Copyright © 2005 Billinis, Knowles, Spyrou, Sofianidis, Birtsas, Psychas, Sofia, Maslarinou, Tontis, Kanteres.
Published by: Portuguese Wildlife Society
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Creative Commons Attribution 2.5 Portugal License

• C. Billinis *
  
• N. Knowles *
  Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey, GU24 0NF, United Kingdom.
• V. Spyrou *
  Department of Animal Production, Technological Educational Institution, 41110 Larissa, Greece.
• G. Sofianidis *
  Laboratory of Pathology Faculty of Veterinary Medicine, Aristotle University, 54124 Thessaloniki, Greece.
• P. Birtsas *
  Hunting Federation of Macedonia and Thrace, Ethnikis Antistasis 173 - 175, 55134 Thessaloniki, Greece.
• V. Psychas *
  Laboratory of Pathology Faculty of Veterinary Medicine, Aristotle University, 54124 Thessaloniki, Greece.
• M. Sofia *
  Laboratory of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Thessaly, Trikalon 224, 43100 Karditsa, Greece; Tel.: +30-2441066011; Fax: +30-2441066092
• O. Maslarinou *
  Hunting Federation of Macedonia and Thrace, Ethnikis Antistasis 173 - 175, 55134 Thessaloniki, Greece.
• D. Tontis *
  Laboratory of Pathology, Faculty of Veterinary Medicine, University of Thessaly, Trikalon 224, 43100 Karditsa, Greece.
• D. Kanteres *
  Laboratory of Pathology, Faculty of Veterinary Medicine, University of Thessaly, Trikalon 224, 43100 Karditsa, 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%).

Keywords: EBHS; genetic analysis; Greece; virus.