5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>an apoptosis event. We also displayed that caspase activity and apoptosisare induced at late during CCHFV infection. Apoptosis is an importantmechanism by which virus infected cells are eliminated from the host.Many viruses have evolved strategies to delay or suppress apoptosis inorder to secure efficient virus replication, assembly and egress <strong>of</strong> infectiousviral particles. In this study, we have been interested to investigate ifCCHFV infection can regulate the apoptosis at early post infection. Ourdata displayed that caspase activation are suppressed/delayed early postinfection both upstream and on executor caspase level, insitu. In addition,we suggest that the multifunctional virus structural protein, NP, is involvedin regulation <strong>of</strong> apoptotic pathways in infected cells. In this study, we alsosuggest that both mitochondrial pathway and death receptors apoptoticpathways are regulated during infection at early and late post infection butin different manner.many cell models such as Vero cells and primary human lung epithelialcells. The activation <strong>of</strong> host innate immune responses by hCoV EMCand SARS CoV have been shown to be relatively modest. In the presentstudy we assessed the capability <strong>of</strong> hCoV EMC to infect and replicate inhuman monocyte derived macrophages and dendritic cells and comparedthese responses with SARS CoV. Preliminary data shows that like SARSCoV, hCoV EMC can infect macrophages but both viruses show impairedability to replicate in these cells. A weak induction <strong>of</strong> interferon(IFN) and IFN 1 gene expression can be seen at 6 h post infection in hCoV EMCinfected macrophages, but at later timepoints this induction is returned tobasal levels. Our data suggests that both <strong>of</strong> these viruses show relativelypoor ability to replicate in primary human antigen presenting cells whichmay be the reason for a weak ability <strong>of</strong> these viruses to induce host cellcytokine gene expression.REF 242Phylogenetic analysis <strong>of</strong> highly pathogenic avian influenza virus H5N1from migratory birds in BangladeshRokshana PARVIN 1,2 , Emdadul H. CHOWDHURY 2 ,Abu.H.M. KAMAL 2 , M. GIASUDDIN 3 , M.R. ISLAM 2 , ThomasW. VAHLENKAMP 11 Institute <strong>of</strong> <strong>Virology</strong>, Faculty <strong>of</strong> Veterinary Medicine, University <strong>of</strong>Leipzig, Leipzig, GERMANY; 2 Department <strong>of</strong> Pathology, Faculty <strong>of</strong>Veterinary Science, Bangladesh Agricultural University, Mymensingh,BANGLADESH; 3 Animal Health Research Division, BangladeshLivestock Research Institute, Dhaka, BANGLADESHSince the first outbreak <strong>of</strong> highly pathogenic avian influenza virus (HPAIV)<strong>of</strong> subtype H5N1 in Bangladesh in 2007 the virus continues to circulateamong domestic poultry causing severe economic losses. To investigatemigratory birds as possible source <strong>of</strong> virus introduction genome characterization<strong>of</strong> two HPAIV H5N1 isolates from migratory birds (A/migratorybird/Bangladesh/P18/2010 and A/migratory bird/Bangladesh/P29/2010)was performed. After full length amplification and sequence analysis <strong>of</strong>the gene segments encoding HA, NA, M and NS a comprehensive phylogeneticanalysis was performed by comparing the gene segments with otherBangladeshi isolates and representative isolates <strong>of</strong> all clades described inthe H5 nomenclature. The Bangladeshi H5N1 isolates from different birdsand poultry were broadly grouped into two clades. The majority <strong>of</strong> isolatesfrom 2007 to 2010 belong to clade 2.2.2 whereas the recent isolates fromchickens and crows <strong>of</strong> 2011 along with the selected migratory bird isolatesdescribed here are grouped into clade 2.3.2.1. The selected isolates revealedmultiple basic amino acids 338QRERRRKR*G346 at their HA cleavagesite similar to other isolates from clade 2.3.2.1. The receptor bindingsite <strong>of</strong> HA molecule contained avian like (SAa 2, 3 Gal) receptor specificity.The results verified the presence <strong>of</strong> HPAIV among migratory birds inBangladesh. Further detailed analyses will reveal whether migratory birdsmay represent the source <strong>of</strong> the newly introduced clade 2.3.2.1 virus whichwas meanwhile diagnosed in Bangladesh from different avian species.REF 243Lack <strong>of</strong> efficient replication and induction <strong>of</strong> host innate immune responsesby the novel betacoronavirus EMC in human monocyte derivedmacrophages and dendritic cellsJanne TYNELL 1 , Esa RÖNKKÖ 1 , Veera ARILAHTI 1 , KristerMELÉN 1 , Bart L. HAAGMANS 2 , Ron A.M. FOUCHIER 2 , PamelaÖSTERLUND 1 , Ilkka JULKUNEN 11 National Institute for Health and Welfare, Helsinki, FINLAND; 2 ErasmusMedical Center, Rotterdam, THE NETHERLANDSThe emergence <strong>of</strong> a new human coronavirus (hCoV EMC) causing severeacute respiratory infection has alerted the community for a possibility <strong>of</strong>another epidemic similar to the one caused by SARS coronavirus (SARSCoV) in 2003. HCoV EMC has been shown to be able to replicate inVi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013REF 244Interaction <strong>of</strong> zoonotic paramyxoviruses with chiropteran cells: Acomparative analysis <strong>of</strong> Nipah virus and bat derived Henipa like virusglycoproteinsNadine KRÜGER 1 , Markus HOFFMANN 1 , Marcel AlexanderMÜLLER 2 , Jan Felix DREXLER 2 , Michael WEIS 3 , Karl HeinzESSER 4 , Andrea MAISNER 3 , Christian DROSTEN 2 , GeorgHERRLER 11 Institute <strong>of</strong> <strong>Virology</strong>/University <strong>of</strong> Veterinary Medicine Hannover,Hannover, GERMANY; 2 Institute <strong>of</strong> <strong>Virology</strong>/Bonn Medical Centre,Bonn, GERMANY; 3 Institute <strong>of</strong> <strong>Virology</strong>/Philipps University Marburg,Marburg, GERMANY; 4 Institute <strong>of</strong> Zoology/University <strong>of</strong> VeterinaryMedicine Hannover, Hannover, GERMANYBats have been reported to be a natural reservoir for paramyxoviruses withzoonotic potential, e.g. the Nipah virus (NiV), a member <strong>of</strong> the genus Henipavirus.This virus can lead to severe and <strong>of</strong>ten fatal infections in humanswhereas bats generally do not show any clinical symptoms. The detection<strong>of</strong> henipa like virus RNA in fecal samples <strong>of</strong> African bats or organ samplesfrom Indonesian bats and the successful isolation <strong>of</strong> a bat borne henipavirusin Australia (Cedar virus) raise the question: how likely is such atransmission from a bat virus to the human population? We investigatedthe entry process <strong>of</strong> an African henipa like virus into immortalized chiropteranand non chiropteran cells and compared it to that <strong>of</strong> NiV. We focusedon the interaction <strong>of</strong> the fusion (F) and glycoprotein (G) <strong>of</strong> NiV and henipalike viruses with cultured cell. To analyze their functional activity we performedfusion assays and screened for syncytia formation following coexpression <strong>of</strong> NiV or henipa like F and G proteins in chiropteran and nonchiropteran cell lines. Syncytia formation was reduced or abolished by thetreatment <strong>of</strong> co transfected cells with different cathepsin inhibitors. Forinvestigation <strong>of</strong> the binding ability <strong>of</strong> henipa like G proteins to the ephrinB2 (Eph B2) receptor we performed a co immunoprecipitation analysis.The fusion assay revealed the functional activity <strong>of</strong> henipa like F and G inchiropteran cells in a cathepsin dependent fashion. Furthermore, co immunoprecipitationanalysis indicated that – similar to NiV G henipa like Ginteracts with the Eph B2 receptor.REF 245A Genetic Survey <strong>of</strong> Crimean Congo Hemorrhagic Fever Virus inTicks from Spain in 2011Ana NEGREDO 1 , Fátima LASALA 1 , Eva RAMÍREZ DEARELLANO 1 , María Dolores FERNÁNDEZ 2,1 , Juan Manuel LUQUE 1 ,Miguel Ángel HABELA 3 , Agustín ESTRADA PEÑA 4 , AntonioTENORIO 11 National Centre for Microbiology, Instituto de Salud Carlos III, Madrid,SPAIN; 2 <strong>European</strong> Public Health Microbiology Training Programme(EUPHEM), <strong>European</strong> Centre for Disease Prevention and Control(ECDC), Solna, SWEDEN; 3 University <strong>of</strong> Extremadura, Cáceres, SPAIN;4 University <strong>of</strong> Zaragoza, Zaragoza, SPAINS187
5 th <strong>European</strong> <strong>Congress</strong> <strong>of</strong> <strong>Virology</strong>Crimean Congo hemorrhagic Fever virus (CCHFV) is a zoonotic virusthat causes outbreaks <strong>of</strong> hemorrhagic fever in human. The viral cycleis maintained by ticks <strong>of</strong> Hyalomma spp., serving as vectors and reservoirs<strong>of</strong> the virus. Vertebral animals suffered an asymptomatic infection.Humans are infected through tick bites or contact with the viremic blood<strong>of</strong> patients or livestock. CCHFV belongs to the Nairovirus genus <strong>of</strong> thefamily Bunyaviridae. It is an enveloped RNA virus with a tri segmentedgenome: S (small), M (medium) and L (Large) segments. The analysis <strong>of</strong>the S, M and L phylogenetic trees topologies document seven lineages,usually coherent between the three segments, although segment re assortmentand recombination events also occur. In Spain, CCHFV genome wasdetected in ticks collected from red deer in a hunting area in Cáceres in2010 and the analysis <strong>of</strong> the amplified fragment suggested the presence <strong>of</strong>an African related strain included in Genotype III (Estrada Peña et al 2011).In order to check the temporal persistence and geographical distributionwe collected ticks from different geographic areas <strong>of</strong> Spain. Eleven from211 ticks collected in 2011 rendered positive results using an unpublishednested RT PCR with degenerated primers designed on S segment. All thepositive results were obtained in ticks collected in Cáceres, demonstratingthe persistence <strong>of</strong> virus circulation in the area and no evidence <strong>of</strong> geographicaldispersion. Nucleotide sequence analysis <strong>of</strong> the amplified fragmentrevealed the same viral genotype detected in 2010.REF 246Nairoviruses chronically infected Hyalomma derived cell linesCristiano SALATA 1,2 , Helen KARLBERG 2 , Lesley BELL SAKYI 3 ,Giorgio PALÙ 1 , Ali MIRAZIMI 2,4,51 Department <strong>of</strong> Molecular Medicine, University <strong>of</strong> Padova, Padova,ITALY; 2 Swedish Institute for Communicable Disease Control, Solna,SWEDEN; 3 The Tick Cell Biobank The Pirbright Institute, Pirbright,Woking, Surrey, UNITED KINGDOM; 4 Department <strong>of</strong> Clinical and ExperimentalMedicine, Linköping University, Linköping, SWEDEN; 5 NationalVeterinary Institute, Uppsala, SWEDENCrimean Congo hemorrhagic fever (CCHF) is a zoonotic viral diseasethat is asymptomatic in infected animals, but a serious threat to humans.Human infections begin with nonspecific febrile symptoms that progress toa serious hemorrhagic syndrome with a high case fatality rate. CCHF virus(CCHFV) is classified as a biosafety level 4 pathogen and is the secondmost widespread <strong>of</strong> all the medically important arboviruses after denguevirus. CCHFV has been found in at least 31 species <strong>of</strong> ticks, in particular,members <strong>of</strong> the genus Hyalomma seem to be the principal vectors asthey play an important role as natural reservoir. In the present work, theinfection susceptibility <strong>of</strong> two Hyalomma derived cell lines to CCHFV andHazara virus (HAZV), an apathogenic virus closely related to the CCHFV,has been investigated in order to compare CCHFV and HAZV virus/hostinteractions in the context <strong>of</strong> arthropod cells. Preliminary results showedthat the tick cell lines tested were susceptible to infection without anycytopathic effect. It was also discovered that infected cells release progenyvirus to supernatant for at least 28 days post infection. The real time PCRdata analysis suggest that these cells are persistently infected. Currently,a transcriptomics approach to study the cellular response to CCHFV orHAZV infection is under investigation. The results <strong>of</strong> the comparativestudies on replication <strong>of</strong> CCHFV and HAZAV in tick cells will be hereindiscussed.REF 247Genetic analysis <strong>of</strong> Crimean Congo hemorrhagic fever virus strainsin Northern TurkeyHarun ALBAYRAK 1 , Emre OZAN 2 , Cafer EROGLU 3 , AbdullahCAVUNT 21 Ondokuz Mayis University, Faculty <strong>of</strong> Veterinary Medicine, Department<strong>of</strong> <strong>Virology</strong>, Samsun, TURKEY; 2 Veterinary Control Institute, Department<strong>of</strong> <strong>Virology</strong>, Samsun, TURKEY; 3 Ondokuz Mayis University, Faculty <strong>of</strong>Medicine, Department <strong>of</strong> Microbiology, Samsun, TURKEYCrimean Congo hemorrhagic fever virus (CCHFV) is causative agent <strong>of</strong> atick borne disease with high mortality rates in human. The distribution <strong>of</strong>CCHFV includes over 30 countries in Asia, the Middle East, southeasternEurope, and Africa. Aim <strong>of</strong> the present study was to investigate molecularepidemiology <strong>of</strong> CCHFV in northern Turkey. The study was performed ona total <strong>of</strong> 23 confirmed CCHFV in ticks from 2008 to 2009. The hard tickswere collected from variety <strong>of</strong> mammalian species (cattle, sheep and goat)in northern Turkey (Amasya, Tokat, Sivas, Samsun, Sinop, Ordu and Giresunprovinces). CCHF viral genoms were obtained from seven tick species,and the most abundant were Rhipicephalus turanicus (10/23), Hyalommamarginatum marginatum (5/23), Rhipicephalus bursa (3/23), Hyalommadetritum (2/23), Ixodes ricinus (1/23), H. anatolicum excavatum (1/23), H.anatolicum anatolicum (1/23). All <strong>of</strong> the CCHF viral strains in northernTurkey were found to belong to the <strong>European</strong> lineage. Local CCHF viralstrains are grouped into two cluster under <strong>European</strong> lineage. The majority<strong>of</strong> the CCHF viral strains (21/23) were found in the cluster 1, two <strong>of</strong> themwhich are obtained from Hyalomma detritum and H. anatolicum anatolicumtick species in Alucra location belong to the cluster 2. Pylogeneticanalysis suggest that replication <strong>of</strong> CCHFV in the tick vectors, whetherHyalomma spp., or other species <strong>of</strong> ticks has no effect on the viral genomicstructure.REF 248Epidemiological, behavioral and environmental risk factors associatedwith CCHFV seropositivity in humans in GreeceAnna PAPA, Persefoni SIDIRA, Andreas TSATSARIS, AnastasiaKONTANA, Katerina TSIOKA, Elpida GAVANADepartment <strong>of</strong> Microbiology, Medical School, Aristotle University <strong>of</strong> Thessaloniki,GREECEBackground: crimean Congo hemorrhagic fever (CCHF) is a viral diseasewith fatality 5 30%. Humans are infected through tick bite or by directcontact with blood or tissues <strong>of</strong> viremic patients or animals. Aim <strong>of</strong> thepresent study was to analyze the epidemiological, behavioral and environmentalrisk factors associated with CCHFV seropositivity in humans inGreece.Materials and Methods: The study was based on serological results on3,152 sera, collected from residents from all peripheries <strong>of</strong> Greece testedfor CCHFV specific IgG antibodies. Age <strong>of</strong> the patients was 1-97 years(median 58 y); 43.3% were male. The association between seropositivityand categorical variables was estimated by the chi squared test orFisher’s exact test. Univariate logistic regression analysis was performedto calculate the odds ratio and the 95% confidence intervals, and to identifypossible risk factors for inclusion into the final multivariate model.S188 Vi<strong>rologie</strong>, Vol 17, supplément 2, septembre 2013
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