The hepatitis C virus (HCV)-related bovine hepacivirus (BovHepV) can cause acute as well as persistent infections in cattle. The true clinical relevance of the virus is not yet known. As reliable antibody detection methods are lacking and prevalence studies have only been conducted in cattle and few countries to date, the true distribution, genetic diversity, and host range is probably greatly underestimated. In this study, we applied several RT-PCR methods and a nano-luciferase-based immunoprecipitation system (LIPS) assay to analyze bovine serum samples from Bulgaria as well as wild ruminant sera from Germany and the Czech Republic. Using these methods, BovHepV infections were confirmed in Bulgarian cattle, with viral genomes detected in 6.9% and serological reactions against the BovHepV NS3 helicase domain in 10% of bovine serum samples. Genetic analysis demonstrated co-circulation of highly diverse BovHepV strains in Bulgarian cattle, and three novel BovHepV subtypes within the genotype 1 could be defined. Furthermore, application of a nested RT-PCR led to the first description of a BovHepV variant (genotype 2) in a wild ruminant species. The results of this study significantly enhance our knowledge of BovHepV distribution, genetic diversity, and host range.

Department of Forest Protection and Wildlife Management Mendel University 61300 Brno Czech Republic

Department of Infectious Diseases Military Medical Academy 1606 Sofia Bulgaria

Department of Microbiology Infectious and Parasitic Diseases Faculty of Veterinary Medicine Trakia University 6000 Stara Zagora Bulgaria

Institute for Terrestrial and Aquatic Wildlife Research University of Veterinary Medicine Hannover 30173 Hannover Germany

Institute of Medical Microbiology Virology and Hygiene University Medical Center Hamburg Eppendorf 20251 Hamburg Germany

Institute of Virology Department of Infectious Diseases University of Veterinary Medicine Hannover 30559 Hannover Germany

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The evolutionary history of hepaciviruses