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Extra-organismal DNA (eoDNA) from material left behind by organisms (noninvasive DNA, e.g., feces, hair) or from environmental samples (eDNA, e.g., water, soil) is a valuable source of genetic information. However, the relatively low quality and quantity of eoDNA, which can be further degraded by environmental factors, results in reduced amplification and sequencing success. This is often compensated for through cost- and time-intensive replications of genotyping/sequencing procedures. Therefore, system- and site-specific quantifications of environmental degradation are needed to maximize sampling efficiency (e.g., fewer replicates, shorter sampling durations), and to improve species detection and abundance estimates. Using 10 environmentally diverse bat roosts as a case study, we developed a robust modeling pipeline to quantify the environmental factors degrading eoDNA, predict eoDNA quality, and estimate sampling-site-specific ideal exposure duration. Maximum humidity was the strongest eoDNA-degrading factor, followed by exposure duration and then maximum temperature. We also found a positive effect when hottest days occurred later. The strength of this effect fell between the strength of the effects of exposure duration and maximum temperature. With those predictors and information on sampling period (before or after offspring were born), we reliably predicted mean eoDNA quality per sampling visit at new sites with a mean squared error of 0.0349. Site-specific simulations revealed that reducing exposure duration to 2–8 days could substantially improve eoDNA quality for future sampling. Our pipeline identified high humidity and temperature as strong drivers of eoDNA degradation even in the absence of rain and direct sunlight. Furthermore, we outline the pipeline's utility for other systems and study goals, such as estimating sample age, improving eDNA-based species detection, and increasing the accuracy of abundance estimates.

The development of methods for individual identification in wild species and the refinement of Capture-Mark-Recapture (CMR) models during the past decades has greatly improved the assessment of population demographic rates to answer ecological and conservation questions. In particular, multistate models, with their flexibility for the analysis of complex study systems, have become popular in the ecological community. However, despite the extensive use of these models, little attention has been paid to the effect of common violations of the CMR model assumptions, such as mark loss and the often-associated recycling of remarked individuals.To explore this knowledge gap we used a wide range of simulation scenarios reflecting frequently encountered real case studies inspired from the survival rates of 700 vertebrates’ species. We estimated the effects of mark loss and recycled individuals on parameter estimates using a multi-state Cormack-Jolly-Seber (MSCJS) framework. We explored parameter bias through simulations of a metapopulation system with different capture and survival rates. We also illustrated how mark loss can be easily estimated and accounted for using an empirical long-term (10 years) CMR dataset of bats, individually identified using both PIT tag technology as marks that can be lost, and multi-locus genotypes as ‘permanent marks’.The results from our simulated scenarios demonstrated that the occurrence of bias and the parameters concerned were highly dependent on the study system, and no general rules on parameter behaviour can be established a priori. The model structure and the interdependency among parameters make it challenging to predict how bias could affect estimates.Our results highlight the need to assess the effect of mark loss when using MSCJS models. Ignoring such violations of model assumptions can have important implications for ecological inferences and conservation policies. In general, the use of permanent marks (e.g. genotype), should always be preferred when modelling of population dynamics and if not possible, combining two types of temporary marks (e.g. PIT tags, bands) should be considered.

Some species are difficult to observe and others, need to be not disturbed because of their vulnerability. In response to the difficulty of studying the dispersal behaviors of these species, some areas of biology have been combined in order to access the information despite practical limitations. Here we present the combination of several methodologies from landscape ecology to non- invasive population genetics that allow us to obtain important information onRinolophus hipposideros, a vulnerable European bat. We genotyped 18 georeferrenced colonies in Brittany (France) from droppings collected in their refuges. We used 6 microsatellite markers in order to obtain the genetic distances between them. On the other hand we calculated Euclidian distances between the refuges occupied by these colonies and some ecological distances with the Pathmatrix module of ArcGis 3.2. We tested hypothesis about the difficulty of dispersal of the species in areas without forest cover or with a low density of hedges. Thanks to the Monmonier algorithm we could infer possible genetic barriers between the colonies and we could compare their location to the presence of landscape barriers (areas with little tree cover). We detected a pattern of isolation by distance that reveals limited dispersal capacities in the species but no pattern linked to ecological distances. We found that some of the neighboring colonies with greater genetic distances between them were located in areas with low density of hedges which could suggest an impact of this landscape element in their movements. Finer studies should allow us to conclude on the need or not of forest cover in the dispersal of this species.Abstract Figure

International audience; Recent theoretical and experimental models have revealed the role played by evolution during species spread, and in particular have questioned the influence of genetic drift at range edges. By investigating the spread of an aquatic invader in patchy habitats, we quantified genetic drift and explored its consequences for genetic diversity and fitness. We examined the interplay of gene flow and genetic drift in 36 populations of the red swamp crayfish, Procambarus clarkii, in a relatively recently invaded wetland area (30 years, Brière, northwest France). Despite the small spatial scale of our study (15 km ), populations were highly structured according to the strong barrier of land surfaces and revealed a clear pattern of colonization through watercourses. Isolated populations exhibited small effective sizes and low dispersal rates that depended on water connectivity, suggesting that genetic drift dominated in the evolution of allele frequencies in these populations. We also observed a significant decrease in the genetic diversity of isolated populations over only a 2-year period, but failed to demonstrate an associated fitness cost using fluctuating asymmetry. This study documents the possible strong influence of genetic drift during the spread of a species, and such findings provide critical insights into the current context of profound rearrangements in species distributions due to global change.

Bats are the longest-lived mammals, given their body size. However, the underlying molecular mechanisms of their extended healthspans are poorly understood. To address this question we carried out an eight-year longitudinal study of ageing in long-lived bats (Myotis myotis). We deep-sequenced ~1.7 trillion base pairs of RNA from 150 blood samples collected from known aged bats to ascertain the age-related transcriptomic shifts and potential microRNA-directed regulation that occurred. We also compared ageing transcriptomic profiles between bats and other mammals by analysis of 298 longitudinal RNA sequencing datasets. Bats did not show the same transcriptomic changes with age as commonly observed in humans and other mammals, but rather exhibited a unique, age-related gene expression pattern associated with DNA repair, autophagy, immunity and tumour suppression that may drive their extended healthspans. We show that bats have naturally evolved transcriptomic signatures that are known to extend lifespan in model organisms, and identify novel genes not yet implicated in healthy ageing. We further show that bats’ longevity profiles are partially regulated by microRNA, thus providing novel regulatory targets and pathways for future ageing intervention studies. These results further disentangle the ageing process by highlighting which ageing pathways contribute most to healthy ageing in mammals. RNA-Seq analysis of an 8-year longitudinal study of long-lived bats reveals unique transcriptomic signatures and novel candidate genes associated with healthy ageing.

Understanding the relationship between habitat quality and population dynamics is fundamental for long-term management and range predictions in ecology. However, habitat quality is generally only investigated at the individual scale, as it is the case for the lesser horseshoe bat (Rhinolophus hipposideros), a species of conservation concern. Using a statistical modelling approach and census data of 94 lesser horseshoe bat colonies located in Brittany (France), we analysed the effect of landscape composition and configuration on the demographic components of surveyed maternity colonies (i.e. colony size, fecundity and growth rate), and compared our result to those provided by individual-based studies. Our results validated that the landscape in a 500-meter buffer around colonies (core foraging area) is crucial for population size and dynamics, and confirmed the positive influence of broadleaved woodland proportion on bat colony size. We revealed a positive effect of lakeshores and riverbanks on colony size and growth rate, underlying the importance of these habitats for the long-term conservation of this non-migratory forest species. Importantly, our results refine previous knowledge concerning the threat posed by the intensification of human activities (e.g. urbanization, agriculture, habitat fragmentation), and highlight the negative effect of large and regular patches of artificial and crop lands and of open land patches shape complexity on all demographic variables investigated. While our results support the dependence of population dynamics and associated conservation management to individual behaviour and sensitivity, environmental responses differed between the population metrics investigated, showing that efficient range prediction will require to fully grasp the complexity of the interaction between landscape and the different population dynamic parameters.

The recent advent of high throughput sequencing and genotyping technologies enables the comparison of patterns of polymorphisms at a very large number of markers. While the characterization of genetic structure from individual sequencing data remains expensive for many non-model species, it has been shown that sequencing pools of individual DNAs (Pool-seq) represents an attractive and cost-effective alternative. However, analyzing sequence read counts from a DNA pool instead of individual genotypes raises statistical challenges in deriving correct estimates of genetic differentiation. In this article, we provide a method-of-moments estimator ofFSTfor Pool-seq data, based on an analysis-of-variance framework. We show, by means of simulations, that this new estimator is unbiased, and outperforms previously proposed estimators. We evaluate the robustness of our estimator to model misspecification, such as sequencing errors and uneven contributions of individual DNAs to the pools. Last, by reanalyzing published Pool-seq data of different ecotypes of the prickly sculpinCottus asper, we show how the use of an unbiasedFSTestimator may question the interpretation of population structure inferred from previous analyses.

International audience; There is a growing literature on the detection of species-specific DNA fragments in water to reveal animals' presence in aquatic environments. Analysis of Rees etal. (Journal of Applied Ecology, 2014, 51, 1450) and additional publications highlight the downside of the method and suggest that some conclusions have to be toned down. Non-detection of species-specific DNA fragments in a water sample does not automatically imply the absence of the target species. A positive signal does not necessarily mean the species is present because the environmental DNA (eDNA) could have been transported or preserved after an animal's death. Detection of aquatic invertebrates is weakly documented. The effect of species abundance on detection efficiency is not always established.Synthesis and applications. Before successfully becoming an efficient tool to assist proper management policies for monitoring of aquatic species, eDNA surveys must implement sampling strategies that allow estimating detection rates and scaling up information from water samples to larger spatial areas. The effects of discharge variations and dendritic organization in running waters require particular consideration. Before successfully becoming an efficient tool to assist proper management policies for monitoring of aquatic species, eDNA surveys must implement sampling strategies that allow estimating detection rates and scaling up information from water samples to larger spatial areas. The effects of discharge variations and dendritic organization in running waters require particular consideration.

International audience; A general pattern in animal behavior is that group-living species tend to bias their sociopositive behavior toward genetic relatives. In nonhuman primates, kin-biased social bonds have been reported in large multimale, multifemale macaque and baboon groups, but little is known for other species. We addressed this with a comparative study on the genetic and social organization of two sympatric forest guenons, Diana (Cercopithecus diana) and Campbell’s monkeys (Cercopithecus campbelli). We conducted long-term observations of social interactions in two groups of each species in their natural West African forest habitat and collected fecal samples for subsequent microsatellite genetic analyses. We found that both formed female-bonded, egalitarian social organizations. We then compared patterns of genetic relatedness, spatial proximity, and key social behaviors and found that females consistently targeted individuals other than their closest relatives to form social bonds. The fact that females did not preferentially favor genetic relatives contributes to a growing literature showing that social bonds, or “friendships,” among unrelated individuals plays a key role in primate social organizations.

The ontogeny and heritability of echolocation, an important sense in echolocating bats, is still not completely understood. Intraspecific variation in echolocation calls can be high, although the importance of possible explanatory variables (e.g. age, sex, social groups) remains largely unknown. Echolocation pulse features may vary among maternity roosts and this can theoretically be caused either by intercolony genetic differences or by vocal dialects learned during ontogeny within a roost (or a combination of both). In the present study, we analyzed intraspecific variation in echolocation parameters in relation to genetic structure at bi-parentally inherited microsatellites and maternally inherited mitochondrial (mt)DNA in maternal colonies of Pipistrellus pipistrellus in Central Europe. We found that individual colonies differ significantly in mtDNA, whereas the structure on nuclear markers is almost absent. This suggests a typical temperate bat social structure pattern, with strong sex-biased dispersal (i.e. philopatric females and dispersing males) (up to 92% of males leave their birth place according to our results). However, we show for the first time that genetic differentiation among mtDNA matrilines is associated with significant intercolony echolocation parameter differences. Because the genetic component of echolocation is not likely to be encoded by mtDNA, the results support the hypothesis of maternal echolocation dialect transmission to offspring, and the role of learning in this process is discussed.

International audience; Climatic variables are often considered when studying environmental impacts on population dynamics of terrestrial species. However, the temporal resolution considered varies depending on studies, even among studies of the same taxa. Most studies interested in climatic impacts on populations tend to average climatic data across timeframes covering life cycle periods of the organism in question or longer, even though most climatic databases provide at least a monthly resolution. We explored the impact of climatic variables on lesser horseshoe bat (Rhinolophus hipposideros) demography based on count data collected at 94 maternity colonies from 2000 to 2014 in Britanny, France. Meteorological data were considered using different time resolutions (month, life cycle period and year) to investigate their adequacy. Model averaging was used to detect significant predictors for each temporal resolution. Our results show that the finest temporal resolution, e.g. month, was more informative than coarser ones. Precipitation predictors were particularly decisive, with a negative impact on colony sizes when rainfall occurred in October, and a positive impact for June precipitations. Fecundity was influenced by April weather. This highlights the strong impact of climatic conditions during crucial but short time periods on the population dynamics of bats. We demonstrate the importance of choosing an appropriate time resolution and suggest that analogous studies should consider fine-scale temporal resolution (e.g. month) to better grasp the relationship between population dynamics and climatic conditions.

This study was funded by the Agence Francaise pour la Biodiversite (research program supervised by J.M.P.) and the Ministere de l'Education Nationale, de l'Enseignement Superieur et de la Recherche (Ph.D. grant to N.B.). This work was carried out under the licenses 08/2015 and 07/2016 delivered by the Prefecture de la Loire-Atlantique. We thank the Parc Natural Regional de Briere, notably J.P. Damien, for supporting our research activities on amphibian ecology. The authors declare that they have no conflict of interest. We are also grateful to two anonymous reviewers for their constructive comments on an earlier version of the manuscript.; International audience; Stable isotope analyses (SIA) are now widely used in ecology research to quantify the trophic implications of a large range of ecological processes. Non-destructive sampling methods have been successfully developed for most vertebrates, but the validation of such methods is missing in amphibians. Filling this methodological gap is critical to reduce the significant lack of knowledge on the trophic ecology of this imperilled group. The relevance of tail fin as a surrogate for muscle, a tissue routinely used for SIA, was assessed in four amphibian species, including tadpoles and adult newts, by (1) testing the link between the isotope ratios of carbon and nitrogen in fin and muscle, and addressing the additional practical issues of (2) the effects of a topical anaesthetic on isotope ratios, and (3) the minimum quantities of fin needed for SIA to limit the severity of sampling. The isotope ratios were highly correlated between the two tissues in all four species. Equations were provided to predict muscle estimates from fin values for each species separately and grouped by their taxonomic order (anurans or urodeles), which represent a valuable solution for any other species. Topical anaesthetics had little influence on isotope ratios, and the surface of tail fin needed for SIA was small. We conclude that the tail fin of amphibians can be used as a non-lethal surrogate for muscle in SIA, and this offers promising prospects for research on the trophic ecology of amphibians, and potentially on their conservation.; Les analyses d'isotopes stables (AIS) sont maintenant largement utilisées en recherche en écologie pour quantifier les implications trophiques d'une large gamme de processus écologiques. Des méthodes d'échantillonnage non-létales ont été développées chez la plupart des vertébrés, mais une telle validation est absente chez les amphibiens. Il est essentiel de combler cette lacune de façon à réduire le manque de connaissances sur l'écologie trophique de ce groupe menacé. La pertinence de la nageoire caudale comme substitut au muscle, un tissu régulièrement utilisé pour les AIS, a été évaluée chez quatre espèces d'amphibiens, incluant des têtards et des tritons, (1) en testant le lien entre les ratios isotopiques du carbone et de l'azote du muscle et de la nageoire, et en répondant à des questions pratiques comme (2) l'effet d'un anesthésique local sur les ratios isotopiques, et (3) les quantités minimales de nageoire nécessaires aux analyses isotopiques, dans le but de limiter la sévérité de l'échantillonnage. Les ratios isotopiques étaient hautement corrélés entre les deux tissus chez les quatre espèces. Des équations ont été fournies pour prédire les valeurs du muscle à partir de celles de la nageoire en considérant chaque espèce séparément et en les regroupant selon leur ordre taxonomique (anoures ou urodèles) ce qui représente une solution intéressante pour d'autres espèces. L'anesthésique topique a eu une faible influence sur les ratios isotopiques, et la surface de nageoire nécessaire pour les AIS était faible. Nous concluons que la nageoire caudale des amphibiens peut être utilisée en tant que substitut au muscle pour conduire des AIS, ce qui offre des perspectives prometteuses pour la recherche en écologie trophique des amphibiens, et potentiellement pour leur conservation.

Animal societies certainly count among the most complex structures that emerged on earth, the latest of the 'major transitions' in evolution. Their study necessarily involves a wide panel of disciplines and a large diversity of approaches. The field is expanding at a rapid pace, which, however, is not going without some turbulence, as testified by recent fierce controversies regarding the concept of inclusive fitness. These battles elicited appeals for 'more cooperation among altruism researchers' (Okasha 2010). In this spirit, we recently organized at the University of Rennes (10-15 October 2010) a workshop on mammalian social systems. The idea was to bring together 'social scientists' working on a diversity of animal models (from bats to elephants and from hyenas to whales) and using a diversity of approaches (from demography to population genetics and from game-theoretical models to behavioural experiments), with the aim of evaluating how key processes (e.g. inclusive fitness and kin selection, versus cooperation and reciprocity, or the interplay between inbreeding, kin competition and kin recognition) might differently affect the emergence and evolution of social systems in a diversity of mammalian lineages, depending on ecological settings or other constraints. This diversity of complementary approaches explains why the several contributions to the workshop collected in the present Special Issue are not all 'pure' molecular ecology.

The impact of ecology and social organization on genetic structure at landscape spatial scales, where gene dynamics shape evolution as well as determine susceptibility to habitat fragmentation, is poorly understood. Attempts to assess these effects must take into account the potentially confounding effects of history. We used microsatellites to compare genetic structure in seven bat species with contrasting patterns of roosting ecology and social organization, all of which are codistributed in an ancient forest habitat that has been exceptionally buffered from radical habitat shifts. Over one thousand individuals were captured at foraging sites and genotyped at polymorphic microsatellite loci. Analyses of spatially explicit genotype data revealed interspecies differences in the extent of movement and gene flow and genetic structure across continuous intact forest. Highest positive genetic structure was observed in tree-roosting taxa that roost either alone or in small groups. By comparison, a complete absence of genetic autocorrelation was noted in the cave-roosting colonial species across the study area. Our results thus reveal measurable interspecies differences in the natural limits of gene flow in an unmodified habitat, which we attribute to contrasting roosting ecology and social organization. The consequences of ecology and behaviour for gene flow have important implications for conservation. In particular, tree-roosting species characterized by lower vagility and thus gene flow will be disproportionally impacted by landscape-scale forest clearance and habitat fragmentation, which are prevalent in the study region. Our method also highlights the usefulness of rapid sampling of foraging bats for assaying genetic structure, particularly where roosting sites are not always known.

The potential for parallel impacts of habitat change on multiple biodiversity levels has important conservation implications. We report on the first empirical test of the 'species-genetic diversity correlation' across co-distributed taxa with contrasting ecological traits in the context of habitat fragmentation. In a rainforest landscape undergoing conversion to oil palm, we show that depauperate species richness in fragments is mirrored by concomitant declines in population genetic diversity in the taxon predicted to be most susceptible to fragmentation. This association, not seen in the other species, relates to fragment area rather than isolation. While highlighting the over-simplification of extrapolating across taxa, we show that fragmentation presents a double jeopardy for some species. For these, conserving genetic diversity at levels of pristine forest could require sites 15-fold larger than those needed to safeguard species numbers. Importantly, however, each fragment contributes to regional species richness, with larger ones tending to contain more species.

Summary 1 Non-invasive genetic data analysed with capture–mark–recapture (CMR) models can be used to estimate population size, particularly for elusive and endangered species. Data generated from non-invasive genetic sampling are different, however, from conventional CMR data because individuals can be contacted several times within a single sampling session. Two methods have been proposed recently to accommodate this type of data, but no study has attempted to compare their estimates and evaluate their reliability compared with independent estimates of population size. 2 We investigated the reliability and accuracy of estimating the abundance of lesser horseshoe bats Rhinolophus hipposideros by genotyping DNA from droppings collected non-invasively at three colonies over 2 consecutive years. The number of times that each individual was ‘contacted’ (i.e. the number of droppings per individual) was used to estimate population size with two different published methods: a maximum likelihood and a Bayesian estimator. 3 Among the 586 samples extracted, 534 provided a complete genotype at six to eight microsatellite loci, which enabled a reliable discrimination of 165 individuals. Statistical estimates of colony sizes often included independent estimates obtained from visual counts, validating the method. Discrepancies appeared when capture heterogeneity was not taken into account while it occurred. 4 Synthesis and applications. We have taken a first step towards improving methods of estimating numbers of bats by demonstrating that genetic data produced from bat faecal DNA are of high quality and can provide accurate estimates of population size even when samples are taken during only one sampling session. Such protocols provide valuable management tools for elusive and rare species in general. The method is relatively easy and cost-efficient because only one sampling session is required.

We studied the noctule bat (Nyctalus noctula), in which the mitochondrial F(ST) is about 10 times that revealed by nuclear markers, to address two questions. We first verified whether random dispersal of one sex is compatible with highly contrasted mitochondrial and nuclear population structures. Using computer simulations, we then assessed the power of multilocus population differentiation tests when the expected population structure departs only slightly from panmixia. Using an island model with sex-specific demographic parameters, we found that random male dispersal is consistent with the population structure observed in the noctule. However, other parameter combinations are also compatible with the data. We computed the minimum sex bias in dispersal (at least 69% of the dispersing individuals are males), a result that would not be available if we had used more classical population genetic models. The power of multilocus population differentiation tests was unexpectedly high, the tests being significant in almost 100% of the replicates, although the observed population structure infered from nuclear markers was extremely low (F(ST) = 0.6%).

Amplification of a mitochondrial DNA fragment was used to compare the efficiency of five methods for extracting DNA from bat droppings. The Qiagen DNA Stool Kit, which yielded > 90% mtDNA amplification success, was chosen to extract DNA from 586 samples taken over two years in three French colonies of the lesser horseshoe bat (Rhinolophus hipposideros). Samples, for which mtDNA amplification was successful, were subject to the multiplex amplification of eight microsatellite loci. This resulted in > 95% amplification success over 12,592 PCRs. Allelic dropout (ADO) and false allele (FA) rates were low, and consequently, sample and locus quality indexes (QI) were high. These results demonstrate that large scale noninvasive studies of bat colonies are possible.

Natal dispersal in a vagile carnivore, the American marten (Martes americana), was studied by comparing radio-tracking data and microsatellite genetic structure in two populations occupying contrasting habitats. The genetic differentiation determined among groups of individuals using F(ST) indices appeared to be weak in both landscapes, and showed no increase with geographical distance. Genetic structure investigated using pairwise genetic distances between individuals conversely showed a pattern of isolation by distance (IBD), but only in the population occurring in a homogeneous high-quality habitat, therefore showing the advantage of individual-based analyses in detecting within-population processes and local landscape effects. The telemetry study of juveniles revealed a leptokurtic distribution of dispersal distances in both populations, and estimates of the mean squared parent-offspring axial distance (sigma2) inferred both from the genetic pattern of IBD and from the radio-tracking survey showed that most juveniles make little contribution to gene flow.

Estimating population size of elusive and rare species is challenging. The difficulties in catching such species has triggered the use of samples collected noninvasively, such as feces or hair; from which genetic analysis yields data similar to capture-mark-recapture (CMR) data. There are, however two differences between classical CMR and noninvasive CMR. First, capture and recapture data are gathered over multiple sampling sessions in classical CMR, whereas in noninvasive CMR they can be obtained from a single sampling session. Second, because of genotyping errors and unlike classical CMR, there is no simple relationship between (genetic) marks and individuals in noninvasive CMR. We evaluated, through simulations, the reliability of population size estimates based on noninvasive CMR. For equal sampling efforts, we compared estimates of population size N obtained from accumulation curves, a maximum likelihood, and a Bayesian estimator. For a closed population and without sampling heterogeneity, estimates obtained from noninvasive CMR were as reliable as estimates from classical CMR. The sampling structure (single or multiple session) did not alter the results, the Bayesian estimator in the case of a single sampling session presented the best compromise between low mean squared error and a 95% confidence interval encompassing the parametric value of N in most simulations. Finally, when suitable field and lab protocols were used, genotyping errors did not substantially bias population size estimates (bias < 3.5% in all simulations). The ability to reliably estimate population size from noninvasive samples taken during a single session offers a new and useful technique for the management and conservation of elusive and rare species.

Metapopulation genetic models consider that colonization and dispersal are distinct behaviours. However, whether colonization and dispersal indeed reflect different biological processes in nature is unclear. One possibility to test this assumption is to assess patterns of autosomal and mitochondrial genetic structure in species with strict female philopatry, such as the communally breeding Bechstein's bat. In this species, mitochondrial DNA can spread only when females establish new colonies, and autosomal DNA is transmitted among colonies only when females mate with solitary males born in foreign colonies. Investigating the genetic structure among 37 colonies, we found that autosomal genes followed an island model on a regional scale and a model of isolation by distance on a larger geographical scale. In contrast, mitochondrial genetic structure revealed no pattern of isolation by distance at a large scale but exhibited an effect of ecological barriers on a regional scale. Our results provide strong empirical evidence that colonization and dispersal do not follow the same behavioural rules in this bat, supporting the assumption of metapopulation genetic models.

The use of noninvasively collected samples greatly expands the range of ecological issues that may be investigated through population genetics. Furthermore, the difficulty of obtaining reliable genotypes with samples containing low quantities of amplifiable DNA may be overcome by designing optimal genotyping schemes. Such protocols are mainly determined by the rates of genotyping errors caused by false alleles and allelic dropouts. These errors may not be avoided through laboratory procedure and hence must be quantified. However, the definition of genotyping error rates remains elusive and various estimation methods have been reported in the literature. In this paper we proposed accurate codification for the frequencies of false alleles and allelic dropouts. We then reviewed other estimation methods employed in hair- or faeces-based population genetics studies and modelled the bias associated with erroneous methods. It is emphasized that error rates may be substantially underestimated when using an erroneous approach. Genotyping error rates may be important determinants of the outcome of noninvasive studies and hence should be carefully computed and reported.

The distribution of mitochondrial control region-sequence polymorphism was investigated in 15 populations of Crocidura russula along an altitudinal gradient in western Switzerland. High-altitude populations are smaller, sparser and appear to undergo frequent bottlenecks. Accordingly, they showed a loss of rare haplotypes, but unexpectedly, were less differentiated than lowland populations. Furthermore, the major haplotypes segregated significantly with altitude. The results were inconsistent with a simple model of drift and dispersal. They suggested instead a role for historical patterns of colonization, or, alternatively, present-day selective forces acting on one of the mitochondrial genes involved in metabolic pathways.

The ability of a PCR-based restriction fragment length polymorphism (RFLP) analysis of the cytochrome b (mtDNA) to distinguish Apodemus alpicola from two other Apodemus species was investigated. The partial sequencing of the cytochrome b allowed the identification of one enzyme as being potentially diagnostic. This was supported by an analysis of 131 specimens previously identified using morphometric and/or allozymic data, indicating that the PCR-based RFLP method provides a rapid and reliable tool for distinguishing A. alpicola from its two co-occurring congenerics. The method is applicable to samples taken in the field for ecological studies, and could easily be adapted to the identification of museum samples.

Genetic structure of a population of the Mediterranean mouflon ( Ovis gmelini ) in the Caroux-Espinouse Massif of southern France was investigated using six microsatellite loci. The population was subdivided into home-range groups of females that corresponded to sociospatial units. Males segregated from females outside rut, but they were faithful to the unit where they were born during rut. Both sexes were philopatric. A test for population differentiation and F -statistics was used to show that the sociospatial units corresponded to genetically differentiated units ( F ST = 0.043, P < 0.01). Pairwise comparisons between sociospatial units indicated that the amount of genetic differentiation coincided with their geographic distribution. Unexpectedly, no deficiencies in heterozygosity were found in the units. Lack of inbreeding is not well understood, but mechanisms of avoidance of incest and philopatry of the mouflon as a spatial attachment rather than a social attachment to relatives are discussed.

The impact of ecology and social organization on genetic structure at landscape spatial scales, where gene dynamics shape evolution as well as determine susceptibility to habitat fragmentation, is poorly understood. Attempts to assess these effects must take into account the potentially confounding effects of history. We used microsatellites to compare genetic structure in seven bat species with contrasting patterns of roosting ecology and social organization, all of which are codistributed in an ancient forest habitat that has been exceptionally buffered from radical habitat shifts. Over one thousand individuals were captured at foraging sites and genotyped at polymorphic microsatellite loci. Analyses of spatially explicit genotype data revealed interspecies differences in the extent of movement and gene flow and genetic structure across continuous intact forest. Highest positive genetic structure was observed in tree-roosting taxa that roost either alone or in small groups. By comparison, a complete absence of genetic autocorrelation was noted in the cave-roosting colonial species across the study area. Our results thus reveal measurable interspecies differences in the natural limits of gene flow in an unmodified habitat, which we attribute to contrasting roosting ecology and social organization. The consequences of ecology and behaviour for gene flow have important implications for conservation. In particular, tree-roosting species characterized by lower vagility and thus gene flow will be disproportionally impacted by landscape-scale forest clearance and habitat fragmentation, which are prevalent in the study region. Our method also highlights the usefulness of rapid sampling of foraging bats for assaying genetic structure, particularly where roosting sites are not always known.

The sensory drive theory of speciation predicts that populations of the same species inhabiting different environments can differ in sensory traits, and that this sensory difference can ultimately drive speciation. However, even in the best-known examples of sensory ecology driven speciation, it is uncertain whether the variation in sensory traits is the cause or the consequence of a reduction in levels of gene flow. Here we show strong genetic differentiation, no gene flow and large echolocation differences between the allopatric Myanmar and Thai populations of the world's smallest mammal, Craseonycteris thonglongyai, and suggest that geographic isolation most likely preceded sensory divergence. Within the geographically continuous Thai population, we show that geographic distance has a primary role in limiting gene flow rather than echolocation divergence. In line with sensory-driven speciation models, we suggest that in C. thonglongyai, limited gene flow creates the suitable conditions that favour the evolution of sensory divergence via local adaptation., Populations of the same species living in different habitats can differ in sensory traits driving speciation, but it is not known if this variation limits gene flow. Here, a genetic and acoustic study of the bumblebee bat suggests that geographic distance, instead of echolocation divergence, limits gene flow.

The potential for parallel impacts of habitat change on multiple biodiversity levels has important conservation implications. We report on the first empirical test of the 'species-genetic diversity correlation' across co-distributed taxa with contrasting ecological traits in the context of habitat fragmentation. In a rainforest landscape undergoing conversion to oil palm, we show that depauperate species richness in fragments is mirrored by concomitant declines in population genetic diversity in the taxon predicted to be most susceptible to fragmentation. This association, not seen in the other species, relates to fragment area rather than isolation. While highlighting the over-simplification of extrapolating across taxa, we show that fragmentation presents a double jeopardy for some species. For these, conserving genetic diversity at levels of pristine forest could require sites 15-fold larger than those needed to safeguard species numbers. Importantly, however, each fragment contributes to regional species richness, with larger ones tending to contain more species.

Population genetic analyses are of considerable importance for conservation strategies to protect endangered primates. We tested microsatellites of human origin with an aim to understand the genetic diversity of a West African forest guenon, Campbell’s monkey, Cercopithecus campbelli. Twelve markers amplified successfully, were polymorphic and were inherited in a Mendelian fashion in a group of 4 individuals kept in captivity. These 12 markers were further amplified from 35 faecal samples collected in Taï National Park. These samples proved to originate from 18 free-ranging monkeys and showed that the 12 markers we developed for this species are polymorphic and suitable for future population genetic and parentage analyses.

International audience; We combined mark-and-recapture studies with genetic techniques of parentage assignment to evaluate the interactions between mating, dispersal, and inbreeding, in a free-ranging population of Crocidura russula. We found a pattern of limited and female-biased dispersal, followed by random mating within individual neighborhoods. This results in significant inbreeding at the population level: mating among relatives occurs more often than random, and FIT analyses reveal significant deficits in heterozygotes. However, related mating partners were not less fecund, and inbred offspring had no lower lifetime reproductive output. Power analyses show these negative results to be quite robust. Absence of phenotypic evidence of inbreeding depression might result from a history of purging: local populations are small and undergo disequilibrium gene dynamics. Dispersal is likely caused by local saturation and (re)colonization of empty breeding sites, rather than inbreeding avoidance.

Although rarely assessed, the population genetics of hibernating colonies can help to understand some aspects of population structure, even when samples from nursery or mating colonies are not available, or in studies of migration when both types of samples are available and can be compared. Here we illustrate both points in a survey of mitochondrial DNA (mtDNA) control region sequences used to study the population genetics of hibernating colonies of a migrating species, the noctule bat (Nyctalus noctula). Lacking samples from Scandinavian nursery colonies, we use a North European hibernacula to suggest that Scandinavian populations are isolated from Central and East European colonies. Then, we compare genetic diversities of nursery and hibernating colonies. We find a significantly higher haplotype diversity in hibernacula, confirming that they consist of individuals from different nursery colonies. Finally, we show that pairwise comparisons of the haplotype frequencies of nursery and hibernating colonies contain some information on the migration direction of the noctule bat.

12 pages; International audience; Background: Emerging infectious diseases in wildlife are major threats for both human health and biodiversity conservation. Infectious diseases can have serious consequences for the genetic diversity of populations, which could enhance the species' extinction probability. The Ebola epizootic in western and central Africa induced more than 90% mortality in Western lowland gorilla population. Although mortality rates are very high, the impacts of Ebola on genetic diversity of Western lowland gorilla have never been assessed. Methodology/Principal Findings: We carried out long term studies of three populations of Western lowland gorilla in the Republic of the Congo (Odzala-Kokoua National Park, Lossi gorilla sanctuary both affected by Ebola and Lossi's periphery not affected). Using 17 microsatellite loci, we compared genetic diversity and structure of the populations and estimate their effective size before and after Ebola outbreaks. Despite the effective size decline in both populations, we did not detect loss in genetic diversity after the epizootic. We revealed temporal changes in allele frequencies in the smallest population. Conclusions/Significance: Immigration and short time elapsed since outbreaks could explain the conservation of genetic diversity after the demographic crash. Temporal changes in allele frequencies could not be explained by genetic drift or random sampling. Immigration from genetically differentiated populations and a non random mortality induced by Ebola, i.e., selective pressure and cost of sociality, are alternative hypotheses. Understanding the influence of Ebola on gorilla genetic dynamics is of paramount importance for human health, primate evolution and conservation biology.

Studying the dispersal behaviour of small, volant, and nocturnal animals such as microchiropterans with direct methods (banding--recapture, telemetry) is a very difficult task. The development of easily scorable and highly variable genetic markers nowadays allows us to study some aspects of dispersal indirectly, using population genetics. Here, we applied these indirect methods to characterize male dispersal behaviour in a European bat species. The eight microsatellite loci analysed were highly variable in all the nursing colonies assessed (h = 0.63-0.93). Contrary to what we found in the mtDNA, an AMOVA and F-statistics showed that the overall European population structure of the noctule bat was very weak, indicating a high male dispersal rate. Nevertheless, the population was not totally panmictic (theta = 0.006, p < 0.001), and neither isolation by distance, nor influence of migration could account for this result. Rather, an analysis of pairwise theta-values showed that the population structure might be explained partly by a geographical barrier to gene flow (the Alps), and partly by the fact that there is some limit to the distance the males can disperse.

During the Pleistocene, the habitat of the noctule bat (Nyctalus noctula) was limited to small refuge areas located in Southern Europe, whereas the species is now widespread across this continent. Using mtDNA (control region and ND1 gene) polymorphisms, we asked whether this recolonization occurred through bottlenecks and whether it was accompanied by population growth. Sequences of the second hypervariable domain of the control region were obtained from 364 noctule bats representing 18 colonies sampled across Europe. This yielded 108 haplotypes that were depicted on a minimum spanning tree that showed a starlike structure with two long branches. Additional sequences obtained from the ND1 gene confirmed that the different parts of the MST correspond to three clades which diverged before the Last Glacial Maximum (18,000 yrC¹⁴ BP), leading to the conclusion that the noctule bat survived in several isolated refugia. Partitioning populations into coherent geographical groups divided our samples (φCT = 0.17; P = 0.01) into a group of highly variable nursing colonies from central and eastern Europe and less variable, isolated colonies from western and southern Europe. Demographic analyses suggest that populations of the former group underwent demographic expansions either after the Younger Dryas (11,000–10,000 yrC¹⁴ BP), assuming a fast mutation rate for HV II, or during the Pleistocene, assuming a conventional mutation rate. We discuss the fact that the high genetic variability (h = 0.69–0.96; π = 0.006–0.013) observed in nursing colonies that are located some distance from potential Pleistocene refugia is probably due to the combined effect of rapid evolution of the control region in growing populations and a range shift of noctule populations parallel to the recovery of forests in Europe after the last glaciations.