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Answerstoourmorecommontechnicalenquiries.
AnaerobicSystemsAntibioticSensitivityTestingCultureMedia,SupplementsandRawMaterialsArethereanyguidelinesforpreparationandreconstitutionofculturemedia?
Arethereanyguidelinesforsterilisingculturemedia?
CanIprepareculturemediainamicrowave?
CanIstoreselectivesupplementsafterreconstitution?
HowcanIremeltsolidmedia?
HowlongcanIstoreculturemediaafterpreparation?
HowshouldIdisposeofusedculturemedia?
HowshouldItestthepHofculturemedia?
Myculturemediumdoesnotlookrightwhathasgonewrong?
Myculturemediumisnotperformingcorrectlywhathasgonewrong?
ShouldIboilmediabeforeautoclaving?
Whatarethemaincomponentsofculturemedia?
Whatexactlyisagar?
WhatgradeofwatershouldIusetoprepareculturemedia?
Whatistheshelflifeofdehydratedculturemediaafterthepothasbeenopened?
WhatqualitycontroltestingshouldIcarryoutonculturemedia?
WhichbloodproductshouldIchoosetomakebloodagar?
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TechnicalSupportFAQs
BLOODPRODUCTSHorseblood,defibrinatedSR0050Horseblood,oxalatedSR0049Horseblood,haemolysedSR0048Sheepblood,defibrinatedSR0051HorseserumSR0035Horseandsheepbloodarethemostwidelyusedanimalbloodproductsinculturemedia.Thechoiceofanimalislargelytraditional,withtheUSAandmuchofcontinentalEuropepreferringsheepblood,whilsttheUKandCommonwealthpartnerspreferhorseblood.
Thehaemolyticreactionsofhorseandsheepbloodarenotidenticalandbloodagarmediadesignedforhorsebloodmaynotbesatisfactorywithsheepbloodandviceversa.SeeBloodAgarBase(Sheep)CM0854.
Itisnotpossibletosterilisewholebloodproductsandthereforetheymustbecollectedaseptically.Freshlydrawnbloodfromhealthyanimalscandestroysmallnumbersofbacteriawhichmaytransientlyinfectitduringcollectionbutconsiderableexpertiseandskillarerequiredtomaintainhighquality,sterileblood.Itisdifficulttoobtainsterilebloodfromabattoirsthereforebleedingiscarriedoutinsuitablepremisesfromanimalswhicharespeciallymaintainedonhighproteindiets.
Defibrinationisnowacceptedasthebestmethodofpreventingbloodclotting.Itmustbecarriedoutimmediatelyafterthedrawingthebloodandtheagitationmustbesufficienttodenaturethefibrinogenbutnottocauseruptureoftheerythrocytesandhaemolysis.SomelysisisinevitableinthisprocessbutthereleaseofNADinhorsebloodfromtherupturederythrocytesstimulatesthegrowthofHaemophilusinfluenzae.Thiseffectisnotseenwithsheepblood.
Theuseofpotassiumoxalateasananticoagulantincreasestheyieldofwholebloodbutitremainsasapotentialchelateofdivalentmetals.Thischelateeffect,whichalsoappliestosodiumcitratetreatedblood,canbeseenintheoverallsmallercolonieswhichgrowonanticoagulantcontainingbloodagar.
Toprolongtheshelflifeofbloodproductsintheuser'shands,bloodiscollected,bottledanddispatchedonthesameday.Thephysicalcharacteristicsofappearance,haemoglobincontentandpackedcellvolumearecheckedbeforereleasebutsterilitytestsarecarriedoutbeyondthetimeofdispatch.Shouldsamplesofindividualanimalbloodcollectionsshowinfectionthenallrecipientsofthesuspectbloodarenotifiedbytelephone,faxortelex.
Haemolysedorlakedhorsebloodisusedforspecialpurposesinculturemedia.IthasbeenusedformanyyearsinCorynebacteriumdiphtheriaemedia,wherebettergrowthwasobservedafterlysisofthehorsebloodbythetelluriteinthemedium.
LysedbloodisstimulatoryformanyClostridiaandHaemophilusspeciesbutFildesPepticDigestofBloodSR0046canalsobeusedforthispurpose.
Inantibioticsusceptibilitytesting,lysedhorsebloodisaddedtothemediumtoimprovethereactionswithtrimethoprimandsulphonamides.Mostculturemedia,unlessspeciallyprocessedforsusceptibilitytesting,containamountsofthymidinewhichcanantagonisetheinhibitoryeffectsoftheseantimicrobials.Whenhorsebloodislysedtheerythrocytesreleaseanenzymethymidinephosphorylasewhichconvertsthymidineintothemuchlessantagonisticcompoundthymine.Thisenzymedoesnotexistinsheeperythrocytes.Thelyticagentusedforlysedhorsebloodiswhitesaponinwhichappearsnottoaffectthegrowthofbacteriaatthelevelrequiredforlysis.
SerumforCultureMediaHorseserumSR0035
Totallyasepticprocessingisnotrequiredforbloodserumproduction.Bloodiscollectedincleanvesselsinacleanenvironment.Afterclottinghastakenplacethebloodisstoredat28Ctoallowtheclottoretractandtheserumseparate.Theseparatedserumisthenfiltersterilisedandfilledoutintosterilebottles.Allserashouldbestoredat28Cbutforlongertermstorageitisbestfrozenat20C.
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DipslidesGeneralToxinDetection
Serumcanbecomecloudyonstoragecausedbylipaseenzymesandlipidchanges.Heatingtheserumto56Cfor30minutes(inactivation)willusuallyovercomethisproblem.
SheepErythrocytesforImmunologicalReactionsSheepblood(inAlseversolution)SR0053
Afterwashingthesheeperythrocytes,bycentrifugingwithphysiologicalsalineuntilthesupernatantsalineisfreefromhaemolysis,thewashedcellsmaybeusedforanyimmunological/serologicaltestwheresheepcellantigensarerequirede.g.RoseWaaler,PaulBunnellandvariousvirologicalcomplementfixationtests.
Thesheepbloodshouldbestoredat28Candusedbeforetheexpirydateonthelabel.
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