5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>In this study, 150 blood sera samples <strong>of</strong> unvaccinated domestic horseswere tested for equine herpesvirus (EHV 1, EHV 4) specific antibodies bycommercially available indirect Enzyme Linked Immunosorbent Assay(ELISA, Svanovir, EHV 1/EHV 4 Ab kit, Svanova Biotech AB, Sweden).EHV 1 and EHV 4 specific antibodies were detected in 52 (34.66%) and101 (67.33%) <strong>of</strong> the 150 tested sera, respectively. The results indicatedthat EHV 1 and EHV 4 infections appear to be widespread in domestichorses in South East Anatolia.Key words: EHV 1, EHV 4, ELISA, horseREF 312Detection <strong>of</strong> antibodies against Blue Tongue Virus in Yaks (BOSGRUNNIENS) in Issyk KulOguzhan AVCI 1 , Orhan YAPICI 2 , Oya BULUT 1 , Mehmet KALE 3 ,Sibel YAVRU 1 , Atilla SIMSEK 11 University <strong>of</strong> Selcuk, Faculty <strong>of</strong> Veterinary Medicine, Department <strong>of</strong> <strong>Virology</strong>,KONYA, TURKEY; 2 University <strong>of</strong> Kyrgyzstan Turkey Manas, Faculty<strong>of</strong> Veterinary Medicine, Department <strong>of</strong> <strong>Virology</strong>, BISHKEK, KYRGYZS-TAN; 3 University <strong>of</strong> Mehmet Akif Ersoy, Faculty <strong>of</strong> Veterinary Medicine,Department <strong>of</strong> <strong>Virology</strong>, BURDUR, TURKEYBluetongue virus (BTV) is the type species <strong>of</strong> the genus Orbivirus withinthe family Reoviridae. There are 26 BTV serotypes that are determinedby the specificity <strong>of</strong> interactions between the virus outer capsid with neutralizingantibodies generated during infection <strong>of</strong> the ruminant host. Theaim <strong>of</strong> this study was to describe the seroprevalence rate <strong>of</strong> bluetonguevirus (BTV) in unvaccinated yaks in Issyk Kul. A total <strong>of</strong> 168 serumsamples were collected from yaks that were randomly selected betweenJuly August 2011. Antibodies to BTV in sera were detected using a commerciallyavailable competitive enzyme linked immunosorbent assay (cELISA, VMRD, USA). The test was performed as per the manufacturer’sinstructions. The results showed that 4 (2.38%) <strong>of</strong> the samples werepositive for BTV antibodies. This study indicates that antibodies againstBTV are uncommon among the yaks in Issyk Kul. In spite <strong>of</strong> the significanteconomic and animal health impacts <strong>of</strong> BTV infection in sheep, todate there has been no assessment <strong>of</strong> the Culicoides spp. involved. Despitethe low results large scale research is necessary for the prevention <strong>of</strong>BTV infection. In conclusion, the seroprevalence <strong>of</strong> BTV in yaks has beeninvestigated for the first time in Issyk Kul.Key words: BTV, yak, ELISAREF 313New subtypes <strong>of</strong> small ruminant lentiviruses (SRLVs) detected inSlovenian goat and sheep flocksDarja BARLIC MAGANJA 3 , Urska KUHAR 1 , Joze GROM 21 University <strong>of</strong> Ljubljana, Veterinary Faculty, Ljubljana, SLOVENIA;2 University <strong>of</strong> Ljubljana, Veterinary Faculty, Ljubljana, SLOVENIA;3 University <strong>of</strong> Primorska, Facutly for Health Sciences, Izola, SLOVENIASmall ruminant lentiviruses (SRLVs) <strong>of</strong> the Retroviridae family infectgoats and sheep worldwide. Phylogenetic analysis divides them into fiveprincipal genotypes (A E). The Slovenian SRLV strains characterized byphylogenetic analysis <strong>of</strong> two genomic regions (1.8 kb gag pol fragmentand 1.2 kb pol fragment) were clustered into A and B genotypes. Theyare highly heterogeneous, with ovine strains belonging to genotype A andcaprine strains to genotypes A and B. A cluster composed <strong>of</strong> four sheepvirus sequences was clearly divergent from all other subtypes in group Aand could not be assigned to any <strong>of</strong> them and two goat virus sequenceswere found to belong to genotype A and could not be assigned to existingsubtypes. The results <strong>of</strong> phylogenetic analysis performed in our studyrevealed two new subtypes within genotype A, subtype A14 and subtypeA15.REF 314Characterization <strong>of</strong> pandemic influenza A(H1N1) 2009 viruses isolatedfrom Pig and Cat in France since 2009Emilie BONIN 1,2 , Séverine HERVÉ 1,2 , Aure SAULNIER 1,2 , StéphaneQUÉGUINER 1,2 , Nicolas BARBIER 1,2 , Stéphane GORIN 1,2 , GaëlleSIMON 1,21 Anses, Laboratoire de Ploufragan Plouzané, Unité Vi<strong>rologie</strong> ImmunologiePorcines, Ploufragan, FRANCE; 2 Université Européenne de Bretagne,FRANCEIn April 2009, a novel influenza A(H1N1) virus (H1N1pdm) was detectedin people in Mexico and the United States. The virus spread rapidly andcaused a flu pandemic in humans within a few months. The new viruscontains a combination <strong>of</strong> genomic segments from swine origin that hadnever been identified previously. Soon after the virus started to spread globallyin humans, its introduction into pig holdings was noticed in manycountries, including <strong>European</strong> member states. Transmission <strong>of</strong> H1N1pdmto French pigs was evidenced to have occurred in Brittany as early asJanuary 2010. However, the virus does not seem to have been maintainedwithin the pig population in this region <strong>of</strong> high pig density where almost50% <strong>of</strong> the herds are infected by <strong>European</strong> enzootic swine influenza viruses(SIVs) for many years. By contrast, H1N1pdm was regularly isolated frompig herds located in central part <strong>of</strong> France since October 2010, demonstratingthat the virus spread in the French pig population, especially inareas with most naive animals. Not only restricted to humans and pigs,H1N1pdm virus was also detected in turkeys, cheetahs, minks, seals, ferrets,dogs and cats. In November 2009, it was isolated from a domesticcat in France, after several members <strong>of</strong> the family contracted the infection.Whole genomes <strong>of</strong> H1N1pdm viruses isolated from pigs and cat inFrance were sequenced. Phylogenetic and polymorphism analyses showedthat all genomic segments were very closely related to respective counterpartsfrom other contemporary pandemic strains isolated in Europe andto reference strain A/California/04/2009. To date, no reassortant virusesbetween H1N1pdm and enzootic SIVs were detected in France unlike inother <strong>European</strong> countries, but may appear. The spread <strong>of</strong> H1N1pdm intopig population is an additional risk for the emergence <strong>of</strong> new influenzaviruses with zoonotic potential.REF 315Serological and virological investigation <strong>of</strong> Canine Adenovirus Infectionin dogsOya BULUT 1 , Orhan YAPICI 2 , Oguzhan AVCI 1 , Atilla SIMSEK 1 ,Kamil ATLI 1 , Irmak DIK 1 , Sibel YAVRU 1 , Sibel HASIRCIOGLU 3 ,Mehmet KALE 3 , Nuri MAMAK 41 University <strong>of</strong> Selcuk, Faculty <strong>of</strong> Veterinary Medicine, Department <strong>of</strong> <strong>Virology</strong>,Konya, TURKEY; 2 University <strong>of</strong> Kyrgyzstan Turkey Manas, Faculty<strong>of</strong> Veterinary Medicine, Department <strong>of</strong> <strong>Virology</strong>, Bishkek, KYRGYZSTAN;3 University <strong>of</strong> Mehmet Akif Ersoy, Faculty <strong>of</strong> Veterinary Medicine, Department<strong>of</strong> <strong>Virology</strong>, Burdur, TURKEY; 4 University <strong>of</strong> Mehmet Akif Ersoy,Faculty <strong>of</strong> Veterinary Medicine, Department <strong>of</strong> Internal Medicine, Burdur,TURKEYAdenoviruses are linear, double stranded DNA viruses which infect a widevariety <strong>of</strong> mammals and birds. Two have been identified in the dog: CanineAdenovirus type 1 (CAV 1) which infects most <strong>of</strong> the major organs causing,amongst other diseases, canine infectious hepatitis, and CAV 2 whichcauses canine infectious laryngotracheitis and enteric diseases. The aim<strong>of</strong> this study is to define the serological and virological status <strong>of</strong> CAVinfection in dogs. In this study, blood serum samples were taken from111 dogs that show clinical signs from the clinics <strong>of</strong> Veterinary MedicineFaculty, Selcuk University. Also 77 dogs were sampled from Ispartaand Burdur dog shelters by random sampling, regardless <strong>of</strong> the clinicalfindings. Clinical findings included a systemic disease, characterized byfever, vomiting, mucopurulent oculonasal discharge and conjunctivitis,Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013S207
5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>severe moist cough, signs <strong>of</strong> pulmonary disease and dehydration. In serologicalstudies, serum samples were investigated for the presence <strong>of</strong> CAVantibodies by indirect ELISA. Antibodies to CAV were detected in 103(54.7%) <strong>of</strong> the examined sera samples. Leukocyte samples were processedand inoculated onto confluent monolayers <strong>of</strong> MDCK cells using standardvirological techniques. Cells were incubated at 37 ◦ C after inoculation andobserved daily for the appearance <strong>of</strong> cytopathogenic effect. After thirdpassage, cells were examined by direct Immun<strong>of</strong>luorescence test (IFT) forCAV antigens. We have not seen any morphological changes in MDCKcells and CAV antigens were not detected in any <strong>of</strong> leukocytes.Key words: CAV, ELISA, IFTREF 316Experimental infection in mice with avian coronavirus (Gammacoronavirus)isolated from chickensMatheus CAVALHEIRO MARTINI 1 , Márcia Mercês AparecidaBIANCHI DOS SANTOS 1 , Leonardo CARDIA CASERTA 1 , RicardoDURÃES CARCAVLHO 1 , Marina AIELLO PADILLA 1 , Helena LAGEFERREIRA 2 , Clarice WEISS ARNS 11 Laboratory <strong>of</strong> Animal <strong>Virology</strong>, Institute <strong>of</strong> Biology, University <strong>of</strong> CampinasUNICAMP, Campinas, BRAZIL; 2 Laboratory <strong>of</strong> Avian Disease,University <strong>of</strong> São Paulo College <strong>of</strong> Animal Science and Food Engineering<strong>of</strong> USP, Pirassununga, BRAZILCoronaviruses, which are single stranded, positive sense RNA viruses, isthe causative agent <strong>of</strong> a wide variety <strong>of</strong> existing and emerging diseasesin humans and other animals. Infectious bronchitis virus (IBV) belongsto the genus Gammacoronavirus <strong>of</strong> the Coronaviridae family and is associatedto a highly contagious upper respiratory tract disease in chickens.IBV has been mainly detected in epithelial cells causing lesions in thenasal turbinates, trachea, kidney, gonads, oviduct, lungs and airsacs. Thisstudy was conducted to investigate if an IBV sample, obtained from anavian host (Gallus gallus), is able to replicate in a murine model. TenBalb/C mice 4 wks old were inoculated with 50 L fluid containgning104.0TCID50/mL <strong>of</strong> IBV/BRAZIL/2007/Unicamp801. Also, a controlgroup with four Balb/C mice was sham inoculated with 50 L <strong>of</strong> tissueculture fluid. Five infected and two control animals were euthanatized at3th and 10th days post inoculation; samples from sinus, tracheas, lung andkidney were collected at necropsy. Viral RNA was detected by real timePCR, based on the UTR gene, with copy numbers varying from 3,97 × 106to 1,21 × 108 (Ct values:35.98 to 38.58) at 3th d.p.i and 4,13 × 103 to1,74 × 108 (Ct values:35,94 to 38.51) at 10th d.p.i; no RNA was detectedin the control group. These results showed that an IBV sample was able toreplicate in a mammal host. To date there is no report <strong>of</strong> avian coronavirusreplication in mammals. A mammal species could constitute a host rangein which the virus can evolve, as well as a space for recombination betweenthe avian and mammal coronavirus. As a consequence, new variantsmight be produced.REF 317Susceptibility <strong>of</strong> Chicken embryonic Stem Cell Derived Keratinocytesto Marek’s Disease Virus infectionMathilde COUTEAUDIER 1 , Katia COURVOISIER GUYADER 1 ,Bertrand PAIN 2 , Caroline DENESVRE 1 , Jean François VAUTHEROT 11 INRA UMR 1282 ISP, Biova, Centre INRA de Tours, Nouzilly, FRANCE;2 INRA UMR 1288 IGFL, (ENSL/CNRS/UCBL1), Centre INRA de ClermontFerrand Theix, Villeurbanne, FRANCEMarek’s Disease Virus (MDV) is a very potent oncogenic alphaherpesviruswhich replicates to high titres in feather follicle epithelium (FFE)cells, from which it is efficiently shed as free virus. Investigations on thereplication <strong>of</strong> MDV in FFE are <strong>of</strong> major importance for a better knowledgeon viral dissemination. No in vitro cell system can afford an environmentthat may reproduce the one found in the cells <strong>of</strong> the differentiated FFE. Toenlarge the array <strong>of</strong> our investigations we explored the possibilities <strong>of</strong>feredby the differentiation <strong>of</strong> pluripotent stem cells from chicken. We thereforeexamined whether chicken embryonic stem cells (cES) could be inducedto differentiate toward the keratinocyte lineage, and whether those keratinocytesor their progenitors at intermediate stages <strong>of</strong> differentiation couldsupport MDV replication. From cES BP25, we derived several homogenouscell populations which showed typical morphological characteristics<strong>of</strong> chicken keratinocytes. In addition, we showed a down regulation <strong>of</strong> pluripotencymarkers over the time, meanwhile the expression <strong>of</strong> late markersgenes <strong>of</strong> epidermal differentiation was upregulated. MDV replication wasexamined in cell populations showing either a typical keratinocyte morphologyor an “epithelial cell” phenotype, by using cell adapted or virulentvirus strains. The permissiveness <strong>of</strong> these differentiated cells was comparedto the one <strong>of</strong> the parental pluripotent cES. The ongoing experimentsare focussed on the characteristics <strong>of</strong> virus replication and morphogenesisin those cell populations.REF 318Antibody prevalence against Caprine Arthritis Encephalitis in SaanenGoats in Adana ProvinceIrmak DIK 2 , Orhan YAPICI 1 , Oguzhan AVCI 2 , Kamil ATLI 2 ,Sibel YAVRU 21 University <strong>of</strong> Kyrgyzstan Turkey Manas, Faculty <strong>of</strong> Veterinary Medicine,Department <strong>of</strong> <strong>Virology</strong>, Bishkek, KYRGYZSTAN; 2 University <strong>of</strong> Selcuk,Faculty <strong>of</strong> Veterinary Medicine, Department <strong>of</strong> <strong>Virology</strong>, Konya, TURKEYCaprine arthritis encephalitis is a viral disease <strong>of</strong> small ruminant that iscaused by the caprine arthritis encephalitis virus (CAEV). CAEV infectionoccurs worldwide and like other lentiviruses, they cause lifelong persistentinfection, which may be essentially symptomless or results in a variety <strong>of</strong>inflammatory, degenerative, or immunosuppressive diseases in a variableproportion <strong>of</strong> infected hosts. This virus causes chronic disease <strong>of</strong> the joints,encephalitis, slowly progressive pneumonia, and mastitis in goat. The aim<strong>of</strong> this study is to define seroprevalence <strong>of</strong> CAEV infection in Saanengoats in Adana in the Mediterranean region <strong>of</strong> Turkey. In the present study,blood serum samples were collected from 150 Saanen goats unvaccinatedagainst CAEV in Adana. Sera samples were analyzed for presence <strong>of</strong>antibodies against CAEV by commercially available enzyme linked immunosorbentassays (ELISA; ID Screen, France). The test was performed asper the manufacturer’s instructions. Antibodies to CAEV were detectedin 4 (2.66%) <strong>of</strong> the examined sera samples. Results <strong>of</strong> this study carryout that CAEV has a low prevalence in Saanen goats in Adana. As a vaccinationprogramme for CAEV infections is not implemented in Turkey.Further studies should focus on explaining the risk factors associated withCAEV prevalence in Saanen goat populations in an attempt to control newinfections.Key words: Caprine arthritis encephalitis virus, ELISA, Saanen goat,SerumS208 Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013
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