Ljungan virus (LV), which belongs to the Parechovirus genus in the Picornaviridae family, was first isolated from bank voles (Myodes glareolus) in Sweden in 1998 and proposed as a zoonotic agent. To improve knowledge of the host association and geographical distribution of LV, tissues from 1685 animals belonging to multiple rodent and insectivore species from 12 European countries were screened for LV-RNA using reverse transcriptase (RT)-PCR. In addition, we investigated how the prevalence of LV-RNA in bank voles is associated with various intrinsic and extrinsic factors. We show that LV is widespread geographically, having been detected in at least one host species in nine European countries. Twelve out of 21 species screened were LV-RNA PCR positive, including, for the first time, the red vole (Myodes rutilus) and the root or tundra vole (Alexandromys formerly Microtus oeconomus), as well as in insectivores, including the bicolored white-toothed shrew (Crocidura leucodon) and the Valais shrew (Sorex antinorii). Results indicated that bank voles are the main rodent host for this virus (overall RT-PCR prevalence: 15.2%). Linear modeling of intrinsic and extrinsic factors that could impact LV prevalence showed a concave-down relationship between body mass and LV occurrence, so that subadults had the highest LV positivity, but LV in older animals was less prevalent. Also, LV prevalence was higher in autumn and lower in spring, and the amount of precipitation recorded during the 6 months preceding the trapping date was negatively correlated with the presence of the virus. Phylogenetic analysis on the 185 base pair species-specific sequence of the 5' untranslated region identified high genetic diversity (46.5%) between 80 haplotypes, although no geographical or host-specific patterns of diversity were detected.

CBGP INRAE CIRAD IRD Montpellier SupAgro Université de Montpellier Montpellier France

Center for Agriculture Food Environment C3A University of Trento and Fondazione E Mach San Michele all'Adige Italy

Centre for Infectious Disease Control National Institute for Public Health and the Environment Bilthoven the Netherlands

Comparative Tropical Medicine and Parasitology Ludwig Maximilians Universität Munich Germany

Department for Terrestrial Ecology Norwegian Institute for Nature Research Trondheim Norway

Department of Biodiversity and Molecular Ecology Research and Innovation Centre Fondazione Edmund Mach San Michele all'Adige Italy

Department of Biodiversity Faculty of Mathematics Natural Sciences and Information Technologies University of Primorska Koper Slovenia

Department of Biomedical Sciences and Veterinary Public Health Section for Parasitology Swedish University of Agricultural Sciences Uppsala Sweden

Department of Botany and Zoology Masaryk University Brno Czech Republic

Department of Cellular Computational and Integrative Biology CIBIO University of Trento Povo Italy

Department of Climate Change and Health Public Health England London United Kingdom

Department of Medical Entomology and Zoonoses Ecology Emergency Response Department Public Health England Salisbury United Kingdom

Department of Theoretical and Applied Sciences Università degli Studi dell'Insubria Varese Italy

Department of Veterinary Biosciences University of Helsinki Helsinki Finland

Department of Veterinary Medicine Università degli Studi di Milano Milan Italy

Department of Virology and Immunology University of Helsinki and Helsinki University Hospital Helsinki Finland

Department of Virology Faculty of Medicine University of Helsinki Helsinki Finland

Department of Viroscience Erasmus University Medical Centre Rotterdam the Netherlands

Department of Wildlife Fish and Environmental Studies Swedish University of Agricultural Sciences Umeå Sweden

Ecology and Vegetation Physiology Group Universidad de los Andes Bogotá Colombia

Friedrich Loeffler Institut Federal Research Institute for Animal Health Institute of Novel and Emerging Infectious Diseases Greifswald Insel Riems Germany

Institute of Animal Hygiene and Veterinary Public Health University of Leipzig Leipzig Germany

Institute of Infectology Friedrich Loeffler Institut Federal Research Institute for Animal Health Greifswald Insel Riems Germany

Institute of Vertebrate Biology Academy of Sciences of the Czech Republic Studenec Czech Republic

Istituto Zooprofilattico Sperimentale della Lombardia e Dell'Emilia Romagna Bruno Ubertini Brescia Italy

Natural Resources Institute Finland Helsinki Finland

School of Biosciences Cardiff University Cardiff United Kingdom

Sezione Zoologia dei Vertebrati MUSE Museo delle Scienze Trento Italy

Slovak Academy of Sciences Institute of Parasitology Košice Slovakia

Slovak Academy of Sciences Institute of Zoology Bratislava Slovakia

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