UNIVERSITY OF THESSALY SCHOOL OF HEALTH SCIENCES FACULTY OF VETERINARY SCIENCE DEPARTMENT OF MICROBIOLOGY AND PARASITOLOGY ECOLOGY AND MANAGEMENT OF EUROPEAN BROWN HARE SYNDROME IN MEDITERRANEAN ECOSYSTEMS A thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy Christos Κ. Sokos MSc, MSc SUPERVISOR: Charalambos Billinis, Professor KARDITSA 2014 Institutional Repository - Library & Information Centre - University of Thessaly 09/12/2017 09:23:25 EET - 137.108.70.7 UNIVERSITY OF THESSALY SCHOOL OF HEALTH SCIENCES FACULTY OF VETERINARY SCIENCE DEPARTMENT OF MICROBIOLOGY AND PARASITOLOGY ECOLOGY AND MANAGEMENT OF EUROPEAN BROWN HARE SYNDROME IN MEDITERRANEAN ECOSYSTEMS Postgraduate student: Christos Κ. Sokos, Wildlife Ecologist MSc, MSc Supervisor: Charalambos Billinis, Professor, Faculty of Veterinary Science, University of Thessaly Advisory Committee: Periklis Birtsas, Associate Professor, Department of Forestry and Management of Natural Environment, Technological Education Institute of Thessaly Leonidas Leontides, Professor, Faculty of Veterinary Science, University of Thessaly Rodi-Burriel Angeliki, Professor, Faculty of Veterinary Science, University of Thessaly Zissis Mamuris, Professor, Department of Biochemistry & Biotechnology, University of Thessaly Athanasios Sfougaris, Associate Professor, Faculty of Agriculture Crop Production and Rural Envinonment, University of Thessaly Vasiliki Spyrou, Associate Professor, Department of Animal Production, Technological Education Institute of Thessaly KARDITSA 2014 Institutional Repository - Library & Information Centre - University of Thessaly 09/12/2017 09:23:25 EET - 137.108.70.7 Dedicated to Prof. Zygmunt Pielowski A pioneer hare researcher in Europe Czempm - Sp. Prof. Zygmunt Pielowski Institutional Repository - Library & Information Centre - University of Thessaly 09/12/2017 09:23:25 EET - 137.108.70.7 The research described in this PhD thesis was performed at the Laboratory of Microbiology and Parasitology, Faculty of Veterinary Science, University of Thessaly, Greece and received partial funding from the European Union Seventh Framework Programme (2007-2013) under grant agreement no. 222633 (Wild Tech). Institutional Repository - Library & Information Centre - University of Thessaly 09/12/2017 09:23:25 EET - 137.108.70.7 ABSTRACT ECOLOGY AND MANAGEMENT OF EUROPEAN BROWN HARE SYNDROME IN MEDITERRANEAN ECOSYSTEMS A Doctor of Philosophy dissertation by Christos Κ. Sokos MSc, MSc Wildlife Ecologist Supervisor: Charalambos Billinis, Professor, Faculty of Veterinary Science Many aspects of wildlife diseases, especially in Mediterranean ecosystems, including ecological parameters and management practices, remain uncertain. The European brown hare (Lepus europaeus) is a major wildlife species in Mediterranean Basin taking into account its importance as a prey for many carnivore species and as a huntable species. The recorded widespread mortality by European Brown Hare Syndrome (EBHS), indicates this viral disease as the most important disease for hares in Europe. This PhD thesis presents a series of studies conducted with the aim to investigate several aspects of the ecology and management of hare and EBHSV in Macedonia, Hellas. Mediterranean ecosystems are characterized by a variety of ecotopes. The free access of Hellenic hunters in countryside gave the opportunity to study hare harvest and EBHSV prevalence in different ecotopes. During the study, 291 hares(Lepus europaeus) were collected by the hunters. The ecotope, where each hare was initially found by the dogs, was recorded and a population index was calculated. Moreover, the PCR was used for the detection of European Brown Hare Syndrome Virus (EBHSV). The greatest harvest in unit area was recorded in areas with mosaic of cereals and woody vegetation; in contrast the lowest harvest took place in forestlands. EBHSV had widespread distribution even in isolated areas and the prevalence was in an endemic stability of 17.8%. EBHSV has higher prevalence in Olive groves, followed by mixed cultivations and shrublands. Population index is related positively with EBHSV prevalence and harvest. Harvest is not related significantly with EBHSV prevalence. In a second step, a more detailed study was taken place with the contribution of multivariate statistics and Geographic Information Systems (GIS). It was found that prevalence increases in areas with higher hare abundance; closer to 1 Institutional Repository - Library & Information Centre - University of Thessaly 09/12/2017 09:23:25 EET - 137.108.70.7 paved road network and in lower altitudes. Secondly, a potential distribution map was constructed to show the relative risk of EBHSV in the study area. Associations of EBHSV infection, sex, age, body condition and spleen mass were studied in 259 hares collected. Data showed a sex-biased prevalence with twice as many males infected with EBHSV than females, indicating a sexual dimorphism in disease exposure or susceptibility. EBHSV infection was not related to hare body condition and a higher body condition was found in males than in females. Adults had a higher ratio of infected animals than young but this difference was not found to be statistically significant. Adults also had higher spleen mass thus indicating past infections. An enlarged spleen was more frequently found in hares positive for EBHSV and a negative relation was found between spleen mass and body condition thus indicating energy consumption for immune defense investment. Ecosystem disturbances, such as wildfires, are driving forces that determine ecological processes, including diseases. Species respond differentially in wildfires, having diverse post-fire population evolution. Hare relative abundance, age ratio, diet quality, body condition and diseases were determined after a wildfire in Kassandra Peninsula of Chalkidikis. The severe habitat disturbance by wildfire caused initially a reduction in hare abundance, although after one year and for four years hares were more abundant in burned area. Wildfire seems not to influence EBHSV prevalence between burned and unburned areas. Nocturnal spotlight survey is a common census technique for many mammal species and is also used for samples collection. The influence of abiotic factors may be essential for the appropriate design of censuses; however this method has not been examined sufficiently in relation to abiotic factors such as weather conditions and the moon. We tested their influence in the case of hare. Wind speed and direction had an influence on hare counts with stronger winds resulted in fewer juvenile hares being observed, and more hares were counted when wind was blowing from the hare towards the vehicle. No significant effects were found by the other factors probably due to the intense relief and thick vegetation of the studied areas. Hare release is a frequent management measure in some countries, and thousands of hares are released annually, mainly for hunting purposes. Survival rates in the wild are low for hares intensively reared in cages but are higher for hares reared extensively in enclosures and wild translocated hares. The benefits of the hare release 2 Institutional Repository - Library & Information Centre - University of Thessaly 09/12/2017 09:23:25 EET - 137.108.70.7 practice are significantly lower than the action's implementation cost. Hare releases have not increased significantly the wild hare population or the hunting harvest in areas where the practice has been applied. The risk of genetic and evolutionary degradation and pathogen transmission is possible in wild populations. The impact of EBHSV on hare populations appears to be highly variable. These studies have provided documentation on this variability and its parameters. Taking into account these ecological parameters of disease prevalence, the necessities on disease prevention are described more efficiently and the proposal of appropriate management measures becomes possible. 3 Institutional Repository - Library & Information Centre - University of Thessaly 09/12/2017 09:23:25 EET - 137.108.70.7 Acknowledgements Acknowledgements appear to be the final difficult part of writing a doctoral thesis. Firstly I want to thank my supervisor, Professor Dr Charalambos Billinis, who first thought that wildlife ecology and virology can find common field and a PhD on this field is a feasible task. Some believe that when two sciences are coming close then knowledge can be acquired more successfully and Charalambos believed that I can respond to this. I thank also Dr Billinis for his valuable guidance during my study and for the ethical support throughout the course of preparing my PhD thesis. Many thanks are also due to Associate Professor Dr Periklis Birtsas, without his contribution this PhD would not be possible, his professional as well as moral support were important for the completion of this thesis. I thank the members of the Advisory Committee, for their valuable comments. Firstly, I would like to to acknowledge Professor Dr Leonidas Leontides for his valuable corrections on manuscript text and statistics. I also owe special thanks to Associate Professor Dr Athanasios Sfougaris for always finding the time for support of my studies and with important comments on ecology and management of hare. I am so grateful to Dr. Alexis Giannakopoulos for his help and guidance on GIS tools. I am also grateful to Dr. Konstantinos Papaspyropoulos for his important contribution on statistics. I thank also the veterinarians