Genetic diversity, gene flow, and landscape resistance in a pond-breeding amphibian in agricultural and natural forested landscapes in Norway.

Genetic diversity is a key part of biodiversity, threatened by human activities that lead to loss of gene flow and reduction of effective population sizes. Gene flow is a result of both landscape connectivity and demographic processes determining the number of dispersing individuals in space and time. Thus, the effect of human impact on processes determining the level of genetic diversity must be interpreted in the context of basic ecological conditions affecting survival and recruitment. When the intensity of human impact and habitat suitability correlate, the effect on genetic diversity and gene flow may be challenging to predict. We compared genetic diversity, gene flow and landscape resistance in two contrasting landscapes in Norway for the pond-breeding amphibian Triturus cristatus: a highly human-impacted, agricultural landscape with ecologically productive habitats, and a forested landscape with less productive habitats and lower levels of human impact. Our results show that genetic diversity was higher and gene flow lower within the forested landscape. Microclimatic moisture conditions and vegetation cover were important determinants of landscape resistance to gene flow within both landscapes. There were indications that landscape resistance was increased by minor roads in the forested landscape, which was not the case for the agricultural landscape, suggesting a higher vulnerability to human interference within the landscape matrix for the populations in less productive habitats. Our findings suggest that the effect of human impact on genetic diversity may not be straightforward but modulated by the ecological conditions underlying local demographic processes. Populations within both landscapes seem to be vulnerable to loss of genetic diversity, but due to different mechanisms. This has implications for the choice of relevant management actions, that is, increasing population stability may be more relevant within an agricultural landscape still permeable for dispersal, while conserving dispersal corridors may be more appropriate in the forested landscape, to avoid isolation and increased genetic drift.

Population connectivity: dam migration mitigations and contemporary site fidelity in arctic char.

BACKGROUND: Animal feeding and spawning migrations may be limited by physical barriers and behavioral interactions. Dam constructions (e.g. hydropower) commonly include gateways for fish migrations to sustain ecological connectivity. Relative genetic impacts of fish passage devices versus natural processes (e.g. hybrid inferiority) are, however, rarely studied. We examined genetic (i.e. microsatellite) population connectivity of highly migrating lake-dwelling Arctic char (Salvelinus alpinus), introduced 20 generations ago, across and within two subalpine lakes separated by a dam with a subterranean tunnel and spill gates after 7 generations. Due to water flow regime, the time window for fish migration is highly restricted. RESULTS: Char populations, with similar genetic structuring and diversity observed across and within lakes, were admixed across the dam with fishways during feeding. For spawning, however, statistically significant, but very low population differentiation (θ; 0.002 - 0.013) was found in nine out of ten reproductive site comparisons, reflecting interactions between extensive migration (mean first generation (F0) = 10.8%) and initial site fidelity. Simulations indicated that genetic drift among relatively small effective populations (mean N(e) = 62) may have caused the observed contemporary differentiation. Novel Bayesian analyses indicated mean contributions of 71% F0 population hybrids in spawning populations, of which 76% had maternal or paternal native origin. CONCLUSIONS: Ecological connectivity between lakes separated by a dam has been retained through construction of fishways for feeding migration. Considerable survival and homing to ancestral spawning sites in hybrid progeny was documented. Population differentiation despite preceding admixture is likely caused by contemporary reduced reproductive fitness of population hybrids. The study documents the beginning stages of population divergence among spatial aggregations with recent common ancestry.

Optimal foraging by a large ungulate in an extreme environment: Wild mountain reindeer select snow-free feeding habitats in winter.

Optimal foraging models predict that individual animals will optimize net energy gain by intensifying forage activity and/or reducing forage energy cost. Then, the free distribution model predicts an animal's distribution in a patchy landscape will match the distribution of the resources. If not modified by other factors, such patterns may be expected to be particularly explicit in variable and extreme, forage-limited, and patchy environments, notably alpine and Arctic environments during winter. The large ungulate wild mountain reindeer (Rangifer tarandus tarandus) surviving in such environments is used as a model during the forage-limited winter season. The largest wild reindeer area in Western Europe (Hardangervidda, 8130 km2) is actively managed to sustain 10,000-12,000 wild reindeer. Since 2001, 104 different individuals have been GPS-tracked at 3-hr intervals. In winter, mountain reindeer may either choose to seek out and forage in patchy snow-free habitats, typically on top of wind-blown ridges, or use energy-demanding digging through the snow to reach ground forage (cratering). We use late April satellite data from Landsat 5 and 8 (30 × 30 m), airborne laser scanning subsampling (processed to 1 × 1 m grid), and topographic information (1 m resolution) derived from digital aerial photographs (0.25 × 0.25 m resolution) to delineate snow-free patches, constituting less than 694 km2. By overlaying recorded wild reindeer GPS positions winters 2001-2017 (188,942 positions), we document a strong positive selection for snow-free patches, which were used about four times more frequently than expected from a "random walk" model. On a daily basis, the preference for snow-free areas was slightly stronger in the evenings. In the sustainable management of wild mountain reindeer, the area of snow-free patches is an important predictor of winter forage availability and important winter source areas. It may be derived from remote sensing data.

Dynamic micro-geographic and temporal genetic diversity in vertebrates: the case of lake-spawning populations of brown trout (Salmo trutta).

Conservation of species should be based on knowledge of effective population sizes and understanding of how breeding tactics and selection of recruitment habitats lead to genetic structuring. In the stream-spawning and genetically diverse brown trout, spawning and rearing areas may be restricted source habitats. Spatio-temporal genetic variability patterns were studied in brown trout occupying three lakes characterized by restricted stream habitat but high recruitment levels. This suggested non-typical lake-spawning, potentially representing additional spatio-temporal genetic variation in continuous habitats. Three years of sampling documented presence of young-of-the-year cohorts in littoral lake areas with groundwater inflow, confirming lake-spawning trout in all three lakes. Nine microsatellite markers assayed across 901 young-of-the-year individuals indicated overall substantial genetic differentiation in space and time. Nested gene diversity analyses revealed highly significant (< or =P = 0.002) differentiation on all hierarchical levels, represented by regional lakes (F(LT) = 0.281), stream vs. lake habitat within regional lakes (F(HL) = 0.045), sample site within habitats (F(SH) = 0.010), and cohorts within sample sites (F(CS) = 0.016). Genetic structuring was, however, different among lakes. It was more pronounced in a natural lake, which exhibited temporally stable structuring both between two lake-spawning populations and between lake- and stream spawners. Hence, it is demonstrated that lake-spawning brown trout form genetically distinct populations and may significantly contribute to genetic diversity. In another lake, differentiation was substantial between stream- and lake-spawning populations but not within habitat. In the third lake, there was less apparent spatial or temporal genetic structuring. Calculation of effective population sizes suggested small spawning populations in general, both within streams and lakes, and indicates that the presence of lake-spawning populations tended to reduce genetic drift in the total (meta-) population of the lake.

Life histories and ecotype conservation in an adaptive vertebrate: Genetic constitution of piscivorous brown trout covaries with habitat stability.

Ecotype variation in species exhibiting different life history strategies may reflect heritable adaptations to optimize reproductive success, and potential for speciation. Traditionally, ecotypes have, however, been defined by morphometrics and life history characteristics, which may be confounded with individual plasticity. Here, we use the widely distributed and polytypic freshwater fish species brown trout (Salmo trutta) as a model to study piscivorous life history and its genetic characteristics in environmentally contrasting habitats; a large lake ecosystem with one major large and stable tributary, and several small tributaries. Data from 550 fish and 13 polymorphic microsatellites (He = 0.67) indicated ecotype-specific genetic differentiation (θ = 0.0170, p < .0001) among Bayesian assigned small riverine resident and large, lake migrating brown trout (>35 cm), but only in the large tributary. In contrast, large trout did not constitute a distinct genetic group in small tributaries, or across riverine sites. Whereas life history data suggest a small, river resident and a large migratory piscivorous ecotype in all studied tributaries, genetic data indicated that a genetically distinct piscivorous ecotype is more likely to evolve in the large and relatively more stable river habitat. In the smaller tributaries, ecotypes apparently resulted from individual plasticity. Whether different life histories and ecotypes result from individual plasticity or define different genetic types, have important consequence for conservation strategies.

Improved work ability and return to work following vocational multidisciplinary rehabilitation of subjects on long-term sick leave.

OBJECTIVE: To evaluate a vocational multidisciplinary rehabilitation programme for patients on long-term sick leave with respect to their work ability and return to work. METHODS: A multidisciplinary rehabilitation programme was administered to an intervention group of 183 patients on long-term sick leave (mean 12.2 months). Effects of the treatment were compared with a control group (n = 96) recruited from the national sickness insurance record of patients on sick leave of 6-12 months duration (mean 11.5 months). Perceived work ability, return to work, background factors and psychosocial aspects of work were assessed on the basis of questionnaires at baseline and after 4 months. RESULTS: Perceived work ability of the intervention group improved significantly after 4 months compared with the control group (p < 0.01). In the intervention group, 80% had returned to work compared with 66% in the control group (p = 0.06). Return to work after 4 months was predicted by good work ability at baseline, improved work motivation, improved work ability at follow-up and increased rumours of change in the workplace (R2 26.1-38.6%, p < 0.05). CONCLUSION: This multidisciplinary rehabilitation programme significantly improved perceived work ability compared with treatment as usual.

Herbivore grazing-or trampling? Trampling effects by a large ungulate in cold high-latitude ecosystems.

Mammalian herbivores have important top-down effects on ecological processes and landscapes by generating vegetation changes through grazing and trampling. For free-ranging herbivores on large landscapes, trampling is an important ecological factor. However, whereas grazing is widely studied, low-intensity trampling is rarely studied and quantified. The cold-adapted northern tundra reindeer (Rangifer tarandus) is a wide-ranging keystone herbivore in large open alpine and Arctic ecosystems. Reindeer may largely subsist on different species of slow-growing ground lichens, particularly in winter. Lichen grows in dry, snow-poor habitats with frost. Their varying elasticity makes them suitable for studying trampling. In replicated factorial experiments, high-resolution 3D laser scanning was used to quantify lichen volume loss from trampling by a reindeer hoof. Losses were substantial, that is, about 0.3 dm3 per imprint in dry thick lichen, but depended on type of lichen mat and humidity. Immediate trampling volume loss was about twice as high in dry, compared to humid thin (2-3 cm), lichen mats and about three times as high in dry vs. humid thick (6-8 cm) lichen mats, There was no significant difference in volume loss between 100% and 50% wetted lichen. Regained volume with time was insignificant for dry lichen, whereas 50% humid lichen regained substantial volumes, and 100% humid lichen regained almost all lost volume, and mostly within 10-20 min. Reindeer trampling may have from near none to devastating effects on exposed lichen forage. During a normal week of foraging, daily moving 5 km across dry 6- to 8-cm-thick continuous lichen mats, one adult reindeer may trample a lichen volume corresponding to about a year's supply of lichen. However, the lichen humidity appears to be an important factor for trampling loss, in addition to the extent of reindeer movement.

Merging and comparing three mitochondrial markers for phylogenetic studies of Eurasian reindeer (Rangifer tarandus).

Phylogenetic analyses provide information that can be useful in the conservation of genetic variation by identifying intraspecific genetic structure. Reconstruction of phylogenetic relationships requires the use of markers with the appropriate amount of variation relative to the timeframe and purpose of the study. Here, genetic structure and clustering are inferred from comparative analyses of three widely used mitochondrial markers, the CR, cytb and the COI region, merged and separately, using Eurasian reindeer as a model. A Bayesian phylogeny and a MJ network, both based on the merged dataset, indicate several distinct maternal haplotype clusters within Eurasian reindeer. In addition to confirm previously described clusters, two new subclusters were found. When comparing the results from the merged dataset with the results from analyses of the three markers separately, similar clustering was found in the CR and COI phylogenies, whereas the cytb region showed poor resolution. Phylogenetic analyses of the merged dataset and the CR revealed congruent results, implying that single sequencing analysis of the CR is an applicable method for studying the haplotype structure in Eurasian reindeer.

Colonizing the High Arctic: Mitochondrial DNA Reveals Common Origin of Eurasian Archipelagic Reindeer (Rangifer tarandus).

In light of current debates on global climate change it has become important to know more on how large, roaming species have responded to environmental change in the past. Using the highly variable mitochondrial control region, we revisit theories of Rangifer colonization and propose that the High Arctic archipelagos of Svalbard, Franz Josef Land, and Novaia Zemlia were colonized by reindeer from the Eurasian mainland after the last glacial maximum. Comparing mtDNA control region sequences from the three Arctic archipelagos showed a strong genetic connection between the populations, supporting a common origin in the past. A genetic connection between the three archipelagos and two Russian mainland populations was also found, suggesting colonization of the Eurasian high Arctic archipelagos from the Eurasian mainland. The age of the Franz Josef Land material (>2000 years before present) implies that Arctic indigenous reindeer colonized the Eurasian Arctic archipelagos through natural dispersal, before humans approached this region.

Molecular responses to toxicological stressors: profiling microRNAs in wild Atlantic salmon (Salmo salar) exposed to acidic aluminum-rich water.

Atlantic salmon (Salmo salar) is among the most sensitive organisms toward acidic, aluminum exposure. Main documented responses to this type of stress are a combination of hypoxia and loss of blood plasma ions. Physiological responses to stress in fish are often grouped into primary, secondary and tertiary responses, where the above mentioned effects are secondary responses, while primary responses include endocrine changes as measurable levels of catecholamines and corticosteroids. In this study we have exposed young (14 months) Atlantic salmon to acid/Al water (pH ≈ 5.6, Al(i) ≈ 80 µg L⁻¹) for 3 days, and obtained clear and consistent decrease of Na⁺ and Cl⁻ ions, and increases of glucose in blood plasma, hematocrit and P(CO2) in blood. We did not measure plasma cortisol (primary response compound), but analyzed effects on microRNA level (miRNA) in muscle tissue, as this may represent initial markers of primary stress responses. miRNAs regulate diverse biological processes, many are evolutionarily conserved, and hundreds have been identified in various animals, although only in a few fish species. We used a novel high-throughput sequencing (RNA-Seq) method to identify miRNAs in Atlantic salmon and specific miRNAs as potential early markers for stress. A total of 18 miRNAs were significantly differentially expressed (FDR<0.1) in exposed compared to control fish, four down-regulated and 14 up-regulated. An unsupervised hierarchical clustering of significant miRNAs revealed two clusters representing exposed and non-exposed individuals. Utilizing the genome of the zebrafish and bioinformatic tools, we identified 224 unique miRNAs in the Atlantic salmon samples sequenced. Additional laboratory studies focusing on function, stress dose-responses and temporal expression of the identified miRNAs will facilitate their use as initial markers for stress responses.

Little impact of hatchery supplementation that uses native broodstock on the genetic structure and diversity of steelhead trout revealed by a large-scale spatio-temporal microsatellite survey.

Artificial breeding programs initiated to enhance the size of animal populations are often motivated by the desire to increase harvest opportunities. The introduction of non-native genotypes, however, can have negative evolutionary impacts. These may be direct, such as introgressive hybridization, or indirect via competition. Less is known about the effects of stocking with native genotypes. We assayed variation at nine microsatellite loci in 902 steelhead trout (Oncorhynchus mykiss) from five rivers in British Columbia, Canada. These samples were collected over 58 years, a time period that spanned the initiation of native steelhead trout broodstock hatchery supplementation in these rivers. We detected no changes in estimates of effective population size, genetic variation or temporal genetic structure within any population, nor of altered genetic structure among them. Genetic interactions with nonmigratory O. mykiss, the use of substantial numbers of primarily native broodstock with an approximate 1:1 male-to-female ratio, and/or poor survival and reproductive success of hatchery fish may have minimized potential genetic changes. Although no genetic changes were detected, ecological effects of hatchery programs still may influence wild population productivity and abundance. Their effects await the design and implementation of a more comprehensive evaluation program.