美国FDA细菌学分析手册第八版(BAM)沙门氏菌(XXXX版)

BAM:SalmonellaNOTICE:IfyouarelookingforBAMChapter5:Salmonella(December2007Edition)thatisincorporatedbyreferencein21CFRParts16and118:FederalRegisterFinalRule1(July9,2009,74FR33030):PreventionofSalmonellaEnteritidisinShellEggsDuringProduction,Storage,andTransportation,pleaseusetheseversionsoftheBAMSalmonellaChapter2(PDF,189Kb)andAppendix1:RapidMethodsforDetectingFoodbornePathogens3(PDF,195Kb).ThemostrecentEditionofBAMChapter5:Salmonellaisavailablebelowthisnotice.November2011VersionBacteriologicalAnalyticalManualChapter5SalmonellaAuthors:WallaceH.AndrewsandThomasHammackRevisionHistory:November2011-AdditiontoSectionC:PreparationoffoodsforisolationofSalmonella:Leafygreenvegetablesandherbs.February2011–RemovedlinktoAppendix1:RapidMethodsforDetectingFoodbornePathogens(nowarchived).December2007–Mameypulpmethodadded,andSectionDrevised.June2006–Eggsmethodrevisedforshelleggsandliquidwholeeggs.April2003–Froglegsmethod,Lacticcasein,Rennetcasein,SodiumcaseinateandRabbitcarcassmethodsrevised,topearsandotherdogchewtoysadded.RemovedsectionA.25,Mechanicalshaker.October25,2001–ExtensionoftheapplicabilityoftheorangejuicemethodinsectionC.19toapplejuiceandapplecider.1999-DEC,2000-MAR,and2000-AUGFinalrevisionon2000-NOV-14(seetheIntroductionforasummaryofchanges).Toobtainacopyofapriorversionnotcurrentlyposted,pleasecontactThomasHammackChapterContentsIntroductionEquipmentandMaterialsMediaandReagentsPreparationoffoodsforisolationofSalmonellaIsolationofSalmonellaIdentificationofSalmonellaReferencesIntroductionSeveralchangesarebeingintroducedinthiseditionofBAM(8thEdition).ThefirstchangeinvolvestheexpandeduseofRappaport-Vassiliadis(RV)medium4forfoodswithbothhighandlowlevelsofcompetitivemicroflora.Inthepreviousedition,RVmediumwasrecommendedonlyfortheanalysisofshrimp.BasedonthecompletionofAOACprecollaborative(5,6)andcollaborative(7,8)studies,RVmediumisnowbeingrecommendedfortheanalysisofhighmicrobialandlowmicrobialloadfoods.RVmediumreplacesselenitecystine(SC)brothfortheanalysisofallfoods,exceptguargum.Inaddition,RVmediumreplaceslauryltryptosebrothforusewithdryactiveyeast.Tetrathionate(TT)5brothcontinuestobeusedasthesecondselectiveenrichmentbroth.However,TTbrothistobeincubatedat43°Cfortheanalysisofhighmicrobialloadfoodsandat35°Cfortheanalysisoflowmicrobialloadfoods,includingguargum.Thesecondchangeinvolvestheoptionofrefrigeratingincubatedpreenrichmentsandselectiveenrichmentsoflow-moisturefoodsforupto72h.Withthisoption,sampleanalysescanbeinitiatedaslateasWednesdayorThursdaywithoutweekendworkbeinginvolved.Thethirdchangeinvolvesreducingtheperiodofincubationofthelysineironagar(LIA)6slants.Intheformeredition(BAM-7),triplesugarironagar(TSI)7andLIAslantswereincubatedat35°Cfor24±2hand48±2h,respectively.Unpublisheddatahavedemonstratedthatthe48hreadingofLIAslantsiswithoutdiagnosticvalue.Of193LIAslantsexamined,allgavedefinitiveresultswithin24±2hofincubation.Nosignificantchangesalteredthefinaltestresultwhentheslantswereincubatedanadditional24h.Thus,boththeTSIandLIAslantsarenowincubatedfor24±2h.Thefourthchangeinvolvestheprocedureforsurfacedisinfectionofshelleggs.Inthepreviousedition(BAM-7),eggshellsweresurface-disinfectedbysoakingin0.1%mercuricchloridesolutionfor1hfollowedbysoakingin70%ethanolfor30min.Mercuricchlorideisclassifiedasahazardouswaste,andisexpensivetodisposeofaccordingtoEnvironmentalProtectionAgencyguidelines.Inthisedition(BAM-8)eggshellsarenowsurface-disinfectedbysoakingforatleast10secina3:1solutionconsistingof3partsof70%alcohol(ethylorisopropyl)to1partofiodine/potassiumiodidesolution.Thefifthchangeinvolvesthesamplepreparationofeggs.Eggcontents(yolkandalbumen)arethoroughlymixedbeforeanalysis.Aftermixingtheeggcontents,25g(ml)areaddedto225mltrypticase(tryptic)soybrothsupplementedwithferroussulfate.Amethodfortheanalysisofguargumhasbeenincluded.Whenguargumispreenrichedata1:9sample/brothratio,ahighlyviscous,nonpipettablemixtureresults.Additionoftheenzymecellulasetothepreenrichmentmedium,however,resultsinareadilypipettablemixture.Amethodfororangejuice(pasteurizedandunpasteurized)hasbeenincludedduetorecentorangejuice-relatedoutbreaks.Thedirectionsforpickingcoloniesfromtheselectiveplatingagarshavebeenmademoreexplicittoreflecttheintentofthemethod.Intheabsenceoftypicalorsuspectcoloniesontheselectiveplatingagars,itisrecommendedthatatypicalcoloniesbepickedtoTSIandLIAslants.Thisrecommendationisbasedonthefactthatupto4%ofallSalmonellaculturesisolatedbyFDAanalystsfromcertainfoods,especiallyseafoods,duringthepastseveralyearshavebeenatypical.Finally,sincethepublicationofBAM-7,a6-waycomparisonwasconductedoftherelativeeffectivenessofthethreeselectiveplatingagarsrecommendedintheBAM(bismuthsulfite8,Hektoenenteric9,andxyloselysinedesoxycholateagars10)andthreerelativelynewagars(EF-18,xyloselysineTergitol4,andRambachagars).Ourresults(9)indicatednoadvantageinreplacinganyoftheBAM-recommendedagarswithoneormoreoftheneweragars.Thus,thecombinationofselectiveplatingagarsrecommendedinBAM-7remainsunchanged.A.EquipmentandMaterials1.Blenderandsterileblenderjars(s