The evolutionary pathways for local adaptation in mountain hares.

Understanding the evolution of local adaptations is a central aim of evolutionary biology and key for the identification of unique populations and lineages of conservation relevance. By combining RAD sequencing and whole-genome sequencing, we identify genetic signatures of local adaptation in mountain hares (Lepus timidus) from isolated and distinctive habitats of its wide distribution: Ireland, the Alps and Fennoscandia. Demographic modelling suggested that the split of these mountain hares occurred around 20 thousand years ago, providing the opportunity to study adaptive evolution over a short timescale. Using genome-wide scans, we identified signatures of extreme differentiation among hares from distinct geographic areas that overlap with area-specific selective sweeps, suggesting targets for local adaptation. Several identified candidate genes are associated with traits related to the uniqueness of the different environments inhabited by the three groups of mountain hares, including coat colour, ability to live at high altitudes and variation in body size. In Irish mountain hares, a variant of ASIP, a gene previously implicated in introgression-driven winter coat colour variation in mountain and snowshoe hares (L. americanus), may underlie brown winter coats, reinforcing the repeated nature of evolution at ASIP moulding adaptive seasonal colouration. Comparative genomic analyses across several hare species suggested that mountain hares' adaptive variants appear predominantly species-specific. However, using coalescent simulations, we also show instances where the candidate adaptive variants have been introduced via introgressive hybridization. Our study shows that standing adaptive variation, including that introgressed from other species, was a crucial component of the post-glacial dynamics of species.

Restoring vertebrate predator populations can provide landscape-scale biological control of established invasive vertebrates: Insights from pine marten recovery in Europe.

Invasive species pose one of the greatest global threats to biodiversity. There has been a long history of importing coevolved natural enemies to act as biological control agents to try to suppress densities of invasive species, with historically limited success and frequent adverse impacts on native biodiversity. Our understanding of the processes and drivers of successful biological control has been focussed on invertebrates and is evidently limited and potentially ill-suited with respect to biological control of vertebrate populations. The restoration of native vertebrate predator populations provides a promising nature-based solution for slowing, halting, or even reversing the spread of some invasive vertebrates over spatial scales relevant to the management of wildlife populations. Here, we first review the growing literature and data from the pine marten-red and grey squirrel system in Europe. We synthesise a multi-decadal dataset to show that the recovery of a native predator has resulted in rapid, landscape-scale declines of an established invasive species. We then use the model system, predator-prey interaction theory, and examples from the literature to develop ecological theory relating to natural biological control in vertebrates and evolutionary processes in native-invasive predator-prey interactions. We find support for the hypotheses that evolutionary naivety of invasive species to native predators and lack of local refuges results in higher predation of naive compared to coevolved prey. We apply lessons learnt from the marten-squirrel model system to examine the plausibility of specific native predator solutions to some of the Earth's most devastating invasive vertebrates. Given the evidence, we conclude that depletion of vertebrate predator populations has increased ecosystem vulnerability to invasions and thus facilitated the spread of invasive species. Therefore, restoration of vertebrate predator populations is an underappreciated, fundamental, nature-based solution to the crisis of invasive species and should be a priority for vertebrate invasive species management globally.

Regime shift tipping point in hare population collapse associated with climatic and agricultural change during the very early 20th century.

Animal populations at northern latitudes may have cyclical dynamics that are degraded by climate change leading to trophic cascade. Hare populations at more southerly latitudes are characterized by dramatic declines in abundance associated with agricultural intensification. We focus on the impact of historical climatic and agricultural change on a mid-latitude population of mountain hares, Lepus timidus hibernicus. Using game bag records from multiple sites throughout Ireland, the hare population index exhibited a distinct regime shift. Contrary to expectations, there was a dynamical structure typical of northern latitude hare populations from 1853 to 1908, during which numbers were stable but cyclic with a periodicity of 8 years. This regime was replaced by dynamics more typical of southern latitude hare populations from 1909 to 1970, in which cycles were lost and numbers declined dramatically. Destabilization of the autumn North Atlantic Oscillation (NAO) led to the collapse of similar cycles in the hare population, coincident with the onset of agricultural intensification (a shift from small-to-large farms) in the first half of the 20th century. Similar, but more recent regime shifts have been observed in Arctic ecosystems and attributed to anthropogenic climate change. The present study suggests such shifts may have occurred at lower latitudes more than a century ago during the very early 20th century. It seems likely that similar tipping points in the population collapse of other farmland species may have occurred similarly early but went undocumented. As northern systems are increasingly impacted by climate change and probable expansion of agriculture, the interaction of these processes is likely to disrupt the pulsed flow of resources from cyclic populations impacting ecosystem function.

Molecular mechanisms of self-regulation in multiannual rodent populations: Experimental test of an updated hypothesis.

IN FOCUS: Edwards, P. D., Frenette-Ling, C., Palme, R., & Boonstra, R. (2021). Social density suppresses GnRH expression and reduces reproductivity in voles: A mechanism for population self-regulation. Journal of Animal Ecology, 90, 784-795. Intrinsic population processes are important in the regulation of populations of small rodents, including those which display multiannual cycles. By measuring reproductive parameters, faecal androgen metabolites, and gene expression and DNA methylation in the CNS of juvenile voles, this paper demonstrates that suppression of reproduction occurs in female voles at high density compared to low density in enclosures, and that this maternal, epigenetic effect is also apparent in their offspring. This suggsests that direct density dependence influences reproduction and, hence, immediate rate of population growth, while gene expression mediated by DNA methylation blocking transcription, may have a delayed density-dependent effect in juveniles. Both direct and delayed density dependence are necessary to generate multiannual population cycles. Edwards et al. (2021) break new ground in demonstrating the molecular and physiological basis of variation in population dynamics of small mammals ranging from multiannual cycles to stability that have fascinated researchers for nearly a century.

No energetic cost of tuberculosis infection in European badgers (Meles meles).

Energy availability and energy use directly influence an organism's life history, fitness and ecological function. In wild animals, abiotic factors such as ambient temperature, season and rainfall, and biotic factors such as body mass, age, social group size and disease status, all potentially influence energy balance. Relatively few studies have examined the effects of disease on the energy expenditure of wild animals. Such studies could further our understanding of factors influencing the transmission of zoonotic diseases. The European badger (Meles meles) is a medium-sized carnivore that occurs in mixed-sex, familial groups across much of its range. In the UK, they are a protected species but are also involved in the epidemiology of bovine tuberculosis (TB) in cattle. We measured the daily energy expenditure (DEE) and resting metabolic rate (RMR) of wild badgers and related this to their TB infection status and a range of other interacting factors including season, group size, disease status, sex, age, body mass and body fat. Individuals were larger and fatter when they were older, and fatter during the winter. Males were also heavier than females during the summer. In addition, individuals from smaller groups that were exposed to TB tended to have lower body mass. There were no direct effects of disease status on DEE or RMR; however, there was a significant interaction whereby DEE increased with body mass in small groups but decreased with body mass in large groups. Results are consistent with the costs of TB infection being met by compensatory mechanisms enabling badgers to survive for extended periods without exhibiting measurable energetic consequences.

Development of a novel immunochromatographic lateral flow assay specific for Mycobacterium bovis cells and its application in combination with immunomagnetic separation to test badger faeces.

BACKGROUND: The European badger is an important wildlife reservoir of Mycobacterium bovis implicated in the spread of bovine tuberculosis in the United Kingdom and Ireland. Infected badgers are known to shed M. bovis in their urine and faeces, which may contaminate the environment. To aid bovine tuberculosis control efforts novel diagnostic tests for detecting infected and shedding badgers are needed. We proposed development of a novel, rapid immunochromatographic lateral flow device (LFD) as a non-invasive test to detect M. bovis cells in badger faeces. Its application in combination with immunomagnetic separation (IMS) to detect Mycobacterium bovis cells in badger faeces is reported here. RESULTS: A novel prototype LFD for M. bovis cells was successfully developed, with unique specificity for M. bovis and a limit of detection 50% (LOD50%) of 1.7 × 104 M. bovis cells/ml. When IMS was employed to selectively capture and concentrate M. bovis cells from badger faeces prior to LFD testing, the LOD50% of the IMS-LFD assay was 2.8 × 105 M. bovis cells/ml faecal homogenate. Faeces samples collected from latrines at badger setts in a region of endemic bovine tuberculosis infection were tested; 78 (18%) of 441 samples tested IMS-LFD assay positive, whereas 140 (32%) tested IMS-qPCR positive (Kappa agreement -0.009 ± 0.044, p = 0.838). Subsequently, when 130 faeces samples from live captured, or captive, badgers of known infection status (on the basis of StatPak, interferon-γ and/or culture results) were tested, the IMS-LFD assay had higher relative diagnostic specificity (Sp 0.926), but poorer relative diagnostic sensitivity (Se 0.081), than IMS-qPCR (Sp 0.706, Se 0.581) and IMS-culture (Sp 0.794, Se 0.436). CONCLUSIONS: The novel IMS-LFD assay, although very specific for M. bovis, has low analytical sensitivity (indicated by the LOD50%) and would only detect badgers shedding high numbers of M. bovis (>104-5 cells/g) in their faeces. The novel LFD would, therefore, have limited value as a non-invasive test for badger TB surveillance purposes but it may have value for alternative veterinary diagnostic applications.

Pure species in a continuum of genetic and morphological variation: sympatric oaks at the edge of their range.

BACKGROUND AND AIMS: Studies on oaks (Quercus spp.) have often been hampered by taxonomic confusion, a situation further compounded by the occurrence of extensive interspecific hybridization. In the present study, a combination of genetic and morphological analyses was used to examine sympatric populations of Q. petraea and Q. robur at the north-western edge of their ranges in Northern Ireland, since it had previously been suggested that hybridization could facilitate the apparent rapid, long-distance dispersal of oaks following the glaciations. METHODS: Samples were collected from 24 sites across Northern Ireland that had been previously designated as ancient or semi-natural woodland. Genotypes were obtained from a total of 950 trees using 12 nuclear microsatellite loci, and admixture coefficients were calculated based on a Bayesian clustering approach. Individuals were also classified as Q. petraea,Q. robur or hybrids based on two objective morphometric characters shown previously to delineate pure individuals effectively. Genetically 'pure' individuals of both species, as defined by the Bayesian clustering, were also genotyped for five chloroplast microsatellites. KEY RESULTS: Genetic and morphological analyses both indicated the presence of pure individuals of both species, as well as a continuum of intermediates. There was a good agreement between the molecular and morphological classification, with a generally clear separation between pure individuals. CONCLUSIONS: Despite millennia of hybridization and introgression, genetically and morphologically pure individuals of both Q. petraea and Q. robur can be found at the edge of their range, where both species occur sympatrically. The high proportion of individuals exhibiting introgression compared with previous studies may reflect the historical role of hybridization in facilitating dispersal following the glaciations. This is further supported by the significantly higher chloroplast diversity in Q. robur compared with Q. petraea.

Initial ecological change in plant and arthropod community composition after wildfires in designated areas of upland peatlands.

Wildfires are an increasing concern due to rising temperatures and incidence of droughts associated with changing climate, poor land management, and direct human interference. Most studies of the impact of fire on temperate heathland and bog examined the consequences of controlled or prescribed burning. Less is known about the impacts of uncontrolled wildfires on sites designated for their conservation value. We examined the initial impact and short-term trajectory (3.5 years) of cool temperate peatland plant and arthropod communities on designated upland sites in Northern Ireland following wildfires, that is, unplanned with respect to where and when they occur, severity, and duration. These near simultaneous wildfires were often due to a failure to control prescribed burns. Wildfires were associated with a loss of blanket bog and heath indicator species. Broad vegetation groups showed initial recovery characterized by a decrease in bare ground and increasing cover of shrub species and bryophytes. However, at a species level, Sphagnum spp and bryophyte communities, which are central to peatland ecosystem functioning, showed no sign of recovery to prefire composition. Rather, bryophyte communities became more divergent over the course of the study and were mainly characterized by increased abundance of the alien pioneer acrocarp Campylopus introflexus. Similarly, composition of arthropod communities (ground beetles and spiders) differed between burnt and unburnt areas and showed no evidence of a return to species composition in unburnt areas. The nationally rare beetle Carabus nitens was more common in the aftermath of wildfire. Synthesis. Whilst, long-term recovery was not investigated, these short-term changes suggest enduring detrimental impacts on the distinctive communities associated with peatlands, primarily through the loss of Sphagnum spp., affecting ecosystem services such as carbon sequestration and water and soil retention. It may not be possible to restore exact prefire species composition of plant and animal communities. We suggest a precautionary approach involving management of upland vegetation, public education, and vigilance, to prevent further wildfires and protect these key upland habitats.

Rodenticide exposure in wood mouse and house mouse populations on farms and potential secondary risk to predators.

We compared capture rates and exposure to SGARs of wood mice (Apodemus sylvaticus) and house mice (Mus domesticus) in autumn/winter on farms that currently used, had previously used, and never used SGARs. 6-10 weeks after baiting programmes began, 15 % of 55 wood mice and 33 % of 12 house mice had detectable liver SGAR residues. Wood mice with residues occurred on farms not using rodenticides, reflecting the high mobility of these animals, and four had multiple liver residues, possibly due to cross-contamination of baits. The winter decline in wood mouse numbers was similar on farms that did and did not use SGARs, suggesting little long-term impact of SGARs on populations on farms. Our results indicate residual levels of rodenticides will be ever present in small mammal prey across the agricultural landscape unless all farms in a locality cease application. The implications for secondary exposure and poisoning of predators are discussed.

Invader abundance and contraction of niche breadth during replacement of a native gammarid amphipod.

The introduction of non-native species to new locations is a growing global phenomenon with major negative effects on native species and biodiversity. Such introductions potentially bring competitors into contact leading to partial or total species replacements. This creates an opportunity to study novel species interactions as they occur, with the potential to address the strength of inter- and intraspecific interactions, most notably competition. Such potential has often not been realized, however, due to the difficulties inherent in detecting rapid and spatially expansive species interactions under natural field conditions. The invasive amphipod crustacean Gammarus pulex has replaced a native species, Gammarus duebeni celticus, in river and lake systems across Europe. This replacement process is at least partially driven by differential parasitism, cannibalism, and intraguild predation, but the role of interspecific competition has yet to be resolved. Here, we examine how abundance of an invasive species may affect spatial niche breadth of a native congeneric species. We base our analyses of niche breadth on ordination and factor analysis of biological community and physical parameters, respectively, constituting a summative, multidimensional approach to niche breadth along environmental gradients. Results derived from biological and environmental niche criteria were consistent, although interspecific effects were stronger using the biological niche approach. We show that the niche breadth of the native species is constrained as abundance of the invader increases, but the converse effect does not occur. We conclude that the interaction between invasive G. pulex and native G. d. celticus under natural conditions is consistent with strong interspecific competition whereby a native, weaker competitor is replaced by a superior invasive competitor. This study indicates a strong role of interspecific competition, alongside other known interactions such as differential intraguild predation, in rapid and expansive species replacements following biological invasions.

Soil δ13C and δ15N baselines clarify biogeographic heterogeneity in isotopic discrimination of European badgers (Meles meles).

Isotopic techniques have been used to study phenomena in the geological, environmental, and ecological sciences. For example, isotopic values of multiple elements elucidate the pathways energy and nutrients take in the environment. Isoscapes interpolate isotopic values across a geographical surface and are used to study environmental processes in space and time. Thus, isoscapes can reveal ecological shifts at local scales, and show distribution thresholds in the wider environment at the macro-scale. This study demonstrates a further application of isoscapes, using soil isoscapes of 13C/12C and 15N/14N as an environmental baseline, to understand variation in trophic ecology across a population of Eurasian badgers (Meles meles) at a regional scale. The use of soil isoscapes reduced error, and elevated the statistical signal, where aggregated badger hairs were used, and where individuals were identified using genetic microarray analysis. Stable isotope values were affected by land-use type, elevation, and meteorology. Badgers in lowland habitats had diets richer in protein and were adversely affected by poor weather conditions in all land classes. It is concluded that soil isoscapes are an effective way of reducing confounding biases in macroscale, isotopic studies. The method elucidated variation in the trophic and spatial ecology of economically important taxa at a landscape level. These results have implications for the management of badgers and other carnivores with omnivorous tendencies in heterogeneous landscapes.

An Annotated Draft Genome of the Mountain Hare (Lepus timidus).

Hares (genus Lepus) provide clear examples of repeated and often massive introgressive hybridization and striking local adaptations. Genomic studies on this group have so far relied on comparisons to the European rabbit (Oryctolagus cuniculus) reference genome. Here, we report the first de novo draft reference genome for a hare species, the mountain hare (Lepus timidus), and evaluate the efficacy of whole-genome re-sequencing analyses using the new reference versus using the rabbit reference genome. The genome was assembled using the ALLPATHS-LG protocol with a combination of overlapping pair and mate-pair Illumina sequencing (77x coverage). The assembly contained 32,294 scaffolds with a total length of 2.7 Gb and a scaffold N50 of 3.4 Mb. Re-scaffolding based on the rabbit reference reduced the total number of scaffolds to 4,205 with a scaffold N50 of 194 Mb. A correspondence was found between 22 of these hare scaffolds and the rabbit chromosomes, based on gene content and direct alignment. We annotated 24,578 protein coding genes by combining ab-initio predictions, homology search, and transcriptome data, of which 683 were solely derived from hare-specific transcriptome data. The hare reference genome is therefore a new resource to discover and investigate hare-specific variation. Similar estimates of heterozygosity and inferred demographic history profiles were obtained when mapping hare whole-genome re-sequencing data to the new hare draft genome or to alternative references based on the rabbit genome. Our results validate previous reference-based strategies and suggest that the chromosome-scale hare draft genome should enable chromosome-wide analyses and genome scans on hares.

Does functional homogenization accompany taxonomic homogenization of British birds and how do biotic factors and climate affect these processes?

Environmental change has reshuffled communities often causing taxonomic homogenization rather than differentiation. Some studies suggest that this increasing similarity of species composition between communities is accompanied by an increase in similarity of trait composition-functional homogenization-although different methodologies have failed to come to any consistent conclusions. Functional homogenization could have a large effect on ecosystem functioning and stability. Here, we use the general definition of homogenization as "reduced spatial turnover over time" to compare changes in Simpson's beta diversity (taxonomic turnover) with changes in Rao's quadratic entropy beta diversity (functional turnover) in British breeding birds at three spatial scales. Using biotic and climatic variables, we identify which factors may predispose a site to homogenization. The change in turnover measures between two time periods, 20 years apart, was calculated. A null model approach was taken to identify occurrences of functional homogenization and differentiation independent of changes in taxonomic turnover. We used conditional autoregressive models fitted using integrated nested Laplace approximations to determine how environmental drivers and factors relating to species distributions affect changes in spatial turnover of species and functional diversity. The measurement of functional homogenization affects the chance of rejection of the null models, with many sites showing taxonomic homogenization unaccompanied by functional homogenization, although occurrence varies with spatial scale. At the smallest scale, while temperature-related variables drive changes in taxonomic turnover, changes in functional turnover are associated with variation in growing degree days; however, changes in functional turnover become more difficult to predict at larger spatial scales. Our results highlight the multifactorial processes underlying taxonomic and functional homogenization and that redundancy in species traits may allow ecosystem functioning to be maintained in some areas despite changes in species composition.

Seasonal and predator-prey effects on circadian activity of free-ranging mammals revealed by camera traps.

Endogenous circadian and seasonal activity patterns are adapted to facilitate effective utilisation of environmental resources. Activity patterns are shaped by physiological constraints, evolutionary history, circadian and seasonal changes and may be influenced by other factors, including ecological competition and interspecific interactions. Remote-sensing camera traps allow the collection of species presence data throughout the 24 h period and for almost indefinite lengths of time. Here, we collate data from 10 separate camera trap surveys in order to describe circadian and seasonal activity patterns of 10 mammal species, and, in particular, to evaluate interspecific (dis)associations of five predator-prey pairs. We recorded 8,761 independent detections throughout Northern Ireland. Badgers, foxes, pine martens and wood mice were nocturnal; European and Irish hares and European rabbits were crepuscular; fallow deer and grey and red squirrels were diurnal. All species exhibited significant seasonal variation in activity relative to the timing of sunrise/sunset. Foxes in particular were more crepuscular from spring to autumn and hares more diurnal. Lagged regression analyses of predator-prey activity patterns between foxes and prey (hares, rabbits and wood mice), and pine marten and prey (squirrel and wood mice) revealed significant annual and seasonal cross-correlations. We found synchronised activity patterns between foxes and hares, rabbits and wood mice and pine marten and wood mice, and asynchrony between squirrels and pine martens. Here, we provide fundamental ecological data on endemic, invasive, pest and commercially valuable species in Ireland, as well as those of conservation importance and those that could harbour diseases of economic and/or zoonotic relevance. Our data will be valuable in informing the development of appropriate species-specific methodologies and processes and associated policies.

Mountain hare transcriptome and diagnostic markers as resources to monitor hybridization with European hares.

We report the first mountain hare (Lepus timidus) transcriptome, produced by de novo assembly of RNA-sequencing reads. Data were obtained from eight specimens sampled in two localities, Alps and Ireland. The mountain hare tends to be replaced by the invading European hare (Lepus europaeus) in their numerous contact zones where the species hybridize, which affects their gene pool to a yet unquantified degree. We characterize and annotate the mountain hare transcriptome, detect polymorphism in the two analysed populations and use previously published data on the European hare (three specimens, representing the European lineage of the species) to identify 4 672 putative diagnostic sites between the species. A subset of 85 random independent SNPs was successfully validated using PCR and Sanger sequencing. These valuable genomic resources can be used to design tools to assess population status and monitor hybridization between species.

Expertly validated models and phylogenetically-controlled analysis suggests responses to climate change are related to species traits in the order lagomorpha.

Climate change during the past five decades has impacted significantly on natural ecosystems, and the rate of current climate change is of great concern among conservation biologists. Species Distribution Models (SDMs) have been used widely to project changes in species' bioclimatic envelopes under future climate scenarios. Here, we aimed to advance this technique by assessing future changes in the bioclimatic envelopes of an entire mammalian order, the Lagomorpha, using a novel framework for model validation based jointly on subjective expert evaluation and objective model evaluation statistics. SDMs were built using climatic, topographical, and habitat variables for all 87 lagomorph species under past and current climate scenarios. Expert evaluation and Kappa values were used to validate past and current models and only those deemed 'modellable' within our framework were projected under future climate scenarios (58 species). Phylogenetically-controlled regressions were used to test whether species traits correlated with predicted responses to climate change. Climate change is likely to impact more than two-thirds of lagomorph species, with leporids (rabbits, hares, and jackrabbits) likely to undertake poleward shifts with little overall change in range extent, whilst pikas are likely to show extreme shifts to higher altitudes associated with marked range declines, including the likely extinction of Kozlov's Pika (Ochotona koslowi). Smaller-bodied species were more likely to exhibit range contractions and elevational increases, but showing little poleward movement, and fecund species were more likely to shift latitudinally and elevationally. Our results suggest that species traits may be important indicators of future climate change and we believe multi-species approaches, as demonstrated here, are likely to lead to more effective mitigation measures and conservation management. We strongly advocate studies minimising data gaps in our knowledge of the Order, specifically collecting more specimens for biodiversity archives and targeting data deficient geographic regions.

Winter Is Coming: Seasonal Variation in Resting Metabolic Rate of the European Badger (Meles meles).

Resting metabolic rate (RMR) is a measure of the minimum energy requirements of an animal at rest, and can give an indication of the costs of somatic maintenance. We measured RMR of free-ranging European badgers (Meles meles) to determine whether differences were related to sex, age and season. Badgers were captured in live-traps and placed individually within a metabolic chamber maintained at 20 ± 1°C. Resting metabolic rate was determined using an open-circuit respirometry system. Season was significantly correlated with RMR, but no effects of age or sex were detected. Summer RMR values were significantly higher than winter values (mass-adjusted mean ± standard error: 2366 ± 70 kJ⋅d(-1); 1845 ± 109 kJ⋅d(-1), respectively), with the percentage difference being 24.7%. While under the influence of anaesthesia, RMR was estimated to be 25.5% lower than the combined average value before administration, and after recovery from anaesthesia. Resting metabolic rate during the autumn and winter was not significantly different to allometric predictions of basal metabolic rate for mustelid species weighing 1 kg or greater, but badgers measured in the summer had values that were higher than predicted. Results suggest that a seasonal reduction in RMR coincides with apparent reductions in physical activity and body temperature as part of the overwintering strategy ('winter lethargy') in badgers. This study contributes to an expanding dataset on the ecophysiology of medium-sized carnivores, and emphasises the importance of considering season when making predictions of metabolic rate.

The validity of the Gammarus:Asellus ratio as an index of organic pollution: abiotic and biotic influences.

In freshwaters. Gammarus spp. are more sensitive to organic pollution than Asellus spp. and the relative abundance of the two taxa has been proposed as a pollution index. We tested the validity of this by examining the relationship between the Gammarus: Asellus (G : A) ratio and (1) a suite of physico-chemical variables. (2) established biotic (average score per taxon, ASPT) and richness (species richness (S) and Ephemeroptera, Plecoptera and Trichoptera families richness (EPT family richness)) indices generated from the macroinvertebrate community. In addition, we investigated a suspected biotic interaction, predation, between Gammarus and Asellus. Both univariate and multivariate analyses showed that the G: A ratio was sometimes responsive to changes in parameters linked to organic pollution, such as BOD5 and nitrate levels. However, the G : A ratio also appeared responsive to variables not directly linked to organic pollution, such as conductivity and distance from source. There were significant positive correlations among the G : A ratio and the ASPT, S and EPT, indicating that changes in the relative abundances of Gammarus and Asellus were reflected in changes in the pollution sensitivity and richness of the wider macroinvertebrate community. A laboratory experiment revealed significant predation of Asellus aquaticus juveniles by Gammarus duebeni celticus adults, but no reciprocal predation. We propose that the G: A ratio may be useful as a crude measure of organic pollution that could supplement more complex indices in a multimetric approach to pollution monitoring or be used for monitoring individual sites, where a simple technique is required for monitoring purposes over a period of time. Also, we urge recognition of the possible role of biotic interactions among taxa used in the generation of pollution indices.

Squirrelpox virus: assessing prevalence, transmission and environmental degradation.

Red squirrels (Sciurus vulgaris) declined in Great Britain and Ireland during the last century, due to habitat loss and the introduction of grey squirrels (Sciurus carolinensis), which competitively exclude the red squirrel and act as a reservoir for squirrelpox virus (SQPV). The disease is generally fatal to red squirrels and their ecological replacement by grey squirrels is up to 25 times faster where the virus is present. We aimed to determine: (1) the seropositivity and prevalence of SQPV DNA in the invasive and native species at a regional scale; (2) possible SQPV transmission routes; and, (3) virus degradation rates under differing environmental conditions. Grey (n = 208) and red (n = 40) squirrel blood and tissues were sampled. Enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qPCR) techniques established seropositivity and viral DNA presence, respectively. Overall 8% of squirrels sampled (both species combined) had evidence of SQPV DNA in their tissues and 22% were in possession of antibodies. SQPV prevalence in sampled red squirrels was 2.5%. Viral loads were typically low in grey squirrels by comparison to red squirrels. There was a trend for a greater number of positive samples in spring and summer than in winter. Possible transmission routes were identified through the presence of viral DNA in faeces (red squirrels only), urine and ectoparasites (both species). Virus degradation analyses suggested that, after 30 days of exposure to six combinations of environments, there were more intact virus particles in scabs kept in warm (25 °C) and dry conditions than in cooler (5 and 15 °C) or wet conditions. We conclude that SQPV is present at low prevalence in invasive grey squirrel populations with a lower prevalence in native red squirrels. Virus transmission could occur through urine especially during warm dry summer conditions but, more notably, via ectoparasites, which are shared by both species.