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Biotic interactions are expected to influence species’ responses to climate change, but they are usually not included when predicting future range shifts. We assessed the importance of biotic interactions to understand future consequences of climate and land use change for biodiversity using as a model system the brown bear (Ursus arctos) in Europe. By including biotic interactions using the spatial variation of energy contribution and habitat models of each food species, we showed that the use of biotic factors considerably improves our understanding of the distribution of brown bears. Predicted future range shifts, which included changes in the distribution of food species, varied greatly when considering various scenarios of change in biotic factors, warning about future indirect climate change effects. Our study confirmed that advancing our understanding of ecological networks of species interactions will improve future scenarios of biodiversity change, which is key for conserving biodiversity and ecosystem services.

Aim: Macroecological studies that require habitat suitability data for many species often derive this information from expert opinion. However, expert- based informa-tion is inherently subjective and thus prone to errors. The increasing availability of GPS tracking data offers opportunities to evaluate and supplement expert- based in-formation with detailed empirical evidence. Here, we compared expert- based habitat suitability information from the International Union for Conservation of Nature (IUCN) with habitat suitability information derived from GPS-tracking data of 1,498 individu-als from 49 mammal species.Location: Worldwide.Time period: 1998–2021.Major taxa studied: Forty- nine terrestrial mammal species.Methods: Using GPS data, we estimated two measures of habitat suitability for each individual animal: proportional habitat use (proportion of GPS locations within a habi-tat type), and selection ratio (habitat use relative to its availability). For each individual we then evaluated whether the GPS- based habitat suitability measures were in agree-ment with the IUCN data. To that end, we calculated the probability that the ranking of empirical habitat suitability measures was in agreement with IUCN's classification into suitable, marginal and unsuitable habitat types.Results: IUCN habitat suitability data were in accordance with the GPS data (> 95% probability of agreement) for 33 out of 49 species based on proportional habitat use estimates and for 25 out of 49 species based on selection ratios. In addition, 37 and 34 species had a > 50% probability of agreement based on proportional habitat use and selection ratios, respectively.Main conclusions: We show how GPS-tracking data can be used to evaluate IUCN habitat suitability data. Our findings indicate that for the majority of species in-cluded in this study, it is appropriate to use IUCN habitat suitability data in macro-ecological studies. Furthermore, we show that GPS-tracking data can be used to identify and prioritize species and habitat types for re-evaluation of IUCN habitat suitability data. expert opinion, GPS, habitat suitability, habitat type, habitat use, IUCN, mammals, movement, selection ratio, telemetry

Quantifying animal movements is necessary for answering a wide array of research questions in ecology and conservation biology. Consequently, ecologists have made considerable efforts to identify the best way to estimate an animal’s home range, and many methods of estimating home ranges have arisen over the past half century. Most of these methods fall into two distinct categories of estimators that have only recently been described in statistical detail: those that measure range distributions (methods such as Kernel Density Estimation that quantify the long-run behavior of a movement process that features restricted space use) and those that measure occurrence distributions (methods such as Brownian Bridge Movement Models and the Correlated Random Walk Library that quantify uncertainty in an animal movement path during a specific period of observation). In this paper, we use theory, simulations, and empirical analysis to demonstrate the importance of applying these two classes of space use estimators appropriately and distinctly. Conflating range and occurrence distributions can have serious consequences for ecological inference and conservation practice. For example, in most situations, home-range estimates quantified using occurrence estimators are too small, and this problem is exacerbated by ongoing improvements in tracking technology that enable more frequent and more accurate data on animal movements. We encourage researchers to use range estimators to estimate the area of home ranges and occurrence estimators to answer other questions in movement ecology, such as when and where an animal crosses a linear feature, visits a location of interest, or interacts with other animals.Open Research StatementTracking data on Aepyceros melampus, Beatragus hunteri, Bycanistes bucinator, Cerdocyon thous, Eulemur rufifrons, Glyptemys insculpta, Gyps coprotheres, Madoqua guentheri, Ovis canadensis, Propithecus verreauxi, Sus scrofa, and Ursus arctos are publicly archived in the Dryad repository (Noonan et al. 2018; https://doi.org/10.5061/dryad.v5051j2), as are data from Procapra gutturosa (Fleming et al. 2014a; https://doi.org/10.5061/dryad.45157). Data on Panthera onca were taken from (Morato et al. 2018). Additional data are publicly archived in the Movebank repository under the following identifiers: Canis latrans, 8159699; Canis lupus, 8159399; Chrysocyon brachyurus, 18156143; Felis silvestris, 40386102; Gyps africanus, 2919708; Lepus europaeus, 25727477; Martes pennanti, 2964494; Panthera leo, 220229; Papio cynocephalus, 222027; Syncerus caffer, 1764627; Tapirus terrestris, 443607536; Torgos tracheliotus, 2919708; and Ursus americanus, 8170674.

Environmental contaminants like arsenic (As), cadmium (Cd), mercury (Hg) or lead (Pb) may disrupt hypothalamic–pituitary–adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axes due to their endocrine toxicity potential. Resulting long-term physiological stress or adverse effects on wildlife reproduction and ontogeny may cause detrimental effects at the individual and population levels. However, data on environmental metal(loid)sʼ impact on reproductive and stress hormones in wildlife, especially large terrestrial carnivores, are scarce. Hair cortisol, progesterone and testosterone concentrations were quantified and modelled with hair As, Cd, total Hg, Pb, biological, environmental and sampling factors to test for potential effects in free- ranging brown bears (Ursus arctos) from Croatia (N=46) and Poland (N=27). Testosterone in males (N=48) and females (N=25) showed positive associations with Hg and an interaction between Cd and Pb, but a negative association with interaction between age and Pb. Higher testosterone was found in hair during its growth phase compared to quiescent phase. Body condition index was negatively associated with hair cortisol and positively associated with hair progesterone. Year and conditions of sampling were important for cortisol variation, while maturity stage for progesterone variation (lower concentrations in cubs and yearlings compared to subadult and adult bears). These findings suggest that environmental levels of Cd, Hg and Pb might influence the HPG axis in brown bears. Hair was shown to be a reliable non-invasive sample for investigating hormonal fluctuations in wildlife while addressing individual and sampling specificities.

The relative importance of environmental factors and species pool in natural regeneration processes is still intensively disputed. Is environmental filtering especially important for species with higher requirements for temperature and soil fertility? Is the seed source limitation especially pronounced among tree species with lower dispersal abilities? Trees, seedlings and saplings measurements were conducted in 600 plots of 0.05 ha distributed in a regular grid in the Tatra Mountains (elevation range 817–1797 m a.s.l.). Boosted regression trees were used to analyse the relationships between the numbers of young trees, factors acting as environmental filters and the potential seed source availability. The most important factor affecting the distribution of young trees of most species was elevation; bedrock type was the second most important factor for Acer pseudoplatanus, while potential solar radiation was important for Sorbus aucuparia. The relationship between the presence of mature trees and abundance of young ones was strong in Fagus sylvatica, moderately strong in A. pseudoplatanus, and weak in Picea abies, Abies alba and S. aucuparia. The role of environmental factors in shaping the abundance of young trees varies strongly among species, while the seed source availability is very important for large-seeded trees.

Sex ratio (ratio between the number of male and female individuals in a population) is an important factor contributing to the survival rates of the population in wild. Imbalanced sex ratio is often the cause of diminished survival rates of endangered species. The analysis of DNA collected through noninvasive methodologies effectively allows to monitor populations of rare species or species that are difficult to observe in the wild. Such method was used to determine the sex ratio in populations of the capercaillie (Tetrao urogallus) and black grouse (Lyrurus tetrix) in the Tatra National Park. We collected over 250 samples (mostly excrement) in the area of the Tatra National Park and identified species via microsatellite genotyping and determined the sex using fragments of the CHD gene. We found that the majority of the samples belonged to male specimens irrespective of the species suggesting a large deviation from an equal sex ratio in wild populations of capercaillie and black grouse in the Tatra National Park. Our results point out the fundamental contribution of genetic assessment of sex ration to monitor the viability of both species in the Tatra National Park.

The European silver fir (Abies alba Mill.) has the largest distribution area of all the European species of fir. It is the only species of this type found in the Carpathian Mts., including their highest range - the Tatras, where it constitutes one of the main components of lower montane forests. In certain sections of the Carpathian Mountains fir stands are affected by the competitive pressure of beech. This may be due to climate change, as well as biotic factors such as plant diseases. One such disease is Herpotrichia needle browning. The goal of the present research was to assess the occurrence of Herpotrichia needle browning in the Tatra National Park and determine the species composition of fungi colonizing symptomatic needles and shoots. Symptoms of the disease were observed across the entire research period, i.e., from May to mid-September 2019. They included the wilting of the needles with subsequent mycelial growth on the remains, as well as shoot deformation. The mycological analysis of the needles and shoots of A. alba resulted in the isolation of 22 fungal taxa. In the case of the needles and shoots, which showed symptoms of infection, the most frequent species of fungus was Sydowia polyspora (Bref. & Tavel) E. Müll., followed by Alternaria alternata (Fr.) Keissl. and Rhizosphaera macrospora Gourb. & M. Morelet. Herpotrichia needle browning seems to be triggered by a complex disease resulting from synergistic interaction of several fungal pathogens

Knowledge about breeding biology is often incomplete in species with complex reproductive strategies. The brown bear Ursus arctos is a polygamous seasonal breeder inhabiting a wide variety of habitats and environmental conditions. We compiled information about brown bear breeding season dates from 36 study areas across their distribution range in the Palearctic and Nearctic regions and investigated how their breeding phenology relates to geographical factors (latitude, photoperiod, altitude and region). Brown bear matings were observed for 8 months, from April to November, with a peak in May%July. We found a 59-day difference in the onset of bear breeding season among study areas, with an average 2.3 days delay for each degree of latitude northwards. The onset of the breeding season showed a strong relationship with photoperiod and latitude, but not with region (i.e. Palearctic vs Nearctic) and altitude. First observa- tions of bear mating occurred earlier in areas at lower latitudes. Photoperiod ranged between 14 and 18 hours at the beginning of the season for most of the study areas. The duration of the breeding season ranged from 25 to 138 days among study areas. None of the investigated factors was related to the length of the breeding season. Our results support the relevance of photoperiod to the onset of breeding, as found in other ursids, but not a shorter breeding season at higher latitudes, a pattern reported in other mammals. Our findings suggest a marked seasonality of bear reproductive behaviour, but also certain level of plasticity. Systematic field observations of breeding behaviour are needed to increase our knowledge on the factors determining mating behaviour in species with complex systems and how these species may adapt to climate change.

The aim of this study was to investigate the temporal trends and geographical differences in 90Sr and stable element (Ba, Ca, Mn, Sr, Pb, Zn) levels in the bones of Croatian and Polish brown bear (Ursus arctos) populations. Experimental data suggest that in the decades after nuclear weapon tests and the Chernobyl accident, 90Sr bone activity concentrations decreased from 352 to 11 Bq kg−1 in the Croatian bear population (period 1982–2015) and from 831 to 27 Bq kg−1 in Polish bears (period 1962–2020). Calculated effective and ecological half-lives were 9 and 13 years for Croatian bears, and 15 and 31 years for Polish bears, respectively. Different temporal trends were noted in levels of Ba, Mn, Pb and Zn between the two countries with majority of bones having lower Pb, Sr and Zn in Croatian than in Polish bears. Estimated values for the soil-to-bear transfer of 90Sr were the same order of magnitude in the studied populations. Contrary to this, the estimated transfer of stable Sr was an order of magnitude lower for the Croatian bear population compared to Polish bears. The observed differences in soil-to-bear transfer between stable Sr and 90Sr found for Croatian bears might suggest the need for careful consideration on the use of stable Sr data as an analogue for 90Sr. To our knowledge, this is the first study that analysed 90Sr activity in tissue of brown bears. As such, it provides insight into the fate and behaviour of one of the most relevant anthropogenic radionuclides at the top of the food chain.

The terrestrial habitat is contaminated with non- essential metal(loid)s (e.g. arsenic, cadmium, mercury, lead) known to have potential for endocrine toxicity, among others adverse health effects. Impairment of the hypothalamic-pituitary- gonadal (HPG) axis, including steroid hormone homeostasis, may heavily impact individuals and populations, while depleting reproduction, development and adaptation to environmental stress. Organic and inorganic (mercury) pollutant- related changes in reproductive hormones in ursids have thus far been investigated and reported only for polar bear populations. We investigated associations of hair arsenic, cadmium, mercury and lead with progesterone and testosterone levels, taking into account biological and sampling- related variables in two populations of free- living European brown bear (Ursus arctos), Dinara- Pindos (N=46) and Carpathian (N=27), captured during the 2010-2019 period. Metal(loid)s were quantified using inductively coupled plasma mass spectrometry (ICP-MS) and hormones were measured via enzyme immunoassay (EIA). The best among generalized linear models (GLM) constructed with the above predictor variables were selected comparing the Akaike Information Criterion (AIC) values. For progesterone in hair, the breeding period and body condition index (BCI), but not metal(loid)s were significant variables explaining 24% of variation. As expected, cubs and yearlings exhibited lower progesterone then subadult and adult bears in both the non-breeding (beta=0.427, 85% confidence interval - CI: 0.207 to 0.646) and breeding period (beta=-0.295, CI: -0.407 to -0.183). Bears with higher BCI appeared to have lower progesterone in hair (beta=-0.062, CI: -0.100 to -0.025). Hair testosterone variation (30%) was explained by the hair growth period (beta=0.065, CI: 0.319 to 0.097 ; higher testosterone in growth phase compared to quiescent phase), Hg (beta=0.036, CI: 0.006 to 0.065), interaction of age and Pb (beta=-0.948, CI: -1.54 to -0.360) and interaction of Pb and Cd (beta=0.903, CI: 0.080 to 1.73). Our findings confirmed the well-known influence of biological variables on reproductive hormones, and pinpointed complex interaction of contaminant metal(loid)s with brown bear testosterone levels.

Dormice monitoring was carried out between 2014 and 2019 at 14 areas located in various regions of the Tatra National Park in Poland. The areas were located in various habitats: beech forests and spruce forests in the low mountain zone; spruce forests, windbreaks and mountain pine (Pinus mugo) thickets in the high mountain zone. The choice of the area for monitoring resulted from the habitat type and technical possibilities enabling regular inspections. Each area was equipped with 30 nest boxes dedicated to dormice. The boxes were hung on trees at a height of 1.5 to 2 meters, only in mountain pine thickets. In habitats with a dense cover of European Red Raspberries (Rubus idaeus) the boxes were hung lower, on branches or on specially constructed poles. The use of boxes was determined on the basis of the presence of dormice or their nests. A comparison of nest box use shows that the Hazel Dormouse has specific habitat preferences in mountainous areas. This species was most frequently found in windbreaks, often those with a high proportion of trees destroyed after an outbreak of the spruce bark beetle (Ips typographus). These habitats are characterized by extensive undergrowth with numerous raspberries, Common Rowan (Sorbus aucuparia), Rosebay Willow Herb (Epilobium angustifolium) and spruce undergrowth (Picea abies). Beech forest (Fagus sylvatica) forms a habitat which is not preferred, but is unavoidable. Negative selectivity was demonstrated in relation to the spruce and mountain pine forests that are dominant in the Tatra National Park. The Forest Dormouse (Dryomys nitedula) was found at only one plot in the fourth year of observation. It was located in a small patch of beech in the vicinity of the site where this species was detected in 2011.

This aim of the survey was to record indirectly using nested-PCR test with faecal samples the presence ofFrom February to June 2019, experienced staff of TNP randomly collected fox and wolf faeces on and off hiking trails at altitudes from 850 m to 2,000 m above sea level. In total, 91 faecal samples from red foxes and 19 from wolves were collected. Genomic DNA was obtained by direct extraction from faecal samples using a commercial kit, and from taeniid eggs retrieved from the same samples after flotation.A nested PCR screening of 91 red fox faeces indicated the prevalence ofThe current results show that humans might be exposed to a risk of fox tapeworm infection in nature, even at high altitude inan alpine zone, in an environment contaminated by roaming red foxes encouraged by food leftovers on mountain trails.

Accurately quantifying species’ area requirements is a prerequisite for effective area‐based conservation. This typically involves collecting tracking data on species of interest and then conducting home‐range analyses. Problematically, autocorrelation in tracking data can result in space needs being severely underestimated. Based on the previous work, we hypothesized the magnitude of underestimation varies with body mass, a relationship that could have serious conservation implications. To evaluate this hypothesis for terrestrial mammals, we estimated home‐range areas with global positioning system (GPS) locations from 757 individuals across 61 globally distributed mammalian species with body masses ranging from 0.4 to 4000 kg. We then applied block cross‐validation to quantify bias in empirical home‐range estimates. Area requirements of mammals 1, meaning the scaling of the relationship changed substantially at the upper end of the mass spectrum., Article impact statement: Due to autocorrelation‐induced bias, conventional methods severely underestimate the area requirements of GPS‐tracked large mammals.

Cambiophagous insects, fires and windthrow cause significant forest disturbances, generating ecological changes and economical losses. The bark beetle (Ips typographus L.), inhabiting coniferous forests and eliminating weakened trees, plays a key role in posing a threat to tree stands, which are dominated by Norway spruce (Picea abies) and covers a large part of mountain areas, as well as the lowlands of Northern, Central and Eastern Europe. Due to the dynamics of the phenomena taking place, the EU recommends constant monitoring of forests in terms of large-area disturbances and factors affecting tree stands’ susceptibility to destruction. The right tools for this are multispectral satellite images, which regularly and free of charge provide up-to-date information on changes in the environment. The aim of this study was to develop a method of identifying disturbances of spruce stands, including the identification of bark beetle outbreaks. Sentinel 2 images from 2015–2018 were used for this purpose; the reference data were high-resolution aerial images, satellite WorldView 2, as well as field verification data. Support Vector Machines (SVM) distinguished six classes: deciduous forests, coniferous forests, grasslands, rocks, snags (dieback of standing trees) and cuts/windthrow. Remote sensing vegetation indices, Multivariate Alteration Detection (MAD), Multivariate Alteration Detection/Maximum Autocorrelation Factor (MAD/MAF), iteratively re-weighted Multivariate Alteration Detection (iMAD) and trained SVM signatures from another year, stacked band rasters allowed us to identify: (1) no changes; (2) dieback of standing trees; (3) logging or falling down of trees. The overall accuracy of the SVM classification oscillated between 97–99%; it was observed that in 2015–2018, as a result of the windthrow and bark beetle outbreaks and the consequences of those natural disturbances (e.g., sanitary cuts), approximately 62.5 km2 of coniferous stands (29%) died in the studied area of the Tatra Mountains.

The composition and structure of forest stands in the Tatra National Park were examined using data gathered in 2016 and 2017 from 617 circular sample plots (0.05 ha each). The diameter at breast height of all living trees, standing dead trees, snags, and wind throws was measured along with diameters and lengths of fallen logs within the plot boundaries. Tree height was measured for all living trees within the core (0.01 ha) of the sample plots. Using the obtained data, height-diameter curves were calculated for all major tree species and in the case of spruce, the height-diameter relationships were also calculated separately for each of the three elevation zones (up to 1200 m, between 1200 and 1400 m, above 1400 m). For each elevation zone and park protection zone, we also determined the volumes of live and dead trees. The volume of living trees in the Tatra National Park amounted to 259 m3/ha, which was higher than the volume of dead trees (176 m3/ha). Snags constituted the largest part of the dead wood whilst over 97% of the standing dead trees were spruce Picea abies. Among living trees, the share of spruce ranged from 81% in the low elevation zone to 98% in the middle zone. Other significant species in the lower zone were Abies alba (11%) and Fagus sylvatica (4.5%), while in the middle and upper elevation zones only Sorbus aucuparia occurred in significant numbers. Furthermore, in the lower elevation zone, Fagus sylvatica was the only species displaying significantly higher volumes in the ‘strict protection’ zone compared to the other park areas. In the ‘landscape protection’ zone, Picea abies was the most dominant species and the share of other species in the lowest elevation zones calculated based on tree density was smaller than calculated based on tree volume, indicating problems with stand conversion from spruce monoculture to mixed forest.

The Swiss stone Pine (Pinus cembra L.) is an alpine species, fairly commonly occurring in the Alps and the Carpathians, close to the timberline. Natural sites of the Swiss stone pine in Poland are found exclusively in the Tatra Mountains, within the area of the Tatra National Park (TNP). In 2017, the health status of the needles of P. cembra was determined and their mycological analysis was carried out. As a result, 11 species of fungi were isolated from the needles showing disease symptoms, manifested by various shades of discoloration. The most frequently observed species was Fusarium oxysporum. It probably does not bring on disease symptoms, but may affect the development of other frequently isolated species (such as Sydowia polyspora, Lophodermium sp. and Lophodermium conigenum) that are pathogenic to Pinus spp.

A.B.L. was supported by a Juan de la Cierva-Incorporación grant (IJCI-2017-31419) from the Spanish Ministry of Science, Innovation and Universities; J.V.L.B. by a Ramón & Cajal research contract (RYC-2015-18932) from the Spanish Ministry of Economy, Industry and Competitiveness (…), Cimatti, M., Ranc, N., Benítez-López, A., Maiorano, L., Boitani, L., Cagnacci, F., Čengić, M., Ciucci, P., Huijbregts, M.A.J., Krofel, M., López-Bao, J.V., Selva, N., Andren, H., Bautista, C., Ćirović, D., Hemmingmoore, H., Reinhardt, I., Marenče, M., Mertzanis, Y., Pedrotti, L., Trbojević, I., Zetterberg, A., Zwijacz-Kozica, T., Santini, L.

The gray wolf Canis lupus range in central Europe is dynamically expanding, reconnecting previously isolated populations. Thus, a recent paper has proposed to merge the current Baltic and Central European (CE) wolf management units, which are no longer isolated by distance. However, recent genetic findings indicate that these two populations are not genetically homogenous. Here we review the most recent data on wolf genetic structure in central Europe and show that even though the CE and Baltic wolves represent the same phylogeographic lineage, their demographic history has resulted in significant genetic structure between these two populations. While the groups are interconnected by moderate gene flow, it is not high enough to reduce the strong founder signal observed in the CE population, suggesting that population dynamics within the CE wolf range are largely independent from those of its source (Baltic) population. Consequently, a management unit combining the CE and Baltic wolves would not form a demographically coherent entity. Thus, we recommend that conservation management units maintain their separate status.

Hybridization and introgression are important processes influencing the genetic diversity and evolution of species. These processes are of particular importance in protected areas, where they can lead to the formation of hybrids between native and foreign species and may ultimately result in the loss of parental species from their natural range. Despite their importance, the contribution of hybridization and introgression to genetic diversity in Sorbus genus remains not fully recognized. We analysed the genetic and morphological variability of several Sorbus species including native (Sorbus aria), foreign (S. intermedia) and potentially hybrid (S. carpatica) individuals from the Polish Carpathian range. Patterns of variation at 13 nuclear microsatellite loci show hybridization between the tested species and confirm the existence of the hybrid form S. carpatica. Biometric analysis on leaves, based of 10 metric features and three parameters, identified several characters for preliminary taxonomic classification; however, none of them could be used as a fully diagnostic marker for faultless annotation of S. intermedia and S. carpatica. The genetic structure analysis indicated complex patterns of population differentiation and its diverse origin. The results allow assessment of genetic variation and identification of parental species participating in hybridization. This knowledge will advance the management of genetic diversity and development of conservation strategies for efficient maintenance of the unique protected ecosystem.

We investigated the level of five non-essential metal(loid)s (As, Cd, Hg, Tl, Pb) and nine essential metals (Mg, Ca, Mn, Fe, Co, Cu, Zn, Se, Mo) in hair and blood components of captive and free-ranging European brown bear populations in Croatia and Poland. Metal(loid) associations with biomarkers of oxidative stress (superoxide dismutase, SOD; glutathione-peroxidase, GSH-Px; malondialdehyde, MDA) and metal exposure (metallothionein, MT) were estimated in this top predatory mammal. Lead was the most abundant non-essential metal(loid) in both blood and hair, with 4 of 35 individuals having blood levels over 100 μg/L. A positive association was found between Pb level and SOD activity in blood. Free-ranging bears had higher blood SOD activity, Mn, Zn and Cd levels, hair Co, Cd, Tl and Pb compared to captive individuals, while the opposite was true for Mg and hair Ca thereby reflecting habitat and diet differences. With increasing age, animals showed lower levels of SOD activity and certain essential metals. Females had higher SOD activity and blood levels of some essential metals than males. Hair showed a higher Fe and Co level when sampled during the growth phase and was not predictive of non-essential metal(loid) blood levels. The established metal(loid) baseline values will enable future risk as- sessment in both captive and wild European brown bear populations.

Local extinction and recolonization events can shape genetic structure of subdivided animal populations. The gray wolf (Canis lupus) was extirpated from most of Europe, but recently recolonized big part of its historical range. An exceptionally dynamic expansion of wolf population is observed in the western part of the Great European Plain. Nonetheless, genetic consequences of this process have not yet been fully understood. We aimed to assess genetic diversity of this recently established wolf population in Western Poland (WPL), determine its origin and provide novel data regarding the population genetic structure of the grey wolf in Central Europe. We utilized both spatially explicit and non-explicit Bayesian clustering approaches, as well as a model-independent, multivariate method DAPC, to infer genetic structure in large dataset (881 identified individuals) of wolf microsatellite genotypes. To put the patterns observed in studied population into a broader biogeographic context we also analyzed a mtDNA control region fragment widely used in previous studies. In comparison to a source population, we found slightly reduced allelic richness and heterozygosity in the newly recolonized areas west of the Vistula river. We discovered relatively strong west-east structuring in lowland wolves, probably reflecting founder-flush and allele surfing during range expansion, resulting in clear distinction of WPL, eastern lowland and Carpathian genetic groups. Interestingly, wolves from recently recolonized mountainous areas (Sudetes Mts, SW Poland) clustered together with lowland, but not Carpathian wolf populations. We also identified an area in Central Poland that seems to be a melting pot of western, lowland eastern and Carpathian wolves. We conclude that the process of dynamic recolonization of Central European lowlands lead to the formation of a new, genetically distinct wolf population. Together with the settlement and establishment of packs in mountains by lowland wolves and vice versa, it suggests that demographic dynamics and possibly anthropogenic barriers rather than ecological factors (e.g. natal habitat-biased dispersal patterns) shape the current wolf genetic structure in Central Europe.

Diet studies are essential to understand animal ecology and ecosystem dynamics, especially in the case of large omnivores. These studies are particularly relevant in areas where human disturbance is intense and, thus, species dietary patterns might change due to the easy accessibility of food resources of human origin, which may hinder the ecosystem services these species provide. We assessed the diet of brown bears (Ursus arctos) by DNA metabarcoding in Tatra National Park (southern Poland), a highly touristic protected area. Brown bears' diet showed a marked seasonality, a characteristic feature of brown bear populations relying on natural foods. Graminoids represented the main food during spring, while fleshy-fruited plants became more important from mid-summer. Fleshy-fruited plants were present in 56% of faeces and during the entire activity period of bears, revealing that fruits play a pivotal role in the feeding ecology of Tatra brown bears. Two berry species, Vaccinium myrtillus and Rubus idaeus, were the most frequently detected (in 42% and 20% of faeces, respectively). The large consumption of fleshy-fruited plants, and particularly of berries, suggests that, despite high levels of human disturbance in the area, brown bears still play a key role as seed dispersers. Management strategies focused on an effective waste management, control of berry picking, strict regulations about human activities in specific areas during sensitive periods in the feeding ecology of bears, and the lack of artificial food provisioning are crucial to ensure the seed dispersal processes and associated ecosystem services that bears and other frugivores provide.

Food availability plays a key role in animal movements. Anthropogenic provisioning of food to wildlife is a common practice of unprecedented magnitude worldwide and is of increasing conservation concern. Ungulate supplementary feeding is widespread in game management; however its effects on non-target species have received little attention. Here, we investigate how ungulate feeding affects the movement behavior of a non-target species, the brown bear ( Ursus arctos ). We tracked bear movements in the Northeastern Carpathians (1500 km 2 ) and inventoried 212 ungulate feeding sites. We analyzed encounter rates of nine GPS-collared bears with ungulate feeding sites (1658 km, n = 49 tracks) and compared them with the corresponding encounter rate of simulated tracks. We also estimated the encounter rate with feeding sites using snow-tracking of unknown bears (232 km, n = 40 tracks). GPS-tracked bears encountered feeding sites three times more frequently (mean ± SE = 0.154 ± 0.022 per km travelled) than would be expected if they were moving randomly (0.054 ± 0.0010 per km random walk). The rate was even higher for snow-tracked bears, which visited on average 0.926 ± 0.271 feeding sites per kilometer travelled. This suggests a link between the winter activity of some individuals and their frequent use of feeding sites. Bears seemed to rely on spatial memory and patrol known sites, independent of whether food was available at the feeding sites. This alteration of the natural behavior of species with behavioral flexibility, such as brown bears, could be interpreted as a sign of environmental degradation. Our results demonstrate an important effect of ungulate feeding on the movement ecology of non-target species. We warn of the impacts of this practice on species and ecosystems and highlight the need to preserve natural movement behaviors and urgently reevaluate management practices involving food provisioning to wildlife.

Baylisascaris transfuga is a roundworm that has been reported worldwide in most bear species. In mammals and possibly humans, the larvae of B. transfuga can migrate in the tissues of aberrant hosts with larva migrans syndrome. The current study was performed to identify B. transfuga in faecal samples from free-ranging brown bears in the Tatra Mountains National Park in southern Poland. A commercial kit was used to extract genomic DNA directly from faecal samples. Additionally, a Chelex resin-based technique was successfully implemented to prepare a PCR template from eggs retrieved by flotation. Based on the flotation results of 32 collected faecal samples, the prevalence of B. transfuga was 15.6%. The parasite was confirmed in samples found to be positive during the initial flotation by a molecular assay using DNA isolated directly from faeces. The retrieved eggs were confirmed as B. transfuga after their DNA was extracted using the Chelex protocol. Based on PCR amplification and sequencing of a 413-bp segment of cytochrome c oxidase 1 (COI), the obtained sequence was 100% identical to the COI segment of B. transfuga after a BLAST comparison to the GenBank™ database. The current study includes the first molecular confirmation of B. transfuga in brown bears in the western part of the Carpathians. We show that direct extraction of parasite DNA from bear faeces is efficient for molecular assays. As an alternative, we present the effectiveness of a Chelex-based technique for fast and convenient DNA isolation from the difficult-to-disrupt eggs of B. transfuga for PCR. Molecular tests of parasite DNA extracted directly from faecal material have limits of detection related to the amount of eggs in the samples. Thus, using classical flotation to obtain eggs for PCR may increase the credibility of the results, particularly in cases with a low number of excreted eggs. The Chelex resin protocol has potential for application in studies of intestinal parasites in wildlife for which conventional flotation is routinely used for microscopy. Electronic supplementary material The online version of this article (10.1007/s00436-017-5660-2) contains supplementary material, which is available to authorized users.

We report on the determination of the spore concentration and the species composition of the airborne fungi in selected caves of the Tatra Mountains, Poland. The following caves were surveyed: Mylna, Oblazkowa, Mroźna, Zimna and Naciekowa. The sampling was carried out in July 2015 and in January 2016. The aeromycological analyses were performed with the impact method, using the Air Ideal 3P apparatus and potato dextrose agar (PDA, Biocorp) culture medium. In the course of the July 2015 analysis, 17 species of fungi were isolated and 11 species were isolated in January 2016. In Mylna and Naciekowa caves, the dominant species were Cladosporium cladosporioides and Stachybotrys cylindrospora. In Oblazkowa cave, Rhizoctonia predominated and in Zimna cave—the colonies of the yeast-like fungi, along with S. cylindrospora. In Mroźna cave, Penicillium notatum was the most abundant taxon. In the winter time, in the majority of the caves Penicillium spp. predominated, with the exception of Mroźna and Naciekowa caves where Aspergillus niger was dominant. We propose that aeromycological monitoring be performed regularly in the following caves: Mroźna, Naciekowa and Zimna.

Most mammals rely upon scent for intraspecific communication. As most bear species have large home ranges and are non-territorial, scent deposit while walking could be an effective way to communicate with conspecifics. Here, we investigate the existence of pedal glands in brown bears and their role in chemical communication from a histological, biochemical and behavioural perspective. We found eccrine glands in footpads, and prominent apocrine and sebaceous glands in the interdigital, metacarpal and metatarsal skin sections. Pedal scent contained 26 compounds including carboxylic acids, important constituents of mammalian secretions. Six of these compounds were exclusive for males. Finally, we describe a specific marking gait recorded in the field, mostly performed by males. Our study supports the existence of chemical communication through pedal marking in brown bears and suggests sex-coding potential of pedal scent.

Knowledge about population size is of high importance for conservationists. We used non-invasively collected samples and microsatellite genotyping to estimate the size of the Tatra National Park population of the endangered capercaillie Tetrao urogallus. This population is one of the most important strongholds of the species in Poland. In 2016 over 150 samples (faeces and feathers) of the capercaillie were collected throughout area of the Tatra National Park. Then, DNA was extracted and genetic profiles were evaluated, using nine microsatellite markers. We obtained 81 reliable genotypes. Among them, 34 unique genotypes were found, corresponding to Minimum Number of individuals Alive in the investigated population. Application of capture-recapture models in the R package Capwire indicated, that the area was inhabited by approx. 54 birds, whereas regression model suggested presence of 36–64 individuals. Previous field surveys suggested that the number of birds in the Tatra National Park is about 50. Hence, we assumed that genetic tagging of non-invasive samples performs well in estimating the abundance of the capercaillie in the investigated population.

V.P. was fnancially supported by (1) the Excellence Project CGL2017-82782-P financed by the Spanish Ministry of Science, Innovation and Universities, the Agencia Estatal de Investigación (AEI) and the Fondo Europeo de Desarrollo Regional (FEDER, EU), and (2) Modalidad Grupos de Investigación Consolidados, Principado de Asturias (IDI/2018/000151). M.M.D. was fnancially supported by the Spanish Ramon y Cajal grant RYC-2014-16263. N.S., C.B. and A.G. were partly supported by the National Centre for Research and Development (GLOBE POL-NOR/198352/85/2013) and the National Science Centre in Poland (DEC-2013/08/M/NZ9/00469; 2016/22/Z/NZ8/00121; 2017/25/N/NZ8/02861). E.R., J.N., A.F., N.S., and C.B. were supported by the Agencia Estatal de Investigación from the Ministry of Economy, Industry and Competitiveness, Spain (project CGL2017-83045-R AEI/FEDER EU, co-fnanced with FEDER)., Bombieri, G., Naves, J., Penteriani, V., Selva, N., Fernández-Gil, A., López-Bao, J.V., Ambarli, H., Bautista, C., Bespalova, T., Bobrov, V., Bolshakov, V., Bondarchuk, S., Camarra, J.J., Chiriac, S., Ciucci, P., Dutsov, A., Dykyy, I., Fedriani, J.M., García-Rodríguez, A., Garrote, P.J., Gashev, S., Groff, C., Gutleb, B., Haring, M., Härkönen, S., Huber, D., Kaboli, M., Kalinkin, Y., Karamanlidis, A.A., Karpin, V., Kastrikin, V., Khlyap, L., Khoetsky, P., Kojola, I., Kozlow, Y., Korolev, A., Korytin, N., Kozsheechkin, V., Krofel, M., Kurhinen, J., Kuznetsova, I., Larin, E., Levykh, A., Mamontov, V., Männil, P., Melovski, D., Mertzanis, Y., Meydus, A., Mohammadi, A., Norberg, H., Palazón, S., Pătrașcu, L.M., Pavlova, K., Pedrini, P., Quenette, P.Y., Revilla, E., Rigg, R., Rozhkov, Y., Russo, L.F., Rykov, A., Saburova, L., Sahlén, V., Saveljev, A.P., Seryodkin, I.V., Shelekhov, A., Shishikin, A., Shkvyria, M., Sidorovich, V., Sopin, V., Støen, O., Stofik, J., Swenson, J.E., Tirski, D., Vasin, A., Wabakken, P., Yarushina, L., Zwijacz-Kozica, T., Delgado, M.M.

Home range estimation is routine practice in ecological research. While advances in animal tracking technology have increased our capacity to collect data to support home range analysis, these same advances have also resulted in increasingly autocorrelated data. Consequently, the question of which home range estimator to use on modern, highly autocorrelated tracking data remains open. This question is particularly relevant given that most estimators assume independently sampled data. Here, we provide a comprehensive evaluation of the effects of autocorrelation on home range estimation. We base our study on an extensive data set of GPS locations from 369 individuals representing 27 species distributed across five continents. We first assemble a broad array of home range estimators, including Kernel Density Estimation (KDE) with four bandwidth optimizers (Gaussian reference function, autocorrelated-Gaussian reference function [AKDE], Silverman´s rule of thumb, and least squares cross-validation), Minimum Convex Polygon, and Local Convex Hull methods. Notably, all of these estimators except AKDE assume independent and identically distributed (IID) data. We then employ half-sample cross-validation to objectively quantify estimator performance, and the recently introduced effective sample size for home range area estimation ((Formula presented.)) to quantify the information content of each data set. We found that AKDE 95% area estimates were larger than conventional IID-based estimates by a mean factor of 2. The median number of cross-validated locations included in the hold-out sets by AKDE 95% (or 50%) estimates was 95.3% (or 50.1%), confirming the larger AKDE ranges were appropriately selective at the specified quantile. Conversely, conventional estimates exhibited negative bias that increased with decreasing (Formula presented.). To contextualize our empirical results, we performed a detailed simulation study to tease apart how sampling frequency, sampling duration, and the focal animal´s movement conspire to affect range estimates. Paralleling our empirical results, the simulation study demonstrated that AKDE was generally more accurate than conventional methods, particularly for small (Formula presented.). While 72% of the 369 empirical data sets had >1,000 total observations, only 4% had an (Formula presented.) >1,000, where 30% had an (Formula presented.)