Search

Dispersal is a key process for the maintenance of intraspecific genetic diversity by ensuring gene flow within and between populations. Despite the ongoing expansion of large carnivores in Europe, lynx populations remain fragmented, isolated, and threatened by inbreeding and loss of genetic diversity. In the course of large carnivore monitoring in the Czech Republic, several biological samples of Eurasian lynx were collected outside the permanent occurrence of this species. Using microsatellite genotyping we identified these as four dispersing lynx males and applied multiple methods (Bayesian clustering in STRUCTURE, Principal Component Analysis (PCA), frequency-based method in GENECLASS2, and machine-learning framework in assignPOP) to assign them to possible source populations. For this we used genotypes from five European lynx populations: the Bohemian-Bavarian-Austrian (N = 36), Carpathian (N = 43), Scandinavian (N = 20), Baltic (N = 15), and Harz (N = 23) population. All four dispersers were successfully assigned to different source populations within Europe and each was recorded at a distance of more than 98 km from the edge of the distribution of the source population identified. Such movements are among the longest described for lynx in Central Europe to this point. The findings indicate the ability of lynx males to disperse in human-dominated landscape thus facilitation of these movements via creation and/or protection of potential migratory corridors together with protection of dispersing individuals should be of high importance in conservation of this iconic predator in Central Europe.

Loss of biodiversity is accelerating, including the loss of genetic diversity. Conservation of small, isolated populations may be important, as they can provide valuable contributions to overall genetic variation and long-term viability of species. Furthermore, such populations may play an essential role in adaptation to new environments following changes in e.g. land-use and climate. Dracocephalum ruyschiana is a threatened plant species throughout its European distribution, but 25% of the European populations are situated within Norway. Therefore, the species has its own action plan in Norway, which includes demographic monitoring. However, this monitoring does not cover genetic variation nor is the selection of monitored populations based on genetic differentiation, therefore this fundamental level of biodiversity is overlooked. We analyzed 43 sites using 96 SNPs developed for D. ruyschiana, to investigate whether the monitored populations cover the genetic variation and differentiation found within the Norwegian distribution. The results show structuring and differentiation between populations and indicate that there are at least four distinct genetic groups, of which only two are covered extensively by current demographic monitoring. We suggest that two sites representing the two other genetic groups should be included in the national monitoring program to better conserve the genetic variation found in the Norwegian population of D. ruyschiana. Overall, our results highlight the importance of an integrated, interdisciplinary framework to better monitor and conserve biodiversity at several levels.

Abstract In birds with extrapair mating, older males usually have higher fertilization success than younger males. Two hypotheses can potentially explain this pattern: 1) females prefer older, and often more ornamented males, or 2) older males invest more in reproduction and fertility than younger males. Here we studied factors associated with age-related male fertilization success in a population of barn swallows Hirundo rustica in Canada. We document that male fertilization success increased gradually up to a minimum age of four-year old. The age effect was especially strong for the number of extrapair offspring obtained and the occurrence of a second brood. The higher fertilization success of older males was also associated with an early start of breeding in spring. The length of the elongated outermost tail feathers, a postulated male ornament preferred by females, also increased with age (in both sexes), but it was not a significant predictor of male fertilization success within age classes. Male fertility traits, especially testis size, but also sperm motility and sperm velocity, increased significantly across age groups. Our results suggest that the higher fertilization success by older males is due to their higher reproductive investments and that their longer tails are an adaptation to early arrival on the breeding grounds. Significance statement The barn swallow is a socially monogamous passerine with extensive extrapair mating. We found that males become more successful in siring both withinpair and extrapair offspring as they become older. Their increased fertilization success was associated with a higher reproductive effort as indicated by larger testes, more motile sperm, and an earlier start of breeding in spring. The length of the outer tail feathers increased with age in both sexes, but long tails did not enhance male fertilization success among males of the same age. Long tails are probably an adaptation to rapid migration and earlier arrival on the breeding grounds. Our findings suggest that the commonly observed age-related increase in male fertilization success in passerine birds is better explained by life history theory than by sexual selection theory.

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.

The occurrence of organohalogenated compounds (OHCs) in wildlife has received considerable attention over the last decades. Among the matrices used for OHCs biomonitoring, feathers are particularly useful as they can be collected in a minimally or non-invasive manner. In this study, concentrations of various legacy OHCs -polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs)-, as well as emerging OHCs -per- and polyfluoroalkyl substances (PFAS) and organophosphate ester flame retardants (OPEs)- were determined in feathers of 72 Eurasian eagle-owls (Bubo bubo) from Norway, with the goal of studying spatiotemporal variation using a non-invasive approach. Molted feathers were collected at nest sites from northern, central and southern Norway across four summers (2013-2016). Additionally, two museum-archived feathers from 1979 to 1989 were included. Stable carbon (δ13C) and nitrogen isotopes (δ15N) were used as dietary proxies. In total, 11 PFAS (sum range 8.25-215.90 ng g

The factors underlying gene flow and genomic population structure in vagile seabirds are notoriously difficult to understand due to their complex ecology with diverse dispersal barriers and extensive periods at sea. Yet, such understanding is vital for conservation management of seabirds that are globally declining at alarming rates. Here, we elucidate the population structure of the Atlantic puffin (Fratercula arctica) by assembling its reference genome and analyzing genome-wide resequencing data of 72 individuals from 12 colonies. We identify four large, genetically distinct clusters, observe isolation-by-distance between colonies within these clusters, and obtain evidence for a secondary contact zone. These observations disagree with the current taxonomy, and show that a complex set of contemporary biotic factors impede gene flow over different spatial scales. Our results highlight the power of whole genome data to reveal unexpected population structure in vagile marine seabirds and its value for seabird taxonomy, evolution and conservation., Kersten et al. sequence a draft genome for the Atlantic puffin and report its population structure, genetic diversity and gene flow among four main clusters of populations across the northern Atlantic. These results identify a secondary contact zone between the puffins from the High Arctic and other colonies and proposes a new population structure from the currently recognized three subspecies.

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

Genome-wide association studies provide good opportunities for studying the genetic basis of adaptive traits in wild populations. Yet, previous studies often failed to identify major effect genes. In this study, we used high-density single nucleotide polymorphism and individual fitness data from a wild non-model species. Using a whole-genome approach, we identified the MC1R gene as the sole causal gene underlying Arctic fox Vulpes lagopus fur colour. Further, we showed the adaptive importance of fur colour genotypes through measures of fitness that link ecological and evolutionary processes. We found a tendency for blue foxes that are heterozygous at the fur colour locus to have higher fitness than homozygous white foxes. The effect of genotype on fitness was independent of winter duration but varied with prey availability, with the strongest effect in years of increasing rodent populations. MC1R is located in a genomic region with high gene density, and we discuss the potential for indirect selection through linkage and pleiotropy. Our study shows that whole-genome analyses can be successfully applied to wild species and identify major effect genes underlying adaptive traits. Furthermore, we show how this approach can be used to identify knowledge gaps in our understanding of interactions between ecology and evolution.

The occurrence of organohalogenated compounds (OHCs) in wildlife has received considerable attention over the last decades. Among the matrices used for OHCs biomonitoring, feathers are particularly useful as they can be collected in a minimally or non-invasive manner. In this study, concentrations of various legacy OHCs –polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs)–, as well as emerging OHCs –per- and polyfluoroalkyl substances (PFAS) and organophosphate ester flame retardants (OPEs)– were determined in feathers of 72 Eurasian eagle-owls (Bubo bubo) from Norway, with the goal of studying spatiotemporal variation using a non-invasive approach. Molted feathers were collected at nest sites from northern, central and southern Norway across four summers (2013–2016). Additionally, two museum-archived feathers from 1979 to 1989 were included. Stable carbon (δ13C) and nitrogen isotopes (δ15N) were used as dietary proxies. In total, 11 PFAS (sum range 8.25–215.90 ng g− 1), 15 PCBs (4.19–430.01 ng g− 1), 6 OCPs (1.48–220.94 ng g− 1), 5 PBDEs (0.21–5.32 ng g− 1) and 3 OPEs (4.49–222.21 ng g− 1) were quantified. While we observed large variation in the values of both stable isotopes, suggesting a diverse diet of the eagle owls, only δ13C seemed to explain variation in PFAS concentrations. Geographic area and year were influential factors for δ15N and δ13C. Considerable spatial variation was observed in PFAS levels, with the southern area showing higher levels compared to northern and central Norway. For the rest of OHCs, we observed between-year variations; sum concentrations of PCBs, OCPs, PBDEs and OPEs reached a maximum in 2015 and 2016. Concentrations from 1979 to 1989 were within the ranges observed between 2013 and 2016. Overall, our data indicate high levels of legacy and emerging OHCs in a top predator in Norway, further highlighting the risk posed by OHCs to wildlife. Keywords: Bird of prey Feathers Isotopes OPE POP PFAS

In species with internal fertilization, the female genital tract appears challenging to sperm, possibly resulting from selection on for example ovarian fluid to control sperm behaviour and, ultimately, fertilization. Few studies, however, have examined the effects of swimming media viscosities on sperm performance. We quantified effects of media viscosities on sperm velocity in promiscuous willow warblers Phylloscopus trochilus . We used both a reaction norm and a character-state approach to model phenotypic plasticity of sperm behaviour across three experimental media of different viscosities. Compared with a standard medium (Dulbecco's Modified Eagle Medium, DMEM), media enriched with 1% or 2% w/v methyl cellulose decreased sperm velocity by up to about 50%. Spermatozoa from experimental ejaculates of different males responded similarly to different viscosities, and a lack of covariance between elevations and slopes of individual velocity-by-viscosity reaction norms indicated that spermatozoa from high- and low-velocity ejaculates were slowed down by a similar degree when confronted with high-viscosity environments. Positive cross-environment (1% versus 2% cellulose) covariances of sperm velocity under the character-state approach suggested that sperm performance represents a transitive trait, with rank order of individual ejaculates maintained when expressed against different environmental backgrounds. Importantly, however, a lack of significant covariances in sperm velocity involving a cellulose concentration of 0% indicated that pure DMEM represented a qualitatively different environment, questioning the validity of this widely used standard medium for assaying sperm performance. Enriching sperm environments along ecologically relevant gradients prior to assessing sperm performance will strengthen explanatory power of in vitro studies of sperm behaviour.

The Changeable Hawk-Eagle Nisaetus cirrhatus complex is represented by two taxa in mainland India: N. c. cirrhatus in the northern plains and peninsula and N. c. limnaeetus in the Himalayan foothills. Traditionally these taxa have been regarded as subspecies of one species, but recently they have been proposed to be different species. Here, we use an integrative taxonomic approach based on considerations of plumage, biometrics, genetics and vocalizations. Several plumage characters are significantly different between the two taxa, but crest length was the only one of 56 characters that was diagnostically different, with no overlap. About 30% of the birds had intermediate crest lengths, suggesting that they are hybrids or backcrosses, as also supported by the microsatellite results. PCAs of adult plumage show many intermediate individuals, irrespective of whether these birds were collected near a putative contact zone. There is restricted gene flow between the two taxa, presumably as a result of their largely allopatric distributions. On current knowledge, reproductive isolation appears to be weak at best, and we therefore recommend continuing to regard limnaeetus morphology, genetics, vocalization, integrative taxonomy

After decades, even centuries of persecution, large carnivore populations are widely recovering in Europe. Considering the recent recovery of the wolverine (Gulo gulo) in Finland, our aim was to evaluate genetic variation using 14 microsatellites and mtDNA control region (579 bp) in order (1) to determine whether the species is represented by a single genetic population within Finland, (2) to quantify the genetic diversity, and (3) to estimate the effective population size. We found two major genetic clusters divided between eastern and northern Finland based on microsatellites (FST= 0.100) but also a significant pattern of isolation by distance. Wolverines in western Finland had a genetic signature similar to the northern cluster, which can be explained by former translocations of wolverines from northern to western Finland. For both main clusters, most estimates of the effective population sizeNewere below 50. Nevertheless, the genetic diversity was higher in the eastern cluster (HE= 0.57,AR= 4.0,AP= 0.3) than in the northern cluster (HE= 0.49,AR= 3.7,AP= 0.1). Migration between the clusters was low. Two mtDNA haplotypes were found: one common and identical to Scandinavian wolverines; the other rare and not previously detected. The rare haplotype was more prominent in the eastern genetic cluster. Combining all available data, we infer that the genetic population structure within Finland is shaped by a recent bottleneck, isolation by distance, human-aided translocations and postglacial recolonization routes.

Background Female promiscuity is highly variable among birds, and particularly among songbirds. Comparative work has identified several patterns of covariation with social, sexual, ecological and life history traits. However, it is unclear whether these patterns reflect causes or consequences of female promiscuity, or if they are byproducts of some unknown evolutionary drivers. Moreover, factors that explain promiscuity at the deep nodes in the phylogenetic tree may be different from those important at the tips, i.e. among closely related species. Here we examine the relationships between female promiscuity and a broad set of predictor variables in a comprehensive data set (N = 202 species) of Passerides songbirds, which is a highly diversified infraorder of the Passeriformes exhibiting significant variation in female promiscuity. Results Female promiscuity was highly variable in all major clades of the Passerides phylogeny and also among closely related species. We found several significant associations with female promiscuity, albeit with fairly small effect sizes (all R2 ≤ 0.08). More promiscuous species had: 1) less male parental care, particularly during the early stages of the nesting cycle (nest building and incubation), 2) more short-term pair bonds, 3) greater degree of sexual dichromatism, primarily because females were drabber, 4) more migratory behaviour, and 5) stronger pre-mating sexual selection. In a multivariate model, however, the effect of sexual selection disappeared, while the other four variables showed additive effects and together explained about 16% of the total variance in female promiscuity. Female promiscuity showed no relationship with body size, life history variation, latitude or cooperative breeding. Conclusions We found that multiple traits were associated with female promiscuity, but these associations were generally weak. Some traits, such as reduced parental care in males and more cryptic plumage in females, might even be responses to, rather than causes of, variation in female promiscuity. Hence, the high variation in female promiscuity among Passerides species remains enigmatic. Female promiscuity seems to be a rapidly evolving trait that often diverges between species with similar ecologies and breeding systems. A future challenge is therefore to understand what drives within-lineage variation in female promiscuity over microevolutionary time scales. Electronic supplementary material The online version of this article (10.1186/s12862-019-1493-1) contains supplementary material, which is available to authorized users.

This study aimed to understand the movement behaviour and utilization distributions of Kori bustards in space and time in the Serengeti ecosystem. A total of 14 individuals were tracked with the aid of GPS (Geographical positioning system) satellite transmitters, and their sexes were identified using DNA analysis. A species utilization distribution was estimated using the Brownian bridge movement model (hereafter dBBMM) in which the probability of being in an area is conditioned by starting and ending (GPS) relocations. Resource selections were analysed by comparing the GPS relocations with locations randomly placed within each individual’s region of utilization in a spatio-temporal approach. Vegetation information was derived from a Serengeti GIS vegetation map and Data Centre and was reclassified as Open grassland, Dense grassland, Shrubbed grassland, Treed grassland, Shrubland, and Woodland. The Shannon diversity index for vegetation was calculated based on the original vegetation classification. Used versus non-used habitats were contrasted using a generalized linear mixed-effects model with a binomial distribution. The results indicated that males were 21.5% more mobile than females, and movements were 6.3% more diffuse during the nonbreeding period compared to the breeding period (7.59 versus 7.14, respectively). Contrasting models indicated that males preferred more open grasslands during the non-breeding period and also preferred closed and shrubbed grassland during the breeding period. Females preferred more woody vegetation during the non-breeding season compared to the breeding season. The most parsimonious model indicated that females preferred to stay closer to rivers and diverse areas during the non-breeding period whereas males preferred areas that were farther from rivers and homogenous. Homogeneous areas were preferred during the breeding period, and heterogeneous areas were preferred during the non-breeding period. We conclude that the movement behaviours of Kori bustards changes with the season and habitat. Further research is needed to understand the factors driving the seasonal movement of Kori bustards in the Serengeti ecosystem. © 2019 Mmassy et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Basic knowledge about the genetic mating system is lacking for the great majority of the approximately 10,000 extant bird species. Filling this knowledge gap is not only critical for a comprehensive understanding of the reproductive ecology of each particular species, but also for increasing the power of comparative approaches to uncover and explain interspecific patterns of variation in avian reproductive traits. Using six polymorphic microsatellite markers, we here present the first parentage study in the socially monogamous Fieldfare Turdus pilaris. In parallel, we also examine variation in sperm morphology and relationships between sperm traits and paternity loss of social males. Across two study years, extra-pair paternity was detected in 46.4% (95% CI: 28.9-64.9%) of 28 broods, and on average 27.6% (95% CI: 16.8-41.9%) of nestlings per brood were extra-pair offspring in a population in central Norway. These observed extra-pair paternity rates fall within the range of reported estimates of extra-pair paternity for four congeneric Turdus species (between 36 and 65% of broods and 27 and 46% of nestlings). Sperm total length was 87.0 +/- 2.9 (SD) mu m (range 79.7-96.8 mu m) and 59.3% (95% CI: 37.1-73.3%) of the total phenotypic variation in sperm total length was explained by differences between sperm samples collected from 17 different males. The among-sample coefficient of variation in mean sperm total length was 2.70% (95% CI: 1.99-3.17%). We found no evidence for effects of sperm total length or relative midpiece length on loss of paternity among broods of 13 males.

Here we describe 16 novel microsatellite loci in the genome of the dragonhead sap beetle Thymogethes norvegicus, which were identifi ed using a next-generation sequencing approach. In 40 dragonhead sap beetles from a population in southern Norway, we found 2–4 alleles per locus. Expected heterozygosity ranged from 0.18–0.65. Three of the loci showed significant deviation from Hardy-Weinberg equilibrium. After correcting for multiple tests, one pairwise locus combination displayed significant linkage disequilibrium. These novel microsatellite loci will be useful for future population structure, genetic diversity and conservation genetic studies of the dragonhead sap beetle. Coleoptera, Nitidulidae, Thymogethes norvegicus, microsatellite loci, genetic diversity

The northern dragonhead Dracocephalum ruyschiana is a plant species experiencing a dramatic population decline that has led to the species being listed on Red Lists for species in many European countries. Here we used restriction-site associated DNA sequencing to isolate and characterize a panel of 96 novel SNP markers from 44 individuals encompassing most of the species range in Norway. The 96 SNPs were adapted for the Fluidigm platform and evaluated by screening another 24 northern dragonheads from a population in southern Norway. The panel of SNP markers developed here are expected to be useful for elucidating genetic diversity and population genetic structure in the northern dragonhead. Dracocephalum ruyschiana · Genetic diversity · Population genetics · SNP

During their flightless summer moult, Taiga Bean Geese Anser fabalis fabalis gather at communal moulting sites. Individuals from the Nord-Trøndelag breeding area in Norway have been observed to join with local individuals on moulting sites in Vilhelmina Municipality, Sweden. These two groups show distinct features in breeding habitat and migratory behaviour, but are they also genetically distinct? We used 12 microsatellite loci for genotyping 109 blood, feather and faecal samples from three sampling areas (Røyrvik in Norway and Stalon and Nästansjö in Sweden) to examine genetic diversity and structure. Clustering and Principal Coordinate analyses of all samples unveiled at least two distinct clusters, which were unevenly distributed over the sampling sites. Grouped by sampling sites, AMOVA and FST analyses showed that samples from the three sites differed genetically. These differences were larger between Røyrvik and Nästansjö than between Stalon and the other two. Relatedness was high among the Røyrvik samples. From our results we conclude that one of the clusters describes the Røyrvik breeding subpopulation, while the other(s) breed mainly in Sweden. Although these subpopulations simultaneously use the same moulting area in Vilhelmina, they appear to be ecologically, behaviourally and genetically distinct, in particular the Røyrvik sub-population. For goose conservation and management, we suggest that the Nord-Trøndelag (Røyrvik) subpopulation is considered a separate flyway management unit. Unravelling the Swedish sub-populations will need further study. For bird conservation is general, we suggest active genetic sampling for detailed population structure analyses and subsequent differentiated conservation and/or management schemes. Birds of different feather flock together - genetic structure of Taiga Bean Goose in Central Scandinavia

Capsule: A description of the methods used for monitoring the Golden Eagle Aquila chrysaetos in Norway Aims: To provide a comprehensive description of monitoring methods. Methods: The intensive monitoring of the Golden Eagle in Norway started in 1991 as part of a national monitoring programme initiated by the Directorate for Nature Management (now the Norwegian Environment Agency). It has since become part of the Norwegian Large Predator Programme, and Golden Eagles are currently being monitored in 12 separate areas. Here we provide a comprehensive description of the current methods used in the intensive monitoring, with definitions, fieldwork and evaluation criteria for the final breeding status. In addition, a description of estimation of annual adult survival by genetic analysis is given. We describe the current methodology used in the intensive part of the Golden Eagle monitoring in Norway. Results: We present some results derived from the Norwegian monitoring system and discuss the potential for further analyses. In addition, we highlight aspects in the monitoring of the Golden Eagles where our methods deviate slightly from those applied in other countries and the potential effects of these. Conclusions: Intensive long-term monitoring programmes, such as this, will become increasingly valuable for monitoring the impact of environmental change, both from natural phenomena and from anthropogenic activities. To facilitate comparisons among the Golden Eagle monitoring programmes, detailed knowledge about the various methods applied is important.

Genetic analysis has shown to be an important and efficient method for monitoring species, but the lack of genetic markers constrains such monitoring for many species. Here, we cross-amplified microsatellite loci in the snowy owl (Bubo scandiacus) using primers previously characterized in other species. Among polymorphic loci, we selected 12 loci that amplified relatively short fragments (

Postcopulatory sexual selection (PCSS) in internally fertilizing vertebrates is a topic of great interest, yet relatively little is known about the characteristics of sperm and ejaculates that confer an advantage in PCSS. In this study, we investigated several measures of sperm morphology that potentially contribute to fertilization success under PCSS. We tested whether sperm morphology related to success in PCSS (via extra-pair paternity) in house wrens (Troglodytes aedon). We found no evidence that sperm morphology differed between extra-pair sires and the within-pair males they cuckolded, nor that sperm morphology correlated with the proportion of within-pair offspring sired, the number of extra-pair offspring sired, or the total annual reproductive success. Male behavioral strategies may affect the probability that their sperm compete with other males’ sperm and that their sperm succeed under competition. Effects of these behavioral strategies, as well as differences between males in sperm number, could mask the effects of sperm morphology on the outcome of PCSS. Despite moderate levels of extra-pair paternity, selection on sperm may be relatively weak in house wrens. Further work is needed to understand general patterns in how sperm morphology relates to fertilization success within species.

Sperm morphometry and motility are important male traits affecting fertility and post-copulatory competitive ability. However, studies examining consistency in sperm traits over time for individual males are rather scarce, particularly in free-living animals. Here, we report our analysis of various measures of sperm morphometry and sperm motility in the Barn Swallow (Hirundo rustica), both within and between breeding seasons. We found high within-season repeatabilities for all measures of sperm morphometry (all r > 0.70) and moderate repeatability of sperm motility (all r > 0.42). Between seasons, repeatabilities were somewhat lower, but total sperm length and flagellum length also showed very high repeatabilities between seasons (r > 0.85). Furthermore, sperm motility showed higher repeatabilities between seasons than within a season (all r > 0.63). Although measures of sperm motility are likely to be more plastic due to, for example, changes in body condition, they still showed moderate repeatabilities within and between seasons. Our study results also imply that a single ejaculate would be sufficient to obtain representative measures of an individual’s sperm quality traits within the same breeding season.

Spermatozoa vary greatly in size and shape among species across the animal kingdom. Postcopulatory sexual selection is thought to be the major evolutionary force driving this diversity. In contrast, less is known about how sperm size varies among populations of the same species. Here, we investigate geographic variation in sperm size in barn swallows Hirundo rustica, a socially monogamous passerine with a wide Holarctic breeding distribution. We included samples from seven populations and three subspecies: five populations of ssp. rustica in Europe (Czech, Italy, Norway, Spain, and Ukraine), one population of ssp. transitiva in Israel, and one population of ssp. erythrogaster in Canada. All sperm traits (head length, midpiece length, tail length, and total length) varied significantly among populations. The variation among the European rustica populations was much lower than the differences among subspecies, indicating that sperm traits reflect phylogenetic distance. We also performed a test of the relationship between the coefficient of between-male variation in total sperm length and extrapair paternity levels across different populations within a species. Recent studies have found a strong negative relationship between sperm size variation and extrapair paternity among species. Here, we show a similar negative relationship among six barn swallow populations, which suggests that the variance in male sperm length in a population is shaped by the strength of stabilizing postcopulatory sexual selection.

It is well documented that sperm size and structure varies considerably among avian species, but we know much less about the extent of intraspecific variation in sperm morphometry and its possible co-variation with somatic traits like body size and condition. Here, we investigate patterns of sperm length variation and co-variation in a population of House Wrens (Troglodytes aedon). Total sperm length showed considerable between-male variation, with high repeatability between seasons indicative of a strong genetic basis for this trait. However, we also detected a seasonal increase in the flagellum:head length ratio, which might indicate phenotypic plasticity or adjustment in the relative size of sperm components. The variation in total sperm length within an ejaculate sample was higher for males sampled very early in the season, which may reflect more heterogeneity in the size of seminiferous tubules when testes are growing. None of the studied sperm morphometry traits correlated significantly with any measures of male body size or physiological condition. Further studies are needed to reveal if the observed individual variation in sperm morphology plays any functional or adaptive role.

Birds breeding on islands often exhibit lower rates of extrapair paternity than their mainland counterparts, perhaps explained by low genetic variation, ‘slower’ life histories and reduced sexual selection in island populations. Extrapair paternity was apparent in 39% (12/19) of broods, and encompassed 15% (21/137) of nestlings, in a population of African Blue Tits Cyanistes teneriffae, in Tenerife, Canary Islands. There were no cases of intraspecific brood parasitism. The incidence of extrapair young lies in the upper range of that reported from mainland populations of the closely related Eurasian Blue Tit Cyanistes caeruleus. We conclude that there is no strong island effect on the genetic mating system in the Cyanistes species group but that extrapair paternity rates in Cyanistes are greater at southern latitudes.

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.

Microsatellites mutate frequently by replication slippage. Empirical evidence shows that the probability of such slippage mutations may increase with the length of the repeat region as well as exposure to environmental mutagens, but the mutation rate can also differ between the male and female germline. It has been hypothesized that more intense sexual selection or sperm competition can also lead to elevated mutation rates, but the empirical evidence is inconclusive. Here, we analyzed the occurrence of germline slippage mutations in the hypervariable pentanucleotide microsatellite locus HrU10 across six species of swallow (Aves: Hirundinidae). These species exhibit marked differences in the length range of the microsatellite, as well as differences in the intensity of sperm competition. We found a strong effect of microsatellite length on the probability of mutation, but no residual effect of species or their level of sperm competition when the length effect was accounted for. Neither could we detect any difference in mutation rate between tree swallows (Tachycineta bicolor) breeding in Hamilton Harbour, Ontario, an industrial site with previous documentation of elevated mutation rates for minisatellite DNA, and a rural reference population. However, our cross-species analysis revealed two significant patterns of sex differences in HrU10 germline mutations: (1) mutations in longer alleles occurred typically in the male germline, those in shorter alleles in the female germline, and (2) male germline mutations were more often expansions than contractions, whereas no directional bias was evident in the female germline. These results indicate some fundamental differences in male and female gametogenesis affecting the probability of slippage mutations. Our study also reflects the value of a comparative, multi-species approach for locus-specific mutation analyses, through which a wider range of influential factors can be assessed than in single-species studies.

1. Why are some common and apparently suitable resources avoided by potential users? This interesting ecological and evolutionary conundrum is vividly illustrated by obligate brood parasites. Parasitic birds lay their eggs into nests of a wide range of host species, including many rare ones, but do not parasitize some commonly co-occurring potential hosts. 2. Attempts to explain the absence of parasitism in common potential hosts are limited and typically focused on single-factor explanations while ignoring other potential factors. We tested why thrushes Turdus spp. are extremely rarely parasitized by common cuckoos Cuculus canorus despite breeding commonly in sympatry and building the most conspicuous nests among forest-breeding passerines. 3. No single examined factor explained cuckoo avoidance of thrushes. Life-history traits of all six European thrush species and the 10 most frequently used cuckoo hosts in Europe were similar except body/egg size, nest design and nestling diet. 4. Experiments (n = 1211) in several populations across Europe showed that host defences at egg-laying and incubation stages did not account for the lack of cuckoo parasitism in thrushes. However, cross-fostering experiments disclosed that various factors during the nestling period prevent cuckoos from successfully parasitizing thrushes. Specifically, in some thrush species, the nest cup design forced cuckoo chicks to compete with host chicks with fatal consequences for the parasite. Other species were reluctant to care even for lone cuckoo chicks. 5. Importantly, in an apparently phylogenetically homogenous group of hosts, there were interspecific differences in factors responsible for the absence of cuckoo parasitism. 6. This study highlights the importance of considering multiple potential factors and their interactions for understanding absence of parasitism in potential hosts of parasitic birds. In the present study, comparative and experimental procedures are integrated, which represent a novel approach that should prove useful for the understanding of interspecific ecological relationships in general.

The evolutionary forces that shape and maintain variation in sperm traits across species in birds have recently attracted increased attention. Much less is known about the patterns, the causes and the consequences of variation in sperm traits within and, in particular, between populations of the same species. We here analyse intraspecific variation in sperm dimensions within and between two populations of the Coal Tit Periparus ater, a socially monogamous passerine with a high frequency of extra-pair paternity. Spermatozoa from a Norwegian population had a greater mean total length compared to those from a German population, mainly as a result of longer sperm heads. Sperm head length in the Norwegian population also accounted for a larger percentage of sperm total length suggesting differences in sperm proportions between populations. Furthermore, spermatozoa from the Norwegian population showed lower mean within-male variation in sperm length and there was significant between-male variation in sperm total length within each of the two populations. We discuss these results in the light of recent comparative evidence suggesting relationships between sperm length, and variation in sperm length, and the frequency of extra-pair paternity across passerine birds.

Sperm competition is widespread among animal taxa and considered a major force in sperm evolution. Recent comparative studies have indicated that sperm competition selects for high sperm production capacity and long and fast-swimming spermatozoa across species. Here, we examine the role of sperm quantity and quality for fertilization success of individual males in a Canadian population of tree swallows Tachycineta bicolor, a socially monogamous, but highly promiscuous passerine. Male fertilization success (the sum of withinpair and extrapair young) was significantly associated with the size of the cloacal protuberance (a proxy for sperm quantity), but not with sperm size or in vitro sperm swimming speed. In a multivariate analysis, both cloacal protuberance volume and relative sperm midpiece size (i.e. high mitochondrial loading) had significant effects on male fertilization success. However, relative sperm midpiece size was not associated with fertilization success in a simple regression. Further, both cloacal protuberance volume and relative midpiece size had significant effects on sperm velocity, both in simple regressions and in a multivariate analysis. The finding that males with large relative midpiece size had both higher fertilization success and faster swimming sperm, suggests an indirect link between sperm morphology and male fertility mediated through sperm velocity. In conclusion, both quantitative and qualitative sperm traits seem to affect male fertilization success in tree swallows.

Melanins are common pigments used for colouration in animal tissue and recent evidence suggests that they may be condition-dependent signals used in mate choice. We examined the hypothesis that individuals moulting during migration produce relatively dull feathers compared to individuals moulting on the breeding grounds prior to migration because of the costs associated with overlapping these two activities. We estimated both the colour and moulting latitude of melanin-based feathers in barn swallows (Hirundo rustica). Moulting latitude was inferred from stable-hydrogen isotope (dD) values of individuals that were known to have bred at the same site the previous year. Contrary to expectations, dD values in feathers suggested that most, if not all, individuals moulted their feathers south of the breeding grounds. Furthermore, we found that males moulting at southern latitudes tended to produce darker, more rufous coloured throat feathers compared to males moulting at more northern latitudes, closer to the breeding grounds. However, we found no such relationship in females. Males could face a trade-off between growing new but relatively dull feathers before or just after the commencement of migration versus waiting to grow more rufous coloured feathers well into fall migration or on the wintering grounds. We suggest that this trade-off could produce an honest signal of male quality if only high quality males can afford to migrate large distances on feathers that are greater than a year old.

We investigated extra-pair paternity (EPP) among 46 young in 32 broods of 19 social pairs of Black Guillemot Cepphus grylle, using a combination of three polymorphic microsatellite markers. The analysis was based on blood samples collected over eight years from a population of individually colour-ringed birds in North Norway. All chicks shared an allele on all three loci with both social parents. Hence, there were no cases of extra-pair parentage. Given our sample sizes, the upper 95% confidence limit of EPP was estimated to 6.2% of the young and 8.9% of the broods. Among seven extra chicks, for which we lacked a DNA sample from the social mother, there was one case of genotype mismatch with the social father, but this could also be because of egg dumping. Low occurrence of EPP has now been documented in five of the six species of Atlantic auks.

Sperm swimming speed is an important determinant of male fertility and sperm competitiveness. Despite its fundamental biological importance, the underlying evolutionary processes affecting this male reproductive trait are poorly understood. Using a comparative approach in a phylogenetic framework, we tested the predictions that sperm swim faster with (1) increased risk of sperm competition, (2) shorter duration of female sperm storage, and (3) increased sperm length. We recorded sperm swimming speed in 42 North American and European free-living passerine bird species, representing 35 genera and 16 families. We found that sperm swimming speed was positively related to the frequency of extrapair paternity (a proxy for the risk of sperm competition) and negatively associated with clutch size (a proxy for the duration of female sperm storage). Sperm swimming speed was unrelated to sperm length, although sperm length also increased with the frequency of extrapair paternity. These results suggest that sperm swimming speed and sperm length are not closely associated traits and evolve independently in response to sperm competition in passerine birds. Our findings emphasize the significance of both sperm competition and female sperm storage duration as evolutionary forces driving sperm swimming speed.

Frequent copulation is assumed to be an important male reproductive strategy for paternity assurance in species with female sexual promiscuity. However, the empirical evidence supporting this hypothesis is limited. We examined copulation behaviour in relation to within-pair paternity in the socially monogamous tree swallow, Tachycineta bicolor, a passerine bird species with frequent copulation, a lack of mate guarding and high levels of extrapair paternity. We found that the frequency of within-pair copulation attempts increased and peaked just before the appearance of the first egg. A marked drop in the frequency of copulation attempts was observed once the first egg was laid. Furthermore, the frequency of successful within-pair copulations was highly repeatable for particular pairs within the 3 days prior to appearance of the first egg. Finally, males that obtained a higher number of successful within-pair copulations during this time also sired a larger proportion of the offspring in their own brood. Our findings suggest that frequent copulations just before the start of egg laying is an adaptation to decrease the risk of being cuckolded in the presence of sperm competition, and thus provide empirical support for the paternity assurance hypothesis. Crown Copyright 2008 Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour. All rights reserved. Paternity assurance is critical to male fitness, particularly in species in which females frequently participate in extrapair copulations (EPCs) and males provide costly parental care. In response to fitness loss due to cuckoldry, several types of paternity assurance behaviour have evolved across taxa. One strategy is constant mate guarding by following, as males can protect their paternity by ensuring that no other male has an opportunity to mate with their paired female (Moller & Birkhead 1991). However, for some species, constant mate guarding may be constrained by territory protection or other activities. The paternity assurance hypothesis (Moller 1987; Simmons 1990), also known as the paternity confidence hypothesis (Eens & Pinxten 1995) and the sperm competition hypothesis (Birkhead et al. 1987), suggests that for species in which EPCs are common, males copulate frequently with their pair females to avoid being cuckolded. Males may minimize cuckoldry through frequent copulation by devaluing EPCs that their mates

Extra-pair paternity is common in birds and much research has focussed on the selective advantage of extra-pair matings for both sexes. In contrast, little attention has been given to the fact that in most species the majority of offspring are sired by the social male. We investigated whether extra-pair matings of female bluethroats (Luscinia svecica) are constrained by the presence of the pair male, by detaining males in cages on their territories for one morning during the peak of female fertility. The proportion of offspring sired by extra-pair males was higher in broods where males had been detained (35%) than in control broods (16%), while the proportion of broods that had at least one extra-pair offspring did not differ significantly between experimental (65%) and control broods (44%). Within the experimental group, levels of extra-pair paternity were not related to the day of experiment in relation to start of egg laying, but males caught early in the morning lost more paternity than males caught later on. Our results show that pair males exert constraints on the frequency of extra-pair paternity by being present during the period of peak fertility, which could be a direct effect of their mate guarding effort and/or due to an advantage in sperm competition for pair males.

Paternity in Atlantic Puffins T. Anker-Nilssen et al. Short communica on No evidence of extra-pair paternity in the Atlantic Puffin Fratercula arctica TYCHO ANKER-NILSSEN, 1 * ODDMUND KLEVEN, 2 TOMAS AARVAK 1 † & JAN T. LIFJELD 2 1 Norwegian Institute for Nature Research, NO-7485 Trondheim, Norway 2 Natural History Museum, Department of Zoology, University of Oslo, P.O. Box 1172 Blindern, NO-0318 Oslo, Norway