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Answerstoourmore
commontechnicalenquiries.

AnaerobicSystems
AntibioticSensitivity
Testing
CultureMedia,
SupplementsandRaw
Materials
Arethereanyguidelinesfor
preparationandreconstitution
ofculturemedia?
Arethereanyguidelinesfor
sterilisingculturemedia?
CanIprepareculturemediain
amicrowave?
CanIstoreselective
supplementsafter
reconstitution?
HowcanIremeltsolidmedia?
HowlongcanIstoreculture
mediaafterpreparation?
HowshouldIdisposeofused
culturemedia?
HowshouldItestthepHof
culturemedia?
Myculturemediumdoesnot
lookrightwhathasgone
wrong?
Myculturemediumisnot
performingcorrectlywhathas
gonewrong?
ShouldIboilmediabefore
autoclaving?
Whatarethemain
componentsofculturemedia?
Whatexactlyisagar?
WhatgradeofwatershouldI
usetoprepareculturemedia?
Whatistheshelflifeof
dehydratedculturemediaafter
thepothasbeenopened?
Whatqualitycontroltesting
shouldIcarryoutonculture
media?
WhichbloodproductshouldI
choosetomakebloodagar?

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TechnicalSupportFAQs
BLOODPRODUCTS
Horseblood,defibrinatedSR0050
Horseblood,oxalatedSR0049
Horseblood,haemolysedSR0048
Sheepblood,defibrinatedSR0051
HorseserumSR0035
Horseandsheepbloodarethemostwidelyusedanimalbloodproductsinculturemedia.Thechoiceof
animalislargelytraditional,withtheUSAandmuchofcontinentalEuropepreferringsheepblood,whilst
theUKandCommonwealthpartnerspreferhorseblood.
Thehaemolyticreactionsofhorseandsheepbloodarenotidenticalandbloodagarmediadesignedfor
horsebloodmaynotbesatisfactorywithsheepbloodandviceversa.SeeBloodAgarBase(Sheep)
CM0854.
Itisnotpossibletosterilisewholebloodproductsandthereforetheymustbecollectedaseptically.
Freshlydrawnbloodfromhealthyanimalscandestroysmallnumbersofbacteriawhichmaytransiently
infectitduringcollectionbutconsiderableexpertiseandskillarerequiredtomaintainhighquality,sterile
blood.Itisdifficulttoobtainsterilebloodfromabattoirsthereforebleedingiscarriedoutinsuitable
premisesfromanimalswhicharespeciallymaintainedonhighproteindiets.
Defibrinationisnowacceptedasthebestmethodofpreventingbloodclotting.Itmustbecarriedout
immediatelyafterthedrawingthebloodandtheagitationmustbesufficienttodenaturethefibrinogen
butnottocauseruptureoftheerythrocytesandhaemolysis.Somelysisisinevitableinthisprocessbut
thereleaseofNADinhorsebloodfromtherupturederythrocytesstimulatesthegrowthofHaemophilus
influenzae.Thiseffectisnotseenwithsheepblood.
Theuseofpotassiumoxalateasananticoagulantincreasestheyieldofwholebloodbutitremainsasa
potentialchelateofdivalentmetals.Thischelateeffect,whichalsoappliestosodiumcitratetreated
blood,canbeseenintheoverallsmallercolonieswhichgrowonanticoagulantcontainingbloodagar.
Toprolongtheshelflifeofbloodproductsintheuser'shands,bloodiscollected,bottledanddispatched
onthesameday.Thephysicalcharacteristicsofappearance,haemoglobincontentandpackedcell
volumearecheckedbeforereleasebutsterilitytestsarecarriedoutbeyondthetimeofdispatch.Should
samplesofindividualanimalbloodcollectionsshowinfectionthenallrecipientsofthesuspectbloodare
notifiedbytelephone,faxortelex.
Haemolysedorlakedhorsebloodisusedforspecialpurposesinculturemedia.Ithasbeenusedfor
manyyearsinCorynebacteriumdiphtheriaemedia,wherebettergrowthwasobservedafterlysisofthe
horsebloodbythetelluriteinthemedium.
LysedbloodisstimulatoryformanyClostridiaandHaemophilusspeciesbutFildesPepticDigestof
BloodSR0046canalsobeusedforthispurpose.
Inantibioticsusceptibilitytesting,lysedhorsebloodisaddedtothemediumtoimprovethereactions
withtrimethoprimandsulphonamides.Mostculturemedia,unlessspeciallyprocessedforsusceptibility
testing,containamountsofthymidinewhichcanantagonisetheinhibitoryeffectsoftheseantimicrobials.
Whenhorsebloodislysedtheerythrocytesreleaseanenzymethymidinephosphorylasewhichconverts
thymidineintothemuchlessantagonisticcompoundthymine.Thisenzymedoesnotexistinsheep
erythrocytes.Thelyticagentusedforlysedhorsebloodiswhitesaponinwhichappearsnottoaffectthe
growthofbacteriaatthelevelrequiredforlysis.
SerumforCultureMedia
HorseserumSR0035
Totallyasepticprocessingisnotrequiredforbloodserumproduction.Bloodiscollectedincleanvessels
inacleanenvironment.Afterclottinghastakenplacethebloodisstoredat28Ctoallowtheclotto
retractandtheserumseparate.Theseparatedserumisthenfiltersterilisedandfilledoutintosterile
bottles.Allserashouldbestoredat28Cbutforlongertermstorageitisbestfrozenat20C.

Dipslides
General
ToxinDetection

Serumcanbecomecloudyonstoragecausedbylipaseenzymesandlipidchanges.Heatingtheserum
to56Cfor30minutes(inactivation)willusuallyovercomethisproblem.
SheepErythrocytesforImmunologicalReactions
Sheepblood(inAlseversolution)SR0053
Afterwashingthesheeperythrocytes,bycentrifugingwithphysiologicalsalineuntilthesupernatant
salineisfreefromhaemolysis,thewashedcellsmaybeusedforanyimmunological/serologicaltest
wheresheepcellantigensarerequirede.g.RoseWaaler,PaulBunnellandvariousvirological
complementfixationtests.
Thesheepbloodshouldbestoredat28Candusedbeforetheexpirydateonthelabel.

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