30～300GB/T4789.35－20036。1.2.3MRS36±1℃16～24hAPI50CHL。1.2.41.2.4.1ClinicalandLaboratoryStandardsInstituteCLSI1。1Table1Theantibioticsusedintheexperimentmgμg/mlpenicillinPEN20088.8%0.25～32gentamicinGEN200638U/mg0.25～128ampicillinAMP20085.7%0.03～32vancomycinVAN1001007U/mg0.125～256clindamycinCLIN10086%0.00755～32erythromycinERY200938U/mg95%0.015～128554391.2.4.2MRS36±1℃16～20h0.5MRS100。96100μlMRS6200μl100μl100μl1196。100μl36±1℃16～20hminimuminhibitoryconcentrationMIC。CLSI。22.12.1.131141×104～1.5×108cfu/mlg1×106cfu/mlg1238.7%6225%450%～100%。1961.3%。。2.1.2523715。2.1.352624、15、7、3、21。1、1、2、2。2.26。114-243-1、18-1、24-1、24-2。27%14/52MIC833-2、13-1、14-2MIC16。26MIC1Table2TheMICof6antibioticsinprobioticsfromyoghourtsMICμg/mlPENGENAMPVANCLINERY3－10.25320.1252560.030.125VAN3－20.51280.510.060.25GEN5－10.25640.12520.1250.125－6－20.25640.0610.060.06－7－10.25640.0610.060.06－9－10.25640.0610.060.03－9－20.25640.0610.060.06－13－10.251280.0610.060.03GEN14－20.251280.06232128GEN-CLIN-ERY150.25640.0620.0150.125－18－10.25640.52560.00750.03VAN190.25640.25220.125－23－10.25640.0610.0150.03－24－10.256412560.030.25VAN24－20.25640.52560.030.25VAN292122/1482－25922/8////－1“－”“/”31/3450%～100%。。523715。31。20min20min。39520109JOURNALOFHYGIENERESEARCHVol.39No.5Sep.201055536±1℃。。、。。。。、1。。。。、、。MRSAβ-7。。3。1.C.200494-97.2.J.2004163262-265.3HUMMELASHERTELCHOLZAPFELWH.AntibioticresistancesofstarterandprobioticstrainsoflacticacidbacteriaJ.ApplEnvironMicrobiol2007733730-739.4.1132J.2006271048-50.5TOSILBERRUTIGDANIELSENMetal.SusceptibilityofStreptococcusthermophilustoantibioticsJ.AntonieVanLeeuwenhoek200792121-28.6.GB/T4789.35—2008S.2008.7.M.2002138-139.2009-10-111000-8020201005-0555-02··11100050R153.2R179A“”1、“”“”2006～2007、1。11.1、2～4、451。1.22397.95±0.612127.84±0.72。“”1。2“”200ml5。20061。1。1.3、、、、、、。