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Abstract

Conservation and management of large carnivores is often hampered by the lack of information of basic biological parameters. This is particularly true for brown bears (Ursus arctos) in the Former Yugoslav Republic (FYR) of Macedonia. The bear population in this country is important, as it links bear populations of the central part of the Dinaric–Pindos population and the endangered population to the south in Greece. The aim of this study was to assess bear presence in FYR Macedonia and to provide the first evaluation of the genetic status of the species in this country. Bear presence was assessed through a questionnaire and sign surveys, while the genetic status of the species was evaluated through noninvasive genetic sampling from power poles and microsatellite analysis. The results of the study indicate the continuous and permanent presence of brown bears in FYR Macedonia from the border to Kosovo in the northwest, along the border to Albania and Greece in the south; bear presence around Mount Kožuf in the south of the country was seasonal. High levels of genetic diversity were recorded, and it appears that this bear population is currently not threatened by low genetic variability. Cross-border movements of bears between FYR Macedonia and Greece were documented, indicating the presence of an interconnected population and outlining the necessity for a coordinated international approach in the monitoring and conservation of the species in southeastern Europe.
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... Usually, when sampling for hair collection, the area was divided into grids so that at least one hair trap was placed in an established grid [33,62,78,81,86,116]. The fixed points were specific for hair-collection methods, while hair samples were also opportunistically collected from areas with bear frequency, such as certain power poles used for rubbing [83][84][85]117], rub trees [78], or buckthorn patches during the berries ripening season [86]. ...
... The relevant studies are described in Table 3, which illustrates the type of bear hair traps and the operating procedures from different study locations. Some studies have used natural rubs, taking advantage of the bears' natural behaviour to rub on wooden and/or power poles [81,[83][84][85]117]. The use of power poles as hair traps is not feasible everywhere, e.g., in Albania; although some of are made of wood, others are concrete, and bears do not rub on them [85]. ...
... Prior to the laboratory analyses, some studies recommend freezing the hair samples [142]. In Macedonia, the envelopes were placed in Ziplock bags with silica gel [84], whereas in Slovakia and Italy, the hair material was stored either in paper envelopes or in 70% [108] or 95% ethanol [54]. In the studies on the Pasvik population and in Banff National Park, the hair material was stored dry in paper envelopes [36,62]; some were labelled with barcodes [36,100]. ...
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Genetic monitoring has proven helpful in estimating species presence and abundance, and detecting trends in genetic diversity, to be incorporated in providing data and recommendations to management authorities for action and policy development. We reviewed 148 genetics research papers conducted on the bear species worldwide retrieved from Web of Science, SCOPUS, and Google Scholar. This review aims to reveal sampling methodology and data collection instructions, and to unveil innovative noninvasively genetic monitoring techniques that may be integrated into the genetic monitoring of a large bear population. In North American studies, hair samples were collected more often than faeces, whereas in Europe, both faeces and hair samples surveys are recommended, usually focusing on faeces. The use of the Isohelix sample collection method, previously tested locally and, if suitable, applied at the national level, could generate numerous advantages by reducing shortcomings. Additionally, dogs trained for faeces sampling could be used in parallel with hunting managers, foresters, and volunteers for sample collection organised during autumn and winter. It was stated that this is the best period in terms of cost-efficiency and high quality of the gathered samples. We conclude that large-scale noninvasive genetic monitoring of a large bear population represents a challenge; nevertheless, it provides valuable insights for biodiversity monitoring and actions to respond to climate change.
... In most studies using a DNA trapping strategy (90%), researchers employed bait or lures to increase the yield of their traps. Very few studies used non-lured DNA traps, for example, barb wire placed at sites used by brown bears (Ursus arctos) (Karamanlidis et al., 2014;Quinn et al., 2014) or modified body snares at otter (Lontra canadensislatin) latrine sites, to collect hair (Godwin et al., 2015). Although it seems perfectly legitimate (and often essential) to increase the attractiveness of DNA traps with food (Cohen et al., 2013), scent marks from other individuals (Anile et al., 2012) or other attractants (e.g. ...
... Pictograms represent a non-exhaustive list of examples for which references are given below. From left to right and top to bottom: whole faeces sampling for species that use faecal territory marking (Modave et al., 2017), hairs collected in snow (Cullingham et al., 2016), hairs collected with unbaited barbed wire (Karamanlidis et al., 2014), DNA trap baited to attract animals (Duenas et al., 2015), skin swabbing in the field without capture (Morinha et al., 2014), capture of reptiles for buccal swabbing (Huang et al., 2014), gun darting of big mammals to collect tissue sample (Proffitt et al., 2015), biopsy on handled invertebrate (López et al., 2007). ...
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The use of DNA data is ubiquitous across animal sciences. DNA may be obtained from an organism for a myriad of reasons including identification and distinction between cryptic species, sex identification, comparisons of different morphocryptic genotypes or assessments of relatedness between organisms prior to a behavioural study. DNA should be obtained while minimizing the impact on the fitness, behaviour or welfare of the subject being tested, as this can bias experimental results and cause long-lasting effects on wild animals. Furthermore, minimizing impact on experimental animals is a key Refinement principle within the ‘3Rs’ framework which aims to ensure that animal welfare during experimentation is optimised. The term ‘non-invasive DNA sampling’ has been defined to indicate collection methods that do not require capture or cause disturbance to the animal, including any effects on behaviour or fitness. In practice this is not always the case, as the term ‘non-invasive’ is commonly used in the literature to describe studies where animals are restrained or subjected to aversive procedures. We reviewed the non-invasive DNA sampling literature for the past six years (380 papers published in 2013-2018) and uncovered the existence of a significant gap between the current use of this terminology (i.e. ‘non-invasive DNA sampling’) and its original definition. We show that 58% of the reviewed papers did not comply with the original definition. We discuss the main experimental and ethical issues surrounding the potential confusion or misuse of the phrase ‘non-invasive DNA sampling’ in the current literature and provide potential solutions. In addition, we introduce the terms ‘non-disruptive’ and ‘minimally disruptive’ DNA sampling, to indicate methods that eliminate or minimise impacts not on the physical integrity/structure of the animal, but on its behaviour, fitness and welfare, which in the literature reviewed corresponds to the situation for which an accurate term is clearly missing. Furthermore, we outline when these methods are appropriate to use.
... used genetic data from 312 brown bears (98 females and 214 males) sampled during the period from 2003 to 2010 in western Greece, which were used in previous studies to assess the size and structure of the local bear population (Karamanlidis et al. 2015b(Karamanlidis et al. , 2018. We also included in the study data from four individual bears that were identified close to the borders of Greece, in southern Albania (Karamanlidis et al. 2014a) and Northern Macedonia (Karamanlidis et al. 2014b). The laboratory procedures and protocols for DNA extraction, microsatellite analysis, errorchecking, and quality assurance, have been previously described (Karamanlidis et al. 2012(Karamanlidis et al. , 2015b(Karamanlidis et al. , 2018. ...
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Despite increasing habitat fragmentation, large carnivore populations in parts of Europe have been recovering and expanding into human-dominated areas. Knowledge of animal dispersal patterns in such areas is important for their conservation, management, and coexistence with humans. We used genetic data based on 15 microsatellite markers from 312 individuals (98 females, 214 males) to assess kinship and dispersal patterns during the recovery and spatial expansion of a wild brown bear (Ursus arctos) population (2003–2010) in the human-dominated landscape of Greece. We hypothesized that bear dispersal in Greece was sex-biased, with females being more philopatric and males dispersing more frequently and over greater distances. Dispersal indeed was sex-biased, with males dispersing more frequently and farther than females. Overall, females were found to be philopatric; males also appeared to be philopatric, but to a lesser degree. However, a high proportion of females displayed dispersal behavior, which may be indicative of a pre-saturation stage of the population in that part of the country. Our results indicate that dispersal may be due to evading competition and avoiding inbreeding. We also documented long-distance dispersal of bears, which is considered to be indicative of a spatially expanding population. Our results highlight the value of using noninvasive genetic monitoring data to assess kinship among individuals and study dispersal patterns in human-dominated landscapes. Brown bears remain threatened in Greece; we therefore recommend systematic genetic monitoring of the species in combination with careful habitat management to protect suitable habitat (i.e., dispersal corridors) and ultimately ensure co-existence with humans and survival of brown bears in the country.
... Recent population recovery and range expansion have likely resulted in population admixture of these two putative subpopulations. In Vitsi-Varnoundas, bears are either another example of a small isolated fragment that survived in the region or a relatively new occurrence; they are genetically distinct from Pindos bears and could have been formed by an expansion of bears from the FYR of Macedonia and/or Albania (Karamanlidis et al. 2014b). In this case the Vitsi-Varnoundas area could be part of a "source-sink" system, where the reduced density of a peripheral population was met with migration from a core population, in this case probably the robust bear populations in the North Dinarics, in Bosnia and Herzegovina, Croatia and Slovenia (Skrbinšek et al. 2012a, c). ...
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Understanding the mechanisms and patterns involved in population recoveries is challenging and important in shaping conservation strategies. We used a recovering rear-edge population of brown bears at their southernmost European range in Greece as a case study (2007-2010) to explore the recovery genetics at a species' distribution edge. We used 17 microsatellite and a mitochondrial markers to evaluate genetic structure, estimate effective population size and genetic diversity, and infer gene flow between the identified subpopulations. To understand the larger picture, we also compared the observed genetic diversity of each subpopulation with other brown bear populations in the region. The results indicate that the levels of genetic diversity for bears in western Greece are the lowest recorded in southeastern Europe, but still higher than those of other genetically depauperate bear populations. Apart from a complete separation of bear populations in eastern and western Greece, our results also indicate a considerable genetic sub-structuring in the West. As bear populations in Greece are now recovering, this structure is dissolving through a "recovery cascade" of asymmetric gene flow from South to North between neighboring subpopulations, mediated mainly by males. Our study outlines the importance of small, persisting populations, which can act as "stepping stones" that enable a rapid population expansion and recovery. This in turn makes their importance much greater than their numeric or genetic contribution to a species as a whole.
... Considering that jackal density in core areas can increase significantly in a very short time (Szabó et al., 2009) and that the species in the region has a positive relationship to human settlements (Giannatos, 2004;Giannatos et al., 2005), golden jackal-human conflicts might increase in the near future and impede the full recovery of the species in the country. The ultimate goal should be the development and implementation of a species Action Plan (Giannatos, 2004) that will be in line with the conservation efforts for the other carnivores in the country (Lescureux and Linnell, 2010;Karamanlidis et al., 2014) and safeguard the future of the golden jackal in FYR Macedonia. ...
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Golden jackals have been expanding across Europe in recent decades. In the former Yugoslav Republic (FYR) of Macedonia they have been considered extinct since the 1960s. Using a questionnaire survey and a camera trap study we provide the first unequivocal evidence for the re-establishment of the species in the country. The questionnaire survey indicated that golden jackals were present in the western and possibly present in the eastern parts of the country. The camera trap study recorded golden jackals on 155 occasions in central and north-western FYR Macedonia; five different individuals were identified, including one pup, while most jackal images were recorded during the night and morning hours. The results of the study are set in context to the limited information on the status and biology of the species and the urgent need for effective management and conservation actions for the golden jackal in FYR Macedonia.
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In the original article,Box 1 was not printed in its entirety, and two references were badly quoted in page 3 of the pdf and were missing in the reference list.
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By the beginning of the twentieth century, many brown bear populations in Europe were on the brink of extinction due to relentless hunting pressure and habitat loss. The situation was critical also in Estonia, where in the 1920s the population went through a severe demographic bottleneck. Thanks to the protective measures implemented in the 1930s, the population started to recover. However, the process has been slow, especially in the western and southern areas of the country. To study the effects of the demographic bottleneck, we analysed 216 brown bear samples from throughout their main range in Estonia. In combination with widely used methods of population genetics, a recently developed spatially explicit analysis (distribution of residual dissimilarity, DResD) was also applied. Three genetic clusters were revealed, of which two were most likely founded by the survivors of the bottleneck. The DResD analysis revealed several contact zones near the Estonian-Russian border, suggesting that the third cluster was influenced by gene flow from the neighbouring population in Russia. The DResD analysis revealed also a male biased movement corridor along the forested south-north axis in the central part of Estonia. In comparison to other European populations, the genetic diversity of the Estonian population is relatively low and is comparable with other populations that have gone through a severe bottleneck. This work has important implications for brown bear conservation and highlights once again the dangers of excessive hunting.
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Technical Report
2018-έργο LIFE15NAT/GR/001108-τεχνική αναφορά δράσης Α3. Θεσσαλονίκη, Νοέμβριος 2018 Ταυτοποίηση και χωροθέτηση τομέων υψηλού κινδύνου σύγκρουσης-αλληλεπίδρασης ανθρώπου-αρκούδας (περιοχή Δ. Αμυνταίου).
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Technical Report
The present technical report is the deliverable from project’s LIFE15NAT/GR/001108 action A2. Action’s 2 main objectives are: (a) the evaluation of presence, distribution and population size of the brown bear (Ursus arctos. L.) population in the areas of Amyndaio and Florina. To answer the aforementioned objectives, (3) combined approaches and methodological protocols have been implemented as follows: (a)evaluation of presence, distribution and circadian activity with the of IR phototraps, (b) evaluation of presence and distribution with the collection of bear biosigns and (c) evaluation of the population size, genetic variablitiy and genetic balance using DNA analyses and fingerprinting. As regards to (a): a network of 12 IR camera traps was deployed over a 4 months sampling period in the area of Amyndaio. After processing 30.078 shots/frames and 17.238 videos we determined the relative abundance of bear presence in the study area as well as the circadian activity of the species which was also correlated to human presence and activity. We observed that the relative abundance of bears appears higher in specific habitat sectors of the study area (a fact that was also validated through the biosigns sampling). This relative abundance appears to increase in relation to human presence and activity which is related to anthropogenic food resources attractive to bears. This relative abundance index decreases as long as the distance from forest edges and rivers and streams increases. Bears showed mostly nocturnal activity patterns in relation to human presence and activity. Humans and bears presence and activity showed a low degree of temporal overlapping (especially as regards to shepherds and livestock raising activity). Regarding approach (c ): genetic analyses and fingerprinting has shown that : the indigenous brown bear sub-population in the study area suffers of mediocre levels of genetic variability, which appear slightly lower compared to other Greek bear sub-populations but slightly higher than certain European brown bear populations. The population size Νc was estimated at 154 individuals which is 3 times higher than the estimated effective minimum population (Νe=54). However this result has to be treated with caution as according to the software creators the estimation accuracy of Nc strongly depends on the capture/recapture rate which has to be around 2 or more. In our case this value hardly exceeds an average of 1,36. A more intensive sampling that will take place under action D5 is expected to better validate with higher accuracy the brown bear population size in the study area. Finally the high index values for inbreeding in combination with the low value of Ne make the targeted brown bear subpopulation vulnerable over time, a fact that should urge conservationists and managers to undertake appropriate measures to slow down and or reverse this trend and secure the viability of this sub-population.
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