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RESEARCH ARTICLE

Evidence for European brown hare syndrome virus introduction with translocated brown hares (Lepus europaeus): implications for management of restocking operations

Vassiliki Spyrou A , Costas Stamatis B , Periklis Birtsas C , Vassilios Psychas D , Katerina Manolakou E , Charalambos Billinis F G H and Zissis Mamuris B H
+ Author Affiliations
- Author Affiliations

A Department of Animal Production, Technological Educational Institute, Larissa, Greece.

B Department of Biochemistry & Biotechnology, University of Thessaly, 26 Ploutonos Street, 41221 Larissa, Greece.

C Department of Forestry and Management of Natural Environment, Technological Education Institute of Larissa, Karditsa, Greece and Hunting Federation of Macedonia and Thrace, Thessaloniki, Greece.

D Laboratory of Pathology, Faculty of Veterinary Medicine, Aristotle University, Thessaloniki, Greece.

E Department of Animal Husbandry and Nutrition, Faculty of Veterinary Medicine, University of Thessaly, Karditsa, Greece.

F Laboratory of Microbiology and Parasitology, Faculty of Veterinary Medicine, University of Thessaly, Karditsa, Greece.

G Department of Biomedicine, Institute for Research and Technology of Thessaly, Larissa, Greece.

H Corresponding author. Email: zmamur@bio.uth.gr; billinis@vet.uth.gr

Wildlife Research 40(7) 545-551 https://doi.org/10.1071/WR12152
Submitted: 27 August 2012  Accepted: 5 November 2013   Published: 5 December 2013

Abstract

Context: Criticisms of wildlife restocking operations typically focus on concerns that translocations can lead to the introduction of pathogens, and risk the integrity of locally adapted genetic diversity. Restocking programs aiming to stabilise population declines of European brown hares using captive-bred individuals have been carried out in several European countries, including Greece.

Aims: To assess the potential for imported hares to introduce novel strains of European brown hare syndrome virus (EBHSV) during restocking operations, by (1) inferring the origin of wild Greek hares on the basis of their mitochondrial DNA (mtDNA) haplotype, (2) screening the hares to detect and characterise EBHSV, and (3) determining whether certain hare origin–EBHSV combinations occur in the wild.

Methods: RNA extraction, polymerase chain reaction (PCR) amplification, and sequence and phylogenetic analyses of EBHSV were performed on 53 hares. Diagnostic RFLP markers of the mtDNA were used to infer the origins of sampled hares.

Key results: Thirty-three hares had ‘typical’ native Greek haplotypes and 20 had mtDNA haplotypes matching those found in imported and released hares. Twelve of the latter and none of the former were positive for EBHSV. Phylogenetic analysis showed that nine virus isolates formed a single genetic lineage distinct from northern–central European ones. Three virus sequences from three imported reared-and-released hares, from Chalkidiki, were closely related to the northern–central European EBHS viruses.

Conclusions: Alien strains of EBHSV are co-introduced with released captive-bred animals, possibly resulting in negative impacts on populations of Greek hares that have not evolved resistance to these novel virus strains.

Implications: The identification of these allopatric EBHSV strains has led the authorities to ban captures and transportations of local brown hares for any restocking operation. We consider it imperative to reinforce microbiological and genetic controls before further releases of captive-bred game species in the wild in Greece.

Additional keywords: disease risk, genetic identification, traceability.


References

Angelici, F. M., Riga, F., Boitani, L., and Luiselli, L. (2000). Fate of captive reared brown hares Lepus europaeus released at a mountain site in central Italy. Wildlife Biology 6, 173–178.

Ballou, J. D. (1993). Assessing the risks of infectious diseases in captive breeding and reintroduction programs. Journal of Zoo and Wildlife Medicine 24, 327–335.

Balmford, A., Mace, G. M., and Leader-Williams, N. (1996). Designing the ark: setting priorities for captive breeding. Conservation Biology 10, 719–727.
Designing the ark: setting priorities for captive breeding.Crossref | GoogleScholarGoogle Scholar |

Billinis, C., Psychas, V., Tontis, D., Spyrou, V., Birtsas, P., Sofia, M., Likotrafitis, F., Maslarinou, O., and Kanteres, D. (2005a). European brown hare syndrome in wild European brown hares from Greece. Journal of Wildlife Diseases 41, 783–786.
European brown hare syndrome in wild European brown hares from Greece.Crossref | GoogleScholarGoogle Scholar | 16456168PubMed |

Billinis, C., Knowles, N. J., Spyrou, V., Psychas, V., Birtsas, P. K., Sofia, M., Maslarinou, O. M., Tontis, D. K., and Kanteres, D. (2005b). Genetic analysis of the first European brown hare syndrome isolates from Greece. Wildlife Biology in Practice 1, 118–127.
Genetic analysis of the first European brown hare syndrome isolates from Greece.Crossref | GoogleScholarGoogle Scholar |

Capucci, L., Scicluna, M. T., and Lavazza, A. (1991). Diagnosis of viral hemorrhagic disease of rabbits and the European brown hare syndrome. Revue Scientifique et Technique de l’Office International des Epizooties 10, 347–370.
| 1:STN:280:DyaK387gtlehsQ%3D%3D&md5=8e839a2caaa73ed44998a5df208f412fCAS |

Chasey, D., Lucas, M., Westcott, D., and Williams, M. (1992). European brown hare syndrome in the UK; a calicivirus related to but distinct from that of viral haemorrhagic disease in rabbits. Archives of Virology 124, 363–370.
European brown hare syndrome in the UK; a calicivirus related to but distinct from that of viral haemorrhagic disease in rabbits.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK383ps12jsQ%3D%3D&md5=43704c5588f9da7123f6c9d79ba07f4aCAS | 1318711PubMed |

Cunningham, A. A. (1991). Bovine spongiform encephalopathy and British zoos. Journal of Zoo and Wildlife Medicine 22, 304–308.

Davidson, W. R., and Nettles, V. F. (1992). Relocation of wildlife: identifying and evaluating disease risks. Transactions of the North American Wildlife and Natural Resources Conference 57, 466–473.

Duff, G. P., Chasey, D., Munro, R., and Wooldridge, M. (1994). European brown hare syndrome in England. The Veterinary Record 134, 669–673.
European brown hare syndrome in England.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK2M%2FislKgtA%3D%3D&md5=303cca5f45791068579bb03fbdb49020CAS |

Feldman, D. B., and Seely, J. C. (1988). Chapter 6. Necropsy of the rabbit (Oryctolagus cuniculus). In ‘Necropsy Guide: Rodents and the Rabbit’. pp. 133–160. (CRC Press: Boca Raton, FL.)

Fiechter, A. (1988). Survie et dispersion de lièvres importés et de levrauts d’élevage lachés. Supplementi alle Ricerche di Biologia della Selvaggina 14, 271–299.

Frölich, K., Meyer, H. H. D., Pielowski, Z., Ronsholt, L., von Seck-Lanzendorf, S., and Stolte, M. (1996). European brown hare syndrome in free-ranging hares in Poland. Journal of Wildlife Diseases 32, 280–285.
European brown hare syndrome in free-ranging hares in Poland.Crossref | GoogleScholarGoogle Scholar | 8722266PubMed |

Frölich, K., Haerer, G., Bacciarini, L., Janovsky, M., Rudolph, M., and Giacometti, M. (2001). European brown hare syndrome in free-ranging European brown and mountain hares from Switzerland. Journal of Wildlife Diseases 37, 803–807.
European brown hare syndrome in free-ranging European brown and mountain hares from Switzerland.Crossref | GoogleScholarGoogle Scholar | 11763744PubMed |

Gavier-Widén, D., and Mörner, T. (1993). Descriptive epizootiological study of European brown hare syndrome in Sweden. Journal of Wildlife Diseases 29, 15–20.
Descriptive epizootiological study of European brown hare syndrome in Sweden.Crossref | GoogleScholarGoogle Scholar | 8383252PubMed |

Ginsberg, J. R. (1994). Captive breeding, reintroduction and the conservation of canids. In ‘Creative Conservation: Interactive Management of Wild and Captive Animals’. (Eds P. J. S. Olney, G. M. Mace and A. T. C. Feistner.) pp. 365–383. (Chapman & Hall: London.)

Gortazar, C., and De Luco, D. F. (1995). La enfermedad hemorrgia de la liebre. Trofeo 295, 30–34.

Green, K. Y., Ando, T., Balayan, M. S., Berke, T., Clarke, I. N., Estes, M. K., Matson, D. O., Nakata, S., Neill, J. D., Studdert, M. J., and Thiel, H. J. (2000). Taxonomy of the caliciviruses. The Journal of Infectious Diseases 181, S322–S330.
Taxonomy of the caliciviruses.Crossref | GoogleScholarGoogle Scholar | 10804145PubMed |

Gulland, F. M. D. (1995). The impact of infectious diseases on wild animal populations – a review. In ‘The Ecology of Infectious Diseases in Natural Populations’. (Eds B. T. Grenfell and A. P. Dobson.) pp. 20–51. (Cambridge University Press: Cambridge, UK.)

Hasegawa, M., Kishino, M., and Yano, T. (1985). Dating the human–ape split by a molecular clock of mitochondrial DNA. Journal of Molecular Evolution 22, 160–174.
Dating the human–ape split by a molecular clock of mitochondrial DNA.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2MXmtFSns7g%3D&md5=3288e186152b4e19db313649f6b5c203CAS | 3934395PubMed |

Henriksen, P., Gavier, D., and Elling, F. (1989). Acute necrotizing hepatitis in Danish farmed hares. The Veterinary Record 125, 486–487.
Acute necrotizing hepatitis in Danish farmed hares.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK3c%2FosFKrsg%3D%3D&md5=eb6e42b1118c6ebaeef1a29dbd8b1f9fCAS | 2595970PubMed |

Hodder, K. H., and Bullock, J. M. (1997). Translocation of native species in the UK: implications for biodiversity. Journal of Applied Ecology 34, 547–565.
Translocation of native species in the UK: implications for biodiversity.Crossref | GoogleScholarGoogle Scholar |

Huelsenbeck, J. P., and Ronquist, F. (2001). Mrbayes: Bayesian inference of phylogeny. Bioinformatics 17, 754–755.
Mrbayes: Bayesian inference of phylogeny.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3MvotV2isw%3D%3D&md5=839ca7f249196ec3121ebf01a650925dCAS | 11524383PubMed |

Hutchins, M., Foose, T., and Seal, U. S. (1991). The role of veterinary medicine in endangered species conservation. Journal of Zoo and Wildlife Medicine 22, 277–281.

Mamuris, Z., Sfougaris, A. I., and Stamatis, C. (2001). Genetic structure of Greek brown hare (Lepus europaeus) populations as revealed by mtDNA–PCR analysis: implications for conserving genetic diversity. Biological Conservation 101, 187–196.
Genetic structure of Greek brown hare (Lepus europaeus) populations as revealed by mtDNA–PCR analysis: implications for conserving genetic diversity.Crossref | GoogleScholarGoogle Scholar |

Mamuris, Z., Sfougaris, A. I., Stamatis, C., and Suchentrunk, F. (2002). Genetic structure of Greek brown hare (Lepus europaeus) populations based on the random amplified polynorphic DNA (RAPD) method. Biochemical Genetics 40, 323–338.
Genetic structure of Greek brown hare (Lepus europaeus) populations based on the random amplified polynorphic DNA (RAPD) method.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XntFSgu70%3D&md5=6b2c15218d208254374a396b588050daCAS | 12392170PubMed |

Marcato, P. S., Benazzi, C., Galeotti, M., and Salda Della, L. (1989). Infective necrotic hepatitis of leporids. Riv Coniglic 26, 41–50.

Mathews, F., Orros, M., McLaren, G., Gelling, M., and Foster, R. (2005). Keeping fit on the ark: assessing the suitability of captive bred animals for release. Biological Conservation 121, 569–577.
Keeping fit on the ark: assessing the suitability of captive bred animals for release.Crossref | GoogleScholarGoogle Scholar |

May, R. M. (1988). Conservation and disease. Conservation Biology 2, 28–30.
Conservation and disease.Crossref | GoogleScholarGoogle Scholar |

McLean, I. G., Lundie-Jenkins, G., and Jarman, P. J. (1996). Teaching an endangered mammal to recognize predators. Biological Conservation 75, 51–62.
Teaching an endangered mammal to recognize predators.Crossref | GoogleScholarGoogle Scholar |

Misiorowska, M., and Wasilewski, M. (2012). Survival and causes of death among released brown hares (Lepus europaeus Pallas, 1778) in central Poland. Acta Theriologica 57, 305–312.
Survival and causes of death among released brown hares (Lepus europaeus Pallas, 1778) in central Poland.Crossref | GoogleScholarGoogle Scholar | 23002287PubMed |

Modesto, P., Cava, P. L., Peletto, S., Colussi, S., Vidus Rosin, A., Meriggi, A., and Acutis, P. L. (2011). Evaluation of genetic variability in brown hare populations from two protected areas in northern Italy. Hystrix, The Italian Journal of Mammalogy 22, 257–268.

Morisse, J. P. (1988). Haemorrhagic septicemia syndrome in rabbits: first observations in France. Le Point Veterinaire 20, 79–83.

Nauwynck, H., Callebaut, P., Peeters, J., Ducatelle, R., and Uyttebroek, E. (1993). Susceptibility of hares and rabbits to Belgian isolate of European brown hare syndrome virus. Journal of Wildlife Diseases 29, 203–208.
Susceptibility of hares and rabbits to Belgian isolate of European brown hare syndrome virus.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK3s3ltlymug%3D%3D&md5=b15335c42ef1d75e129cea07c22c34dfCAS | 8387610PubMed |

Palumbi, S. R., Martin, A., Romano, S., McMillan, W. O., Stice, L., and Grabowski, G. (1991). ‘The Simple Fool’s Guide to PCR.’ (Department of Zoology, University of Hawaii: Honolulu, HI.)

Posada, D., and Crandall, K. A. (1998). Modeltest: testing the model of DNA substitution. Bioinformatics 14, 817–818.
Modeltest: testing the model of DNA substitution.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXktlCltw%3D%3D&md5=0aa3ac3f27e7fa504a0b65962714c6a6CAS | 9918953PubMed |

Salmela, P., Belak, K., and Gavier-Widén, D. (1993). The occurrence of European brown hare syndrome in Finland. Acta Veterinaria Scandinavica 34, 215–217.
| 1:STN:280:DyaK2c%2FptFaltw%3D%3D&md5=744013ea3c27868af7a5985716d9aaffCAS | 8266901PubMed |

Scott, M. E. (1988). The impact of infection and disease on animal populations: implications for conservation biology. Conservation Biology 2, 40–56.
The impact of infection and disease on animal populations: implications for conservation biology.Crossref | GoogleScholarGoogle Scholar |

Smith, R. K., Vaughan Jennings, N., and Harris, S. (2005). A quantitative analysis of the abundance and demography of European hares Lepus europaeus in relation to habitat type, intensity of agriculture and climate. Mammal Review 35, 1–24.
A quantitative analysis of the abundance and demography of European hares Lepus europaeus in relation to habitat type, intensity of agriculture and climate.Crossref | GoogleScholarGoogle Scholar |

Sostaric, B., Lipej, Z., and Fuchs, R. (1991). The disappearance of free living hares in Croatia: European brown hare syndrome. Veterinarski Arhiv 61, 133–150.

Stamatis, C., Suchentrunk, F., Sert, H., Triantaphyllidis, C., and Mamuris, Z. (2007). Genetic evidence for survival of released captive-bred brown hares (Lepus europaeus) during restocking operations in Greece. Oryx 41, 548–551.
Genetic evidence for survival of released captive-bred brown hares (Lepus europaeus) during restocking operations in Greece.Crossref | GoogleScholarGoogle Scholar |

Stamatis, C., Suchentrunk, F., Moutou, K. A., Giacometti, M., Haerer, G., Djan, M., Vapa, L., Vukovic, M., Tvrtković, N., Sert, H., Alves, P. C., and Mamuris, Z. (2009). Phylogeography of the brown hare (Lepus europaeus) in Europe: a legacy of south-eastern Mediterranean refugia? Journal of Biogeography 36, 515–528.
Phylogeography of the brown hare (Lepus europaeus) in Europe: a legacy of south-eastern Mediterranean refugia?Crossref | GoogleScholarGoogle Scholar |

Studdert, M. J., and Symes, S. J. (2008). Caliciviruses. In ‘Encyclopedia of Virology’. 3rd edn. (Eds B. W. J. Mahy and M. H. V. van Regenmortel). pp. 410–419. (Elsevier: Oxford, UK.)

Swofford, D. L. (1998). ‘PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods). Version 4.’ (Sinauer Associates: Sunderland, MA.)

Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., and Higgins, D. G. (1997). The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25, 4876–4882.
The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXntFyntQ%3D%3D&md5=833a7f47d93b6a88a34f7c8d9b5c9b77CAS | 9396791PubMed |

Viggers, K. L., Lindenmayer, D. B., and Spratt, D. M. (1993). The importance of disease in reintroduction programmes. Wildlife Research 20, 687–698.
The importance of disease in reintroduction programmes.Crossref | GoogleScholarGoogle Scholar |

Woodford, M. H. (2001). ‘Quarantine and Health Screening Protocols for Wildlife Prior to Translocation and Release into the Wild.’ (IUCN Species Survival Commission’s Veterinary Specialist Group, Office International des Epizooties, Care for the Wild and the European Association of Zoo and Wildlife Veterinarians: Gland, Paris, UK, Switzerland.)

Woodford, M. H., and Rossiter, P. B. (1993). Disease risks associated with wildlife translocation projects. Revue Scientifique et Technique de l’Office International des Epizooties 12, 115–135.
| 1:STN:280:DyaK3s3pvV2qsw%3D%3D&md5=b9f244edb3afec502c1763d852efdb28CAS |