Tula Virus Infections in the Eurasian Water Vole in Central Europe Mathias Schlegel, 1 Eveline Kindler, 2 Sandra S. Essbauer, 3 Ronny Wolf, 4 Jo ¨ rg Thiel, 5 Martin H. Groschup, 1 Gerald Heckel, 2,7 Rainer M. Oehme, 6 and Rainer G. Ulrich 1 Abstract Recent reports of novel hantaviruses in shrews and moles and the detection of rodent-borne hantaviruses in different rodent species raise important questions about their host range and specificity, evolution, and host adaptation. Tula virus (TULV), a European hantavirus, is believed to be slightly or non-pathogenic in humans and was initially detected in the common vole Microtus arvalis, the East European vole M. levis (formerly rossiaemeridionalis), and subsequently in other Microtus species. Here we report the first multiple RT-PCR de- tection and sequence analyses of TULV in the Eurasian water vole Arvicola amphibius from different regions in Germany and Switzerland. Additional novel TULV S-, M-, and L-segment sequences were obtained from M. arvalis and M. agrestis trapped in Germany at sites close to trapping sites of TULV-RT-PCR-positive water voles. Serological investigations using a recombinant TULV nucleocapsid protein revealed the presence of TULV-reactive antibodies in RT-PCR-positive and a few RT-PCR-negative water voles. Phylogenetic analyses revealed a geographical clustering of the novel S-, M-, and L-segment sequences from A. amphibius with those of M. arvalis- and M. agrestis-derived TULV lineages, and may suggest multiple TULV spillover or a potential host switch to A. amphibius. Future longitudinal studies of sympatric Microtus and Arvicola populations and exper- imental infection studies have to prove the potential of A. amphibius as an additional TULV reservoir host. Key Words: Arvicola amphibius—Central Europe—Hantavirus—Tula virus. Introduction H antaviruses represent a genus of the family Bu- nyaviridae with emerging human pathogenic represen- tatives in Europe, Asia, and the Americas. Despite the problem of isolating and propagating these viruses, in the last three decades the knowledge of European and Asian hantaviruses has broadened significantly. Thus many new hantavirus spe- cies and strains, their host association, and prevalence and the frequency of human infection and disease have been charac- terized. Most importantly, besides the rodent-borne hanta- viruses during recent years a large number of novel hantaviruses were detected in different shrew and mole species in Europe and Asia, with thus far unknown human pathoge- nicity (Song et al. 2007a,2007b,2007c,2009; Arai et al. 2008; Kang et al. 2009). Transmission of hantaviruses is believed to be mainly indirect by inhalation of virus-contaminated rodent- excreta derived aerosols, whereas bites seem to represent a very rare mode of transmission (Scho ¨nrich et al. 2008). On the Eurasian continent hantaviruses may cause hemorrhagic fever with renal syndrome (HFRS) of different severity and case fatality rates of up to 10% (Kru ¨ ger et al. 2011). One representative is the bank vole Myodes glareolus- transmitted Puumala virus (PUUV) distributed in almost all parts of Europe and causing a mild to moderate form of HFRS, designated nephropathia epidemica, with a case fatality rate of < 0.1% (Brummer-Korvenkontio et al. 1999). Second, Dobrava-Belgrade virus (DOBV) genetic lineages DOBV- Af, DOBV-Ap, DOBV-Aa, and Saaremaa associated with the yellow-necked mouse Apodemus flavicollis, Caucasian wood 1 Friedrich-Loeffler-Institut, Institute for Novel and Emerging Infectious Diseases, Greifswald–Insel Riems, Germany. 2 Computational and Molecular Population Genetics (CMPG), Institute of Ecology and Evolution, University of Bern, Bern, Switzerland. 3 Bundeswehr Institute of Microbiology, Department of Virology and Rickettsiology, Munich, Germany. 4 Institute for Biology, University of Leipzig, Leipzig, Germany. 5 Thu ¨ ringer Landesanstalt fu ¨ r Wald, Jagd und Fischerei, Gotha, Germany. 6 Landesgesundheitsamt Baden-Wuerttemberg, Stuttgart, Germany. 7 Swiss Institute of Bioinformatics, Genopode, Lausanne, Switzerland. VECTOR-BORNE AND ZOONOTIC DISEASES Volume 12, Number 6, 2012 ª Mary Ann Liebert, Inc. DOI: 10.1089/vbz.2011.0784 503