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The contents of the digestive tracts of 21 loggerhead turtles (20 juveniles and 1 adult) collected along the French Mediterranean coast were extracted during necropsies. Hard and soft items were collected and identified by visual observation. A 15-mL sample of the liquid from the rinsed contents of each digestive tract was collected and dietary DNA (dDNA) was extracted and amplified with a 18S universal eukaryotic primers. The obtained reads were compared with taxonomic DNA reference database. Ten taxonomic groups (annelids, cnidarians, crustaceans, echinoderms, fish, insects, molluscs, plants, poriferans and tunicates) were identified with visual observation and eight (ctenophores, cnidarians, crustaceans, fish, molluscs, poriferans, tunicates and plants) with dDNA metabarcoding. Annelids, echinoderms and insects were detected only with visual observation, whereas ctenophores were detected only with dDNA. The two methods were complementary to cover the range of prey, with more soft-bodied organisms detected by dDNA and more hard-shelled organisms detected by visual observation. The increased use of the dDNA metabarcoding method will help compare sea turtle diets of the different stages such as juveniles and adults, or specimens living in different areas, despite the limitations and complexity associated with its use.

Sea turtles are considered as bio-indicators for monitoring the efficiency of restoration measures to reduce marine litter impacts on health. However, the lack of extended and standardised empirical data has prevented the accurate analysis of the factors influencing litter ingestion and the relationships with individual health. Historic data collected from 1988 and standard data collected from 2016 were harmonised to enable such analyses on necropsied loggerhead turtles (Caretta caretta) in eight Mediterranean and North-East Atlantic countries. Litter was found in 69.24 % of the 1121 individuals, mostly single-use and fishing-related plastics. Spatial location, sex and life history stage explained a minor part of litter ingestion. While no relationships with health could be detected, indicating that all individuals can be integrated as bio-indicators, the mechanistic models published in literature suggest that the high proportion of plastics in the digestive contents (38.77 % per individual) could have long-term repercussions on population dynamics. © 2022

The presence of marine litter is a complex, yet persistent, threat to the health and biodiversity of the marine environment, and plastic is the most abundant, and ubiquitous type of marine litter. To monitor the level of plastic waste in an area, and the prospect of it entering the food chain, bioindicator species are used extensively throughout Northern European Seas, however due to their distribution ranges many are not applicable to the Mediterranean Sea. Guidance published for the Marine Strategy Framework Directive suggests that the contents of fish stomachs may be analyzed to determine trends of marine plastic ingestion. In order to equate transnational trends in marine plastic ingestion, the use of standardized fish species that widely occur throughout the basin is favoured, however for the Mediterranean Sea, specific species are not listed. Here we propose a methodology to assess how effective Mediterranean fish species, that are known to have ingested marine plastic, are as bioindicators. A new Bioindicator Index (BI) was established by incorporating several parameters considered important for bioindicators. These parameters included species distribution throughout the Mediterranean basin, several life history traits, the commercial value of each species, and the occurrence of marine litter in their gut contents. By collecting existing data for Mediterranean fish, ranked scores were assigned to each trait and an average value (BI value) was calculated for each species. Based on their habitat preferences, Engraulis encrasicolus (pelagic), Boops boops (benthopelagic), three species of Myctophidae (Hygophum benoiti, Myctophum punctatum and Electrona risso) (mesopelagic), Mullus barbatus barbartus (demersal) and Chelidonichthys lucerna (benthic), were identified as currently, the most suitable fish for monitoring the ingestion of marine plastics throughout the Mediterranean basin. The use of standardized indicator species will ensure coherence in the reporting of marine litter ingestion trends throughout the Mediterranean Sea.

[EN] The infectious chytrid fungus Batrachochytrium salamandrivorans (Bsal) has been responsible for severe population declines of salamander populations in Europe. Serious population declines and loss of urodelan diversity may occur if appropriate action is not taken to mitigate against the further spread and impact of Bsal. We provide an overview of several potential mitigation methods, and describe their possible advantages and limitations. We conclude that long-term, context-dependent, multi-faceted approaches are needed to successfully mitigate adverse effects of Bsal, and that these approaches should be initiated pre-arrival of the pathogen. The establishment of ex situ assurance colonies, or management units, for species threatened with extinction, should be considered as soon as possible. While ex situ conservation and preventive measures aimed at improving biosecurity by limiting amphibian trade may be implemented quickly, major challenges that lie ahead are in designing in situ disease containment and mitigation post-arrival and in increasing public awareness., This work was supported by the European Commission (Tender ENV.B.3/SER/2016/0028, Mitigating a new infectious disease in salamanders to counteract the loss of biodiversity). E.V. and S.C. are supported by the Research Foundation Flanders (FWO grants 12E6616N and FWO16/PDO/019 respectively). D.S.S. and A.L. were supported by the Belmont Forum (DFG-SCHM 3059/6-1) and the Axa Research Fund through the project GloMEC. M.C.F. was supported by NERC (NE/K014455/1), the Leverhulme Trust (RPG-2014-273), the Morris Animal Foundation (D16ZO-022) and the Canadian CIFAR “Fungal Kingdom” programme. T.W.J.G. was supported by NERC (NE/S000992/1, NE/N009967/1). S.L., S.S., M. Vences, M. Veith and N.W. were supported by the Federal Agency for Nature Conservation (BfN) (R+D project “Monitoring und Entwicklung von Vorsorgemaßnahmen zum Schutz vor der Ausbreitung des Chytridpilzes Batrachochytriumsalamandrivorans (“Bsal”) im Freiland”). S.L., N.W., Michael Veith and Miguel Vences were also supported by a seed grant from Deutsche Bundesstiftung Umwelt (DBU).

This study aims to characterise the genetic structure and composition of 245 individuals of loggerhead sea turtles collected from stranding and bycatch events along the French Mediterranean coasts (Gulf of Lion, Provence and Corsica). We obtained sequences of the mitochondrial control region for two fragments (683 bp and 241 bp for 170 and 51 individuals, respectively). The analysis of the long fragment revealed that 163 samples (95.9%) are attributed to the haplogroup II (mainly Mediterranean) whereas only seven individuals are included in haplogroup IB (Atlantic Ocean). The mixed stock analysis performed on the same dataset indicated that the biggest rookeries from the eastern Mediterranean mainly contributed to the French stock, with major contributions being from Greece (36% and 56% for adults and juveniles, respectively), Crete (12% and 18%) and Western Turkey (14% and 4%). The thirteen microsatellite nuclear markers that have been analysed for 81 specimens did not reveal much genetic structure within sampled individuals, thus suggesting that the studied individuals could belong to the same genetic group. The microsatellite analyses revealed however that the nine individuals issuing from two nests sampled in Gulf of Lion and Provence are clearly differentiated from the remaining samples, thus suggesting a long-distance colonisation of the western Mediterranean. Our results allowed addressing the question of loggerhead turtle conservation in the western Mediterranean basin that until now is not considered as a Management Unit despite high densities of juveniles and recent nesting observed on French, Italian and Spanish coasts.

Background Mitigating the effect of linear transport infrastructure (LTI) on fauna is a crucial issue in road ecology. Wildlife crossing structures (tunnels or overpasses) are one solution that has been implemented to restore habitat connectivity and reduce wildlife mortality. Evaluating how these crossings function for small wildlife has often been recommended but, mainly due to technical limitations, is not often conducted in practice or only as short-term monitoring (less than 1 year). In this study, we developed and tested an automated device that records the detailed behaviour of animals when using wildlife tunnels. The method is based on marking and detecting individuals with RFID (radio-frequency identification) tags and allows small animals to be tracked. Composed of four antennas (detectors) placed at roughly 2 m intervals, the system was tested in a tunnel in northern France in the summer of 2017. One species of amphibian (the toad Bufo spinosus, n = 13) and two carabids (the ground beetles Carabus coriaceus and Carabus nemoralis, respectively, n = 3 and n = 4), marked beforehand with PIT (passive integrated transponder) tags, were recorded when they crossed these detectors. This allowed individual trajectories, including crossing speed, to be estimated. Results We found that 12 of the 13 toads and 3 of the 7 ground beetles successfully crossed the entire wildlife passage of 7 m long. The detection rate of each detector varied from 8.33 to 100%, with a mean of 52.08%. All individuals were recorded by at least one detector. We observed high variability in the crossing characteristics of toads (mean transit duration = 41 min and 15 s ± 25 min) and ground beetles (6 h 11 min ± 3 h 30 min). The system provided information on precise trajectories (e.g., crossing speed, U-turns, distance travelled in the tunnel, proportion of individuals reaching the exit, etc.) for each individual, in a context of tunnel crossing. Conclusion The system allowed us to record small animal behaviour in the context of tunnel crossing in which other types of tracking (e.g. radio-tracking) or detection (e.g. camera traps) are not effective. It also opens the possibility for a range of experiments that would contribute to a better understanding of the behaviour of small animals in tunnels, allowing a comparison of tunnel characteristics (such as size, building material, substrate, etc.) with the aim of increasing wildlife use of the tunnel and proposing guidelines for the construction and maintenance of these mitigation measures.

The following protocol is intended to respond to the requirements set by the European Union’s Marine Strategy Framework Directives (MSFD) for the D10C3 Criteria reported in the Commission Decision (EU), related to the amount of litter ingested by marine animals. Standardized methodologies for extracting litter items ingested from dead sea turtles along with guidelines on data analysis are provided. The protocol starts with the collection of dead sea turtles and classification of samples according to the decomposition status. Turtle necropsy must be performed in authorized centers and the protocol described here explains the best procedure for gastrointestinal (GI) tract isolation. The three parts of the GI (esophagus, stomach, intestine) should be separated, opened lengthways and contents filtered using a 1 mm mesh sieve. The article describes the classification and quantification of ingested litter, classifying GI contents into seven different categories of marine litter and two categories of natural remains. The quantity of ingested litter should be reported as total dry mass (weight in grams, with two decimal places) and abundance (number of items). The protocol proposes two possible scenarios to achieve the Good Environmental Status (GES). First: “There should be less than X% of sea turtles having Y g or more plastic in the GI in samples of 50-100 dead turtles from each sub-region”, where Y is the average weight of plastic ingested and X% is the percentage of sea turtles with more weight (in grams) of plastic than Y. The second one, which considers the food remain versus plastic as a proxy of individual health, is: “There should be less than X% of sea turtles having more weight of plastic (in grams) than food remains in the GI in samples of 50-100 dead turtles from each sub-region”. © 2019 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.

This work aims to illustrating how it is possible to measure different modalities of adult dispersal in two subdivided populations of the alpine newt (Triturus alpestris). Recent developments in capture-mark-recapture methods make it possible to estimate transience rates from individuals captured only once. In the context of subdivided newt populations, transience is assumed to express nomadic behaviour that contribute to breeding dispersal. Skeletochronology and recaptures within each pond system also made it possible to estimate emigration rates and local dispersal. Two subdivided populations of alpine newts were monitored over 4 and 5 years, respectively. Whereas population A is suspected to have been established for more than 100 years, population B was monitored when colonizing a newly created archipelago of ponds. Transience was detected in each population at similar rates (37% in population A and 35% in the population B). Annual apparent survival rates were estimated as 82% in population A vs. 33% in population B. Similarity of age structures between populations leads us to consider such low survival rates in population B as resulting from emigration. Emigration was thus negligible in population A and was estimated to reach 57·3% in population B. Conversely, high local dispersal (movements within a pond system) was detected in population A, but not in population B. Even though the causation of dispersal in newts (genetic polymorphism vs. phenotypic plasticity) remains unexplored, our study succeeded in identifying several dispersal components that could result from different selective pressures (habitat heterogeneity at different temporal scales). Experimental approaches are needed to investigate the causative bases of these traits.

Este artículo contiene 9 páginas, 3 figuras, 2 tablas., Range expansions are an intrinsic part of the biogeography of most species. Among them, some invasive species represent a paradigmatic case in which local introductions are followed by rapid expansions in several directions. Theoretically, this creates a recursive founder effect, leading to sustained losses of genetic diversity and – through stochastic changes in allele frequency – to a population structure that mirrors invasion history. Here, we use restriction site-associated DNA (RAD) markers to detect these patterns in the invasive range of the Mediterranean Painted Frog (Discoglossus pictus) in Europe. We identify and genotype a large panel of loci using genotyping-by-sequencing in several sites along its two main directions of invasion – northwards and southwards. Although the frog has a few translocated isolated populations, most of the invasive range is contiguous, allegedly corresponding to a self-sustained expansion from a single introduction point in Banyuls de la Marenda (Southern France) more than a century ago. As expected, the highest genetic diversity was found in this location, with progressive losses of genetic diversity north- and southwards. Genetic similarities among sampling sites also coincided with predictions according to the documented process of expansion, showing a latitudinal population structure matching predictions according to geographic distance among locations. This poses the two opposite expansion directions as independent processes of loss of genetic diversity and creation of population structure, that is, two different cases of range expansion which are ready for further examinations., Part of the fieldwork for this study was done during the support of the Spanish Ministerio de Educaci on y Deporte to EPB (FPU grant: AP2010-5563).

The American bullfrog, native to the eastern half of North America including southern Quebec, is considered one of the most invasive species in the world. It has been introduced in America, Asia and Europe, mainly for food purposes. A study on the age and body size of this species was carried out on individuals obtained from four introduced populations (one from Western Canada and three from France), using the skeletochronological method. Adults did not differ between sexes in mean body size or mean age, with the exception of one population in France where males were younger than females. Several differences in the mean age and body size were observed, with the individuals from the population in Canada exhibiting the highest values for both sexes. The scarcity of studies in the native range prevents the comparison of introduced versus native populations, and we focus on comparisons between introduced populations.

Invasive species can be a threat to native species in several ways, including transmitting lethal infections caused by the parasites they carry. However, invasive species may also be plagued by novel and lethal infections they acquire when invading, making inferences regarding the ability of an invasive host to vector disease difficult from field observations of infection and disease. This is the case for the pathogenic fungus Batrachochytrium dendrobatidis (Bd) in Europe and one invasive host species, the North American bullfrog Lithobates catesbeianus, hypothesized to be responsible for vectoring lethal infection to European native amphibians. We tested this hypothesis experimentally using the alpine newt Ichthyosaura alpestris as our model native host. Our results show that infected bullfrog tadpoles are effective vectors of Bd. Native adult newts co-housed with experimentally infected bullfrog tadpoles became Bd infected (molecular and histological tests). Moreover, the exposed adult newts suffered mortality while the majority of infected bullfrog tadpoles survived until metamorphosis. These results cannot resolve the historical role of alien species in establishing the distribution of Bd across Europe or other regions in the world where this species was introduced, but they show its potential role as a Bd reservoir capable of transmitting lethal infections to native amphibians. Finally, our results also suggest that the removal of infected bullfrogs from aquatic environments may serve to reduce the availability of Bd in European amphibian communities, offering another justification for bullfrog eradication programmes that are currently underway or may be considered.

Detection is crucial in the study and control of invasive species but it may be limited by methodological issues. In amphibians, classical survey techniques exhibit variable detection probability depending on species and are often constrained by climatic conditions often requiring several site visits. Furthermore, detection may be reduced at low density because probability capture (passive traps), or activity (acoustic surveys) drop. Such limits may impair the study of invasive species because low density is typical of the onset of colonisation on a site. In the last few years, environmental DNA (eDNA) methods have proved their ability to detect the presence of aquatic species. We developed here an eDNA method to detectXenopus laevisin ponds. This austral African species is now present worldwide because of its use in biology and as a pet. Populations have settled and expanded on several continents so that it is now considered as one of the major invasive amphibians in the World. We detected the presence ofX. laevisat density as low as 1 ind/100 m2and found a positive relationship between density in ponds and rate of DNA amplification. Results show that eDNA can be successfully applied to survey invasive populations ofX. laeviseven at low density in order to confirm suspected cases of introduction, delimit the expansion of a colonized range, or monitor the efficiency of a control program.

International audience; 1. In the Northern Hemisphere, an increase in both the frequency and magnitude of violent flooding events has been reported due to climate change. According to life history theory, one might postulate that in ‘slow’ species: (i) environmental canalisation may act as a selective force that minimises to some extent adult survival variations caused by catastrophic flood and (ii) extreme flooding events would cause important variations in recruitment and young survival. Hence, it may be hypothesised that (iii) the population growth rate of ‘slow’ species might be relatively insensitive to changes in the frequency of extreme climatic events if adult survival remains largely unaffected.2. In this study, we investigated how extreme rainfall events resulting in severe flood impact popula- tion dynamics of a long-lived endangered amphibian, the yellow-bellied toad (Bombina variegata: Bombinatoridae). To address this issue, we used capture–recapture (CR) data collected on two populations (768 and 1154 individuals identified) in southern France and developed multi-event CR models.3. Our results indicated that extreme flooding did not cause any variation in sub-adult or adult survival, whereas recruitment and juvenile survival were negatively impacted. Furthermore, our simulations indicated that the population growth rate was only marginally sensitive to potential changes in the frequency of extreme flooding in the future.4. Hence, we suggest that extreme flooding does not appear to be a proximal factor of extinction risk for this endangered amphibian species.

Age and size at sexual maturity are major life-history traits that affect growth rate and reproductive output. The differences in these life-history traits can be better assessed in syntopic populations of related species, where environmental variability in time and space is removed. The Spadefoot Toads (Pelobates spp.) are highly specialized burrowing and nocturnal species with a narrow ecological niche. We tested if age-related parameters are responsible for sexual size dimorphism and size differences in two syntopic populations from the Danube Delta Biosphere Reserve (Romania). The two populations differed significantly in size and body mass, and showed distinct sexual dimorphism, with females significantly larger in P. fuscus, and males slightly larger in P. syriacus. We also found differences in age structure: P. fuscus was on average 5.0 yr old, whereas P. syriacus was on average 7.4 yr old. Both species reached sexual maturity at a similar age. Different growth rates before sexual maturity...

International audience; Variation in the environment can induce different patterns of genetic and phenotypic differentiation among populations. Both neutral processes and selection can influence phenotypic differentiation. Altitudinal phenotypic variation is of particular interest in disentangling the interplay between neutral processes and selection in the dynamics of local adaptation processes but remains little explored. We conducted a common garden experiment to study the phenotypic divergence in larval life-history traits among nine populations of the common toad (Bufo bufo) along an altitudinal gradient in France. We further used correlation among population pairwise estimates of quantitative trait (QST) and neutral genetic divergence (FST from neutral microsatellite markers), as well as altitudinal difference, to estimate the relative role of divergent selection and neutral genetic processes in phenotypic divergence. We provided evidence for a neutral genetic differentiation resulting from both isolation by distance and difference in altitude. We found evidence for phenotypic divergence along the altitudinal gradient (faster development, lower growth rate and smaller metamorphic size). The correlation between pairwise QSTs–FSTs and altitude differences suggested that this phenotypic differentiation was most likely driven by altitude-mediated selection rather than by neutral genetic processes. Moreover, we found different divergence patterns for larval traits, suggesting that different selective agents may act on these traits and/or selection on one trait may constrain the evolution on another through genetic correlation. Our study highlighted the need to design more integrative studies on the common toad to unravel the underlying processes of phenotypic divergence and its selective agents in the context of environmental clines.

The early detection and identification of pathogenic microorganisms is essential in order to deploy appropriate mitigation measures. Viruses in the Iridoviridae family, such as those in the Ranavirus genus, can infect amphibian species without resulting in mortality or clinical signs, and they can also infect other hosts than amphibian species. Diagnostic techniques allowing the detection of the pathogen outside the period of host die-off would thus be of particular use. In this study, we tested a method using environmental DNA (eDNA) on a population of common frogs (Rana temporaria) known to be affected by a Ranavirus in the southern Alps in France. In six sampling sessions between June and September (the species’ activity period), we collected tissue samples from dead and live frogs (adults and tadpoles), as well as insects (aquatic and terrestrial), sediment, and water. At the beginning of the breeding season in June, one adult was found dead; at the end of July, a mass mortality of tadpoles was observed. The viral DNA was detected in both adults and tadpoles (dead or alive) and in water samples, but it was not detected in insects or sediment. In live frog specimens, the virus was detected from June to September and in water samples from August to September. Dead tadpoles that tested positive for Ranavirus were observed only on one date (at the end of July). Our results indicate that eDNA can be an effective alternative to tissue/specimen sampling and can detect Ranavirus presence outside die-offs. Another advantage is that the collection of water samples can be performed by most field technicians. This study confirms that the use of eDNA can increase the performance and accuracy of wildlife health status monitoring and thus contribute to more effective surveillance programs.

Pathogenic fungi and viruses cause mortality outbreaks in wild amphibians worldwide. In the summer of 2012, dead tadpoles and adults of the European common frog Rana temporaria were reported in alpine lakes in the southwestern Alps (Mercantour National Park, France). A preliminary investigation using molecular diagnostic techniques identified a Ranavirus as the potential pathogenic agent. Three mortality events were recorded in the park, and samples were collected. The amphibian chytrid fungus Batrachochytrium dendrobatidis was not detected in any of the dead adult and juvenile frogs sampled (n=16) whereas all specimens were positive for a Ranavirus. The genome sequence of this Ranavirus was identical to previously published sequences of the common midwife toad virus (CMTV), a Ranavirus that has been associated with amphibian mortalities throughout Europe. We cultured virus from the organs of the dead common frogs and infecting adult male common frogs collected in another alpine region where no frog mortality had been observed. The experimentally infected frogs suffered 100% mortality (n=10). The alpine die-off is the first CMTV outbreak associated with mass mortality in wild amphibians in France. We describe the lesions observed and summarize amphibian populations affected by Ranaviruses in Europe. In addition, we discuss the ecologic specificities of mountain amphibians that may contribute to increasing their risk of exposure to and transmission of Ranaviruses.

International audience; Dispersal is a central component of life history evolution. An increasing number of studies suggest that spatiotemporally variable environments may promote the evolution of “dispersal syndromes,” consisting of covariation patterns between dispersal and morphologi- cal, physiological, behavioral, and life history traits. At the interspecific scale, the “colonizer syndrome” appears to be one of the most frequently recorded associations between dispersal and life history traits, linking a high dispersal rate, high fecundity, and a short lifespan as sys- tematically combined adaptations in spatiotemporally varying environments. However, few studies have highlighted the existence of a “colonizer syndrome” at the intraspecific scale, and none have investigated how different degrees of habitat stochasticity might shape covariation patterns between dispersal and life history traits. In this study, we examined this issue in free- ranging metapopulations of the yellow-bellied toad (Bombina variegata) using capture–recap- ture data. Combining the results of this study with another recent study, we found that a high dispersal rate, high fecundity, and a short lifespan are associated in metapopulations experi- encing unpredictable environments. In contrast, a very low dispersal rate (close to zero), low fecundity and a long lifespan are associated in metapopulations occupying predictable environ- ments. We discuss these results as well as their demographic and evolutionary consequences.

Summary 1. Alien invasive species (AIS) are one of the major causes of biodiversity loss and global homogenization. Once an AIS becomes established, costs of control can be extremely high and complete eradication is not always achieved. The ability to detect a species at a low density greatly improves the success of eradication and decreases both the costs of control and the impact on ecosystems. 2. In this study, we compare the sensitivity of traditional field methods, based on auditory and visual encounter surveys, with an environmental DNA (eDNA) survey for the detection of the American bullfrog Rana catesbeiana = Lithobates catesbeianus, which is invasive in south-western France. 3. We demonstrate that the eDNA method is valuable for species detection and surpasses traditional amphibian survey methods in terms of sensitivity and sampling effort. The bullfrog was detected in 38 sites using the molecular method, compared with seven sites using the diurnal and nocturnal surveys, suggesting that traditional field surveys have strongly underestimated the distribution of the American bullfrog. 4. Synthesis and applications. The environmental DNA approach permits the early detection of alien invasive species (AIS), at very low densities and at any life stage, which is particularly important for the detection of rare and ⁄ or secretive aquatic species. This method can also be used to confirm the sensitivity of control operations and to better identify the distributions of vulnerable species, making this a very relevant tool for species inventory and management.

The delimitation of the spatial equivalent of isolated populations or interacting sets of local populations is crucial for conservation management. In Amphibia, spatial organization in metapopulations requires knowledge on the individual variation of annual migratory capacity within local populations to delimit core habitats around the breeding site and to estimate the connectivity among neighbouring populations. The migratory behaviour of 143 adult Bufo calamita was monitored using radio-telemetry at eight localities covering a latitudinal range from 41° to 54°N. Aims of the study were to assess the geographic variation of migratory distance moved by males and females and to model their capacity for dispersal assuming exclusively unidirectional movements. Migratory range was not sex-biased, but was three times lower in population inhabiting sandy areas than in those on clay soils, probably because of the scarcity of moist shelters causing more frequent and more distant movements. For conservation management of local natterjack populations, we propose to use the migratory capacity of the 50% most sedentary individuals to delimit the core area around a given breeding site. To estimate the potential genetic connectivity between neighbouring local population, we propose to use the minimum migratory capacity of the 5% of individuals that moved most. Estimates obtained for populations in central Europe and the UK indicate a core area of 600 m around the breeding site and a maximum distance of 2250 m between the breeding ponds to maintain connectivity. Thus, the principal conservation problem in the UK is that most populations are isolated by distance and prone to local extinction. In contrast, core areas of populations in Spain extend to distances of about 5 km and connectivity is maintained up to a distance 12 km between neighbouring breeding ponds.

Large-scale patterns of body size variation are described by well-known generalizations such as Bergmanns rule; the generality and underlying causes of these patterns have been much debated. Intraspecific extension of this rule was tested in various ectotherms, and evidence was found for both Bergmann and converse Bergmann clines. In this study, we explored spatial patterns of variation in a widespread amphibian, the Common toad (Bufo bufo), along a 2240 km latitudinal gradient across Europe. We tested for covariation of adult body size, age and growth parameters with latitude, altitude, length of activity period and mean temperature during this period using both original and literature data. We selected 13 European populations, representing a latitudinal range from 43 to 63� N and altitudinal range from 15 to 1850 m a.s.l. The length of activity period (12–33 weeks) and Tmean (6.6–15.6� C) significantly decreased as latitude and altitude of these populations increased. Mean body size decreased as latitude increased (not with altitude), and increased with Tmean (not with length of activity period). Mean and minimal adult age increased with latitude and altitude, longevity increased with altitude only. Age increased as length of activity period decreased (not with Tmean). The growth coefficient (0.32–0.92 in males, 0.18–0.74 in females, available for six populations) decreased as altitude increased, and increased as both length of activity period and Tmean increased; latitudinal trend was non-significant. Our analysis shows that B. bufo clearly exhibited a converse Bergmann cline along latitudinal gradient, but not along altitudinal gradient; the main effect of elevation was on age. The effects of ecological conditions also differed: body size increased with Tmean, while age parameters were related to the length of activity period. This study highlights that, to identify causal factors underlying general ecogeographical rules, we have to take into account different phases of the life cycle, co-variation among life history traits and ecological factors acting on each of these traits. In amphibians with complex life cycles, lack of appropriate demographical or ecological data may affect our understanding of the variety of observed body size patterns.

International audience; Solar ultraviolet-B (UV-B) radiation can be harmful for developing amphibians. As the UV-B dose increases with altitude, it has been suggested that high-altitude populations may have an increased tolerance to high levels of UV-B radiation as compared to lowland populations. We tested this hypothesis with the common frog (Rana temporaria) by comparing populations from nine altitudes (from 333 to 2450 in above sea level). Eggs collected in the field were used for laboratory experiments, i.e., exposed to high levels of artificial UV-B radiation. Eggs were reared at 14 +/- 2 degrees C and exposed to UV treatments until hatching. Embryonic developmental rates increased strongly and linearly with increasing altitude, suggesting a genetic capacity for faster development in highland than lowland eggs. Body length at hatching varied significantly with UV-B treatments, being lower when eggs developed under direct UV-B exposure. Body length at hatching also increased as the altitude of populations increased, but UV-B exposure times were shorter as altitude of population increased. However, the body length difference between exposed and non-exposed individuals in each population decreased as altitude of populations increased, suggesting a costly effect of UV exposure on growth. Type of UV exposure did not influence the mean rates of embryonic mortality and deformity, but both mortality and deformity rates increased as the altitude of populations increased (while UV-B exposure duration decreased). The effect of UV-B on body length at hatching, mortality, and deformities suggests that the sensitivity to UV-B varied among populations along the altitudinal gradient. These results are discussed in evolutionary terms, specifically the potential of R. temporaria high-altitude populations to develop local genetic adaptation to high levels of UV-B. (c) 2008 Elsevier GmbH. All rights reserved.

Propagule pressure is considered the main determinant of success of biological invasions: when a large number of individuals are introduced into an area, the species is more likely to establish and become invasive. Nevertheless, precise data on propagule pressure exist only for a small sample of invasive species, usually voluntarily introduced. We studied the invasion of the American bullfrog, Rana catesbeiana, into Europe, a species that is considered a major cause of decline for native amphibians. For this major invader with scarce historical data, we used population genetics data (a partial sequence of the mitochondrial cytochrome b gene) to infer the invasion history and to estimate the number of founders of non-native populations. Based on differences between populations, at least six independent introductions from the native range occurred in Europe, followed by secondary translocations. Genetic diversity was strongly reduced in non-native populations, indicating a very strong bottleneck during colonization. We used simulations to estimate the precise number of founders and found that most non-native populations derive from less than six females. This capability of invasion from a very small number of propagules challenges usual management strategies; species with such ability should be identified at an early stage of introduction.

We used skeletochronology to determine the ages of 149 (74 females, 44 males, and 31 juveniles) Mountain Yellow-legged Frogs (Rana muscosa) from 13 locations (elevation 1509–3501 m) throughout their current range in the Sierra Nevada mountains of California. Lines of arrested growth (LAGs) from excised toe bones were distinct in these high elevation frogs, and each LAG was assumed to represent one year of age. Females ranged in age from 0–10 years (mean = 4.1 years) and males from 0–8 years (mean = 4.0 years). The skeletochronological age was that of the post-metamorphic frog and did not include the tadpole stage. Mountain Yellow-legged Frogs spend 3–4 years as tadpoles, but no age markers are found in their cartilaginous skeletons; thus, their total age, if both tadpole and post-metamorphic stages were included, would range up to 14 years. Females were significantly longer (snout–vent length: SVL) than males and had greater mean mass, but there was no difference in the mean ages. Juvenile fr...

International audience; Predicting the probability of successful establishment and invasion of alien species at global scale, by matching climatic and land use data, is a priority for the risk assessment. Both large- and local-scale factors contribute to the outcome of invasions, and should be integrated to improve the predictions. At global scale, we used climatic and land use layers to evaluate the habitat suitability for the American bullfrog Rana catesbeiana, a major invasive species that is among the causes of amphibian decline. Environmental models were built by using Maxent, a machine learning method. Then, we integrated global data with information on richness of native communities and hunting pressure collected at the local scale. Global-scale data allowed us to delineate the areas with the highest suitability for this species. Predicted suitability was significantly related to the invasiveness observed for bullfrog populations historically introduced in Europe, but did not explain a large portion of variability in invasion success. The integration of data at the global and local scales greatly improved the performance of models, and explained > 57% of the variance in introduction success: bullfrogs were more invasive in areas with high suitability and low hunting pressure over frogs. Our study identified the climatic factors entailing the risk of invasion by bullfrogs, and stresses the importance of the integration of biotic and abiotic data collected at different spatial scales, to evaluate the areas where monitoring and management efforts need to be focused.

Times Cited: 1; International audience; In conservation biology genetic diversity is recognized as an important criterion to consider when prioritizing populations for protection. Today, population genomics offers the opportunity to evaluate both neutral and adaptive components of genetic diversity directly at the genome level with molecular tools. By screening the genome with many genetic markers, it is possible to detect loci supposedly under natural selection and thus of adaptive significance. We devised a new diversity index, the population adaptive index (PAI), which accounts for the adaptive value of the population it refers to. To estimate this index, we performed a genome scan with amplified fragment length polymorphism markers to identify neutral and selected loci in several populations of a widespread amphibian (common frog, Rana temporaria) and a threatened plant (Austrian dragonhead, Dracocephalum austriacum L.). We then investigated four different conservation strategies aimed at protecting the maximum amount of genetic diversity (neutral or selected). In particular we explored the relevance of the principle of complementarity, usually applied to the protection of species, in the management of intraspecific diversity. This principle advocates the conservation of sets of units that together maximize the species' orgenetic diversity, which is in opposition to the traditional approach of targeting populations that are the most diverse individually. Four major conclusions emerged from these results. First, the PAI seemed to be a valuable index to evaluate the adaptive diversities within populations. Second, in the two species, the neutral and adaptive diversities within and among populations were not correlated, so conservation strategies based on the neutral and adaptive indexes would not select the same populations for protection. Third, because of its efficiency in conserving genetic diversity, the principle of complementarity deserves to be used more often for this purpose. Fourth, when neutral and adaptive results conflict, additional arguments (e.g., demography, ecology, and geographic proximity) should be considered together with levels of genetic diversity to determine a conservation strategy.

International audience; Because it modulates the fitness returns of possible options of energy expenditure at each ontogenetic stage, environmental stochasticity is usually considered a selective force in driving or constraining possible life histories. Divergent regimes of envi- ronmental fluctuation experienced by populations are expected to generate differences in the resource allocation schedule between survival and reproductive effort and outputs. To our knowledge, no study has previously examined how different regimes of stochastic variation in environmental conditions could result in changes in both the temporal variation and mean of demographic parameters, which could then lead to intraspecific variation along the slow–fast continuum of life history tactics. To investigate these issues, we used capture–recapture data collected on five populations of a long-lived amphibian (Bombina variegata) experiencing two distinct levels of stochastic environmental variation: (1) constant availability of breeding sites in space and time (predictable environment), and (2) variable spatio-temporal availability of breeding sites (unpredictable environment). We found that female breeding propensity varied more from year to year in unpredictable than in pre- dictable environments. Although females in unpredictable environments produced on average more viable offspring per year, offspring production was more variable between years. Survival at each ontogenetic stage was slightly lower and varied significantly more from year to year in unpredictable environments. Taken together, these results confirm that increased environmental stochasticity can modify the resource allocation schedule between survival and reproductive effort and outputs and may lead to intraspecific variation along the slow–fast continuum of life history tactics.

Abstract The isolation of animal populations due to urban activities provides a useful framework for studying the consequences of landscape fragmentation. We studied a population of natterjack toads (Bufo calamita) in an urban park near Paris, France. In 2001 and 2002 we used radio-tracking to estimate the terrestrial movements of adults around their breeding sites. Twenty-four toads were equipped with internal transmitters in 2001 to record movements during and after the breeding period. In 2002, 19 males were released at 300 and 380 meters from their breeding ponds. Natterjack toad movements around and outside their breeding ponds were reduced compared to previous observations on this species. The only exchanges that were observed occurred between closely neighbouring breeding sites. During a translocation experiment in 2002, 58% of the displaced males returned to their site of capture and this happened mainly during the breeding period. The remaining 42% stayed close to the release site. There was no exchange of males between distant breeding sites. Natterjack toad conservation needs to take into account the high fidelity to a breeding site and the reduced breeding dispersal and homing ability of these animals. Conservation biology in urban landscapes constitutes a specific urban ecology with specific concepts such as 'population area'. Information from this study can assist land managers in establishing protected areas of high habitat quality around breeding ponds in urban areas, and managing parks for the protection of amphibian populations, particularly by facilitating exchanges between available areas.

International audience; We investigated the ontogeny of responses to scent marks in immature terrestrial salamanders (Salamandra lanzai) that inhabit the southwest Alps. In this species, sexual maturity is usually reached at about 8 years, and adults exhibit territoriality. One should expect territoriality to take place largely before the acquisition of sexual maturity if sexual competition is not the main force driving territoriality. However, both the difficulties for inexperienced animals to find a suitable territory and size-related competition may delay the acquisition of territoriality in juveniles. We performed choice tests with juveniles belonging to two age groups (1+/-1 years old versus 4+/-1 years old). Each focal animal was offered, in random order, the choice between two shelters, one without scent and the other containing one of the following scents: own, same-aged animal, a juvenile belonging to the other age group, or adult female. We also performed choice tests with adult females for which the scents of two juveniles belonging to a different age group were successively offered in a random order. Older juveniles were strongly attracted toward their own shelter and mostly avoided the shelters that contained the scent of juveniles of about the same age and of adult females. Adult females avoided the scents of older juveniles but not younger juveniles. These results therefore suggest that older juveniles use territorial marking. Conversely, young juveniles behaved randomly with respect to their own scents and to those of a same-aged juvenile, and they were significantly attracted towards the odor of an older animal, especially adult females. Both older juveniles and adult females displayed a random behavior toward the scents of young juveniles. Our results suggest that young juveniles do not defend territories but use spaces occupied by older individuals.

International audience

Conservation plans of European amphibians usually take into account: (1) species in dramatic decline due to a series of humaninduced factors, and (2) species with a narrow distribution area and/or particular ecological requirements. Salamandra lanzai belongs to the second group, being limited to the SW Alps. Furthermore, it is a high altitude amphibian, living in sites above 1200 m, and it is also a unique endemic vertebrate of the Alps. We here summarise information regarding its distribution and ecology to be interpreted for conservation. In this sense, we also provide new data regarding the age structure of a recently studied low altitude population in the Germanasca Valley. We then compare this population (exemplified as “low altitude population”) to a population living at 2200 m in the Guil Valley (example of a “high altitude population”). The knowledge obtained on these different life-history traits has allowed us to devise an age-structured model useful in identifying the main demographic parameter...

We studied age at maturity, longevity, and growth in a marbled newt (Triturus marmoratus) population in southwestern France. Parameters of the life cycle such as date of migration, reproductive period, and terrestrial activity were documented for three neighbouring temporary ponds. This Mediterranean marbled newt population was characterized by aestivation and hibernation periods, a prolonged aquatic phase in adults, and a presence of juveniles in water. Aestivation and hibernation lead to distinct growth periods as determined by skeletochronology. Minimum age at maturity was 2 and 3 years, whereas maximum age observed was 8 and 9 years in males and females, respectively. A higher growth rate and a lower survival rate were found in this Mediterranean population compared with other populations exposed to variable environmental conditions in western Europe.

Summary 1. This work aims to illustrating how it is possible to measure different modalities of adult dispersal in two subdivided populations of the alpine newt ( Triturus alpestris ). Recent developments in capture‐mark‐recapture methods make it possible to estimate transience rates from individuals captured only once. In the context of subdivided newt populations, transience is assumed to express nomadic behaviour that contribute to breeding dispersal. Skeletochronology and recaptures within each pond system also made it possible to estimate emigration rates and local dispersal. 2. Two subdivided populations of alpine newts were monitored over 4 and 5 years, respectively. Whereas population A is suspected to have been established for more than 100 years, population B was monitored when colonizing a newly created archipelago of ponds. 3. Transience was detected in each population at similar rates (37% in population A and 35% in the population B). Annual apparent survival rates were estimated as 82% in population A vs. 33% in population B. Similarity of age structures between populations leads us to consider such low survival rates in population B as resulting from emigration. Emigration was thus negligible in population A and was estimated to reach 57·3% in population B. Conversely, high local dispersal (movements within a pond system) was detected in population A, but not in population B. 4. Even though the causation of dispersal in newts (genetic polymorphism vs. phenotypic plasticity) remains unexplored, our study succeeded in identifying several dispersal components that could result from different selective pressures (habitat heterogeneity at different temporal scales). Experimental approaches are needed to investigate the causative bases of these traits.

International audience; Scent marking is often related to activities associated with the occupation of a home range or territory. It is widely agreed that when an animal sends signals, this influences the decisions made by the animal that receives them. Scent marks are mainly used as a warning, in order to decrease the cost of territorial behaviour. They are deposited directly on a substrate by rubbing them on but also through urination and defecation. European terrestrial salamanders show high site fidelity and move very little around their shelter. Risks and costs are compared before an animal decides whether or not to leave its shelter, and a rapidly detectable and unambiguous cue in the direction of the home shelter reduces risk. Furthermore, territoriality has also been observed in amphibians, both in Anurans (frogs and toads) and Urodela (newts and salamanders). We tested the use of faecal pellets in alpine salamanders in the laboratory (Salamandra lanzai and Salamandra atra) with regard to homing and territorial behaviour. Our experimental design allowed us to distinguish attraction to from avoidance of shelters containing varying olfactory information. Our results clearly argue for the role faecal pellets play in shelter recognition. Furthermore, contrary to the results observed in other terrestrial salamanders such as Plethodontids, the faecal pellets of S. lanzai and S. atra enable intruders to identify the residents, thus fulfilling the requirements of a genuine territorial warning, as observed in mammals. Excrement marking is used as an economic tactic to identify and warn about the occupation of a territory in these two species of alpine salamanders.

Fifty individuals of the endemic Alpine salamander, Salamandra atra, representing 13 populations throughout the range of the two currently recognized subspecies, atra and aurorae, were examined for sequence variation in a large portion (1050 bp) of the mitochondrial cytochrome b gene. We revealed a large number of mitochondrial DNA (mtDNA) haplotypes (10). Interpopulation sequence divergence was very low, ranging from 0 to 3.1%. The relationships among haplotypes were poorly resolved. The divergence time estimate between several mtDNA haplotypes suggested a pre-Pleistocene differentiation approximately 3 million years ago. Moreover, the impact of the Pleistocene glaciations on the phylogeographical patterns appears to have been secondary, although a somewhat reduced genetic variability was found in populations living in areas that were directly affected by the glaciation.

The black salamander, Salamandra lanzai, exists in isolated and fragmented populations in the Cottian Alps. To examine the degree of intra- and interpopulation divergence, mitochondrial cytochrome b sequences were obtained from samples representing all populations of S. lanzai. No genetic variation was observed either within or among populations. The presence of a single lineage suggests historical explanations closely related to Pleistocene events.

International audience; We describe the life-history traits (body size, age at maturity, longevity, and survivorship) of a terrestrial salamander, Mertensiella luschani, from a population living in the Mediterranean environment of southwestern Turkey. We used phalangeal skeletochronology to estimate the age of juveniles and adults. Lines of arrested growth, deposited during the winter cessation of bone growth, allowed us to estimate the age at maturity to be 3 years old in both sexes. Males attained a maximum age of 8, whereas females attained a maximum age of 10. Juveniles ranged from 1 to 3 years old. Males were smaller than females. Growth curves fitted to von Bertalanffy's equation revealed similar growth coefficients in adults and larger average maximum body lengths in males. Adult annual survivorship and adult life expectancy were 0.79 and 5.4 years, respectively. Knowledge of age, growth, and survivorship in this species under the constraints of a seasonal Mediterranean climate, and a comparison of similar traits in other salamandrid species, allow us to test predictions regarding optimal timing of growth and reproduction in these species.

International audience; We describe the age structures of two neighbouring terrestrial salamander populations. The skeletochronological method was also used on larvae in utero and on new-born individuals. The age of adults was 8-24 years in population A, while males reached maturity at 3-5 years old and the youngest females were 6 years old in population B. Males and females from population B were also larger than those in population A. For the first time, lines of arrested growth (LAGs) were also found in the humerus of intra-uterine larvae and new-born individuals, indicating that young can spend up to 3 years in utero (population B) and up to 4 years (population A) before hatching. Growth of adults (fitted by the Bertalanffy model) also exhibited differences in growth coefficient (k) and mean asymptotic length (SVLmax) between sexes and populations. Local climatic conditions differed between the two areas of these populations and we hypothesize that the number of rainy days directly influences foraging during the short period of activity (< 3 months), leading to a delay in age at maturity, smaller length and growth rate, and increased gestation duration in the drier environment. The discussion is focused on proximate environmental influences on the variation of length and associated life-history traits in ectotherms, especially in terrestrial salamanders.

In farmlands, the population viability of many amphibians is suspected to depend on the resistance the matrix of crop fields presents to movements between ponds and terrestrial sites and movements among ponds. Over recent decades the increase in cereal growing at the expense of cattle breeding has caused a drastic change in habitat matrix in many European regions. We investigated the effect of such change on populations of three newt species ( Triturus helveticus, T. alpestris, and T. cristatus) by comparing their abundances in sites that varied in amount of cultivated ground. A multivariate regression analysis of the relationship of newt abundance to both pond and landscape variables demonstrated the negative influence of cultivated ground on abundance. The width of the uncultivated sector linking the pond to the forest was a good predictor of abundance after the influences of both pond area and fish presence were removed. Moreover, newt presence was positively related to the number of ponds within that 50-ha surrounding area, highlighting the role of metapopulation functioning in newt occupancy of ponds. The relationship between newt abundance and width of uncultivated sectors agrees with present knowledge of the orientation mechanisms that underlie migration movements in urodeles. Such a relationship between connectedness and sector width shows that narrow, linear corridors such as hedgerows may not be useful in newt conservation. Our study also highlights the need to incorporate a behavioral component of habitat use into models of connectivity in conservation biology. Resumen: Se considera que en tierras de cultivo la disponibilidad poblacional de muchos anfibios depende de la resistencia que la matriz de las tierras de cultivo presentan a los movimientos entre estanques y los sitios terrestres, y entre estanques. Durante las decadas recientes el incremento en la produccion de cereal a cambio de la cria de ganado ha ocasionado un cambio drastico en la matriz del habitat en muchas regiones de Europa. Investigamos los efectos de estos cambios en las poblaciones de tres especies de tritones ( Triturus helveticus, T. alpestris y T. cristatus) comparando sus abundancias en sitios que variaron en la cantidad de tierra cultivada. Un analisis de regresion multivariado de las relaciones entre la abundancia de tritones y las variables de los estanques y del paisaje demostro las influencias negativas de las tierras de cultivo sobre la abundancia. La amplitud de sectores sin cultivar que conecto a los estanques con el bosque fue un buen pronosticador de la abundancia despues de remover las influencias del area del estanque y la presencia de peces. Mas aun, la presencia de tritones estuvo positivamente relacionada con el numero de estanques dentro de un area adyacente de 50 ha, resaltando el papel del funcionamiento de la metapoblacion en la ocupacion de estanques. La relacion entre la abundancia de tritones y la amplitud de sectores sin cultivar concuerda con el conocimiento actual sobre los mecanismos de orientacion que delimitan los movimientos de migracion en urodelos. Esta relacion entre la conectividad y la amplitud del sector muestra que los corredores estrechos y lineales como lo son los cercos podrian no ser utiles para la conservacion de tritones. Nuestro estudio tambien resalta la necesidad de incorporar un componente conductual del uso del habitat cuando se modela la conectividad en estudios de conservacion biologica.

The movements of 19 natterjack toads (Bufo calamita) were monitored by telemetry in a semi-arid agricultural landscape in NE Spain. Toads reproduced in two water pools in fields of barley and an ancient clayquarrying zone filled by rainfall. Thirteen toads were followed during the breeding season (weeks 5-17 of 1997) and 11 toads were followed after reproduction (weeks 18 to 46). During the breeding season four toads moved distances of up to 500 m between breeding sites. No significant differences in movement patterns were found between the sexes, but a tendency was observed for males to show higher terrestrial site fidelity than females. Distances moved after the breeding season were substantial. The distance between the furthest positions recorded was from 164 to 1201 m, the maximum distance observed between daily recordings ranged from 125 to 353 m and the maximum overall distances ranged from 567 to 4411 m. No significant differences were observed between males and females or between small and large toads. The movements of these toads in a semi-arid agricultural landscape in Spain are compared with those observed in England and Germany.

Life-history traits of Rana temporaria were studied in an alpine French population and in the literature. In the living frogs, mean adult body length was greater in females than in males. Sexual dimorphism in body length was 0.109 using Lovich & Gibbon's (1992) formula, but tended to decrease with age. Age of adult frogs was assessed by skeletochronology, and age distribution was not significantly different between the sexes (range 4–15 years in males, 5–12 in females). Adult survival rate was about 0.80 in both sexes. Once maturity was reached, the total expected longevity was 6.1 years in males and 5.5 years in females. Age and body length were positively correlated in both sexes. The growth coefficient (K) was 0.47 in males, and 0.55 in females, mainly reflected as faster female growth between metamorphosis and maturation. Growth rate generally decreased before sexual maturity was reached. On average, females matured 1 year later than males. Newly metamorphosed froglets averaged 16.1 mm. When combined with published data from 12 European populations of R. temporaria, the following general patterns emerge. Mean adult body length is significantly greater in females than in males, and mean body length at maturity shows the same trend. Variation in mean age at maturity and in longevity are considerable among populations, but there is no consistent trend of difference between the sexes. Body length and age are correlated between males and females, i.e. populations with long and old males also have long and old females. Mean adult body length, mean body length at maturity, age at maturity, and longevity all increase with decreasing activity period. Adults exposed to a short activity period grow slower but attain a greater final length. Sexual dimorphism in body length generally increases as activity period gets shorter. Polygons describing norms of reaction for maturation in an age–body length space are similarly oriented in both sexes, but with a wider range in age for females. This is due to an older age at maturity for females in populations with a short activity season. Mean age and length at maturity are significantly correlated in females, but not in males, partly supporting the hypothesis that this species has a flexible pattern of development. Observed patterns are compared with predictions from life-history theory, paying attention to all life stages and environmental variation.

Oviposition site selection in Triturus helveticus, T. alpestris and T. cristatus was studied both in natural and laboratory situations. In a natural breeding site, eggs of the three species were not laid equally on several plants: T. cristatus laid practically all their egges on only one plant (Nasturtium officinale) while T. helveticus eggs were collected on four plant species. In laboratory experiments, plastic and cotton fabric supports made it possible to test ovipositing female selectivity according to flexibility and shape of support and egg-laying distance below the surface. In all three species, a high proportion of eggs were laid on supports of linear shape (as opposed to arborescent shape) and egg frequency decreased as a function of depth (from 0 to 40 cm below the surface). Selection according to flexibility was also observed, the smallest species (T. helveticus) laying more eggs on the thinner supports. The role of support selectivity during oviposition in Triturus is discussed, in term of proximate (i.e. female protection against predators, breathing necessity) and ultimate (i.e. egg survival) factors, and related to new ecology.

The fauna of the Aralkum has been studied only partly. But the lists of mammals of the Kazakhstan part of the Aral Sea region, the migratory breeding bird species and rare winter visitors, the resident breeding bird species, passage visitors (birds), vagrant birds, reptiles collected around Aral Sea in 2002–2004, and taxonomical diversity of insect orders and other groups are documented. The ecological disaster of the Aral Sea reduced the faunistic diversity of the area in a selective manner. Aquatic and semiaquatic animal species such as fish-eating birds, waterfowl, Amphibia, water snakes and aquatic insects suffered dramatic reductions in numbers. Some freshwater species and species of riverine forest died out or left the area completely. On the other hand, desert species and certain eurybionts were able to extend their ranges into the Aralkum. Further monitoring of the fauna of the Aralkum is strongly recommended to document the very active migrations to and invasions of the dynamic new ecosystems.

Anthropogenic sources of nitrogen that pollute bodies of water can have toxic and sub-lethal effects on amphibians. It has been hypothesized that such exposure may promote local adaptation, that is, selection for higher tolerance in individuals in populations exposed to pollutants. We tested this hypothesis with respect to the Natterjack toad (Bufo calamita Laurenti, 1768), by comparing the nitrate dose response of tadpoles from eight populations (doses: 0, 50, 100, 500 and 1000 mg/l nitrate) from relatively unpolluted and intensively farmed environments. We evaluated the effect of nitrate exposure by observing the behavior (movements) of tadpoles exposed to different concentrations of nitrates. Exposure to high nitrate levels did not cause tadpole mortality in the populations used in our experiments; however, we did observe changes in activity for all populations, with these changes being either dose-related responses (decreased activity after exposure to 500 or 1000 mg/l), or more complex responses (increased activity when exposed to 50 or 100 mg/l nitrate, followed by decreased activity at higher concentrations). Natterjack toad tadpoles exhibited variable behavioural responses among the tested populations. Although these populations were selected on the basis of their potential agrochemical contamination, the observed variation in population tolerance was not related to the parameters used to estimate this contamination in these breeding sites. Possible explanations for this apparent lack of local adaptation in B. calamita tadpoles include inadequate estimates of the toads’ actual nitrate exposure in the field, and the biological characteristics of B. calamita, which may limit the effects of exposure or favor phenotypic plasticity.

Predation on newt eggs (Triturus helveticus and T. alpestris) was studied in both natural (pond) and control (aquaria) conditions. Eggs were laid on artificial supports and set in a pond near Bourg-en-Bresse (south-east France). Survival from segmentation to pre-hatching stages was similar in these two species and estimated at 0·16 ± 0·07. Survival of eggs protected in a closed bag of plastic netting was significantly higher (0·79 ± 0·08), suggesting predation as the major mortality risk. Eggs of T. alpestris were offered to the following potential predators: newts Triturus alpestris and T. helveticus (males and females), adult aquatic insects (Dytiscus marginalis, Acilius suleatus. Notonecta glauca, Ranatra linearis. Ilyocoris hermanni). a snail (Lunnaea stugnalis) and a tadpole (Rana temporaria). Only the newts and water beetles were observed to cat the eggs. Females wrapped their eggs in a fold made with the support during oviposition. D. marginalis fed on wrapped eggs and those we unwrapped in the same proportion. but newts (especially females) and A. sulcatus clearly ate fewer wrapped than unwrapped eggs. These results are discussed in terms of mortality risk and antipredator adaptations.

Changes in age structure of local populations were analyzed to determine the migrations of adult crested newts (Triturus cristatus) in a network of ponds. An individual's age was estimated by applying a skeletochronological method to the phalanges. The local populations studied bred in four neighbouring cattle ponds, the distance between any two of which did not exceed 150 m. These ponds belong to the larger network. Though the size of the whole population doubled from one year to the next, one local population became extinct due to lack of recruitment of young breeders. The larger local population attracted not only young breeders but also older newts, which probably came from other ponds. In ponds inhabited by fish, the newts were mainly young animals (2–3 years old), suggesting that they leave such ponds after their first breeding season. The results are discussed with regard to the model proposed by Gill for metapopulations of Notophthalmus viridescens. In the crested newt, contrary to Gill's predictions, all the sites do not recruit young breeders, and fidelity to the breeding site may be broken. The two species differ in longevity and age at maturity, hence it is suggested that functional models of metapopulations must integrate life-history parameters.

The African clawed frog Xenopus laevis is by far the most widely used amphibian species in laboratories. In the wild, X. laevis is an asymptomatic carrier of an emerging infectious disease called chytridiomycosis. The vector is the chytrid fungus Batrachochytrium dendrobatidis ( Bd), which has devastating effects on wild amphibian populations around the world. The impact of Bd on the metabolism of X. laevis has not been comprehended yet. However, even if asymptomatic, an infection is likely to affect the individual's physiology, immunology, development, reproduction and overall response to stress from a purely medical point of view, which will introduce noise and therefore increase variance within experimental groups of X. laevis. This could have implications on the scientific results from studies using this species. Here, we review the current knowledge on treatments of infected amphibians and propose a hygiene protocol adapted to laboratory populations and amphibian husbandry. Following the presented sanitation guidelines could further prevent the spread of Bd and probably of other amphibian pathogens. The sanitation guidelines will help to reduce the impact of amphibian husbandry on natural populations and must be considered a crucial contribution to amphibian conservation, as today 32% of all amphibians are considered threatened.