Noninvasive Genetic Studies of Brown Bears Using Power Poles
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Eur J Wildl Res DOI 10.1007/s10344-010-0363-3 ORIGINAL PAPER Noninvasive genetic studies of brown bears using power poles Alexandros A. Karamanlidis & Elena Drosopoulou & Miguel de Gabriel Hernando & Lazaros Georgiadis & Lambros Krambokoukis & Stavri Pllaha & Andreas Zedrosser & Zacharias Scouras Received: 27 August 2009 /Revised: 13 December 2009 /Accepted: 13 January 2010 # Springer-Verlag 2010 Abstract One difficulty in the conservation of endangered samples, evenly distributed throughout six study areas. wildlife is the lack of reliable information on its status. This During 87 sampling sessions in the main study area, we lack of knowledge can often be attributed to financial and collected 191 samples and identified six microsatellite loci logistic constraints as well as the lack of trained personnel to that were variable enough for individual bear identification. collect data. We test a simple method to study bears in the The most and best-quality hair samples were collected during southern Balkans by inspecting power poles, which are used the mating period, and DNA was most successfully extracted by bears for marking and rubbing purposes. We created a from samples remaining <4 weeks in the field. In the six network of barbed-wire fitted poles for the collection of hair study areas, we identified 47 bears. An advantage of using power poles for hair sampling is their availability and accessibility; no bait is required, and the network can be Communicated by H. Kierdorf easily set up. A drawback may be an unequal capture * : : A. A. Karamanlidis: ( ) M. de Gabriel Hernando probability of sex and age classes of bears. Despite this L. Georgiadis L. Krambokoukis limitation, using power poles proved to be a simple and ARCTUROS, Civil Society for the Protection and Management of Wildlife and the Natural Environment, cheap method for the noninvasive genetic study of bears that Roggoti Str. 3, did not require any prior knowledge on habitat use and 54624 Thessaloniki, Greece activity patterns. The method is suitable for large-scale e-mail: [email protected] surveys to estimate distribution and relative densities of bears E. Drosopoulou : Z. Scouras and could also be applied for studying other species. Department of Genetics, Development and Molecular Biology, School of Biology, Faculty of Sciences, Keywords Conservation . DNA . Endangered species . Aristotle University of Thessaloniki, Greece . Management . Methodology. Southern Balkans . 54124 Thessaloniki, Greece Ursus arctos S. Pllaha Transborder Wildlife Association, Rr. Pandeli Cale Nr. 26, Introduction Korçë, Albania : A. A. Karamanlidis A. Zedrosser Several species of terrestrial mammals are critically Department of Ecology and Natural Resource Management, endangered, and some will perish unless effective conser- Norwegian University of Life Sciences, vation measures are implemented soon (Ceballos and P.O. Box 5003, 1432 Ås, Norway Ehrlich 2002; Schipper et al. 2008). Large carnivores are A. Zedrosser often species of particular conservation concerns (Boitani Department of Integrative Biology and Biodiversity Research, 2000; Breitenmoser et al. 2000; Swenson and Sandegren Institute of Wildlife Biology and Game Management, 2000). Despite costly recovery efforts, our general knowl- University of Natural Resources and Applied Life Sciences, Gregor-Mendel Str. 33, edge over the status and biology of many large carnivores 1180 Vienna, Austria still remains incomplete (Karanth and Chellam 2009), and Eur J Wildl Res wildlife ecologists and conservation biologists are constant- study area of 630 km2 in the Prefecture of Grevena, in the ly in search of new methods to monitor and protect them central part of the Pindos Mountain Range in northwestern more efficiently (Long et al. 2008). Greece (Fig. 1). Major forest vegetation types consisted of Noninvasive genetic monitoring has emerged as a sensitive, oak (Quercus sp.) and black pine (Pinus nigra). The area reliable, and time- and cost-efficient tool to deal with the was characterized by a mosaic of dense forests, openings, inherent difficulties of studying rare, elusive, and often and small-scale agricultural cultivations. Elevations ranged endangered animals (Schwartz et al. 2006). Traps for collect- between 500 and 2,200 m, and human activity was intense ing hairs for subsequent genetic analysis have been success- at lower elevations. This pilot study area was located at fully used across a wide range of species and geographical the center of the species’ western nucleus of distribution in locations (Belant 2003;Longetal.2007; Castro-Arellano et Greece (Mertzanis 1994) and is the construction site of al. 2008). To increase the reliability of methods and results of two major highways (Karamanlidis and Mertzanis 2003; genetic studies, Waits and Paetkau (2005) recommended Karamanlidis and Georgiadis 2009). carrying out pilot studies prior to initiating a new study in order to determine the optimal study method. Study area in the southern Balkans for the large-scale The brown bear (Ursus arctos) is the most widespread testing of the method bear species in the world, with a Holarctic distribution in Europe, Asia, and North America (Servheen et al. 1999). In We tested the protocol on a wider geographical scale in Europe, human persecution and habitat loss have led to the areas of known and expected bear distribution within disappearance of the species from large parts of its original Greece, Albania, and FYROM (Mertzanis 1994; Zedrosser range (Zedrosser et al. 2001). Bear populations in the et al. 2001; Karamanlidis et al. 2007; Fig. 1). In addition to southern Balkans, i.e., in Albania, the Former Yugoslav resampling our pilot study area, we selected areas at the Republic of Macedonia (FYROM), and Greece, are prefectures of Florina and Trikala and at Mount Grammos, considered to be particularly endangered, not only because within the western nucleus of the distribution of the species of their small sizes but also due to the lack of systematic in the country (Mertzanis 1994; Fig. 1). The study area in and coordinated efforts to study and protect them Florina was located north of the pilot study area, was (Zedrosser et al. 2001). Considering the socioeconomic 1,017 km2 large, and was densely populated by humans. changes currently taking place throughout the region and Mount Grammos is a sparsely populated, high mountain their potential effects on habitat connectivity (Karamanlidis massif northwest of the pilot study area; the study site there and Mertzanis 2003; Karamanlidis and Georgiadis 2009), was 740 km2. The study site in Trikala was 708 km2 and was there is an urgent need for a method to rapidly and reliably separated from the pilot study area by an interstate road. obtain data on the status and distribution of the species. Study areas in Albania (496 km2)andFYROM Marking and rubbing behavior of brown bears on poles of (5,615 km2) were selected in the southern parts of both the electricity and telephone network (hereafter referred to as countries adjacent to the borders with Greece (i.e., the study power poles) was recorded in Greece in 2003, and a method areas in both countries constituted an extension of the study was developed to document the presence of the species in the areas at Mount Grammos and the prefecture of Florina, and country. As a result of the marking and rubbing behavior of the habitat was very similar), in order to collect information the species, bear hair was commonly found on power poles that could eventually be used in the transborder manage- (Karamanlidis et al. 2007). We have used the hair left from ment and protection of the species (Fig. 1). this behavior to study brown bears with a noninvasive genetic method. The specific aims of our study were to: Sampling procedures of the pilot study 1. Design a protocol for the systematic noninvasive genetic Power poles are made of wood (approximately 30–50 cm in study of brown bears using hairs found on power poles. diameter and 10 m high) and processed with a wood 2. Test the protocol on a large geographic scale in the preservative (i.e., coal tar creosote, a substance derived southern Balkans. from tars) to resist decay and insect damage. Depending on topography, poles are usually placed 50 to 100 m apart, and vegetation is cleared 5 m from each side of the power pole Materials and methods line. At the beginning of our pilot study, we inspected all 841 power poles present in the pilot study area and Pilot study area classified them according to the intensity of their use (i.e., how intensively they were used for marking or/and rubbing The protocol for our systematic noninvasive genetic study by bears) by applying the classification system developed of brown bears was developed and initially tested in a pilot by Karamanlidis (2008; Table 1). Power poles (N=39) Eur J Wildl Res Fig. 1 Map of the southern Balkans indicating the locations of the six study areas in Albania, the Former Yugoslav Republic of Macedonia (FYROM), and Greece (Florina, Grammos, Grevena, Trikala), the power poles inspected, and the poles that were used as sampling stations for genetic sampling evenly distributed across the study area were selected for the number of existing guard hairs (sample category 0: no the creation of a noninvasive, genetic sampling network, guard hairs in the sample, I: one to five guard hairs in the and whenever possible we used poles with a high intensity sample, II: six to ten guard hairs in the sample, III: more of use. Each power pole/sampling station