Genetic Analysis of the First European Brown Hare Syndrome Virus Isolates from Greece

Abstract

Eight European brown hares ( Lepus europaeus ) found dead throughout Greece were examined for the presence of European brown hare syndrome (EBHS). The diagnosis of the disease was established by macro- and microscopical lesions, and RT-PCR analysis. The most common lesions, such as necrotic hepatitis, were observed in the liver, while congestion and haemorrhages were present mainly in lungs and tracheal mucosa. To determine the extent of genetic heterogeneity of the EBHSV isolates, a 265 bp fragment of the capsid protein (VP60) gene of the eight isolates were amplified by RT-PCR and sequenced. Phylogenetic analysis showed that the Greek isolates were indeed EBHS viruses and that EBHSV and RHDV differed by an average of approximately 39%. The maximum nucleotide variation amongst all the EBHS viruses was 15%, whereas variation within the RHDV group amounts to 11%. Apart from GRE-8, all the Greek viruses fell on a single genetic lineage and differed between 1 and 7%; however, bootstrap confidence limits were particularly low on all the branches leading from the base of the EBHSV lineage to GRE-8 (viz. 20, 13 and 9%). The grouping to the remaining Greek isolates was better supported (65%).