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Harvest can disrupt wildlife populations by removing adults with naturally high survival. This can reshape sociospatial structure, genetic composition, fitness, and potentially affect evolution. Genetic tools can detect changes in local, fine-scale genetic structure (FGS) and assess the interplay between harvest-caused social and FGS in populations. We used data on 1614 brown bears, Ursus arctos, genotyped with 16 microsatellites, to investigate whether harvest intensity (mean low: 0.13 from 1990 to 2005, mean high: 0.28 from 2006 to 2011) caused changes in FGS among matrilines (8 matrilines; 109 females ≥4 years of age), sex-specific survival and putative dispersal distances, female spatial genetic autocorrelation, matriline persistence, and male mating patterns. Increased harvest decreased FGS of matrilines. Female dispersal distances decreased, and male reproductive success was redistributed more evenly. Adult males had lower survival during high harvest, suggesting that higher male turnover caused this redistribution and helped explain decreased structure among matrilines, despite shorter female dispersal distances. Adult female survival and survival probability of both mother and daughter were lower during high harvest, indicating that matriline persistence was also lower. Our findings indicate a crucial role of regulated harvest in shaping populations, decreasing differences among “groups,” even for solitary-living species, and potentially altering the evolutionary trajectory of wild populations. anthropogenic, dispersal, hunting, male mating, maternal, predator, survival

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 humandominated 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. genetic relationship, Greece, kinship, spatial autocorrelation, wildlife conservation

Knowledge about the connectivity among natural populations is essential to identify management units for effective conservation actions. Conservation-minded management has led to the recovery of large carnivore populations in northern Europe, possibly restoring connectivity between the two separated, but expanding brown bear (Ursus arctos) populations on the Scandinavian peninsula to the west and Karelia, a part of the large Eurasian population, to the east. The degree of connectivity between these populations has been poorly understood, therefore we investigated the extent of connectivity between the two populations using autosomal microsatellites and Y chromosome haplotypes in 924 male bears (the dispersing sex), sampled during a period of 12 years (2005–2017) across the transborder area where these two populations meet. Our results showed that the two populations are not genetically isolated as reported in earlier studies. We detected recent asymmetrical gene flow at a rate (individuals per generation) of 4.6–5.5 (1%) from Karelia into Scandinavia, whereas the rate was approximately 27.1–34.5 (8%) in the opposite direction. We estimated historical gene flow of effective number of migrants to be between 1.7 and 2.5 between the populations. Analyses of Y chromosome markers supported these results. Successful recovery and expansion of both populations led to the restoration of connectivity, however, it is asymmetric, possibly due to different recovery histories and population densities. By aligning monitoring between neighboring countries, we were able to better understand the biological processes across the relevant spatial scale. Brown bear Genetic structure Male gene flow Microsatellites Migration Recovery Ursus arctos Wildlife monitoring Y chromosome

The degree of gene flow within and among populations, i.e. genetic population connectivity, may closely track demographic population connectivity. Alternatively, the rate of gene flow may change relative to the rate of dispersal. In this study, we explored the relationship between genetic and demographic population connectivity using the Scandinavian brown bear as model species, due to its pronounced male dispersal and female philopatry. Thus, we expected that females would shape genetic structure locally, whereas males would act as genetic mediators among regions. To test this, we used eight validated microsatellite markers on 1531 individuals sampled noninvasively during country-wide genetic population monitoring in Sweden and Norway from 2006 to 2013. First, we determined sex-specific genetic structure and substructure across the study area. Second, we compared genetic differentiation, migration/gene flow patterns, and spatial autocorrelation results between the sexes both within and among genetic clusters and geographic regions. Our results indicated that demographic connectivity was not a reliable indicator of genetic connectivity. Among regions, we found no consistent difference in long-term gene flow and estimated current migration rates between males and females. Within regions/genetic clusters, only females consistently displayed significant positive spatial autocorrelation, indicating male-biased small-scale dispersal. In one cluster, however, males showed a dispersal pattern similar to females. The Scandinavian brown bear population has experienced substantial recovery over the last decades; however, our results did not show any changes in its large-scale population structure compared to previous studies, suggesting that an increase in population size and dispersal of individuals does not necessary lead to increased genetic connectivity. Thus, we conclude that both genetic and demographic connectivity should be estimated, so as not to make false assumptions about the reality of wildlife populations.

We present data on the species composition of helminths in brown bears (Ursus arctos) from the Murmansk Region, Russia. The absence of any information about helminths of brown bear in the region necessitated the conduct of these studies. Samples were collected in 2014 and 2015 in the southern part of the Kola Peninsula from the White Sea coastal habitats. Annually, in the study area, 1-3 bears are legally hunted and biological samples for examination are very difficult to obtain. Therefore, we used fecal samples. We studied 93 feces and identified parasite eggs identified in 43 of them by morphometric criteria. The surveys revealed eggs of the following helminths: Dicrocoelium sp., Diphyllobothrium sp., Anoplocephalidae, Capillariidae, Baylisascaris sp., Strongylida 1, and Strongylida 2. These results represent the first reconnaissance stage, which allowed characterizing the taxonomic diversity and prevalence of parasites of brown bears of the Kola Peninsula.

The number of effective breeders (N b) and effective population size (N e) are population parameters reflective of evolutionary potential, susceptibility to stochasticity, and viability. We have estimated these parameters using the linkage disequilibrium‐based approach with LDNE through the latest phase of population recovery of the brown bears (Ursus arctos) in Finland (1993–2010; N = 621). This phase of the recovery was recently documented to be associated with major changes in genetic composition. In particular, differentiation between the northern and the southern genetic cluster declined rapidly within 1.5 generations. Based on this, we have studied effects of the changing genetic structure on N b and N e, by comparing estimates for whole Finland with the estimates for the two genetic clusters. We expected a potentially strong relationship between estimate sizes and genetic differentiation, which should disappear as the population recovers and clusters merge. Consistent with this, our estimates for whole Finland were lower than the sum of the estimates of the two genetic clusters and both approaches produced similar estimates in the end. Notably, we also found that admixed genotypes strongly increased the estimates. In all analyses, our estimates for N e were larger than N b and likely reflective for brown bears of the larger region of Finland and northwestern Russia. Conclusively, we find that neglecting genetic substructure may lead to a massive underestimation of N b and N e. Our results also suggest the need for further empirical analysis focusing on individuals with admixed genotypes and their potential high influence on N b and N e.

We here describe 34 novel microsatellite loci from the brown bear (Ursus arctos) identified using a next-generation sequencing approach. While we found allelic variation in 30 of these, we selected 16 polymorphic microsatellites (11 di- and 5 tetranucleotide repeats) for four multiplex-PCR assays based on genetic variability, PCR performance and interpretability. Applying these assays for the 16 novel microsatellites, we screened 31 brown bears from a northern European population. The number of alleles encountered at single loci was between 4 and 10, and expected and observed heterozygosities ranged between 0.66–0.85 and 0.60–0.97, respectively. Seven of 120 pairwise locus combinations showed linkage disequilibrium and one locus deviated from Hardy–Weinberg equilibrium, while no null-alleles were detected. Our novel multiplex-PCR assays may be useful in forensics, parentage and population genetic studies.

Loss of connectivity and habitat destruction may lead to genetic depletion of wild animal populations, especially in species requiring large, connected territories as the brown bear (Ursus arctos). Brown bear populations of North Western Russia, Finland and Northern Norway have been assumed to form one large, continuous population; however this hypothesis has not been tested sufficiently. We have genotyped 1,887 samples from 2005 to 2008 from four distinct areas and used the resulting DNA profiles from 146 different individuals to analyze the genetic diversity, population structure, and the migration rates among groups. In addition, we have tested for traces of previous genetic bottlenecks. Individuals from Eastern Finland and Russian Karelia were grouped in the same cluster (“Karelia”), while distinctive subpopulations of brown bears were detected in the north (“Pasvik”), and the east (“Pinega”). All three subpopulations displayed high genetic variation, with expected heterozygosities (HE) of 0.77–0.81, but differentiation among the clusters was relatively low (average FST = 0.051, P < 0.001). No evidence of genetic bottlenecks in the past was found. We detected a highly significant isolation-by-distance (IBD) pattern. For Pasvik, self-recruitment was found to be very high (96%), pointing to the possibility of genetic isolation. In contrast, between Karelia and Pinega we detected high, bi-directional migration rates (~30%), indicating genetic exchange. Conclusively, despite of a substantial influence of IBD on the genetic structure in the region, we detected considerable variation in connectivity among the identified clusters that could not be explained solely by the distance between them.

A set of 13 dinucleotide STR loci (G1A, G10B, G1D, G10L, MU05, MU09, MU10, MU15, MU23, MU26, MU50, MU51, MU59) were selected as candidate markers for a DNA forensic profiling system for Northern European brown bear ( Ursus arctos ). We present results from validation of the markers with respect to their sensitivity, species specificity and performance (precision, heterozygote balance and stutter ratios). All STRs were amplified with 0.6ng template input, and there were no false bear genotypes in the cross-species amplification tests. The validation experiments showed that stutter ratios and heterozygote balance was more pronounced than in the tetranucleotide loci used in human forensics. The elevated ratios of stutter and heterozygote balance at the loci validated indicate that these dinucleotide STRs are not well suited for interpretation of individual genotypes in mixtures. Based on the results from the experimental validations we discuss the challenges related to genotyping dinucleotide STRs in single source samples. Sequence studies of common alleles showed that, in general, the size variation of alleles corresponded with the variation in number of repeats. The samples characterized by sequence analysis may serve as standard DNA samples for inter laboratory calibration. A total of 479 individuals from eight Northern European brown bear populations were analyzed in the 13 candidate STRs. Locus MU26 was excluded as a putative forensic marker after revealing large deviations from expected heterozygosity likely to be caused by null-alleles at this locus. The remaining STRs did not reveal significant deviations from Hardy–Weinberg equilibrium expectations except for loci G10B and MU10 that showed significant deviations in one population each, respectively. There were 9 pairwise locus comparisons that showed significant deviation from linkage equilibrium in one or two out of the eight populations. Substantial genetic differentiation was detected in some of the pairwise population comparisons and the average estimate of population substructure ( F ST ) was 0.09. The average estimate of inbreeding ( F IS ) was 0.005. Accounting for population substructure and inbreeding the total average probability of identity in each of the eight populations was lower than 1.1×10 −9 and the total average probability of sibling identity was lower than 1.3×10 −4 . The magnitude of these measurements indicates that if applying these twelve STRs in a DNA profiling system this would provide individual specific evidence.

The development of wildlife genetics combined with non-invasive sampling might be both an economic benefit for the society and a benefit for the survival of the threatened species. The aims of this study are to develop a quality assured approach for DNA profiling of brown bears ( Ursus arctos ) in Northern Europe using material from non-invasive sampling and to generate a population database that can be used for conservation management as well as a reference database for forensic purposes. Non-invasive sampling was performed by the collection of scats in the field and by using sets of hair traps in a grid pattern in specified geographical areas. Genotypes from 12 STR loci were determined for 232 Norwegian bears. Initial analysis of the entire sample indicated a high level of substructure. Thus, the sample was divided into four geographically different populations consisting of 206 individuals for further validation of the markers. Ten STRs (G1D, G10L, Mu05, Mu09, Mu10, Mu15, Mu23, Mu50, Mu51, and Mu59) conformed to Hardy–Weinberg equilibrium (HWE) expectations with only minor deviations, while the remaining two STR loci (Mu26 and G10B) were excluded from our set of putative forensic profiling system markers after revealing significant deviations from HWE in all four sub-populations. The average estimate of population substructure for Norwegian bears using 10 STRs ( F ST ) was determined to be 0.1, while the estimate for inbreeding ( F IS ) was −0.02. Accounting for the F ST -value, the average probability of identity (PI ave ) was 5.67×10 −10 and the average probability of sibling identity (PI sib ) was 1.68×10 −4 . In Norway, this brown bear DNA profiling system has been applied to forensic casework.