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1、ArchivesofVirologyArchivesofVirology57,43--51(1978)©bySpringer-Verlag1978ReplicationofanEntericBovineCoronavirusinIntestinalOrganCulturesByJA~ICEC.BRIDGER1,E.0.CAUL2,andS.I.EGGLESTO:NE3InstituteforResearchonAnimalDiseases,Compton,Berkshire,UnitedKingdomPublicHealthLaboratory,Bristol,U
2、nitedKingdom3SchoolofEnvironmentalSciences,PlymouthPolytechnic,Pl~wnouth,UnitedKingdomWith9FiguresAcceptedOctober18,1977SummaryAcoronavirusisolatedintrachealorganculturesfromthefaecesofacalfwithdiarrhoeareadilymultipliedonpassageinintestinalorgancultures.Evidenceformultiplicationwasob
3、tainedbytheproductionofviralhaemagglutinininorganculturefluidsandthepresenceofimmunofluorescencoandviralparticlesinthecolumnarepithelialcellsofthevilli.Thusvirusmultiplicationwasstudiedinvitrointhecelltypeinwhichitmultipliesnaturally.IntroductionThecultivationincellculturesofvirusesas
4、sociatedwithdiarrhoeaofmanandanimalshasprovedtobedifficultorimpossible(1,6,9,10,12).However,organculturesofintestinaltissueprovidealternativeinvitroculturesystemswhicharesimilartothetissuesinwhichentericvirusesmultiplynaturally.Theyhavebeenusedforanumberofhumanentericviruses(2,3,4,7,1
5、1)andanimalentericviruses(8).Thispaperdescribesthereplicationoftheneonatalcalfdiarrhoeacoronavirusinbovineintestinalorgancultures.MaterialsandMethodsSourceo]VirusInoculumThebovinecoronavirus('TheBritishIsolate')usedinthisstudyhadbeenisolatedfromthefaecesofadiarrhoeiccalfbyinoculationo
6、fafaecalfiltrateintobovinetrachealorganculture(Bridger,~Toode&Meyling,tobepublished).ThefluidfromtheseprimaryisolationcultureshadahaemagglutinatingtitreofI/sandwasstoredat--70°Cbeforeinoculationintogutorgancultures.0304-8608/78/0057/0043/$0t.8044JANICEC.BI~IDGER,E.O.CAUL,andS.I.EGGLES
7、TONE:PreparationandInoculationo/OrganCulturesOrganculturesofuppersmallintestinefrom4.5to6monthbovinefoetuseswerepreparedasdescribedpreviously(2),Themaintenancemediumwasreplenishedevery24hours.Cultureswereinfected24hoursafterpreparationbydropping0.05mlofinoculumontoeachpieceoftissue.Af
8、terad
