Integrating chytrid fungal parasites into plankton ecology: research gaps and needs.

Chytridiomycota, often referred to as chytrids, can be virulent parasites with the potential to inflict mass mortalities on hosts, causing e.g. changes in phytoplankton size distributions and succession, and the delay or suppression of bloom events. Molecular environmental surveys have revealed an unexpectedly large diversity of chytrids across a wide range of aquatic ecosystems worldwide. As a result, scientific interest towards fungal parasites of phytoplankton has been gaining momentum in the past few years. Yet, we still know little about the ecology of chytrids, their life cycles, phylogeny, host specificity and range. Information on the contribution of chytrids to trophic interactions, as well as co-evolutionary feedbacks of fungal parasitism on host populations is also limited. This paper synthesizes ideas stressing the multifaceted biological relevance of phytoplankton chytridiomycosis, resulting from discussions among an international team of chytrid researchers. It presents our view on the most pressing research needs for promoting the integration of chytrid fungi into aquatic ecology.

The Ninth Plenary of the Intergovernmental Platform on Biodiversity and Ecosystem Services (IPBES-9): sustainable use, values, and business (as usual).

The Intergovernmental Science-Policy Platform for Biodiversity and Ecosystem Services (IPBES) held its ninth plenary session in hybrid form at Bonn, Germany in July 2022. The plenary had a packed agenda with assessments on the Sustainable Use of Wild Species and Values of Biodiversity to consider, as well as discuss a "Nature Future Framework" and a scoping document for business and biodiversity. Here, we present key issues that emerged from the plenary and suggest some matters Platform members need to consider for the future.

Biofilm community composition is changing in remote mountain lakes with a relative increase in potentially toxigenic algae.

Mountain lakes provide clear drinking water to humankind but are strongly impacted by global change. Benthic biofilms are crucial for maintaining water quality in these oligotrophic lakes, yet little is known about the effects of global change on mountain biofilm communities. By combining analyses of metabarcoding data on 16S and 18S rRNA genes with climatic and environmental data, we investigated global change effects on the composition of biofilm prokaryotic and micro-eukaryotic assemblages in a five-year monitoring program of 26 Pyrenean lakes (2016-2020). Using time-decay relationships and within-lake dissimilarity modelling, we show that the composition of both prokaryotic and micro-eukaryotic biofilm communities significantly shifted and their biodiversity declined from 2016 to 2020. In particular, analyses of temporal trends with linear mixed models indicated an increase in the richness and relative abundance of cyanobacteria, including potentially toxigenic cyanobacteria, and a concomitant decrease in diatom richness and relative abundance. While these compositional shifts may be due to several drivers of global change acting simultaneously on mountain lake biota, water pH and hardness were, from our data, the main environmental variables associated with changes for both prokaryotic and micro-eukaryotic assemblages. Water pH and hardness increased in our lakes over the study period, and are known to increase in Pyrenean lakes due to the intensification of rock weathering as a result of climate change. Given predicted climate trends and if water pH and hardness do cause some changes in benthic biofilms, those changes might be further exacerbated in the future. Such biofilm compositional shifts may induce cascading effects in mountain food webs, threatening the resilience of the entire lake ecosystem. The rise in potentially toxigenic cyanobacteria also increases intoxication risks for humans, pets, wild animals, and livestock that use mountain lakes. Therefore, our study has implications for water quality, ecosystem health, public health, as well as local economies (pastoralism, tourism), and highlights the possible impacts of global change on mountain lakes.

The role of abiotic variables in an emerging global amphibian fungal disease in mountains.

The emergence of the chytridiomycete fungal pathogen Batrachochytrium dendrobatidis (Bd), causing the disease chytridiomycosis, has caused collapse of amphibian communities in numerous mountain systems. The health of amphibians and of mountain freshwater habitats they inhabit is also threatened by ongoing changes in environmental and anthropogenic factors such as climate, hydrology, and pollution. Climate change is causing more extreme climatic events, shifts in ice occurrence, and changes in the timing of snowmelt and pollutant deposition cycles. All of these factors impact both pathogen and host, and disease dynamics. Here we review abiotic variables, known to control Bd occurrence and chytridiomycosis severity, and discuss how climate change may modify them. We propose two main categories of abiotic variables that may alter Bd distribution, persistence, and physiology: 1) climate and hydrology (temperature, precipitation, hydrology, ultraviolet radiation (UVR); and, 2) water chemistry (pH, salinity, pollution). For both categories, we identify topics for further research. More studies on the relationship between global change, pollution and pathogens in complex landscapes, such as mountains, are needed to allow for accurate risk assessments for freshwater ecosystems and resulting impacts on wildlife and human health. Our review emphasizes the importance of using data of higher spatiotemporal resolution and uniform abiotic metrics in order to better compare study outcomes. Fine-scale temperature variability, especially of water temperature, variability of moisture conditions and water levels, snow, ice and runoff dynamics should be assessed as abiotic variables shaping the mountain habitat of pathogen and host. A better understanding of hydroclimate and water chemistry variables, as co-factors in disease, will increase our understanding of chytridiomycosis dynamics.

Complex chemical cocktail, containing insecticides diazinon and permethrin, drives acute toxicity to crustaceans in mountain lakes.

Mountain lakes have long been perceived as pristine environments. However, atmospheric deposition of persistent organic pollutants (POPs) have been shown to expose these sensitive ecosystems to chemical pollution. Little is known on how this pollution impacts aquatic ecosystems at high altitudes. We combined passive sampling with liquid and gas chromatography high resolution mass spectrometry (LC- and GC-HRMS) to screen the water of eight lakes in three different regions of the French Pyrenees. In total, we screened for 479 organic chemicals including POPs, polycyclic aromatic hydrocarbons (PAHs), legacy and current-use pesticides, biocides, and musk fragrances. We detected a complex cocktail of 151 individual chemicals and used their toxic unit summation (ΣTU) to assess toxicity for crustaceans and algae. While risks for algae never reached chronic risks, this was always the case for crustaceans. Acute toxic risk thresholds for crustaceans were even exceeded in several of our sites. At sites with acute toxic risk levels (> 0.1 ΣTU) crustaceans were completely absent or showed a low abundance. We conclude that crustaceans were at least partly impacted by the high toxic risks driven by the insecticides diazinon and permethrin. These drugs are widely used to protect livestock from blue tongue disease transmitted by sucking insects, suggesting free roaming livestock as local source. Our results provide important evidence on toxic chemical pollution in relatively remote mountain areas, with important consequences for aquatic mountain ecosystems.

Environment is associated with chytrid infection and skin microbiome richness on an amphibian rich island (Taiwan).

Growing evidence suggests that the origins of the panzootic amphibian pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal) are in Asia. In Taiwan, an island hotspot of high amphibian diversity, no amphibian mass mortality events linked to Bd or Bsal have been reported. We conducted a multi-year study across this subtropical island, sampling 2517 individuals from 30 species at 34 field sites, between 2010 and 2017, and including 171 museum samples collected between 1981 and 2009. We analyzed the skin microbiome of 153 samples (6 species) from 2017 in order to assess any association between the amphibian skin microbiome and the probability of infection amongst different host species. We did not detect Bsal in our samples, but found widespread infection by Bd across central and northern Taiwan, both taxonomically and spatially. Museum samples show that Bd has been present in Taiwan since at least 1990. Host species, geography (elevation), climatic conditions and microbial richness were all associated with the prevalence of infection. Host life-history traits, skin microbiome composition and phylogeny were associated with lower prevalence of infection for high altitude species. Overall, we observed low prevalence and burden of infection in host populations, suggesting that Bd is enzootic in Taiwan where it causes subclinical infections. While amphibian species in Taiwan are currently threatened by habitat loss, our study indicates that Bd is in an endemic equilibrium with the populations and species we investigated. However, ongoing surveillance of the infection is warranted, as changing environmental conditions may disturb the currently stable equilibrium.

Microbiome function predicts amphibian chytridiomycosis disease dynamics.

BACKGROUND: The fungal pathogen Batrachochytrium dendrobatidis (Bd) threatens amphibian biodiversity and ecosystem stability worldwide. Amphibian skin microbial community structure has been linked to the clinical outcome of Bd infections, yet its overall functional importance is poorly understood. METHODS: Microbiome taxonomic and functional profiles were assessed using high-throughput bacterial 16S rRNA and fungal ITS2 gene sequencing, bacterial shotgun metagenomics and skin mucosal metabolomics. We sampled 56 wild midwife toads (Alytes obstetricans) from montane populations exhibiting Bd epizootic or enzootic disease dynamics. In addition, to assess whether disease-specific microbiome profiles were linked to microbe-mediated protection or Bd-induced perturbation, we performed a laboratory Bd challenge experiment whereby 40 young adult A. obstetricans were exposed to Bd or a control sham infection. We measured temporal changes in the microbiome as well as functional profiles of Bd-exposed and control animals at peak infection. RESULTS: Microbiome community structure and function differed in wild populations based on infection history and in experimental control versus Bd-exposed animals. Bd exposure in the laboratory resulted in dynamic changes in microbiome community structure and functional differences, with infection clearance in all but one infected animal. Sphingobacterium, Stenotrophomonas and an unclassified Commamonadaceae were associated with wild epizootic dynamics and also had reduced abundance in laboratory Bd-exposed animals that cleared infection, indicating a negative association with Bd resistance. This was further supported by microbe-metabolite integration which identified functionally relevant taxa driving disease outcome, of which Sphingobacterium and Bd were most influential in wild epizootic dynamics. The strong correlation between microbial taxonomic community composition and skin metabolome in the laboratory and field is inconsistent with microbial functional redundancy, indicating that differences in microbial taxonomy drive functional variation. Shotgun metagenomic analyses support these findings, with similar disease-associated patterns in beta diversity. Analysis of differentially abundant bacterial genes and pathways indicated that bacterial environmental sensing and Bd resource competition are likely to be important in driving infection outcomes. CONCLUSIONS: Bd infection drives altered microbiome taxonomic and functional profiles across laboratory and field environments. Our application of multi-omics analyses in experimental and field settings robustly predicts Bd disease dynamics and identifies novel candidate biomarkers of infection. Video Abstract.

Scientists' warning of threats to mountains.

Mountains are an essential component of the global life-support system. They are characterized by a rugged, heterogenous landscape with rapidly changing environmental conditions providing myriad ecological niches over relatively small spatial scales. Although montane species are well adapted to life at extremes, they are highly vulnerable to human derived ecosystem threats. Here we build on the manifesto 'World Scientists' Warning to Humanity', issued by the Alliance of World Scientists, to outline the major threats to mountain ecosystems. We highlight climate change as the greatest threat to mountain ecosystems, which are more impacted than their lowland counterparts. We further discuss the cascade of "knock-on" effects of climate change such as increased UV radiation, altered hydrological cycles, and altered pollution profiles; highlighting the biological and socio-economic consequences. Finally, we present how intensified use of mountains leads to overexploitation and abstraction of water, driving changes in carbon stock, reducing biodiversity, and impacting ecosystem functioning. These perturbations can provide opportunities for invasive species, parasites and pathogens to colonize these fragile habitats, driving further changes and losses of micro- and macro-biodiversity, as well further impacting ecosystem services. Ultimately, imbalances in the normal functioning of mountain ecosystems will lead to changes in vital biological, biochemical, and chemical processes, critically reducing ecosystem health with widespread repercussions for animal and human wellbeing. Developing tools in species/habitat conservation and future restoration is therefore essential if we are to effectively mitigate against the declining health of mountains.

The eighth plenary of the Intergovernmental Science-Policy Platform for Biodiversity and Ecosystem Services (IPBES-8): online, nexus, and transformative change.

The Intergovernmental Science-Policy Platform for Biodiversity and Ecosystem Services held its 8th plenary session online in June 2021, during which a new budget was approved up to 2023, continuing work plans were agreed, and scoping documents for two new assessments, the nexus and transformative change assessments, were accepted.

Topsoil microbial community structure responds to land cover type and environmental zone in the Western Pacific region.

Soil encompasses diverse microbial communities that are essential for fundamental ecosystem functions such as biogeochemical cycling. To better understand underlying biogeochemical processes, it is necessary to know the structure of soil archaeal and bacterial communities and their responses to edaphic and climate variables within and across various land cover types (LCTs) and environmental zones (ENZs). Here we sampled eighty-nine sites across five ENZs and four LCTs within the Western Pacific region. Through leveraging the second-generation sequencing of topsoil samples, we showed that α-diversity (taxonomic diversity) of archaea strongly varied within LCTs, whereas bacterial α-diversity was significantly controlled by both LCT and ENZ. Soil archaea and bacteria showed global niche differentiation associated with contrasting diversity responses to latitude and differential responses of microbial diversity patterns to edaphic and climate variables within LCTs and ENZs. In contrast to α-diversity, microbial ß-diversity (the compositional dissimilarity between sites) was majorly governed by ENZs, particularly for archaea (P < 0.01). Our results highlight the importance of LCTs and ENZs for understanding soil microbial contributions to nutrient dynamics and ecosystem resilience under land-use intensification and climate change.

Factors driving pathogenicity vs. prevalence of amphibian panzootic chytridiomycosis in Iberia.

Amphibian chytridiomycosis is a disease caused by the fungus Batrachochytrium dendrobatidis (Bd). Whether Bd is a new emerging pathogen (the novel pathogen hypothesis; NPH) or whether environmental changes are exacerbating the host-pathogen dynamic (the endemic pathogen hypothesis; EPH) is debated. To disentangle these hypotheses we map the distribution of Bd and chytridiomycosis across the Iberian Peninsula centred on the first European outbreak site. We find that the infection-free state is the norm across both sample sites and individuals. To analyse this dataset, we use Bayesian zero-inflated binomial models to test whether environmental variables can account for heterogeneity in both the presence and prevalence of Bd, and heterogeneity in the occurrence of the disease, chytridiomycosis. We also search for signatures of Bd-spread within Iberia using genotyping. We show (1) no evidence for any relationship between the presence of Bd and environmental variables, (2) a weak relationship between environmental variables and the conditional prevalence of infection, (3) stage-dependent heterogeneity in the infection risk, (4) a strong association between altitude and chytridiomycosis, (5) multiple Iberian genotypes and (6) recent introduction and spread of a single genotype of Bd in the Pyrenees. We conclude that the NPH is consistent with the emergence of Bd in Iberia. However, epizootic forcing of infection is tied to location and shaped by both biotic and abiotic variables. Therefore, the population-level consequences of disease introduction are explained by EPH-like processes. This study demonstrates the power of combining surveillance and molecular data to ascertain the drivers of new emerging infections diseases.

Biodiversity loss, emerging pathogens and human health risks.

Outbreaks of emerging infectious diseases are occurring with increasing frequency and consequences, including wildlife diseases and zoonoses. Those have potentially long-lasting effects on human and wildlife populations, with inevitable direct and indirect effects on ecosystems. The intensifying emergence of infectious pathogens has many underlying reasons, all driven by the growing anthropogenic impact on nature. Intensifying pathogen emergence can be attributed to climate change, biodiversity loss, habitat degradation, and an increasing rate of wildlife-human contacts. All of these are caused by synergies between persisting intense poverty and a growing human population. Improved global management of the human-driven biological degradation and international dispersal processes that exacerbate those pandemic threats are now long overdue. It is vital that we act decisively in the aftermath of the COVID-19 crisis to radically change how we collectively manage the planet as a whole.

Environmental Factors and Host Microbiomes Shape Host-Pathogen Dynamics.

Microorganisms are increasingly recognized as ecosystem-relevant components because they affect the population dynamics of hosts. Functioning at the interface of the host and pathogen, skin and gut microbiomes are vital components of immunity. Recent work reveals a strong influence of biotic and abiotic environmental factors (including the environmental microbiome) on disease dynamics, yet the importance of the host-host microbiome-pathogen-environment interaction has been poorly reflected in theory. We use amphibians and the disease chytridiomycosis caused by the fungal pathogen Batrachochytrium dendrobatidis to show how interactions between host, host microbiome, pathogen, and the environment all affect disease outcome. Our review provides new perspectives that improve our understanding of disease dynamics and ecology by incorporating environmental factors and microbiomes into disease theory.

A GIS-based policy support tool to determine national responsibilities and priorities for biodiversity conservation.

Efficient biodiversity conservation requires that limited resources be allocated in accordance with national responsibilities and priorities. Without appropriate computational tools, the process of determining these national responsibilities and conservation priorities is time intensive when considering many species across geographic scales. Here, we have developed a computational tool as a module for the ArcGIS geographic information system. The ArcGIS National Responsibility Assessment Tool (NRA-Tool) can be used to create hierarchical lists of national responsibilities and priorities for global species conservation. Our tool will allow conservationists to prioritize conservation efforts and to focus limited resources on relevant species and regions. We showcase our tool with data on 258 bird species and various biophysical regions, including Environmental Zones in 58 Asian countries and regions. Our tool provides a decision support system for conservation policy with attractive and easily interpretable visual outputs illustrating national responsibilities and priorities for species conservation. The graphical output allows for smooth integration into assessment reports, such as the European Article 17 report, the Living Planet Index report, or similar regional and global reports.

Advantages of volunteer-based biodiversity monitoring in Europe.

Without robust and unbiased systems for monitoring, changes in natural systems will remain enigmatic for policy makers, leaving them without a clear idea of the consequences of any environmental policies they might adopt. Generally, biodiversity-monitoring activities are not integrated or evaluated across any large geographic region. The EuMon project conducted the first large-scale evaluation of monitoring practices in Europe through an on-line questionnaire and is reporting on the results of this survey. In September 2007 the EuMon project had documented 395 monitoring schemes for species, which represents a total annual cost of about 4 million euro, involving more than 46,000 persons devoting over 148,000 person-days/year to biodiversity-monitoring activities. Here we focused on the analysis of variations of monitoring practices across a set of taxonomic groups (birds, amphibians and reptiles, mammals, butterflies, plants, and other insects) and across 5 European countries (France, Germany, Hungary, Lithuania, and Poland). Our results suggest that the overall sampling effort of a scheme is linked with the proportion of volunteers involved in that scheme. Because precision is a function of the number of monitored sites and the number of sites is maximized by volunteer involvement, our results do not support the common belief that volunteer-based schemes are too noisy to be informative. Just the opposite, we believe volunteer-based schemes provide relatively reliable data, with state-of-the-art survey designs or data-analysis methods, and consequently can yield unbiased results. Quality of data collected by volunteers is more likely determined by survey design, analytical methodology, and communication skills within the schemes rather than by volunteer involvement per se.

Biological Concepts for the Control of Aquatic Zoosporic Diseases.

Aquatic zoosporic diseases are threatening global biodiversity and ecosystem services, as well as economic activities. Current means of controlling zoosporic diseases are restricted primarily to chemical treatments, which are usually harmful or likely to be ineffective in the long term. Furthermore, some of these chemicals have been banned due to adverse effects. As a result, there is a need for alternative methods with minimal side-effects on the ecosystem or environment. Here, we integrate existing knowledge of three poorly interconnected areas of disease research - amphibian conservation, aquaculture, and plankton ecology - and arrange it into seven biological concepts to control zoosporic diseases. These strategies may be less harmful and more sustainable than chemical approaches. However, more research is needed before safe application is possible.

Pathogen invasion history elucidates contemporary host pathogen dynamics.

Amphibians, the most threatened group of vertebrates, are seen as indicators of the sixth mass extinction on earth. Thousands of species are threatened with extinction and many have been affected by an emerging infectious disease, chytridiomycosis, caused by the fungal pathogen, Batrachochytrium dendrobatidis (Bd). However, amphibians exhibit different responses to the pathogen, such as survival and population persistence with infection, or mortality of individuals and complete population collapse after pathogen invasion. Multiple factors can affect host pathogen dynamics, yet few studies have provided a temporal view that encompasses both the epizootic phase (i.e. pathogen invasion and host collapse), and the transition to a more stable co-existence (i.e. recovery of infected host populations). In the Sierra Nevada mountains of California, USA, conspecific populations of frogs currently exhibit dramatically different host/ Bd-pathogen dynamics. To provide a temporal context by which present day dynamics may be better understood, we use a Bd qPCR assay to test 1165 amphibian specimens collected between 1900 and 2005. Our historical analyses reveal a pattern of pathogen invasion and eventual spread across the Sierra Nevada over the last century. Although we found a small number of Bd-infections prior to 1970, these showed no sign of spread or increase in infection prevalence over multiple decades. After the late 1970s, when mass die offs were first noted, our data show Bd as much more prevalent and more spatially spread out, suggesting epizootic spread. However, across the ~400km2 area, we found no evidence of a wave-like pattern, but instead discovered multiple, nearly-simultaneous invasions within regions. We found that Bd invaded and spread in the central Sierra Nevada (Yosemite National Park area) about four decades before it invaded and spread in the southern Sierra Nevada (Sequoia and Kings Canyon National Parks area), and suggest that the temporal pattern of pathogen invasion may help explain divergent contemporary host pathogen dynamics.

Essential biodiversity variables for mapping and monitoring species populations.

Species distributions and abundances are undergoing rapid changes worldwide. This highlights the significance of reliable, integrated information for guiding and assessing actions and policies aimed at managing and sustaining the many functions and benefits of species. Here we synthesize the types of data and approaches that are required to achieve such an integration and conceptualize 'essential biodiversity variables' (EBVs) for a unified global capture of species populations in space and time. The inherent heterogeneity and sparseness of raw biodiversity data are overcome by the use of models and remotely sensed covariates to inform predictions that are contiguous in space and time and global in extent. We define the species population EBVs as a space-time-species-gram (cube) that simultaneously addresses the distribution or abundance of multiple species, with its resolution adjusted to represent available evidence and acceptable levels of uncertainty. This essential information enables the monitoring of single or aggregate spatial or taxonomic units at scales relevant to research and decision-making. When combined with ancillary environmental or species data, this fundamental species population information directly underpins a range of biodiversity and ecosystem function indicators. The unified concept we present links disparate data to downstream uses and informs a vision for species population monitoring in which data collection is closely integrated with models and infrastructure to support effective biodiversity assessment.

National responsibilities in European species conservation: a methodological review.

One particular challenge in reducing the loss of biodiversity by 2010, as agreed on at the Earth Summit in 2002, is to assign conservation tasks to geographic or administrative entities (e.g., countries or regions) on different geographical scales. To identify conservation tasks, it is imperative to determine the importance of a specific area for the global survival of a species. So far, these national or subnational responsibilities for the conservation of species have been included differently in methods prioritizing conservation. We reviewed how 12 European and 3 non-European methods determined national conservation responsibilities and evaluated the international importance of a biological population. Different countries used different methodologies, which made a direct comparison of assessments of national responsibilities among countries extremely difficult. Differences existed in the importance criteria used. Criteria included population decline, range reduction, rarity status, degree of isolation of a population, endemism, proportional distribution, and geographic location. To increase comparability, it is imperative to develop criteria for which data are generally available and to standardize the methodology among countries. A standardized method would allow conservation decisions to be based on the conservation status of a species and on the responsibility of a geographic or administrative entity for the survival of a species. We suggest that such a method should use a scalable index of proportional distribution, taxonomic status, and the distribution pattern of a taxon or species as key elements. Such a method would allow for the creation of hierarchical lists and would be highly relevant for parts of the world with multiple political jurisdictions or state unions and for nations with regional governmental structures. Conservation priorities could then be reasonably set by combining national responsibility assessments with the international conservation status of a species.

Jailed in the mountains: Genetic diversity and structure of an endemic newt species across the Pyrenees.

The Pyrenees represent a natural laboratory for biogeographic, evolutionary and ecological research of mountain fauna as a result of the high variety of habitats and the profound effect of the glacial and interglacial periods. There is a paucity of studies providing a detailed insight into genetic processes and better knowledge on the patterns of genetic diversity and how they are maintained under high altitude conditions. This is of particular interest when considering the course of past climate conditions and glaciations in a species which is considered site tenacious, with long generation times. Here we analyzed the genetic patterns of diversity and structure of the endemic Pyrenean brook newt (Calotriton asper) along its distribution range, with special emphasis on the distinct habitat types (caves, streams, and lakes), and the altitudinal and geographical ranges, using a total set of 900 individuals from 44 different localities across the Pyrenean mountain range genotyped for 19 microsatellite loci. We found evidence for a negative longitudinal and positive altitudinal gradient of genetic diversity in C. asper populations. The fact that genetic diversity was markedly higher westwards is in accordance with other Pyrenean species. However, the impact of altitudinal gradient on the genetic diversity seems to differ from other species, and mostly from other amphibians. We found that lower altitudes can act as a barrier probably because the lowlands do not provide a suitable habitat for C. asper. Regarding the distinct habitat types, caves had significantly lower values of genetic diversity compared to streams or lakes. The mean FST value was relatively high (0.304) with maximum values as high as 0.771, suggesting a highly structured total population. Indeed, populations were grouped into five subclusters, the eastern populations (cluster 1) remained grouped into two subclusters and the central-western Pyrenees (cluster 2) into three subclusters. The increase of isolation with geographical distance is consistent with the population structure detected. In conclusion, C. asper seems to be adapted to high altitude mountain habitats, and its genetic diversity is higher in the western Pyrenees. In terms of conservation priority, we consider more relevant the populations that represent a reservoir of genetic diversity.