5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>natural reservoir <strong>of</strong> influenza A virus, and are likely to play the “mixingvessel” epidemiological role traditionally assigned to domestic pigs. Seroprevalencefor influenza in <strong>European</strong> wild boar seems to be relatively lowin most published studies, ranging from 0% to 7.8%. In the context <strong>of</strong>increasing populations <strong>of</strong> wild boars in Europe, the aim <strong>of</strong> this study wasto determine the apparent seroprevalence <strong>of</strong> influenza in Belgian wildboars and to identify the potential risk factors associated with seropositivity.We identified foci <strong>of</strong> high H1 influenza seroprevalence (>45%) inspecific hunting territories. If this high prevalence is the result <strong>of</strong> recurrenttransmission from domestic pigs or reflects an endemic circulation inlocal wild boar population remains to be determined. Viral isolation andphylogenetic studies would be helpful to answer this question.REF 256Hepatitis e virus (HEV): genetic relationship <strong>of</strong> the first identifiedhuman strain and swine strains in CroatiaLorena JEMERSIC 1 , Jelena PRPIC 1 , Nenad PANDAK 2 , DraganBRNIC 1 , Oktavija DAKOVIC RODE 3 , Tomislav KEROS 1 , Besi ROIC 1 ,Andreja JUNGIC 1 , Tomislav BEDEKOVIC 1 , Ivana LOJKIC 11 Croatian Veterinary Institute, Zagreb, CROATIA; 2 General HospitalSlavonski Brod, Slavonski Brod, CROATIA; 3 3Universitiy Hospital forInfectious Diseases ”Dr. Fran Mihaljevic”, Zagreb, CROATIAHepatitis E is an important health problem with a possibility <strong>of</strong> zoonotictransmission through contact with infected animals. Hepatitis E virus(HEV) belongs to the genus Hepevirus, family Hepeviridae. Four HEVgenotypes have been recognized so far. Genetic typing <strong>of</strong> swine HEVstrains, indicate their close relationship to human HEV strains and provethat swine could be recognised as reservoirs <strong>of</strong> HEV in nature. The aim <strong>of</strong>our study was to investigate the potential genetic relationship <strong>of</strong> a humanHEV strain identified for the first time in Croatia in a patient showingsigns <strong>of</strong> acute hepatitis, with strains detected in swine from the samegeographical area. Swine samples (blood, spleen and liver) were collectedfrom different age categories and breeding systems (two large commercialfarms and small farms up to 20 animals). In total, 152 swine were includedin this study. All samples were tested by a nested RT PCR protocol inorder to amplify a fragment within the metil transferase gene. Representativepositive samples including the human HEV strain were sequencedand phylogenetically analysed. HEV RNA was detected in swine samplesshowing a high viral prevalence (31.5%) regardless <strong>of</strong> the breeding systemor age category. The human and swine RNA sequences clustered into phylogeneticgroup 3 proving their close genetic relationship. However, swinesequences further clustered into subgroup 3a and 3e, whereas the humanHEV strain clustered into subgroup 3f showing the highest similarity witha reference swine strain identified in The Netherlands. Even though ourresults support the fact that swine in Croatia are reservoires <strong>of</strong> HEV theirtrue role in the chain <strong>of</strong> viral spread is still unknown. The source <strong>of</strong> thefirst reported human infection in Croatia still remains unidentified. Weconclude that further epidemiological data is needed, including testing <strong>of</strong>swine products.are responsive to West Nile Virus. There have been described the sicknesscases <strong>of</strong> alligators at crocodile farms, and there have been documentedthe cases <strong>of</strong> people among the staff <strong>of</strong> the farms being infected. For thecrocodilians the West Nile Virus is a typical emergent infection. The coldblooded vertebrata may serve as reservoir other dangerous arboviral infections.Mosquitoes <strong>of</strong> Culicidae family (prevalent in all the continents)that live on the warm blooded animals as well as on the cold bloodedones are a component in the encephalomyelitis viral shedding chain. Newemergent infections pose a serious potential hazard not only to the artificiallycreated reptilian and amphibian populations but also to the naturalones. Reptiles and amphibias turn out to be in the environment <strong>of</strong> newpathogens after they are taken from their wildlife and trapped. Stress,transportation and inappropriate maintenance conditions result in significantdecrease in immunity which makes animals more vulnerable toinfections. Pathogenic agents can evolve and infect new hosts in whichthey have not been present previously. This may lead to extension <strong>of</strong> thecausative pathogens <strong>of</strong> dangerous diseases among responsive animals andpeopleREF 258Screening <strong>of</strong> Influenza H1N1P antibodies in bats from different geographicalregionsElizanbeth LOZA RUBIO 1 , Edith ROJAS ANAYA 1 , Catalina TUFIÑOLOZA 1 , Oscar RICO 21 INIFAP, CENID Microbiología Animal, Mexico City, MEXICO; 2 FMVZUNAM, Mexico City, MEXICOBats are potentially reservoir <strong>of</strong> many emergent and re emergent zoonosisall over the world. Mexico has so far 138 species <strong>of</strong> bats in which antibodieshave been detected against rabies, dengue and swine Rubulavirus.Recently, at least 60 new species <strong>of</strong> paramyxoviruses have been identifiedin bats. Additionally, it was discovered that some <strong>of</strong> these paramyxovirusesare genetically similar to those affecting human mankind. Concerning toinfluenza virus, is an Orthomixovirus which affects a wide range <strong>of</strong> hostsincluding birds and mammals. Following the 2009 outbreak <strong>of</strong> H1N1, thescientific community has reported the presence <strong>of</strong> the virus in several species,both domestic and wild. So, it is <strong>of</strong> interest to conduct studies on thesubject, recently identified as a new hemagglutinin into a bat at two locationsin Guatemala. In our country there are no reports on the presence <strong>of</strong>antibodies in bats. The objective <strong>of</strong> this study was to detect antibodies insera against pandemic in different bats species and geographic region. Wecaught bats in the state <strong>of</strong> Mexico, Puebla, Hidalgo and Morelos, wheredifferent number <strong>of</strong> samples was taken (29, 23, 5 and 2, respectively). Serawere analyzed by hemagglutination inhibition test using 4 hemagglutinatingunits (HU). The species tested were Desmodus rotundus, Sturniralilium, Sturnira ludovici, Carollia perspicillata, Pteronotus parneli, Artibeusliteratus, Artibeus intermedius, and Myotis microtis Leptonycterisnivalis. All sera were negative. We conclude that apparently this variantvirus is not circulating in the populations analyzed; however we suggestto sample more individuals.REF 257Research <strong>of</strong> exotic animals, which are a reservoir <strong>of</strong> infectious diseases,as a component <strong>of</strong> biosafetyZinaida KLESTOVA, Irina SAVINOVAThe Institute <strong>of</strong> Veterinary Medicine NAAS <strong>of</strong> Ukraine, Kiev, UKRAINEThe question <strong>of</strong> virus diseases <strong>of</strong> the cold blooded animals and <strong>of</strong> theirbeing virus carriers or reservoirs has not been adequately studied yet andis a topic <strong>of</strong> interest. The representatives <strong>of</strong> all the reptilian classes mayserve as bridging hosts or reservoirs for different viruses, which mayinfect people, mammals and birds. It has been established that reptilesREF 259Seoul hantavirus in Rattus norvegicus in Lyon, FranceLorraine MCELHINNEY 2,4 , Kieran POUNDER 1 , Denise MARSTON 2 ,Florence AYRAL 3 , Andrew BREED 2 , Michael BEGON 1 , CharlotteFEATHERSTONE 2 , Marc ARTOIS 3 , Anthony FOOKS 2,41 University <strong>of</strong> Liverpool, Liverpool, UNITED KINGDOM; 2 AnimalHealth and Veterinary Laboratories Agency (AHVLA), Weybridge, UNI-TED KINGDOM; 3 Université de Lyon, Marcy L’Étoile, FRANCE;4 National Consortium for Zoonosis Research, Leahurst, UNITED KING-DOMVi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013S191
5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>Hantaviruses (family: Bunyaviridae, genus Hantavirus) are single strandedRNA viruses, which are transmitted to humans primarily via inhalation <strong>of</strong>aerosolised virus in contaminated rodent urine and faeces. Whilst infectedreservoir hosts are asymptomatic, human infections can lead to two clinicalmanifestations, haemorrhagic fever with renal syndrome (HFRS) andhantavirus cardiopulmonary syndrome (HCPS), with varying degrees <strong>of</strong>morbidity and mortality. Surveillance in Europe has detected six rodentborne hantaviruses; Dobrava Belgrade Virus (DOBV), Saaremaa virus(SAAV), Seoul virus (SEOV), Puumala virus (PUUV), Tatenale virus(TATV) and Tula virus (TULV). There were over 1900 HFRS cases reportedin France between 2005 and 2010. However, the prevalence <strong>of</strong> rodentand human cases associated with SEOV in Europe are considered to below, and speculated to be driven by the sporadic introduction <strong>of</strong> infectedbrown rats (Rattus norvegicus) via ports. Between October 2010and March 2012, 128 brown rats were caught at sites across the Lyonregion in France. SEOV RNA was detected in the lungs <strong>of</strong> 18 brown rats(14%, 95% CI 8.6–21.3) using a nested pan hantavirus RT PCR (polymerasegene). We did not detect any evidence <strong>of</strong> a genetic differencebetween infected and non infected rats (cytochrome b gene). Our findingsand the isolation <strong>of</strong> Seoul virus in UK brown rats, suggest thatSEOV is more prevalent in <strong>European</strong> brown rats and may contribute toa greater number <strong>of</strong> the reported HFRS cases in Europe than previouslybelieved.REF 260Cell culture isolation and molecular analysis <strong>of</strong> HEV strains fromUruguay: evidence <strong>of</strong> a recent history <strong>of</strong> the infectionSantiago MIRAZO 1 , Natalia RAMOS 1 , José RUSSI 2 , GustavoCASTRO 3 , Juan ARBIZA 11 Sección Virología. Facultad de Ciencias. UdelaR, Montevideo,URUGUAY; 2 British Hospital, Montevideo, URUGUAY; 3 Ministerio deGanadería, Agricultura y Pesca, Montevideo, URUGUAYHepatitis E virus (HEV) infection is an important public health concernin many developing countries causing waterborne outbreaks and sporadicautochthonous hepatitis. Aultough it is transmitted primarily by the fecal– oral route, zoonotic trasmission from animal reservoirs has also beensuggested. Data regarding the molecular characterization <strong>of</strong> HEV isolatesfrom South America lacks and further investigation is needed. We recentlyreported the first cases <strong>of</strong> autochtonous acute HEV infection in Uruguay.Molecular analysis and phylogenetic reconstruction <strong>of</strong> the viral hypervariableregion and capsid gene showed that they clustered together withinGenotype 3 and were closely related to a set <strong>of</strong> <strong>European</strong> strains but weredissimilar to reported South American isolates. The co circulation <strong>of</strong> subtypes3i and 3 h was observed. Here, we isolated selected Uruguayan HEVstrains in a cell culture based system by blind serial passages in A549 cellline. In vitro replication was confirmed by immun<strong>of</strong>luorescence assay andRT PCR. To better understand the molecular epidemiology <strong>of</strong> the HEVisolates we analyzed a larger region <strong>of</strong> the capsid gene and a conservedfragment <strong>of</strong> the methyltransferase gene. Additionally, we investigated thecirculation <strong>of</strong> HEV in domestic pig population in nationwide herds byRT PCR. We confirmed that the HEV infection in Uruguay has probablya <strong>European</strong> origin and several sources have existed. According to phylogeneticanalysis and since no swine HEV strain was detected in thisstudy, we also suggest that the history <strong>of</strong> HEV in our country is veryrecent.REF 261Hantavirus circulating among Rattus rattus in Mayotte Island, IndianOceanSéverine MURRI 1 , Frédérik BEAULIEUX 2 , Philippe MARIANNEAU 1 ,Amélie DESVARS 3 , Lénaïg HALOS 4 , Gwenaël VOURC’H 3 , NoëlTORDO 51 <strong>Virology</strong> Unit, Anses Laboratoire de Lyon, Lyon, FRANCE; 2 Centrede recherche clinique, Hôpital Croix Rousse, Lyon, FRANCE; 3 UR346Animal Epidemiology, INRA, Saint Genès Champanelle, FRANCE;4 Mérial, Lyon, FRANCE; 5 Unit Antiviral Strategies, Institut Pasteur, Paris,FRANCEFollowing the Chikungunya virus epidemics in the Indian Ocean, morethan 3500 animals (primates, bats, rodents, insectivores) were capturedand sampled (blood, serum, organs). The serum <strong>of</strong> 539 rodents (Rattusrattus, R. norvegicus, Mus musculus) and insectivores (Suncus murinus,Tenrec ecaudatus) captured in 2006 7 in La Réunion island (379), in 2007in Mayotte island (160) were checked for arenaviruses and hantaviruses.RNA was extracted, pooled by groups <strong>of</strong> 5, screened by nested RT PCR forhantaviruses (Klempa, 2007, EID 13: 520 2; Kang, 2009, <strong>Virology</strong> 388: 814) and arenaviruses (Vieth, 2007, Trans R Soc Trop Med 101: 1253 64).For each positive pool, samples were screened individually, then sequenced.None <strong>of</strong> the samples from La Reunion Island was positive for eitherarenavirus or hantavirus. In Mayotte Island, no sample was positive forarenavirus. However 29/160 (i.e. 18%) serums from Rattus rattus testedpositive for hantavirus. The sequence <strong>of</strong> 246 nucleotides within the polymeraseL coding region showed a limited genetic diversity and a perfectconservation <strong>of</strong> the amino acid sequence. The phylogenetic analysis indicatedthat these new hantaviruses clustered closer but clearly distinct fromthe hantaviruses Serang virus and Jurong virus. A phylogeographic analysissuggests that several variants seem to have been largely spread andmixed across the Mayotte Island. This work invites to a better surveillance<strong>of</strong> hantaviruses in rodents <strong>of</strong> Indian Ocean islands and to assess the risk<strong>of</strong> transmission <strong>of</strong> Haemorrhagic Fever with Renal Syndrome (HFRS) tothe population.REF 262Bat rabies surveillance in France. First report <strong>of</strong> Isolation <strong>of</strong> theBokeloh Bat Lyssavirus in France in Myotis nattereri in 2012Evelyne PICARD MEYER 1 , Alexandre SERVAT 1 , EmmanuelleROBARDET 1 , Marie MOINET 1 , Dorothée JOUAN 2 , ChristopheBOREL 2 , Florence CLIQUET 11 ANSES Nancy Laboratory for Rabies and Wildlife, French agency forfood, environmental and occupational health safety, Malzeville, FRANCE;2 CPEPESC Lorraine, Velaine en Haye, FRANCEPassive surveillance <strong>of</strong> bat rabies has been improved in France since 2000,thanks to the surveillance network constituted by local veterinary servicesand bat handlers from the Chiroptera group (SFEPM, French nationalbat conservation network). Since its inception in 1989, the network hasreported 61 cases <strong>of</strong> EBLV 1 infection for 2,457 tested bats. All casesinvolved the same species, Eptesicus serotinus in different areas <strong>of</strong> thecountry. In July 2012, we reported for the first time the presence <strong>of</strong> theBokeloh Bat Lyssavirus (BBLV), isolated in Myotis nattereri in a forestin North Eastern <strong>of</strong> France. The Bokeloh Bat Lyssavirus was previouslyS192 Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013
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