rologie i - European Congress of Virology

5 th European Congress of Virologywere only 52 54%. The phylogenetic analysis of the entire polyproteinperformed on altogether 37 amino acid sequences confirmed the classificationof cardioviruses into the two species Encephalomyocarditis virusand Theilovirus, and revealed that the two virus isolates from marmosetscan be considered novel EMCV strains with the closest genetic relationshipto the “old” Mengo virus strain “M”, which was already isolated 50years before.REF 327Prevalence of hepatitis E virus in population of free living and gameenclosure wild boarsMonika KUBANKOVA 1,4 , Jiri LAMKA 2 , Vladimir ZAKOVCIK 3 , PetraVASICKOVA 11 Veterinary Research Institute, Brno, CZECH REPUBLIC; 2 Charles Universityin Prague, Faculty of Pharmacy in Hradec Kralove, HradecKralove, CZECH REPUBLIC; 3 Central Military Veterinary Institute atHlucin, Hlucin, CZECH REPUBLIC; 4 University of Veterinary and PharmaceuticalSciences Brno, Faculty of Veterinary Medicine, Brno, CZECHREPUBLICHepatitis E virus (HEV) is a small, non enveloped RNA virus, which isclassified in the genus Hepevirus, family Hepeviridae. Isolates of HEVare divided into four major genotypes and several subtypes accordingto sequence comparisons and phylogenetic analysis. Mammalian HEVstrains have been obtained from domestic pigs, wild boars, rabbits, rats,deer and mongoose and possibly cattle and sheep. The existence of HEVantibodies have been identified in other animal species as well. HepatitisE is considered to be as a zoonotic disease and domestic pigs, wild boarsand perhaps other animal species are natural reservoirs of the virus. In theCzech Republic, HEV RNA has been detected by triplex real time reversetranscription polymerase chain reaction (qRT PCR) in free living wildboars (17.3%) and wild boars originated from game enclosures (28.6%).In one game enclosure was detected prevalence up to 62.5% in wild boars.The HEV prevalence in game enclosures is significantly higher than infree living wild boars, since the breeding conditions are different from thenatural environment. Subsequently, sequence and phylogenetic analysesof obtained HEV isolates suggested a potential cross species transmissionand circulation of HEV in the Czech Republic. The discoveries of pathogenesis,clinical manifestation epizootology and epidemiology of HEV inwild animals are still missing.This work was supported by grants: NT13884 4/2012 and AdmireVetCZ1.05/2.1.00/01.0006 ED0006/01/01.software, phylogenic trees were constructed by neighbor joining accordingto Kimura 2 parameter model in MEGA 5.1 software.Analysis of phylogenic trees (P4b and fpv140) showed a wide diversity ofstrains. Most of isolates cluster in Canarypox virus clade (84%), mainlyin subclades B1 (Canarypox) and B2 (Starlingpox). The others clusterin Fowlpox virus clade, mainly in subclade 2 (Turkeypox). This studyshows that a wide diversity of avipoxviruses can affect a unique hostspecies, even in a restricted geographic area (most of the observed diversityrefers to birds from Morocco) and suggests that vaccination alone canhardly protect Houbara bustard against avipoxvirus infections. Finally itquestions the taxonomy of avipoxviruses, which classically attributes thehost name to the virus species and provides clues for a better understandingof avipoxviruses epidemiology.REF 329Monoclonal antibodies against accessory protein 7b of feline coronavirusTanja LEMMERMEYER 1 , Benjamin LAMP 1,2 , Heinz Jürgen THIEL 11 Institute of Virology, Department for Veterinary Medicine, Justus LiebigUniversität, Giessen, GERMANY; 2 present adress: Institute of Virology,Department for Pathobiology, University of Veterinary Medicine, Vienna,AUSTRIAFeline infectious peritonitis virus (FIPV), the virulent biotype of felinecoronavirus (FCoV), causes feline infectious peritonitis (FIP), a lethaldisease. FIPV is thought to arise by mutation(s) during persistent infectionwith feline enteric coronavirus (FECV). Nucleotide sequence analysesindicated that mutations within the genes S, 3abc and 7ab are involved inthe transition from FECV to FIPV. This study aims at the generation ofmonoclonal antibodies (mAbs) against the 7b protein of FCoV in order tocharacterize this viral antigen and to develop novel diagnostic reagents.First the 7b protein was expressed as GST fusion protein and purified byaffinity chromatography. After immunization of mice with the 7b proteinmAb producing hybridomas were generated. Two reactive mAbs termed5B6 and 14D8 were identified by ELISA. Both mAbs detected bacteriallyexpressed 7b by immunoblot. The specific binding region was mappedwithin the amino acids 50 65 of 7b (total length: 206 aa). The isotypesof 5B6 and 14D8 were IgG2a and IgG1, respectively. Both mAbs detectedthe authentic viral protein in FIPV (strain 79 1146) infected cells byimmunoblot and immunofluorescent assay (IFA). The mAbs also showedcross reactivity with bacterially expressed 7b of FCoV strain Black. Itwill be interesting to study the biosynthesis of this protein in FCoV infectedcells which is expected to be glycosylated and secreted. Moreover itsintracellular localization will be determined.REF 328Avipoxvirus diversity in Houbara bustard in captivityGuillaume LE LOC’H 1,2,3 , Christelle CAMUS BOUCLAINVILLE 1,2 ,Mariette DUCATEZ 1,2 , Stéphane BERTAGNOLI 1,21 INRA, UMR 1225, Toulouse, FRANCE; 2 Université de Toulouse, INPENVT, Toulouse, FRANCE; 3 Emirates Center for Wildlife Propagation,Missour, MOROCCOAvipoxvirus sp. is a serious threat for the success of reinforcementprograms of some endangered birds species such as Houbara bustard(Chlamydotis sp.). Avipoxvirus infections can lead to high morbidity andcompromise the survival of wild release birds. In order to better understandits epidemiology and adapt the prophylaxis strategy, the diversityof Avipoxvirus sp. strains was studied in Houbara bustard. From 2008to 2012, 70 pox lesions from houbara bustards in 3 breeding projects(Morocco, UAE and Uzbekistan) were sampled and two regions of theviral genome (P4b: 578pb and fpv140: 2 3 kb) were amplified and sequenced.After alignment of obtained and published sequences with ClustalXREF 330Infection of cats with a new recombinant FCoVI/CCoVI strain harboringspecific forms of non structural protein gp3Anne Laure PHAM HUNG D’ALEXANDRY D’ORENGIANI 1 , LidiaDUARTE 1 , Cristina HORHOGEA 2 , Carine PINHAS 1 , AstridVABRET 3 , Nicole PAVIO 1 , Sophie LE PODER 11 UMR 1161 Virologie, INRA, ENVA, ANSES, Maisons Alfort, FRANCE;2 Université des Sciences Agricoles et Médecine Vétérinaire Ion Ionescude la Brad Iasi, Iasi, ROMANIA; 3 Laboratoire de Virologie, EA 2128,CHU de Caen, Caen, FRANCEOne of the main characteristics of Coronaviruses is their rapid genetic evolution,allowing cross species transmission, sometimes leading to humanepidemic such as SARS in 2002 and the new hCoV EMC strain in theMiddle East. Despite these new threats originating from animal reservoirs,mechanisms of these inter specific host transmissions remain unclear. Bothcanine (CCoV) and feline (FCoV) Coronaviruses are subdivided into twoVirologie, Vol 17, supplément 2, septembre 2013S211