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Annex 4e Draft scientific protocol for the inventory and monitoring of large carnivores populations in the Poloniny National Park (the European Diploma holding area) Target species: brown bear (Ursus arctos), Eurasian lynx (Lynx lynx), wolf (Canis lupus), ungulates (indirectly). Methods: bait stations, snow tracking, photo-trapping, radio-tracking Collaboration terms: State Nature Conservancy of the Slovak Republic is not directly aimed to the research and scientific activities, thus the organization has long time cooperation with different universities and research institutions, mostly in Slovakia, but in the other countries as well. On the national level bilateral agreements of the cooperation, including research are signed with the relevant institutions. With some others official agreement does not exists, but cooperation is ongoing or planned and could be supported from various EU structural funds. For example – within the frame of the project aimed to the management of European Bison in NP Poloniny was started collaboration between Universities in Italy, of Udine, Padova and Perugia Some of the proposed research and scientific activities is depending on the relevant budget and thus is conditioned by othe sources. Some of the activities are continuously realised and some are included in the management plans for large carnivores. BAIT STATIONS Aim of the study: collect information on bear and lynx presence in different areas, also in function of season, and obtain a genetic map of the population. Genetic characteristics of the brown bear population, through organic material collection (hair) at hair trap sites. Hair traps are made of a barbed wire polygon of about 4-7 m2, nailed at 50 cm high around 3-5 tree trunks. The food bait made of fish is put inside the polygon, both on the soil and hung at 3 m high. The fish bait is covered with dead wood and big stones, in order to estimate the size of the animal visiting the bait from the entity of the bait damage. A handful of corn is spread inside the polygon as reward. Baits are controlled and refreshed every 15 days, for 4 times per session, two sessions/year (spring-summer and summer-autumn). Density of stations: at least 3/10 km2. Genetic characteristics of the lynx population through hair collection at hair trap sites. Lynx hair traps exploit the natural cheek rubbing behaviour of felines. Substances (synthetic feline pheromones and vegetal extracts) are used to elicit this behaviour on the hair traps. Those are made of a carpet square (10 cm2) on which substances are adsorbed and Velcro® (30 cm x 5 cm) to collect hair. The hair trap is nailed on tree trunks at 50 cm high and strengthen by a visual attractant (aluminium pie) hung on a branch at 1,5 m high. A small piece of carpet with the attractants is also hung together with the visual bait in order to increase the spatial effectiveness of the bait station. Bait stations are checked and refreshed at 15 days interval, in two sessions/year (spring-summer and summer-autumn), 4 times/session. Density of stations: at least 3/10 km2. 1 Hair analysis. Hair samples are submitted to a preliminary screening with tricomicroscopy methods, in order to maximize genetic analysis effort. Microscopy slides of each hair samples are prepared and digital photos taken. Characteristic measures are taken from medullar and cuticolar photos. DNA extraction and analysis (species, sex and individual recognition) will be done for all sure and doubt samples (regarding target species). SNOW TRACKING Aim of the study: collect information on animal movements, habitat use, habitual pathways, predation strategy. Useful information about track and footprints characteristic measures can be obtained, in order to discriminate age-sex classes or even individuals. It is a necessary preliminary work for captures and radio-tracking sessions. All species of big carnivores can be detected with snow tracking. Transects are decided a priori and each is covered at least 4 times. The ideal interval between transects and their length is calculated considering the target species, the home range area and the entity of estimated movements between foraging areas. Transect are chosen in order to intercept those movements, therefore are usually along valleys and/or through mountain passes. As en example, regarding the Eurasian lynx in eastern Italy, for an estimated home range of 10 000 ha, the transect should be minimum 8 km long and repeated every 12 days in order to maximize the probability of finding lynx tracks which are not older than 3 days. PHOTO-TRAPPING Aim of the study: collect information on animal site use, food habits, carcass consuming rates, necrofagy and cleptoparassitism. Target species: all mammals and occasionally birds. Photo traps are positioned on fresh predation sites, on bait stations, on other foraging sites, along wildlife trails or in random way. Both digital and 35 mm cameras are used. The photo trap is activated by a movement and infrared sensor. In case of predation, the camera is left on site until the carcass is still present, limiting the visits to the site to the minimum necessary for changing the batteries. For the other cases a priori time of field permanence is decided (i.e. 10 days) for each site. Two examples of photos are reported, one taken at a prey site (the lynx) and the other at a bait station site (the brown bear). 2 Photos: examples of results obtained with 35 mm photo traps, on prey site (lynx) and bait station (brown bear). 3 RADIO-TRACKING Aim of the study: information collection on animal movement and habitat use. Captures are defined after a preliminary monitoring work (bait station, snow-tracking and photo-trapping sessions) in order to maximize capture effort. Lynxes can be captured with box traps on passages or with telenarcosis on predation sites. Bears can be captured with foot snares and telenarcosis. Wolves can be captured with telenarcosis on feeding sites or with box traps. Also capture methods are defined in function of habitat features and in order to minimize animal injuries. GPS system will be used. Number of trapped animals and tracking sessions will be defined in function of personal availability, in order to maximize the quality of collected information. WOLF-HOWLING AND LYNX CALLING Aim of the study: collect information on wolf and lynx presence. For wolves, obtain information on pack composition and presence of juveniles. For lynx, obtain information on territorial habits. Trails and emission points are defined in function of preliminary results of the other monitoring sessions and the local topography. For wolf-howling two sessions/year will be considered, in winter and summer. For the lynx only one session/year will be organized, in winter during the reproductive season (February). Animal answers can be recorded and submitted to sound analysis. OTHER DATA COLLECTED During all field work sessions, in particular during snow tracking, photo trapping sessions and carnivore’s prey analysis, also data on ungulates will be taken. The kind of data collected can be useful to estimate population density, localize foraging areas and evaluate body condition (if necroscopic observations are done on preys). Those data are useful in the understanding of carnivore strategies and habitat use. 4