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Our understanding of animal adaptations to human pressure is limited by the focus on rare taxa, despite that common species are more significant in shaping structure, function and service provision of ecosystems. Thus better understanding of their ecology and behavioural adjustments is central for drafting conservation actions. In this study, we used radiotelemetry on 21 individuals (10 females, 11 males) to provide data on spatial ecology, habitat selection and use of roosts of one of the commonest species, the whiskered bat (Myotis mystacinus), inhabiting the Carpathian Mountains (southern Poland). We tested, whether this species prefers natural over human-modified landscapes to seek prey and roosts. Mean home range size of the whiskered bat in the Carpathian Mountains was 26.3 ha (SE ± 3.2, Local Convex Hull) and 110 ha (SE ± 22.1, Minimum Convex Polygon with all locations), and included between one and three patches, among which bats moved along linear environmental features, such as scrubby banks of streams or lines of trees. During foraging whiskered bats selected small woodlands within agricultural landscapes, avoided large mountain forests and open areas, and used built-up areas proportionally to their availability. Whiskered bats occupied roosts located mainly in buildings (>97%), at an average altitude of 547.9 m above sea level (SE ± 8.3). Roosts were used for 5.4 days, on average. Our study shows that whiskered bats adapted well to the mosaic of semi-natural and anthropogenic habitats. It highlights the importance of buildings serving as roosts and small woodlands used as foraging areas in human-dominated montane landscapes.

Molecular methods are routinely used to estimate the effective size of populations (Ne). However, underlying model assumptions are frequently violated to an unknown extent. Although simulations can detect sources of bias and help to adjust sampling strategies and analyses methods, additional information from empirical data can also be used to calibrate methods and improve molecular Ne estimation methods. Here, we take advantage of long‐term genetic and ecological monitoring data of the grey wolf (Canis lupus) in Germany, and detailed population genetic studies in Poland, Spain and Portugal to improve Ne estimation strategies in this species, and species with similar life history traits. We first calculated Ne from average lifetime reproductive success and detailed census data from the German population, which served as a baseline to compare to molecular estimates based on linkage disequilibrium and sibship frequency. This yielded a robust Ne/Nc estimation that we used to calibrate molecular estimates of German, Polish and Iberian wolf populations. The linkage disequilibrium method was strongly influenced by spatial genetic structure, much more than the sibship frequency method. When Ne was estimated in local neighbourhoods, both methods yielded comparable results. Estimates of the metapopulation effective size seemed to correspond generally well with the sum of the estimates of local neighbourhoods. Overall, we found that the number of packs is a good proxy of the effective population size. Using this as a rule of thumb, we evaluated for all European wolf populations the Ne 500 indicator and concluded that half of the European wolf populations do not yet fulfil this criterion.

If protected areas are to remain relevant in our dynamic world they must be adapted to changes in species ranges. In the EU one of the most notable such changes is the recent recovery of large carnivores, which are protected by Natura 2000 at the national and population levels. However, the Natura 2000 network was designed prior to their recent recovery, which raises the question whether the network is sufficient to protect the contemporary ranges of large carnivores. To investigate this question we evaluated Natura 2000 coverage of the three wolf Canis lupus populations in Poland. Wolf tracking data showed that wolves have recolonised almost all suitable habitat in Poland (as determined by a recent habitat suitability model), so we calculated the overlap between the Natura 2000 network and all wolf habitat in Poland. On the basis of published Natura 2000 criteria, we used 20% as the minimum required coverage. At the national level, wolves are sufficiently protected (22% coverage), but at the population level, the Baltic and Carpathian populations are far better protected (28 and 47%, respectively) than the endangered Central European Lowland population (12%). As Natura 2000 insufficiently protects the most endangered wolf population in Poland, we recommend expansion of Natura 2000 to protect at least an additional 8% of wolf habitat in western Poland, and discuss which specific forests are most in need of additional coverage. Implementation of these actions will have positive conservation implications and help Poland to fulfil its Habitats Directive obligations. As it is likely that similar gaps in Natura 2000 are arising in other EU member states experiencing large carnivore recoveries, particularly in Central Europe, we make the case for a flexible approach to Natura 2000 and suggest that such coverage evaluations may be beneficial elsewhere.

Background Canine hepatozoonosis caused by Hepatozoon canis is a common infection in dogs, with frequent case reports from the Mediterranean region and more recently from several Central European countries, such as Hungary and Germany. Despite the high prevalence of H. canis in red foxes, no infections have been reported to date in dogs in Poland. We describe here the first autochthonous cases of H. canis infection in dogs, including their clinical features, and report the prevalence of H. canis in grey wolves from different regions of Poland. Methods Thin smears prepared from blood samples collected from dogs were evaluated by microscopic examination. A total of 60 wolves and 47 dogs were tested. Infections were confirmed by PCR and sequencing. Results Gamonts of H. canis were found in > 50% of the neutrophils of two dogs and in < 10% of the neutrophils in another five dogs. Molecular typing by PCR sequencing of the 18S ribosomal RNA gene fragment confirmed infections in 11 dogs from different regions of Poland, in 2.7% of dogs attending veterinary practices in central Poland and in 35% of wolves from various geographical regions of Poland. Clinical features manifested mostly in older dogs, and the most common signs were anaemia and apathy. Young dogs usually remained asymptomatic. Conclusions This is the first report of H. canis infection in dogs and wolves in Poland. Although the exact vector of the parasite is not known, veterinary practitioners should be aware of this new parasitosis and should consider appropriate diagnostics to confirm/exclude this infection. Further studies are needed to understand the transmission routes of H. canis in domestic and wild canids in Poland.

We assessed changes in the population size, density, and diet composition of wolves inhabiting the Romincka Forest (RF), an area of 480 km2 situated along the state border between Poland, Russian Federation (Kaliningrad), and Lithuania. We compared the results of our research in 2020–2021 with data from other projects conducted since 1999. We found that both packs living in RF had transboundary territories. The number of packs was stable over 21 years; the average pack size almost doubled (from 4–4.5 to 7.5–8 wolves per pack); the total wolf numbers increased 1.8 times, reaching 15–16 wolves; and the density increased 1.5 times up to 3.1–3.3 wolves/100 km2 in winter 2020/2021. Our analyses of 165 scats revealed that beavers Castor fiber made up 45.6% of food biomass in the wolf diet in 2020, which was 3.4 times more than in 1999–2004 (n = 84 scats,13.4%). Wild ungulates constituted 44.8% of the wolf food biomass in 2020, 1.6 times less than before (71.1%). In our study, among wild ungulates, wolves primarily consumed roe deer Capreolus capreolus (22.6% of food biomass), then wild boars Sus scrofa (13.7%), and red deer Cervus elaphus (5.0%), while moose Alces alces was eaten rarely (0.4%). We also recorded domestic dogs (4.9% of food biomass) and cattle (3.1%). The food niche breadth was wider (B = 2.31) than in the earlier period (B = 1.84), and the Pianka index showed moderate similarity in food composition between both periods (α = 0.816). In November 2022, due to the migration crisis, a 199 km impermeable fence along the state border with Kaliningrad was erected, which blocked access to 48% of the RF area that was regularly used by the resident wolf packs. This may cause wolf numbers to decrease and isolation from the central part of the Baltic wolf population to which they belong, according to our DNA analyses.

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.

Aim: To compare predictions of the habitat suitability model (HSM) for wolves Canis lupus in Poland with actual wolf distribution in western Poland after 15 years of recolonization. Location: Western Poland (WPL, ca. 136,000 km²), west of the 18°48'E meridian. Methods: Data on wolf occurrence (8,057 records) were gathered in 2001-2016. Wolf presence in 10 × 10 km cells was classified as follows: (1) permanent occurrence with reproduction, (2) permanent occurrence with no reproduction and (3) sporadic occurrence (interpreted as dispersing individuals). These cells were compared to all 10 × 10 km cells in WPL with respect to the probability of wolf occurrence as predicted by the HSM and habitat variables important for wolves. For temporal analysis, data were divided into two 8-year subsets: the initial and later phases of wolf recovery. Results: Wolves were recorded in 259 cells (19.8% of the study area). The pairs and packs settled in areas predicted by the HSM to have good and very good habitat, in cells characterized by high forest cover and low densities of roads. Wolf groups that reproduced were found in the best-quality habitats characterized by denser forest cover and markedly lower shares of anthropogenic structures. Dispersing individuals were mostly recorded in unsuitable and suboptimal habitats, and they avoided both the poorest and the best habitats. In the initial phase of wolf recovery, cells selected by wolves for settling down and those used by dispersing wolves did not differ in their habitat parameters. However, in the later phase, as WPL became more saturated with wolf packs, dispersing individuals were recorded in less suitable habitats. Main conclusions: The HSM for Polish wolves predicted with high accuracy the areas later occupied by wolf groups in the western part of the country. A similar approach may also be useful to predict the future distribution of wolves in the lowlands of central and western Europe where environmental conditions are comparable and recolonizing wolves originate from the same source population.

Aim The aim of the study was to assess the effect of changes in population size and migration on variation in functional immunity genes in the previously bottlenecked population of the grey wolf, Canis lupus. Location Eastern/Central Europe: Poland, Czechia, Slovakia. Methods We genotyped 7 immunity genes: three MHC‐DLA genes (dog leukocyte antigen) and four Toll‐like receptor (TLR) genes among 130 wolves originating from three populations: two lowland (Baltic and Central European), and highland Carpathian. We contrasted the population structure in immunity genes with a neutral structure based on 13 microsatellites, and we analysed signatures of selection in the immunity loci. Results We found high overall genetic variance in immunity genes and no evidence for decreased diversity in the recently established populations. The population structure in immunity loci was weak, with pairwise FST lower than for neutral markers. Although the results of neutrality tests were not significant, we identified codons under selection, both positive and negative. Main Conclusions We demonstrated that despite recent population expansion which is expected to result in decreased genetic diversity, the diversity of immunity genes in the newly established wolf population is similar to those in the source population. This suggests that migrations do not cause allele loss in grey wolf. Signatures of selection on codon level, but not in tests using allele frequencies, suggest the contrasting effects of demography and selection.

The European grey wolf (Canis lupus) and domestic dog (Canis lupus familiaris) share not only a common origin but also many similarities in behaviour. Due to the introduction of legal protection, wolves have recolonised large parts of Europe. They are increasingly inhabiting human‐dominated landscapes, leading to a growing potential for interactions between wolves and domestic dogs. However, these interactions between wolves and dogs are still poorly understood. Scent marking is one of the main forms of communication in canids and is crucial for marking territories, synchronising reproduction, establishing hierarchies within groups, and forming new breeding pairs. We hypothesised that the presence of domestic dog scent markings in wolf territories may induce a behavioural response and therefore interfere with wolf behaviour. To test this, we experimentally scent‐marked objects within known wolf home ranges in Kampinos National Park, Poland, to simulate the presence of “unknown dogs” (dog urine from outside the area) and water as a control. To test whether and how wolves' behavioural response to the scents of domestic dogs and wolves differs, we additionally left scent marks of “unknown wolves” (wolf urine from outside the area). Using camera traps, we studied the behavioural responses of local wolf families exposed simultaneously to all three scent stimuli. Juveniles spent significantly more time sniffing wolf scent (37.1 ± 8.9 s) than dog scent (7.1 ± 3.4 s), while breeding pairs displayed more diverse marking behaviours, including overmarking and ground scratching, particularly in response to wolf scent. Wolves spent a longer time responding to wolf scent marks than to dog scent marks, indicating they may distinguish between them, but inexperienced juveniles spent much more time exploring dog scent marks than adults. Our results indicate that domestic dog scent marks trigger a behavioural response in wild wolves. This suggests that the increasing occurrence of dogs inside wolf territories could affect and potentially disturb the scent‐marking behaviour of wolves. Wolves and Dogs Coexist in Human‐Dominated Landscapes: As wolves recolonise parts of Europe, they increasingly encounter domestic dogs, leading to interactions that are poorly understood, especially in terms of scent marking and communication. Scent Marking Experiment: In an experiment in Kampinos National Park, Poland, wolf territories were scent‐marked with urine from “unknown dogs” and “unknown wolves” to observe behavioural responses via camera traps. Water was used as a control. Wolves Respond More to Wolf Scent Marks: Wolves spent significantly more time sniffing scent marks from “unknown wolves” compared to “unknown dogs”, indicating that wolves may differentiate between these scents and prioritise responses to conspecifics. Age‐Related Differences in Behaviour: Juvenile wolves spent more time exploring dog scent marks than adult wolves, suggesting inexperience or greater curiosity. Adults primarily focused on wolf scent marks, displaying behaviours like overmarking and ground scratching. Implications for Wolf Behaviour: The presence of domestic dogs in wolf territories could interfere with wolves' natural scent‐marking behaviours, especially for juveniles. These findings highlight the need for further research on how dog presence might disrupt wolf territoriality and communication.

Dirofilaria repens is an expanding vector-borne zoonotic parasite of canines and other carnivores. Sub-clinically infected dogs constitute the most important reservoir of the parasite and the source of infection for its mosquito vectors. However, occurrence of D. repens infection in wild animals may contribute to the transmission of the parasite to humans and may explain the endemicity of filariae in newly invaded regions. The aim of the current study was to determine the occurrence of D. repens in 511 blood and spleen samples from seven species of wild carnivores (wolves, red foxes, Eurasian badgers, raccoons, raccoon dogs, stone martens, and pine martens) from different regions of Poland by means of a PCR protocol targeting the 12S rDNA gene. Dirofilaria repens–positive hosts were identified in seven of fourteen voivodeships in four of the seven regions of Poland: Masovia, Lesser Poland, Pomerania and Warmia-Masuria. The highest prevalence was found in Masovia region (8%), coinciding with the highest previously recorded prevalence in dogs in Central Poland. The DNA of Dirofilaria was detected in 16 samples of three species (total prevalence 3.13%). A low and similar percentage of positive samples (1.9%, 4.2% and 4.8%) was recorded among badgers, red foxes, and wolves, respectively. Dirofilaria repens–positive hosts were identified in seven of fourteen voivodships. Based on detection in different voivodeships, D. repens–positive animals were recorded in four out of the seven regions of Poland: in Masovia, Lesser Poland, Pomerania, and Warmia-Masuria. The highest prevalence of filariae was found in Masovia region (8%), reflecting the highest previously recorded prevalence in dogs (12–50%) in Central Poland. In summary, we conducted the first comprehensive study on the epidemiology of D. repens in seven species of wild hosts in all seven regions of Poland and identified the first case of D. repens infection in Eurasian badgers in Poland and the second in Europe.