Looking into the past – the reaction of three grouse species to climate change over the last million years using whole genome sequences

Tracking past population fluctuations can give insight into current levels of genetic variation present within species. Analysing population dynamics over larger timescales can be aligned to known climatic changes to determine the response of species to varying environments. Here, we applied the Pairwise Sequentially Markovian Coalescent (psmc ) model to infer past population dynamics of three widespread grouse species; black grouse, willow grouse and rock ptarmigan. This allowed the tracking of the effective population size (N_e) of all three species beyond 1 Mya, revealing that (i) early Pleistocene cooling (~2.5 Mya) caused an increase in the willow grouse and rock ptarmigan populations, (ii) the mid‐Brunhes event (~430 kya) and following climatic oscillations decreased the N_e of willow grouse and rock ptarmigan, but increased the N_e of black grouse and (iii) all three species reacted differently to the last glacial maximum (LGM) – black grouse increased prior to it, rock ptarmigan experienced a severe bottleneck and willow grouse was maintained at large population size. We postulate that the varying psmc signal throughout the LGM depicts only the local history of the species. Nevertheless, the large population fluctuations in willow grouse and rock ptarmigan indicate that both species are opportunistic breeders while black grouse tracks the climatic changes more slowly and is maintained at lower N_e. Our results highlight the usefulness of the psmc approach in investigating species’ reaction to climate change in the deep past, but also that caution should be taken in drawing general conclusions about the recent past.     

Range shifts or extinction? Ancient DNA and distribution modelling reveal past and future responses to climate warming in cold‐adapted birds

Global warming is predicted to cause substantial habitat rearrangements, with the most severe effects expected to occur in high‐latitude biomes. However, one major uncertainty is whether species will be able to shift their ranges to keep pace with climate‐driven environmental changes. Many recent studies on mammals have shown that past range contractions have been associated with local extinctions rather than survival by habitat tracking. Here, we have used an interdisciplinary approach that combines ancient DNA techniques, coalescent simulations and species distribution modelling, to investigate how two common cold‐adapted bird species, willow and rock ptarmigan (Lagopus lagopus and Lagopus muta), respond to long‐term climate warming. Contrary to previous findings in mammals, we demonstrate a genetic continuity in Europe over the last 20 millennia. Results from back‐casted species distribution models suggest that this continuity may have been facilitated by uninterrupted habitat availability and potentially also the greater dispersal ability of birds. However, our predictions show that in the near future, some isolated regions will have little suitable habitat left, implying a future decrease in local populations at a scale unprecedented since the last glacial maximum.     

Northern glacial refugia for the pygmy shrew Sorex minutus in Europe revealed by phylogeographic analyses and species distribution modelling

The southern European peninsulas (Iberian, Italian and Balkan) are traditionally recognized as glacial refugia from where many species colonized central and northern Europe after the Last Glacial Maximum (LGM). However, evidence that some species had more northerly refugia is accumulating from phylogeographic, palaeontological and palynological studies, and more recently from species distribution modelling (SDM), but further studies are needed to test the idea of northern refugia in Europe. Here, we take a rarely implemented multidisciplinary approach to assess if the pygmy shrew Sorex minutus, a widespread Eurasian mammal species, had northern refugia during the LGM, and if these influenced its postglacial geographic distribution. First, we evaluated the phylogeographic and population expansion patterns using mtDNA sequence data from 123 pygmy shrews. Then, we used SDM to predict present and past (LGM) potential distributions using two different training data sets, two different algorithms (Maxent and GARP) and climate reconstructions for the LGM with two different general circulation models. An LGM distribution in the southern peninsulas was predicted by the SDM approaches, in line with the occurrence of lineages of S. minutus in these areas. The phylogeographic analyses also indicated a widespread and strictly northern‐central European lineage, not derived from southern peninsulas, and with a postglacial population expansion signature. This was consistent with the SDM predictions of suitable LGM conditions for S. minutus occurring across central and eastern Europe, from unglaciated parts of the British Isles to much of the eastern European Plain. Hence, S. minutus likely persisted in parts of central and eastern Europe during the LGM, from where it colonized other northern areas during the late‐glacial and postglacial periods. Our results provide new insights into the glacial and postglacial colonization history of the European mammal fauna, notably supporting glacial refugia further north than traditionally recognized.     

Pyrenean ptarmigans decline under climatic and human influences through the Holocene

In Europe, the Quaternary is characterized by climatic fluctuations known to have led to many cycles of contraction and expansion of species geographical ranges. In addition, during the Holocene, historical changes in human occupation such as colonization or abandonment of traditional land uses can also affect habitats. These climatically or anthropically induced geographic range changes are expected to produce considerable effective population size change, measurable in terms of genetic diversity and organization. The rock ptarmigan (Lagopus muta) is a small-bodied grouse occurring throughout Northern hemispheric arctic and alpine tundra. This species is not considered threatened at a continental scale, but the populations in the Pyrenees are of concern because of their small population size, geographical isolation and low genetic diversity. Here, we used 11 microsatellites to investigate genetic variations and differentiations and infer the overall demographic history of Pyrenean rock ptarmigan populations. The low genetic variability found in these populations has been previously thought to be the result of a bottleneck that occurred following the last glacial maximum (i.e., 10 000 years ago) or more recently (i.e., during the last 200 years). Our results clearly indicate a major bottleneck affecting the populations in the last tenth of the Holocene. We discuss how this decline can be explained by a combination of unfavorable and successive events that increased the degree of habitat fragmentation.     

Spring density and summer habitat use of alpine rock ptarmigan Lagopus muta helvetica in the southeastern Alps

In the Alps tourism, land-use practices and climate change may cause a loss of suitable habitat of rock ptarmigan (Lagopus muta helvetica). Hence, there is a need for basic research on population densities and habitat requirements of this species as a basis for long-term monitoring studies. So far, in the Austrian Alps, no accurate data on densities and trends of rock ptarmigan populations are available. We carried out counts of calling cocks in the southeastern part of the Austrian Alps (Nockberge National Park, Carinthia, Austria) between 31 May and 1 July 2003 and evaluated the distribution of male rock ptarmigan territories. The spring density of 5.4 territorial cocks per square kilometre was similar to densities in the Swiss Alps but higher than spring population densities in the Italian Alps and the Pyrenees. To investigate summer habitat use, we established a grid mapping of habitat parameters and rock ptarmigan signs. In total, 38 squares with signs of rock ptarmigan presence were recorded (i.e. 32% of all recorded squares). The presence of rock ptarmigan was significantly correlated with cover of rock and rock ptarmigan preferred habitat patchiness. Our study provides a useful reference for future monitoring of this species in the alpine distribution range considering the potential decrease and fragmentation of alpine habitats resulting from climate change and human impacts.     

Willow bud and shoot foraging by ptarmigan in relation to snow level in NW Finnish Lapland

Willow buds and twigs compose the main part of the willow ptarmigan’s (Lagopus lagopus) diet during winter. This study evaluated how position of willow ramets in relation to snow level affects the browsing of willow ptarmigan in NW Finland. Ramet position in relation to snow level affected the browsing intensity of willow ptarmigan: 5% of shoots were cut and more than 8% of buds were eaten by willow ptarmigan when ramet was staked at the maximum snow level, whereas ramets bent down were unbrowsed and browsing of ramets staked upright was negligible. Browsing significantly decreased the number of vegetative buds and catkins. The results show that snow level and ramet characteristics are important determinants of willow ptarmigan foraging.     

Variable reproductive effort for two ptarmigan species in response to spring weather in a northern alpine ecosystem

Predicting how animal populations respond to climate change requires knowledge of how species traits influence the response of individuals to variation in anuual weather. Over a four‐year study with two warm and two cold years, we examined how sympatric rock ptarmigan Lagopus muta and white‐tailed ptarmigan L. leucura in the southern Yukon Territory respond to spring weather in terms of breeding phenology and the allocation of reproductive effort. The onset of breeding was approximately synchronous; for each one‐degree rise in spring temperature, mean breeding dates of rock and white‐tailed ptarmigan advanced by about 2.7 and 4 days respectively. Although onset of breeding was similar, the two species differed in their reproductive effort. As breeding was delayed, average first clutch sizes of rock ptarmigan declined from 9.4 to 5.8 eggs over the breeding period, while those of white‐tailed ptarmigan only declined from an average of 7.8 to 6.8. Rock ptarmigan were also less likely to re‐nest if their first clutch was lost to predators and as a consequence they had shorter breeding seasons. White‐tailed ptarmigan produced about 25% more offspring annually than rock ptarmigan and contributed more young through re‐nesting. While white‐tailed ptarmigan had higher annual reproductive output, adult rock ptarmigan had a 20–25% higher annual survival rate, which may indicate a reproduction–survival trade‐off for the two species. These results show that even within the same location, closely related species can differ in how they allocate effort as environmental conditions fluctuate.     

Small-scale habitat use of black grouse (Tetrao tetrix L.) and rock ptarmigan (Lagopus muta helvetica Thienemann) in the Austrian Alps

We investigated the small-scale habitat use of two grouse species, black grouse (Tetrao tetrix L.) and rock ptarmigan (Lagopus muta helvetica Thienemann) in a study area in the Austrian Central Alps in summer. To build habitat suitability models, we applied multiple logistic regression using presence–absence data from fieldwork as the response variable and a set of habitat characteristics as explanatory variables, respectively. To gain a better understanding of the mechanisms that drive habitat selection, we tested for two-way interaction terms before excluding any variables from the initial variable set. Four explanatory variables significantly contributed to the black grouse model: dwarf shrub cover, dwarf shrub height, patchiness and ant hills. The final model for rock ptarmigan contained three explanatory variables: dwarf shrub cover, rock cover and dwarf shrub height. Most notably, the interaction terms dwarf shrub cover × patchiness in the black grouse model and dwarf shrub cover × dwarf shrub height, rock cover × dwarf shrub height in the rock ptarmigan model point out trade-off mechanisms between food, cover and overview providing features. Thus, our models do not only identify the parameters that mainly drive habitat selection, but also deepen our understanding about the causal relationships between these factors. Therefore, the information gained in this study allows for a deduction of appropriate habitat management strategies and supports conservation efforts of local stakeholders.     

TEMPORAL ALLOZYME FREQUENCY CHANGES IN DENSITY FLUCTUATING POPULATIONS OF WILLOW GROUSE (LAGOPUS LAGOPUS L.)

The population density of willow grouse (Lagopus lagopus L.) in northern Scandinavia changes in synchrony with the cyclic density variations in populations of microtine rodents. To assess the genetic changes accompanying the variations in population number, allozyme variation was studied at 23 loci in 640 willow grouse, representing four mainland and one island locality sampled during high and low population density. The average heterozygosity (H = 8.3%) and proportion of polymorphic loci (P = 26%) is not lower in willow grouse than in avian species with a more stable demography; the recurrent population density changes do not appear to affect drastically the long term effective population size, presumably because of extensive migration. Significant allele frequency differences were found both between populations and between different density phases. The genetic distance (D; Nei, 1972) was, in about 50% of the cases, larger between two consecutive time periods than between two localities in a certain year. Spatial and temporal allele frequency variation each represented around 3% of the gene diversity. The temporal heterogeneity may be caused by nonrandom sampling of family groups, rather than drift of allele frequencies between generations due to small effective population size, as has been suggested for microtine species.     

Territory occupancy rate of goshawk and gyrfalcon: no evidence of delayed numerical response to grouse numbers

Two recent studies on territory occupancy rates of goshawk Accipiter gentilis and gyrfalcon Falco rusticolus report a 2–3-year-delayed numerical response to grouse numbers, which is a requirement for a hypothesis of predator-generated grouse cycles. The time lags were assumed to reflect the average age of sexual maturity in the raptor species. In southern Norway, however, subadult (two-year-old) goshawk hens occupied only 18–25% of territories where occupancy was not recorded in the preceding year, and there was no significant relationship between the proportion of subadults among recruits and grouse indices two years earlier. We argue that territory occupancy rates are not appropriate indices of total raptor population levels, but rather reflect the proportion of territorial pairs that attempt to nest. Because this depends on the body condition of the hens, fluctuations in other important winter resident prey species (most important for the goshawk) and winter weather (most important for the gyrfalcon) should also be addressed. During 1988–2006, the annual proportion of goshawk territories with recorded nesting attempts in southern Norway was most closely related to the preceding autumn’s population indices of black grouse Tetrao tetrix and mountain hare Lepus timidus, whereas the annual proportion of gyrfalcon territories with observations of falcons or with confirmed breeding attempts in central Norway were best explained by population indices of willow grouse Lagopus lagopus and ptarmigan L. mutus from the previous autumn, and by December temperatures. Hence, our studies do not support the predation hypothesis for grouse cycles.     

Demographic consequences of age-structure in extreme environments: population models for arctic and alpine ptarmigan

Organisms living in arctic and alpine environments are increasingly impacted by human activities. To evaluate the potential impacts of global change, a better understanding of the demography of organisms in extreme environments is needed. In this study, we compare the age-specific demography of willow ptarmigan (Lagopus lagopus) breeding at arctic and subalpine sites, and white-tailed ptarmigan (L. leucurus) breeding at an alpine site. Rates of egg production improved with age at the alpine and subalpine sites, but the stochastic effects of nest and brood predation led to similar rates of annual fecundity among 1-, 2-, and 3+-year-old females. All populations had short generation times (T<2.7 years) and low net reproductive rates (R ₀<1.2). Stable age distributions were weighted towards 1-year-old females in willow ptarmigan (>59%), and to 3+-year-old females in white-tailed ptarmigan (>47%). High damping ratios (ρ>3.2) indicated that asymptotic estimates were likely to match natural age distributions. Sensitivity and elasticity values indicated that changes in juvenile survival would have the greatest impact on the finite rate of population change (λ) in willow ptarmigan, whereas changes to the survival of 3+-year-old females would have a greater effect in white-tailed ptarmigan. High survivorship buffers white-tailed ptarmigan in alpine environments against the potential effects of climate change on annual fecundity, but may make the species more sensitive to the effects of pollutants or harvesting on adult survival. Conversely, processes that reduce annual fecundity would have a greater impact on the population viability of willow ptarmigan in arctic and subalpine environments. If these same demographic patterns prove to be widespread among organisms in extreme environments, it may be possible to develop general recommendations for conservation of the biological resources of arctic and alpine ecosystems.     

A TEST OF THE GLACIAL REFUGIUM HYPOTHESIS USING PATTERNS OF MITOCHONDRIAL AND NUCLEAR DNA SEQUENCE VARIATION IN ROCK PTARMIGAN (LAGOPUS MUTUS)

The glacial refugium hypothesis (GRH) proposes that glaciers promoted differentiation and generation of intraspecific diversity by isolating populations in ice-free refugia. We tested three predictions of this hypothesis for the evolutionary divergence of rock ptarmigan (Lagopus mutus) during the Wisconsin glaciation of the late Pleistocene. To do this, we examined subspecies distributions, population genetic structure, and phylogenetic relationships in 26 populations across North America and the Bering Sea region. First, we analyzed sequence variation in the mitochondrial control region, in a nuclear intron (Gapdh), and in an internal transcribed spacer (ITS1). Control region sequences of 154 rock ptarmigan revealed strong population and phylogeographic structure. Variation in intron sequences of 114 rock ptarmigan also revealed significant population structure compatible with results for the control region. Rock ptarmigan were invariant for ITS1. Second, we show that five known Nearctic refugia and an Icelandic refugium are concordant with the current distribution of morphologically distinct subspecies; five of these six refugia are geographically concordant with the distribution of closely related control region haplotypes. Third, our estimates of the time since phylogenetic lineages diverged predated the last glacial maximum for all but two lineages. In addition, all lines of evidence suggest that two unknown refugia in the Bering Sea region supported rock ptarmigan during the Wisconsin glaciation. Overall, our results are most consistent with the hypothesis that isolated populations of rock ptarmigan diverged in multiple refugia during the Wisconsin and that geographic variation reflects patterns of recolonization of the Nearctic after the ice receded. The GRH may therefore offer the most plausible explanation for similar biogeographic patterns in a variety of Nearctic vertebrates.     

Where is the pulse to have the finger on? A retrospective analysis of two decades of Alpine Galliforms (Aves: Galliformes) census and game bag data in Italy

Information on the abundance of the Italian populations of black grouse (Lyrurus tetrix), Alpine rock ptarmigan (Lagopus muta helvetica) and Alpine rock partridge (Alectoris graeca saxatilis) rely only on extrapolations of local data to the national scale, since there is no national standardized survey. Consequently, their status is virtually unknown. We performed a first-ever assessment of a medium-term (1996–2014) population trend of these species using and comparing post-breeding count and bag data at hunting district scale. These data were collected from various authorities in charge of wildlife management and allowed us to test the influence of hunting policies on the estimated trends. Rock partridge showed a stable trend with numbers fluctuating between years, while there was evidence of a severe decline for rock ptarmigan. No general conclusion could be drawn for the black grouse, as we detected lack of consistency of count and bag data. Counts were greatly overdispersed as a result of an uneven count effort among hunting districts. Adding the game management authority as model covariate resulted in more robust trend estimations, suggesting a significant effect of different policies that emerged also as similar hunting pressure across species within authorities. Hunting effort variation over the time was instead negligible. Species-specific game management bias is discussed. Our results highlight the need for a survey scheme or guidelines to be applied uniformly at a national scale.     

Rethinking long‐term vegetation dynamics: multiple glacial refugia and local expansion of a species complex

Evidence is accumulating that some arcto‐boreal plant taxa persisted through the last glacial maximum (LGM) in Alaska and adjacent Canada. However, the spatial patterns of glacial persistence and associated postglacial colonization remain largely unknown. In this study, we investigated the LGM refugia of an alder (Alnus) species complex (n = 3 taxa) and assess the spatiotemporal dynamics of Alnus in this vast region. Specifically, we conducted high‐throughput DNA sequencing (ddRADseq) on Alnus foliar samples collected from a dense population network to investigate patterns of genetic structure and infer the presence of glacial lineages. Species distribution modeling (SDM) was used to investigate the probability and possible locations of glacial persistence. These analyses were integrated and then compared with fossil pollen data to identify the locations of refugial populations and spatial patterns of postglacial colonization. Our genetic analyses revealed two glacial lineages with separate geographic origins for each Alnus taxon, suggesting that the genus persisted in multiple LGM refugia. Non‐overlapping hindcast distributions based on SDMs further support the presence of multiple, spatially distinct refugia. These ddRADseq and SDM results, in conjunction with reassessment of fossil pollen records, suggest that Alnus expanded from several population nuclei that existed during the LGM and coalesced during the Holocene to form its present range. These results challenge the unidirectional model for postglacial vegetation expansion, implying that climate buffering associated with landscape heterogeneity and adaptation to millennial‐scale environmental variability played important roles in driving late‐Quaternary population dynamics.     

Weak compensation of harvest despite strong density-dependent growth in willow ptarmigan

Ptarmigan and grouse species (Lagopus spp.) are thought to be able to compensate for a modest harvest because there is a surplus of breeding birds that are prevented from breeding by territory holders. To estimate the degree of harvest-mortality compensation reliably we experimentally harvested 0%, 15% and 30% of the willow ptarmigan (Lagopus lagopus) on 13 estates ranging from 20 to 54 km2 in size during four hunting seasons in Norway according to a regional block design. Population overwinter growth rate was strongly negatively density dependent, but despite this, and contrary to earlier findings, only 33% of the harvest was compensated for. The lack of compensation was probably caused by long-distance juvenile dispersal that was unaffected by the harvest. The need for large-scale management experiments to detect the effects of harvest was clearly demonstrated: lack of compensation was found only when we used the whole dataset and not when the data were analysed by year or block. Our study shows that it is very difficult to demonstrate a population's lack of harvest compensation and warns against using small-scale, out-of-season or poorly replicated studies as a basis for future harvest-management decisions.     

Parasites and Scottish ptarmigan numbers

Summary Unlike red grouse (Lagopus lagopus scoticus), which have almost 100% prevalence of the parasitic threadworm Trichostrongylus tenuis and frequently high tapeworm numbers, 70% (n=71) of Scottish ptarmigan (Lagopus mutus) had no threadworms and only 4% had tapeworms. Other parasites and pathogenic bacteria were infrequent. Threadworms occurred in 12% (n=25) of birds on granite hills where mean ptarmigan densities were low, and in 43% (n=46) of birds over schists and limestones where mean densities were high. The notion that parasites cause the cyclic-type ptarmigan declines observed on the granite Cairngorms massif is unlikely (ptarmigan fluctuations over the richer rocks are irregular).     

Phylogeography of willow grouse (Lagopus lagopus) in the Arctic: taxonomic discordance as inferred from molecular data

Using independently segregating nuclear single nucleotide polymorphisms (SNPs) and mitochondrial control region sequences, we found an east–west division among sampled willow grouse Lagopus lagopus subspecies. This division cut across the range of the subspecies with the largest distribution (lagopus) and thus contradicted existing taxonomic classifications. Russian Lagopus lagopus lagopus tended to cluster with North American willow grouse partly classified as other subspecies. Scandinavian willow grouse (L. l. lagopus) clustered with red grouse from Britain and Ireland (Lagopus lagopus scoticus and Lagopus lagopus hibernicus) but substructuring confirmed the monophyly of the latter. In North America, we could not detect any major genetic divisions apart from two birds described as alexandrae from the Heceta Island (Alaska) when using mitochondrial sequences. Other samples from North America were intermingled regardless of whether they were described as muriei, alexandrae or lagopus. A specimen described as alexandrae was to some extent distinct when analysing the SNP data. The genetic analyses indicated some concordance between genetics and taxonomy but not complete congruence. This is particularly evident for mitochondrial DNA network analyses. We suggest that the taxonomy of this species would benefit by a careful re‐examination of the available evidence for subspecies. It appears as if subspecies status is a poor proxy for assigning evolutionary significant units and management units in this species. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 77–90.     

The occurrence of gapeworm (Syngamus trachea) in willow grouse.

21.4% of the willow grouse (Lagopus lagopus) examined from a coastal locality in northern Norway were infected with Syngamus trachea. None of the willow grouse examined from an inland locality with colder and longer winters were infected with the parasite. It is suggested that the parasite may influence survival of the grouse population.     

Population genetic structure of rock ptarmigan in the 'sky islands' of French Pyrenees: implications for conservation

Expected consequences of global warming include habitat reduction in many cool climate species. Rock ptarmigan is a Holarctic grouse that inhabits arctic and alpine tundra. In Europe, the Pyrenean ptarmigan inhabits the southern edge of the species' range and since the last glacial maximum its habitat has been severely fragmented and is restricted to high-alpine zones or 'sky islands'. A recent study of rock ptarmigan population genetic in Europe found that the Pyrenean ptarmigan had very low genetic diversity compared with that found in the Alps and Scandinavia. Habitat fragmentation and reduced genetic diversity raises concerns about the viability of ptarmigan populations in the Pyrenees. However, information on population structuring and gene flow across the Pyrenees, which is essential for designing a sound management plan, is absent. In this study, we use seven microsatellites and mitochondrial control region sequences to investigate genetic variation and differentiation among five localities across the Pyrenees. Our analyses reveal the presence of genetic differentiation among all five localities and a significant isolation-by-distance effect that is likely the result of short dispersal distances and high natal and breeding philopatry of Pyrenean ptarmigan coupled with severe habitat fragmentation. Furthermore, analysis of molecular variance, principal component analysis and Bayesian analysis of genetic structuring identified the greatest amount of differentiation between the eastern and main parts of the Pyrenean chain separated by the Sègre Valley. Our data also show that the Canigou massif may host an isolated population and requires special conservation attention. We propose a management plan which includes the translocation of birds. If a sky island structure affects genetic divergence in rock ptarmigan, it may also affect the genetic structure of other sky island species having low dispersal abilities.     

Genetic diversity and differentiation among Lagopus lagopus populations in Scandinavia and Scotland: evolutionary significant units confirmed by SNP markers

Single Nucleotide Polymorphism in four Scandinavian populations of willow grouse (Lagopus lagopus) and two Scottish populations of red grouse (Lagopus lagopus scoticus) were assessed at 13 protein‐coding loci. We found high levels of diversity, with one substitution every 55 bp as an average and a total of 76 unlinked parsimony informative SNPs. Different estimators of genetic diversity such as: number of synonymous and non‐synonymous sites, average number of alleles, number and percentage of polymorphic loci, mean nucleotide diversity (π_s, π_a) and gene diversity at synonymous and non‐synonymous sites showed higher diversity in the northern populations compared to southern ones. Strong levels of purifying selection found in all the populations together with neutrality tests conforming to neutral expectations agree with large effective population sizes. Assignment tests reported a clear distinction between Scandinavian and Scottish grouse suggesting the existence of two different evolutionary significant units. The divergence time between willow and red grouse ranging between 12 500 and 125 000 years, in conjunction with the presence of ‘specific’ markers for each subspecies prompt a reassessment of the taxonomical status of the Scottish red grouse.