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Genetic monitoring of populations currently attracts interest in the context of the Convention on Biological Diversity but needs long-term planning and investments. Genetic diversity has been largely neglected in biodiversity monitoring, and when addressed is treated separately, detached from other conservation issues, such as habitat alteration due to climate change. Genetic monitoring supports the conservation and management of fisheries, game, and threatened populations. It also can contribute to the assessment of predicted and realized impacts of climate change, and their management. We report the first accounting of genetic monitoring efforts among countries in Europe (their ‘genetic monitoring capacity’, GMC) to determine where GMC suggests the combination of national infrastructure, political support and resources for continued and expanded monitoring. Overlaying GMC with areas where species ranges approach current and future climate niche limits (i.e., niche marginality) helps identify whether GMC coincides with anticipated climate change effects on biodiversity. Our analysis suggests that country area extent, financial resources, and conservation policy influence GMC, high values of which inconsistently match joint species patterns of climate niche marginality. Populations at niche margins likely hold genetic diversity that is important to adaptation to changing climate, and our results illuminate the need in Europe for expanded genetic monitoring across the climate gradients occupied by species, a need arguably greatest in southeastern European countries.

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

The species we have studied the spatiotemporal genetic change in the northern dragonhead, a plant species that has experienced a drastic population decline and habitat loss in Europe. We have added a temporal perspective to the monitoring of northern dragonhead in Norway by genotyping herbarium specimens up to 200 years old. We have also assessed whether northern dragonhead has achieved its potential distribution in Norway. To obtain the genotype data from 130 herbarium specimens collected from 1820 to 2008, mainly from Norway (83) but also beyond (47), we applied a microfluidic array consisting of 96 SNP markers. To assess temporal genetic change, we compared our new genotype data with existing data from modern samples. We used sample metadata and observational records to model the species' environmental niche and potential distribution in Norway. Our results show that the SNP array successfully genotyped all included herbarium specimens. Hence, with the appropriate design procedures, the SNP array technology appears highly promising for genotyping old herbarium specimens. The captured genetic diversity correlates negatively with distance from Norway. The historical-modern comparisons reveal similar genetic structure and diversity across space and limited genetic change through time in Norway, providing no signs of any regional bottleneck (i.e., spatiotemporal stasis). The regional areas in Norway have remained genetically divergent, however, both from each other and more so from populations outside of Norway, rendering continued protection of the species in Norway relevant. The ENM results suggest that northern dragonhead has not fully achieved its potential distribution in Norway and corroborate that the species is anchored in warmer and drier habitats. Spatiotemporal monitoring of the rare northern dragonhead (Dracocephalum ruyschiana, Lamiaceae) — SNP genotyping and environmental niche modeling herbarium specimens

Aim: We have studied population genetic change through time in the Northern dragonhead, Dracocephalum ruyschiana (Lamiaceae); a plant species that has experienced a drastic population decline and habitat loss in Europe. We aimed at adding a historic level to the monitoring of dragonhead by testing a microfluidic SNP array approach on herbarium specimens up to 200 years old and comparing the genomic results with that of modern populations in Norway. We also aimed to gain a more holistic species knowledge to guide monitoring efforts by combining herbarium genomics with ecological niche modelling (ENM). Location: Europe (mainly Norway) Methods: We have applied a microfluidic array consisting of 96 SNP markers on 130 herbarium specimens collected from 1820 to 2008. Obtained genotype data were compared with SNP data from modern samples using various population genetic analyses. We used sample metadata and observational records to model the species’ environmental niche. Results: The SNP array successfully genotyped all included herbarium specimens but was less capable of capturing diversity outside of Norway, which was genetically highly divergent from the Norwegian dragonheads. The historic-modern comparison revealed similar genetic structure in space and limited change through time in Norway. The ENM suggests that dragonhead has not fully achieved its potential distribution in Norway, which is anchored in warmer and drier regions, including areas where it does not occur today. Main conclusions: With the appropriate design procedures, the SNP array technology is promising for genotyping old herbarium specimens; an invaluable source of information from the past. We found no signs of the severe reduction in population size in our temporal genomic data of Norwegian dragonhead. Regardless, the regional populations in Norway are genetically divergent, both from each other and more so from populations outside of Norway, rendering continued protection of all existing populations of the species relevant.

The species we have studied the spatiotemporal genetic change in the northern dragonhead, a plant species that has experienced a drastic population decline and habitat loss in Europe. We have added a temporal perspective to the monitoring of northern dragonhead in Norway by genotyping herbarium specimens up to 200 years old. We have also assessed whether northern dragonhead has achieved its potential distribution in Norway. To obtain the genotype data from 130 herbarium specimens collected from 1820 to 2008, mainly from Norway (83) but also beyond (47), we applied a microfluidic array consisting of 96 SNP markers. To assess temporal genetic change, we compared our new genotype data with existing data from modern samples. We used sample metadata and observational records to model the species' environmental niche and potential distribution in Norway. Our results show that the SNP array successfully genotyped all included herbarium specimens. Hence, with the appropriate design procedures, the SNP array technology appears highly promising for genotyping old herbarium specimens. The captured genetic diversity correlates negatively with distance from Norway. The historical-modern comparisons reveal similar genetic structure and diversity across space and limited genetic change through time in Norway, providing no signs of any regional bottleneck (i.e., spatiotemporal stasis). The regional areas in Norway have remained genetically divergent, however, both from each other and more so from populations outside of Norway, rendering continued protection of the species in Norway relevant. The ENM results suggest that northern dragonhead has not fully achieved its potential distribution in Norway and corroborate that the species is anchored in warmer and drier habitats.

Gray wolf populations have been recovering recently across Europe, a fact that poses serious challenges to the management of the species. We investigate the population genetics of wolves at the south-eastern edge of their European range, in Greece, and identify conservation priorities for the species in the country. During population monitoring efforts (1998–2014) 48 tissue and hair samples were collected and genotyped at 14 microsatellite loci. Eight samples were discarded from further downstream analysis because of possible dog-wolf admixture. Unlike many other edge populations, wolves in Greece showed high levels of genetic variation (HE = 0.73; HO = 0.66). We detected two genetic clusters of wolves but no genetic bottleneck, which suggests that the human-caused population reduction of wolves in Greece may not have been as severe as previously assumed and that enough wolves may have survived in inaccessible areas and/or neighboring countries to maintain genetic diversity. Two of the main conservation priorities identified for gray wolves in Greece are non-invasive genetic monitoring and ensuring the functional connectivity of the Natura 2000 network through the identification and protection of ecological corridors and road-less areas.

Human—bear conflicts occur frequently in the Pasvik Valley, Norway. We used a variant of the hair-trapping method with higher densities of traps (2.5 × 2.5 km grid) to detect brown bears moving near human settlements and livestock. We distributed 20 hair traps for one month close to a farm with frequent observations of grazing bears. The study area consisted of one area close to the farm, and one adjacent area without settlements. We collected 85 hair samples and identified 13 different individuals by STR analysis. In the farm area, we detected 4 different males once, and a female that was detected in both areas. In comparison, nine bears (2 males and 7 females) were detected for more than one week in the area without settlements, suggesting lower roaming activity. Conclusively, hair trapping has the potential to survey bears at specific locations of importance to the wildlife management.

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.

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.

In framework of an international project of brown bear population monitoring, the bear population size and structure were analyzed on the Russian side of the Pasvik-Inari Trilateral Park (Russia, Norway, Finland) in 2015. The paper presents data gathered by two noninvasive methods: DNA analysis of hair samples and feces and the camera traps method. The material was obtained with 5 hair traps of made of barbed wire and scent lure and 4 Boskon Guard camera traps. In addition, feces were collected, as well as hair samples from national border fencing by staff of the Russian Border Authority. In total, 54 hair samples and 10 feces samples were collected. On the basis of the results the brown bear population size was estimated at 20 individuals in the study area: 13 individuals were detected by DNA analysis only, the remaining 7 animals – with the help of camera traps. The records include 9 females and 7 males; 13 adults, 4 yearlings and 3 cubs. The social structure consisted of three family groups, 10 singles, including 1 possible dominant male. Average brown bear density was 100 animals/1000 km². The combined usage of hair and feces collection, camera traps and GIS proved the techniques to be complementary in the analysis of the size and structure of the brown bear population.

Noninvasively collected genetic data can be used to analyse large-scale connectivity patterns among populations of large predators without disturbing them, which may contribute to unravel the species' roles in natural ecosystems and their requirements for long-term survival. The demographic history of brown bears (Ursus arctos) in Northern Europe indicates several extinction and recolonization events, but little is known about present gene flow between populations of the east and west. We used 12 validated microsatellite markers to analyse 1580 hair and faecal samples collected during six consecutive years (2005-2010) in the Pasvik Valley at 70°N on the border of Norway, Finland and Russia. Our results showed an overall high correlation between the annual estimates of population size (N(c) ), density (D), effective size (N(e) ) and N(e) /N(c) ratio. Furthermore, we observed a genetic heterogeneity of ∼0.8 and high N(e) /N(c) ratios of ∼0.6, which suggests gene flow from the east. Thus, we expanded the population genetic study to include Karelia (Russia, Finland), Västerbotten (Sweden) and Troms (Norway) (477 individuals in total) and detected four distinct genetic clusters with low migration rates among the regions. More specifically, we found that differentiation was relatively low from the Pasvik Valley towards the south and east, whereas, in contrast, moderately high pairwise F(ST) values (0.91-0.12) were detected between the east and the west. Our results indicate ongoing limits to gene flow towards the west, and the existence of barriers to migration between eastern and western brown bear populations in Northern Europe.

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.

Recovery of natural populations occurs often with simultaneous or subsequent range expansions. According to population genetic theory, genetic structuring emerges at the expansion front together with decreasing genetic diversity, owing to multiple founder events. Thereupon, as the expansion proceeds and connectivity among populations is established, homogenization and a resurgence of genetic diversity are to be expected. Few studies have used a fine temporal scale combined with genetic sampling to track range expansions as they proceed in wild animal populations. As a natural experiment, the historical eradication of large terrestrial carnivores followed by their recovery and recolonization may facilitate empirical tests of these ideas. Here, using brown bear (Ursus arctos) as model species, we tested predictions from genetic theory of range expansion. Individuals from all over Finland were genotyped for every year between 1996 and 2010 using 12 validated autosomal microsatellite markers. A latitudinal shift of about 110 km was observed in the distribution and delineation of genetic clusters during this period. As the range expansion proceeded, we found, as theory predicts, that the degree of genetic structure decreased, and that both genetic variation and admixture increased. The genetic consequences of range expansions may first be detected after multiple generations, but we found major changes in genetic composition after just 1.5 generations, accompanied by population growth and increased migration. These rapid genetic changes suggest an ongoing concerted action of geographical and demographic expansion combined with substantial immigration of bears from Russia during the recovery of brown bears within the large ecosystem of northern Europe.

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.