Genetic analysis of the first European brown hare syndrome virus isolates from Greece

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 ofgenetic heterogeneity of the EBHSVisolates, a 265 bp fragment of the capsidprotein (VP60) gene of the eight isolates were amplified by RT-PCR and sequenced. Phylogenetic analysis showed that the Greek isolates were indeedEBHS viruses and that EBHSVand RHDVdiffered by an average ofapproximately 39%. The maximum nucleotide variation amongst all the EBHS viruses was 15%, whereas variation within the RHDVgroup amountsto 11%. Apart from GRE-8, all the Greek viruses fell on a single geneticlineage and differed between 1 and 7%; however, bootstrap confidence limits were particularly low on all the branches leading from the base of the EBHSVlineage to GRE-8 (viz. 20, 13 and 9%)isolates was better supported (65%).