MOD N ASP ENT OF GAME E PO DERN NABL PECT LE MA OPULA LATIO GEME ANAG. din. ngs. University

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ww ww.huntssymposiium.com m M MOD DERN N ASP PECT TS OF F SUSTAIN NABL LE MA ANAG GEME ENT OF GAME G E PO OPULA LATIO ON Proceeed din ngs No ovi Saad, Serrbia, 17-20 Octob O ber, 20013. U University y of Novi Sad, Facculty of Agricultur A re Novvi Sad, Seerbia

2 nd INTERNATIONAL SYMPOSIUM ON HUNTING MODERN ASPECTS OF SUSTAINABLE MANAGEMENT OF GAME PROCEEDINGS Novi Sad, Serbia, 17-20 October, 2013. University of Novi Sad, Faculty of Agriculture Novi Sad, Serbia

ISBN: 978-86-7520-279-0 UDC: 639.1(082) International symposium on hunting Modern aspects of sustainable management of game population Publisher University of Novi Sad, Faculty of Agriculture, Trg Dositeja Obradovića 8, 21000, Novi Sad, Serbia Phone: +38121 485 35 00; www.polj.uns.ac.rs Editor in chief Prof. dr Miloš Beuković, University of Novi Sad, Faculty of Agriculture, Serbia Editors Prof. dr Zoran Popović, University of Belgrade, Faculty of Agriculture, Zemun, Serbia Prof. dr Nenad Đorđević, University of Belgrade, Faculty of Agriculture, Zemun, Serbia Prof. dr Milutin Đorđević, University of Belgrade, Faculty of Veterinary Medicine, Serbia Prof. dr Mihajla Đan, University of Novi Sad, Faculty of Sciences, Serbia mr Dejan Beuković, University of Novi Sad, Faculty of Agriculture, Serbia MScEF Vukan Lavadinović, Albert Ludwig University of Freiburg, Faculty of Environment and Natural Resources, Germany Paper Review All papers reviewed by the international Board of Reviewers Cover mr Dejan Beuković Printed by Stojkov Novi Sad Number of copies 100 copies SUPPORTED BY

Organizing Board: President Prof. dr Miloš BEUKOVIĆ ~ University of Novi Sad, Faculty of Agriculture, Serbia Prof. dr Zoran POPOVIĆ ~ University of Belgrade, Faculty of Agriculture, Zemun, Serbia Prof. dr Nenad ĐORĐEVIĆ ~ University of Belgrade, Faculty of Agriculture, Zemun, Serbi Prof. dr Milutin ĐORĐEVIĆ ~ University of Belgrade, Faculty of Veterinary Medicine, Serbia Prof. dr Mihajla ĐAN ~ University of Novi Sad, Faculty of Sciences, Serbia Symposium secretary: mr Dejan BEUKOVIĆ ~ University of Novi Sad, Faculty of Agriculture, Serbia MScEF Vukan LAVADINOVIĆ ~ Albert Ludwig University of Freiburg, Faculty of Environment and Natural Resources, Germany M.Sc Nevena VELIČKOVIĆ ~ University of Novi Sad, Faculty of Sciences, Serbia M.Sc Nikola PUVAČA ~ University of Novi Sad, Faculty of Agriculture, Serbia kj Organizers University of Novi Sad, Faculty of Agriculture, Serbia University of Novi Sad, Faculty of Sciences, Serbia University of Belgrade, Faculty of Agriculture, Zemun, Serbia University of Belgrade, Faculty of Veterinary Medicine, Serbia Ss Cyril and Methodius University in Skopje, Faculty of Forestry, F.Y. Republic of Macedonia Albert Ludwig University of Freiburg, Faculty ofj Environment and Natural Resources, Germany

Honorary Board: Prof dr. Dragan GLAMOČIĆ Dipl. ecc Danilo GOLUBOVIĆ Prof. dr Radivoje MITROVIĆ mr Nenad KATANIĆ Prof. dr Dragoslav PETROVIĆ Prof. dr Miroslav VESKOVIĆ Prof. dr Milan POPOVIĆ Prof. dr Milica PETROVIĆ Prof. dr Vlado TEODOROVIĆ Prof.dr Neda Mimica DUKIĆ Prof. dr Milan MEDAREVIĆ dr. Al Mahmoud Daghistani MASSOUD Dipl ecc. Igor BRAUNOVIĆ dipl ing. Milan PAŽIN Ministry of Agriculture, Trade, Forestry and Water management, Department of Forestry Government of Republic of Serbia Minister Ministry of Agriculture, Trade, Forestry and Water management, Department of Forestry Government of Republic of Serbia State secretary Ministry of Education and Science Development, Government of Republic of Serbia State secretary Ministry of Agriculture, Trade, Forestry and Water management, Department of Forestry Government of Republic of Serbia Deputy Minister Provincial Secretariat for Science And Technological Development, Autonomous Province of Vojvodina University of Novi Sad, Rector of the University University of Novi Sad, Faculty of Agriculture, Serbia, Dean University of Belgrade, Faculty of Agriculture, Serbia, Dean University of Belgrade, Faculty of Veterinary Medicine, Serbia, Dean University of Novi Sad, Faculty Science, Serbia, Dean Faculty of Forestry, University of Belgrade, Serbia, Dean General secretary, Kennel Club of Republic Serbia State Enterprise for Forest Management "Srbijašume" Belgrade., General Manager Hunt Association of Vоjvodina, Novi Sad, Serbia Symposium supported by: Ministry of Agriculture, Trade, Forestry and Water management Ministry of Education and Science Provincial Secretariat for Science And Technological Development, Autonomous Province of Vojvodina

Scientific Board: Prof. dr Miloš BEUKOVIĆ Prof. dr Rory PUTMAN, professor emeritus Prof. dr Jon SWENSON Prof. dr Zoran POPOVIĆ Prof. dr Milutin ĐORĐEVIĆ Prof. dr Nenad ĐORĐEVIĆ Prof. dr Vladimir MALETIĆ Prof. dr Algimantas PAULAUSKAS Prof. dr Ulrich SCHRAML Prof. dr Ladislav PAULE Prof. dr Sándor CSÁNYI Prof. dr Dragan GVOZDIĆ Prof. dr Duško ĆIROVIĆ Prof. dr Ivan KOS Prof. dr Tihomir FLORIJANČIĆ Prof. dr Hristo MIHAILOV Prof. dr Mile POČUČA Prof. dr Rajko TEPAVAC Prof dr Zoran RISTIĆ Prof. dr Zdravko JANICKI Doc. Ing. Jaroslav SLAMEČKA, CSc Asst. prof. dr. Yusuf Ziya OGRAK Asst. prof. dr. Mihajla ĐAN Asst. prof.dr. Ivan STANČIĆ (United Kingdom) (Norway) (F.Y. Republic of Macedonia) (Lithuania) (Germany) (Slovakia) (Hungaria) (Slovenia) (Croatia) (Bulgaria) (Croatia) (Slovakia) (Turkey) Symposium supported by: Kennel Club of Serbia State Enterprise for Forest Management "Srbijašume" Vectornic Aerospace

2 nd International Symposium on Hunting,»Мodern aspects of sustainable management of game populations«novi Sad, Serbia, 17 20. October, 2013. Original scientific paper UDC: 616.98:599.731.1 THE BACTERIAL INFECTIONS OF RESPIRATORY TRACT OF WILD BOARS Prodanov-Radulović, J. 1, Došen, R., 1 Pušić, I., 1 Stojanov, I., 1 Petrović, T., 1 Urošević, M., 2 Summary: In our country a certain number of wild boars are reared in controlled and enclosed hunting grounds, while a number of free-ranging populations are mainly unknown. The control of health status of wild boar population is quite demanding and it is not easy to achieve. Also, often is not possible entirely to perform a complete diagnostic examination in wild boars in each evaluated case. The aim of this research was to evaluate the significance of bacterial infections on occurrence of respiratory tract disease of wild boars. The material for this research included three hunting grounds in Vojvodina, where clinical signs, health disorders and dead of young categories of wild boars were recorded. The following research methods were applied: epidemiological, clinical and pathological examination of live and shot wild boars, standard laboratory testing for detection the presence of aerobic and anaerobic bacteria in tissue samples (lympho nodes, lung), hystological examination of the lung tissue samples and parasitological examination of the lung tissue samples and feces (isolation, identification and parasite determination). In all examined cases, macropathogical examination revealed changes dominantly in the respiratory tract. By pathohystological and parasitological examination the presence of lung worms in the trachea, bronchi and in posteroventral parts of the diaphragmatic lung lobes were detected. By bacteriological testing on tissue samples derived from diseased shot or dead wild boars the presence of Streptococcus dysgalactiae, Actinobacillus sp., Mannheimiahaemolytica (Pasteurellahaemolytica), Pasteurellamultocidaand Escherichia coli haemolytica was detected. In conclusion, the achieved results strongly suggest that bacterial infections of respiratory tract represent economically important diasease of wild boars in our climatic conditions. In wild boars special attention should be given to the presence of lung worms since this parasite is a factor which favours secondary respiratory infections by bacteria. Key words: wild boars, lung bacterial infection Introduction Number of wild boars (Susscrofascrofa) has dramatically increased over the past 60 years. Because of their wide distribution (Wu et al., 2011) transmission of diseases is possible, but also a higher contact rate between hosts (Ruiz-Fons et al., 2008). Increasing food availability and climatic change provide optimal conditions for a rapid wild boar multiplication and expansion (Wu et al., 2011). The population of wild boar in 15 member states of European Union (EU) has been roughly estimated between 800,000 and 1 milion heads, but its density varies from country to country (Laddomada, 2000). The size of wild boar population is regulated greatly by man and larger predators such as the wolf (Järvis et al., 2007). The increase in population density of wild boars raises concerns regarding the welfare and an increasing prevalence of infectious diseases and parasites (Prodanov-Radulović et al., 2010; Ruiz-Fons et al., 2008). Domestic pigs and wild boars belong to the same species and they share the same pathogens (Wu et al., 2011).Wild boar pathogens are highly relevant not only for the livestock industry but also for wildlife conservation and for the hunting industry. Because several million wild boar are harvested and consumed yearly in Europe, wild boar meat and derivates are a likely source of human infections (Boadella et al., 2012). The parallel increase of outdoor piggeries has led to a higher risk of contacts, and thus of disease transmission, between wild boars and domestic pigs (Prodanov et al., 2009; Wu et al., 2011). 1 Jasna Prodanov-Radulović, PhD Reasearch Associate; Radoslav Došen, MSc, Advisor Specialist; Ivan Pušić, MSc, Advisor Specialist; Igor Stojanov, PhD Senior Reasearch Associate, Scientific Veterinary Institute Novi Sad, Novi Sad, Serbia; 2 Miroslav Urošević, DVM, PhD, Institute of Food Technology, University of Novi Sad, Novi Sad. Corresponding author: Jasna Prodanov-Radulović, Scientific Veterinary Institute Novi Sad, Rumenački put 20, 21000 Novi Sad,; e-mail: jasna.prodanov@gmail.com; phone:++381 21 4895 314 220

2 nd International Symposium on Hunting,»Мodern aspects of sustainable management of game populations«novi Sad, Serbia, 17 20. October, 2013. Material and Methods The material for this research included three hunting grounds in Vojvodina, where clinical signs of health disorders and dead of young categories of wild boars were recorded. The following research methods were applied: epidemiological, clinical and pathological examination of live and dead wild boars, standard laboratory testing for detection the presence of aerobic and anaerobic bacteria in tissue samples (lympho nodes, lung) deriving from dead wild boars, hystological examination of the lung tissue samples and parasitological examination of the lung tissue samples and feces (isolation, identification and determination). Results and Discussion In two hunting grounds (I+II), by epizootiological and clinical examination distinct clinical signs of respiratory infection in 15 young wild boars were detected: dispnea, nasal discharge, intensive coughing ( thumping ). Beside this, the wild boars were emaciated andsigns of slow gait, lagging behind the pack were noticed. The wild boars were with shrunken eyes and tough dry hair. As part of health control programm of clinically diseased wild boars shot by hunters, pathological examination of trunci and internal organs deriving from 10 wild boars was performed. The pathomorphological examination of clinically suspected i.e. all diseased animals shot by hunters revealed changes dominantly in the respiratory tract: purulent nasal discharge, effervescent content in bronchi and bronchioles, mixed with a large number of lung worms, which were like mucoid plugs filling the respiratory pathways. All lobes of the lungs were diffusely swollen, edematous and reddened with marginal emphysema and consolidation. A large amount of clear, foamy fluid and numerous slender, white nematodes 4-7 cm long were visible in the trachea and bronchial trees.the presence of lung worms (Metastrongylus spp.) in the trachea, bronchi and in posteroventral parts of the diaphragmatic lobes were detected(metastrongylosis pulmonum summ). In several cases, pathomorphologically the presence of numerous abscesses (Pneumonia apostematosadisseminata) in lung tissue were discovered. By pathohystological examination of macroscopically changed lung tissue Pneumonia interstitialis was diagnosed. In the most of the cases, applying hystological examination the presence of lung worms in the trachea, bronchi and in posteroventral parts of the diaphragmatic lobes were detected(pneumonia verminosa). Applying parasitological control of the faecal material extracted from the rectum the presence of the several parasites was discovered: Trichuris suis, Oesophagostomum sp. and Hyostrongylus sp. By bacteriological testing on tissue samples (lungs, mediastinal lymphonodes) derived from shot clinically diseased wild boars the presence of Streptococcus dysgalactiae, and Escherichia coli haemolytica was detected. Lung parasites of the genus Metastrongylus are found in wild and domestic pigs worldwide (Garcia- González et al., 2013). They are considered to be one of the most important selective factors acting on wild boar population, increasing the mortality of weaker young and adult animals. Clinically, they may cause dyspnea, bronchopneumonia and permanent weight loss in addition to inflicting tissue damages which allow opportunistic infections of viruses and bacteria (Da Silva and Muller, 2013).Lung worms can significantly undermine the lung tissue integrity and consequently contribute to genesis of observed bacterial lung infections (Prodanov-Radulović et al., 2010). Verminous migration through lung tissue may induce a small hemorrhagic foci, abcesses, or firm granulomas. The verminous processes are mainly located dorsocaudally in the lung (Christensen et al., 1999). This parasite has an indirect life cycle and its transmission occurs through ingestion of intermediate hosts (annelids) (Da Silva and Muller, 2013). Because earthworms are required for their transmission, parasitism is less frequent in indoor pig-rearing facilities. Young wild boars are thought to ingest a higher number of earthworms than adults and therefore may have a higher level of parasitism (Garcia-González et al., 2013). It is considered that the prevalence of infection in wild boars is approximately 50% to 100% whenever it is found (Yoon et al., 2010). An important factor that intensifies the infection of animals with helminths is the density of population. In the population with high density, contacts between wild boars and other definitive and intemediate hosts of helminths become more frequent, facilitating transmission of helminths (Järvis et al., 2007; Prodanov-Radulović et al., 2010). 221

2 nd International Symposium on Hunting,»Мodern aspects of sustainable management of game populations«novi Sad, Serbia, 17 20. October, 2013. The achieved results in our study suggests that lung worms are common in wild boars in our region. In Europe, these parasites also have high prevalence, affecting more than 80% of pigs created in extensive system and considered one of the main causes of respiratory changes of these animals (Da Silva and Müller, 2013). On the third examined hunting ground (III), the case history data revealed the problem in piglets category with clinical signs of growth retardation. Clinically, in 8 diseased piglets the following signs were detected: reluctance to move, fatigued, easy to catch. Recently, a large number of dead piglets was discovered by hunters. As part of health control program 8clinically diseased wild boars (staggering gait, with long bristling hair and arched back)were shot by hunters. Patomorphological examination revealed changes dominantly in the respiratory tract: severe necrotizing pleuropneumonia, the presence of multiple abscesses in the lung tissue (Pneumonia apostemosa disseminata). In few animals, purple to gray areas of consolidation of lung tissue were detected (Pneumonia fibrinosa in statu hepatisatiois rubrae et griseae). Macroscopically the lung lobes were very similar to the hepato or pancreatic tissues. In addition, the trachea and bronchi were filled with a foamy, mucous exsudate mixed with small number of lung worms in the respiratory pathways.by standard bacteriological testing on tissue samples (lungs, mediastinal lympho nodes) Mannheimiahaemolytica (Pasteurellahaemolytica), Pasteurellamultocidaand bacteria from genusactinobacillus (Actinobacillus pleuropneumoniae)were detected. Table 1. Summary of clinical and pathological control in investigated hunting grounds Hunting ground Total number of clinically examined wild boars Total number of clinically diseased wild boars Total number of pathologically examined wild boars I+II 55 15 10 III 25 8 8 Number of detected dominant pathological changes Metastrongylosis pulmonum summ (10) Pneumonia apostematosadisseminata (4) Pleuropneumoniaactinobacillosa (2) Pneumonia apostemosa disseminata (4) Pneumonia fibrinosa (2) Metastrongylosis pulmonum summ (6) The upper respiratory tract is the natural habitat for myriads of commensal microorganismswhich may have a favourable competitive effect for their host in outnumbering pathogenic agents (Christensen et al., 1999; Dosen et al., 2007). Pasteurella multocida probably is the most frequent and damaging invader in the lung. However, these bacteria are tipically secondary invadersand was never isolated in the bronchial tree of healthy pigs. Even the most pathogenic strains are not capable of infecting a healthy lung, unlike Actinobacillus pleuropneumoniae (Pijoan, 1999).It is considered that A. pleuropneumoniaeis an obligate parasite of the porcine respiratory tract and there are no other natural hosts.the clinical and pathologic outcome of infection depends on serovar and virulence factors, but also on age, immunity, hygiene, infectious pressure, breed, and stress factors (Reiner et al., 2010). Pleuropneumonia caused by A. pleuropneumoniae is one of the important bacterial diseases of the respiratory tract of the pig and occurs in most pig-keeping countries (Boadella et al., 2012). The course of pleuropneumonia can range from peracute to chronic, including subclinical cases. Survivors of acute infections become carriers, and the infectious agent is located mainly in necrotic lung lesions and/or in the tonsils, less frequently in the nasal cavity (Pijoan, 1999). Reiner et al. (2010) discovered a wide distribution ofa. pleuropneumoniae among wild boars, with more than one-third of the tested animals infected. 222

2 nd International Symposium on Hunting,»Мodern aspects of sustainable management of game populations«novi Sad, Serbia, 17 20. October, 2013. Picture 1. Mucoido-purulent content filling the respiratory pathways Picture 2. Lung worms in posteroventral parts of the diaphragmatic lobes In our country a certain number of wild boars are reared in controlled and enclosed hunting grounds, while a number of free-ranging populations are mainly unknown. One of the characteristics of outdoor swine production in some regions is raising free-roaming domestic pigs, where they share forest habitat with wild boars (Prodanov et al., 2009). In areas in which traditionally raising free-roaming domestic pigs has been introduced in the woods, hybridization with the wild boars has led to crossbreeding, production which is often reffered to as a feral pig or feral hog (Ruiz-Fons et al., 2008). In the research Prodanov et al. (2009) during the epizootiological and clinical examination, the existance of this type of animal hybrids in the backyards of the owners who practice exstensive grasing was discovered. Monitoring wildlife diseases faces a number of wildlife-specific constaints, including sampling difficulties regarding proper sample and site stratification, consistent samling of the same sites, and limitations of the diagnostic test available for wildlife (Boadella et al., 2012;Morita et al., 2007). The health control of the population of wild boars is very complex and requires applying the specific plan and strategy which takes into consideration the disease characteristics, the wild boar ethology, interaction which occurs between wild boars and free-roaming domestic animals and the interfering human activities (Prodanov et al., 2009; Ruiz-Fons et al., 2008). Conclusion On the bases of necropsy and result of patomorphological examination, it was concluded that the bacterial diseases of the respiratory tract in shot wild boars had characteristics of subacute and chronic course. Pathological examination discovered that in the largest number of animals the health problems were mainly connected to the parasitic infestations of respiratory organs (Pneumonia verminosa), which favoured secondary respiratory infections by bacteria. Animal health surveillance is routinely applied to domestic animals, but limited data exist on the prevalence and distribution of infectious agents of wild boars. To better understanding the role of this species in serving as a reservoir of these diseases, additional epidemiologic data are needed, as well as the isolation and molecular characterization of etiologic agents. 223

2 nd International Symposium on Hunting,»Мodern aspects of sustainable management of game populations«novi Sad, Serbia, 17 20. October, 2013. Acknowledgement This paper is a result of the research within the project TR31084 Wild animal health monitoring and introduction of new biotechnology procedures in detection of infectious and zoonotic agents risk analysis for human health, domestic and wild animal health and for environmental contamination, financed by the Ministry of Education, Science and Technological Development, Republic of Serbia. References 1. Boadella, M., Ruiz-Fons, J., F., Vicente, J., Martin, M., Segalés, J., Gortazar, C., 2012, Transboundary and Emerging Diseases, 59, 395-404. 2. Christensen G., Sorenson V., and Mousing J.: Diseases of the Respiratory System, in: Diseases of Swine. 1999, Eds. B., Straw, S., DʼAllaire, W., L., Mengeling, D., J., Taylor, Iowa State University Press, p 913-940. 3. Da Silva, D., Müller, G., 2013, Parasitol Res, 112:1353-1356. 4. Došen, R., Prodanov, J., Milanov D., Stojanov, I., Pušić, I., 2007, Biotechnol Anim Husb, 23 (5-6), 237 243. 5. Garcia-González, A., M., Pérez-Martin, J., E., Gamito-Santos, J., A., Calero-Bernal, R., Alcaide M., Frontera Carrión, E., M., 2013, Journal of Wildlife Diseases,49(1), 157-162. 6. Higgins R. and Gottschalk M.: Streptococcal Diseases, in: Diseases of Swine. 1999, Eds. B., Straw, S., DʼAllaire, W., L., Mengeling, D., J., Taylor, Iowa State University Press, p 563-578. 7. Järvis, T., Kapel, Ch., Moks, E., Talvik, H., Mägi, E., 2007, Veterinary Parasitology, 150, 366-369. 8. Laddomada, A., 2000, Vet Microbiol., 73, 121-130. 9. Montagnaro, S., Sasso, S., De Martino, L., Longo, M., Iovane, V., Ghiurmino, G., Pisanelli, G., Nava, D., Baldi, L., Pagnini, U., 2010, Journal of Wildlife Diseases, 46(1), 316-319. 10. Morita, T., Haruta, K., Shibata-Haruta, A., Kanda, E., Imai, S., Ike, K., 2007, J. Vet. Med. Sci., 69(4): 417-420. 11. Pijoan C.: Pneumonic Pasteurellosis, in: Diseases of Swine. 1999, Eds. B., Straw, S., DʼAllaire, W., L., Mengeling, D., J., Taylor, Iowa State University Press, p 511-520. 12. Prodanov, J., Došen, R., Pušić, I., Petrović, T., Orlić, D., Maljković, M., Lupulović, D., 2009, Biotechnol Anim Husb 25 (5-6), 879-885. 13. Prodanov-Radulović, J., Došen, R., Pušić, I., Orlić, D., Stojanov, I., Radulović, G., 2010, Arhiv vet med, 3(2), 19-25. 14. Reiner, G., Fresen, C., Bronnert, S., Haak, I., Willems H., 2010, Journal of Wildlife Diseases, 46(2), 551-555. 15. 16. Ruiz-Fons, F., Segales, J., Gortazar, C., 2008, The Veterinary Journal, 158-169. 17. Wu, N., Abril, C., Hinić, V., Brodard, I., Thűr, B., Fattebert, J., Hüssy, D., Ryser-Degiorgis, M-P., 2011, Journal of Wildlife Diseases, 47(4), 868-879. 18. Yoon, B., Kim, H.-C., Kim, J.-T., 2010, Journal of Wildlife Diseases, 46(3), 1052-1054. 224