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Climate change has amplified the threat posed by aquatic invasive species as potential disruptors of biodiversity and ecosystem functioning. Species Distribution Models (MaxEnt) based on original data and ecological variables have identified contemporary seven global centers of the oriental rivel prawn Macrobrachium nipponense distribution: the native range in East Asia, Northern, Western and Eastern Europe, the Irano-Turanian region, and North and South America. By 2050, further expansion in Europe is expected, likely due to climate change, particularly temperature changes (Bio1) and rain precipitation during the warmest quarter (Bio18). However, the species may see a range reduction in southern Europe due to lower precipitation and increased droughts related to climate change. Therefore, a northward shift in the range of the species is also predicted. In the context of global change, and especially biological invasions, this study highlights the risks of introducing aquaculture based on M. nipponense and recommends controlling such economic activities, which are associated with a high risk for native species and ecosystems. Further, long-term monitoring is needed to assess impacts and to efficiently manage M. nipponense populations that are already present in their non-native habitats, for mitigating their negative effects on native species and ecosystems worldwide. [ABSTRACT FROM AUTHOR]

Aquaculture's role in addressing food security has grown, with a spotlight on Asian carp species. Polyculture, i.e. cultivating multiple fish species in a single system, is being increasingly adopted for its resource efficiency and economic benefits. This practice for Asian fish for food and ornamental purposes is gaining traction in Europe despite their invasive potential. Rising temperatures due to climate change offer an opportunity for thermophilic Asian carps (Hypophthalmichthys molitrix and Aristichthys nobilis). Using GIS modeling (Maxent), we identified the possible settlement of Asian carp in Northern Europe amidst climate change. We analyzed carp global distribution centers, assessed the potential carp spread in Europe, and evaluated their potential suitability for polyculture systems. By 2050, H. molitrix may extend its range to 58–62° N latitude, with a potential 1.7-fold habitat increase, while A. nobilis, which are more heat-tolerant, may move north to 52–58° N latitude, with a 1.3-fold potential increase. Despite the slight ecological differences in their native habitats, niche modeling indicates that these carp can occupy similar niches in Europe (proven statistically). The eventuality of using Asian species for polyculture in Europe presents both opportunities and challenges in the face of a changing climate as long as invasion risks are prevented. Envisaging such polyculture, yet very carefully for the protection of ecosystems, can help food security. [ABSTRACT FROM AUTHOR]

Crayfish plague is a devastating disease of European freshwater crayfish and is caused by the oomycete Aphanomyces astaci (Ap. astaci), believed to have been introduced to Europe around 1860. All European species of freshwater crayfish are susceptible to the disease, including the white-clawed crayfish Austropotamobius pallipes. Ap. astaci is primarily spread by North American crayfish species and can also disperse rapidly through contaminated wet gear moved between water bodies. This spread, coupled with competition from non-indigenous crayfish, has drastically reduced and fragmented native crayfish populations across Europe. Remarkably, the island of Ireland remained free from the crayfish plague pathogen for over 100 years, providing a refuge for A. pallipes. However, this changed in 1987 when a mass mortality event was linked to the pathogen, marking its introduction to the region. Fortunately, crayfish plague was not detected again in Ireland until 2015 when a molecular analysis linked a mass mortality event in the Erne catchment to Ap. astaci. Since then, the pathogen has appeared across the island. Between 2015 and 2023, Ap. astaci was detected in 18 water catchments, revealing multiple genotypes. Intriguingly, the pathogen in Ireland is present without its natural host species. The uneven distribution of various genetic lineages strongly suggests the human-mediated transport of zoospores via contaminated water equipment as a primary cause of spread. This review details the timeline of these events, Ap. astaci's introduction into Ireland, and its rapid spread. As well, this review references the genotypes that have been determined, and discusses the issue of non-indigenous crayfish species in Ireland and management efforts. [ABSTRACT FROM AUTHOR]

Repetitive elements are a major component of DNA sequences due to their ability to propagate through the genome. Characterization of Metazoan repetitive profiles is improving; however, current pipelines fail to identify a significant proportion of divergent repeats in non-model organisms. The Decapoda order, for which repeat content analyses are largely lacking, is characterized by extremely variable genome sizes that suggest an important presence of repetitive elements. Here, we developed a new standardized pipeline to annotate repetitive elements in non-model organisms, which we applied to twenty Decapoda and six other Crustacea genomes. Using this new tool, we identified 10% more repetitive elements than standard pipelines. Repetitive elements were more abundant in Decapoda species than in other Crustacea, with a very large number of highly repeated satellite DNA families. Moreover, we demonstrated a high correlation between assembly size and transposable elements and different repeat dynamics between Dendrobranchiata and Reptantia. The patterns of repetitive elements largely reflect the phylogenetic relationships of Decapoda and the distinct evolutionary trajectories within Crustacea. In summary, our results highlight the impact of repetitive elements on genome evolution in Decapoda and the value of our novel annotation pipeline, which will provide a baseline for future comparative analyses. [ABSTRACT FROM AUTHOR]

Genomics provides effective tools to characterize biodiversity, but the full implementation of genomic techniques in practical conservation is still limited. We review some of the main approaches in biodiversity genomics available to conservationists and genomicists. High-quality, long-read sequencing and bioinformatic technologies facilitate genome sequencing and assembly for any species. We summarize how reference genomes, in conjunction with population genomic data, can contribute to biodiversity monitoring, conservation, and restoration efforts. Over the past decade, many initiatives to generate reference genomes spanning the tree of life have emerged worldwide. We call for increased integration of reference genomes and population genomics data into interdisciplinary conservation efforts to fully unlock the potential of genomics in safeguarding global biodiversity. The availability of public genomic resources can greatly assist biodiversity assessment, conservation, and restoration efforts by providing evidence for scientifically informed management decisions. Here we survey the main approaches and applications in biodiversity and conservation genomics, considering practical factors, such as cost, time, prerequisite skills, and current shortcomings of applications. Most approaches perform best in combination with reference genomes from the target species or closely related species. We review case studies to illustrate how reference genomes can facilitate biodiversity research and conservation across the tree of life. We conclude that the time is ripe to view reference genomes as fundamental resources and to integrate their use as a best practice in conservation genomics. [ABSTRACT FROM AUTHOR]

Wetlands are of great importance for biodiversity and nature conservation, especially geographically isolated wetlands (GIW). Yet literature about the ecological value of such GIW is missing, especially at the edge of the distribution of endangered species such as amphibians. In 2018 and 2022, we monitored amphibian communities in 15 isolated (GIW) and 12 non-isolated (nGIW) ponds by counting individuals using three methods: (1) capturing with hand nets, (2) visual counting, and (3) capturing with fyke traps. The three methods provided similar results, showing the great importance of GIW for amphibians, especially newts, whose abundance was 5–13 times greater in GIW compared to nGIW. The largest numbers of species and individuals (adults and larvae) were found in isolated wetlands (GIW). In non-isolated water bodies (nGIW) where more than 10 individuals of the Chinese sleeper Perccottus glenii, an alien invasive fish, were found, amphibians were not found at all. Importantly, between 2018 and 2022, the northern crested newt, T. cristatus, dramatically decreased in the nGIW. As a result of our work, it was revealed that the reconstruction of geographically isolated wetlands is very important for the conservation of amphibian biodiversity in a changing climate. One of the most effective measures aimed at protecting amphibians from negative factors—the spread of alien invasive fish species and diseases—is the reconstruction of wetlands in historically exploited landscapes with the creation of a wide range of water bodies yet broadly dominated by geographically isolated ponds. [ABSTRACT FROM AUTHOR]

In the modern world, when the problems of the environment are most acutely associated with climate change, amphibians are considered to be the most vulnerable group of anamniotes as an indicator of the state of wetlands. Along with a decrease of numbers among amphibians in Europe, nowadays newts especially suffer from the impact of invasive species, in particular predators such as the Chinese sleeper, Perccottus glenii. This predatory fish species has recently spread to areas of primary relevance for newt reproduction. This fish consumes eggs, larvae, and even adult newt individuals. Using an ecological niche approach and climate based species distribution models (SDM), we applied the coefficient of determination (R2) for comparing the level of similarity of the built SDM for the newts Triturus cristatus and Lissotriton vulgaris, and the Chinese sleeper. We show that by 2050, the level of climatic niche similarity for these native and invasive species will increase from 12% to 22% throughout Europe, and from 44% to 66% in Eastern Europe. This study highlights the expansion of the Chinese sleeper as a real threat to European biodiversity of wetlands in the near future, especially in their most northeastern distribution range. [ABSTRACT FROM AUTHOR]

Objectives: Crayfish plague disease, caused by the oomycete pathogen Aphanomyces astaci represents one of the greatest risks for the biodiversity of the freshwater crayfish. This data article covers the de novo transcriptome assembly and annotation data of the noble crayfish and the marbled crayfish challenged with Ap. astaci. Following the controlled infection experiment (Francesconi et al. in Front Ecol Evol, 2021, https://doi.org/10.3389/fevo.2021.647037), we conducted a differential gene expression analysis described in (Boštjančić et al. in BMC Genom, 2022, https://doi.org/10.1186/s12864-022-08571-z) Data description: In total, 25 noble crayfish and 30 marbled crayfish were selected. Hepatopancreas tissue was isolated, followed by RNA sequencing using the Illumina NovaSeq 6000 platform. Raw data was checked for quality with FastQC, adapter and quality trimming were conducted using Trimmomatic followed by de novo assembly with Trinity. Assembly quality was assessed with BUSCO, at 93.30% and 93.98% completeness for the noble crayfish and the marbled crayfish, respectively. Transcripts were annotated using the Dammit! pipeline and assigned to KEGG pathways. Respective transcriptome and raw datasets may be reused as the reference transcriptome assemblies for future expression studies. [ABSTRACT FROM AUTHOR]

Ponds in agricultural landscapes are often used by amphibians as breeding habitat. However, the characteristics of agricultural ponds and especially the surrounding area are usually said to be suboptimal for many amphibian species. Using suboptimal habitats might allow a species’ survival and reproduction, but can have negative consequences at the individual and population level. In the present study, we investigated Palmate Newt (Lissotriton helveticus) populations from an intensive wine-growing region in southern Germany and compared them with populations located in a nearby forested area in terms of biometric traits, age and genetic structure. By analyzing over 900 adult newts from 11 ponds, we could show that newts reproducing in forest ponds were larger than newts reproducing in agricultural ponds. We did not find differences in the newt age and growth rate between habitat types. Therefore, differences in the body size of newts might already existed in larvae and/or juveniles, what might be related to a lower habitat quality for larvae and/or juveniles in the agricultural landscape. Body mass, body condition and sexual dimorphic traits (length of the caudal filament and max. height of the tail) correlated with body size, but no additional effect of the habitat type was found. The analysis of microsatellites revealed a higher genetic diversity in forest ponds. However, no clear sign of inbreeding was observed in any agricultural population, suggesting some degree of gene flow between them. We conclude, that agricultural ponds can be suitable habitats for the Palmate Newt and that conservation effort should aim to preserve them. The observed effects on body size indicate the need to increase the quality of the aquatic and terrestrial habitat for early life stages of this newt species in agricultural landscapes. [ABSTRACT FROM AUTHOR]

Pesticides are one of the main drivers of the worldwide amphibian decline. Their actual toxicity depends on a number of factors, like the species in focus or the developmental stage of exposed individuals. As ectothermic species, the metabolism of amphibians is influenced by ambient temperature. Therefore, temperature also affects metabolic rates and thus processes that might enhance or reduce toxic effects. Studies about the interactive effect of temperature and toxicity on amphibians are rare and deliver contrasting results. To investigate the temperature-dependent pesticide sensitivity of larvae of two European species we conducted acute toxicity tests for the viticultural fungicide Folpan® 500 SC with the active ingredient folpet at different temperatures (6°C, 11°C, 16°C, 21°C, 26°C). Sensitivity of Rana temporaria and Bufotes viridis was highly affected by temperature: early larvae (Gosner stage 20) were about twice more sensitive to Folpan® 500 SC at 6°C compared to 21°C. Next to temperature, species and developmental stage of larvae had an effect on sensitivity. The most sensitive individuals (early stages of R. temporaria at 6°C) were 14.5 times more sensitive than the least sensitive ones (early stages of B. viridis at 26°C). Our results raise concerns about typical ecotoxicological studies with amphibians that are often conducted at temperatures between 15°C and 20°C. We suggest that future test designs should be performed at temperatures that reflect the temperature range amphibians are exposed to in their natural habitats. Variations in the sensitivity due to temperature should also be considered as an uncertainty factor in upcoming environmental risk assessments for amphibians. [ABSTRACT FROM AUTHOR]

Invasive alien crayfish threaten the diversity of freshwater ecosystems and native crayfish fauna. In Europe, this is largely due to transmission of the crayfish plague to susceptible native crayfish. Many invasive species tolerate crayfish plague, but the infection still has the potential to reduce the fitness of a tolerant host due to energy trade-offs between immune response maintenance and life-history traits, such as growth and reproduction. In combination with other unfavourable conditions, such a response could alter further invasion success of an otherwise successful crayfish invader. We examined whether repeated infection with one of the most virulent haplogroups of crayfish plague agent (Aphanomyces astaci) affects growth or survival of the juvenile marbled crayfish (Procambarus virginalis). Juveniles were exposed to i) two levels of pathogen concentrations, and ii) two different feeding regimes under the higher pathogen concentration. In all performed trials, repeated infection reduced growth rates, while the combination of recurring infection and food limitation significantly increased mortality. The average energy cost of the immune response was estimated at 12.07 J/day for individuals weighing 0.3 grams. Since infections were frequent and pathogen concentrations high, results suggest that marbled crayfish is resistant to A. astaci pathogen and its survival is only affected by adding the stress of food limitation. The survival of almost half of the individuals exposed to high pathogen loads and extreme food limitation indicates that chronic infection by crayfish plague is unlikely to be an important factor impeding invasion success of the marbled crayfish, even under harsh conditions. Our results add to the growing body of evidence that marbled crayfish has potential to become one of the most successful freshwater invaders. [ABSTRACT FROM AUTHOR]

Most agricultural soils are expected to be contaminated with agricultural chemicals. As the exposure to pesticides can have adverse effects on non-target organisms, avoiding contaminated areas would be advantageous on an individual level, but could lead to a chemical landscape fragmentation with disadvantages on the metapopulation level. We investigated the avoidance behavior of juvenile common toads (Bufo bufo) in response to seven pesticide formulations commonly used in German vineyards. We used test arenas filled with silica sand and oversprayed half of each with different pesticide formulations. We placed a toad in the middle of an arena, filmed its behavior over 24 hours, calculated the proportion of time a toad spent on the contaminated side and compared it to a random side choice. We found evidence for the avoidance of the folpet formulation Folpan® 500 SC, the metrafenone formulation Vivando® and the glyphosate formulation Taifun® forte at maximum recommended field rates for vine and a trend for avoidance of Wettable Sulphur Stulln (sulphur). No avoidance was observed when testing Folpan® 80 WDG (folpet), Funguran® progress (copper hydroxide), SpinTorTM (spinosad), or 10% of the maximum field rate of any formulation tested. In the choice-tests in which we observed an avoidance, toads also showed higher activity on the contaminated side of the arena. As video analysis with tracking software is not always feasible, we further tested the effect of reducing the sampling interval for manual data analyses. We showed that one data point every 15 or 60 minutes results in a risk of overlooking a weak avoidance behavior, but still allows to verify the absence/presence of an avoidance for six out of seven formulations. Our findings are important for an upcoming pesticide risk assessment for amphibians and could be a template for future standardized tests. [ABSTRACT FROM AUTHOR]

The European weather loach (Misgurnus fossilis) is classified as highly endangered in several countries of Central Europe. Populations of M. fossilis are predominantly found in ditches with low water levels and thick sludge layers and are thus hard to detect using conventional fishing methods. Therefore, environmental DNA (eDNA) monitoring appears particularly relevant for this species. In previous studies, M. fossilis was surveyed following eDNA water sampling protocols, which were not optimized for this species. Therefore, we created two full factorial study designs to test six different eDNA workflows for sediment samples and twelve different workflows for water samples. We used qPCR to compare the threshold cycle (Ct) values of the different workflows, which indicate the target DNA amount in the sample, and spectrophotometry to quantify and compare the total DNA amount inside the samples. We analyzed 96 water samples and 48 sediment samples from a pond with a known population of M. fossilis. We tested several method combinations for long‐term sample preservation, DNA capture, and DNA extraction. Additionally, we analyzed the DNA yield of samples from a ditch with a natural M. fossilis population monthly over one year to determine the optimal sampling period. Our results showed that the long‐term water preservation method commonly used for eDNA surveys of M. fossilis did not lead to optimal DNA yields, and we present a valid long‐term sample preservation alternative. A cost‐efficient high salt DNA extraction led to the highest target DNA yields and can be used for sediment and water samples. Furthermore, we were able to show that in a natural habitat of M. fossilis, total and target eDNA were higher between June and September, which implies that this period is favorable for eDNA sampling. Our results will help to improve the reliability of future eDNA surveys of M. fossilis. [ABSTRACT FROM AUTHOR]

Identification of the geographic extent of population boundaries, the distribution of genetic lineages, and the amount of genetic exchange among breeding groups is needed for effective conservation of vulnerable marine migratory species. This is particularly true of the flatback turtle (Natator depressus), which only breeds in Australia but has extensive migrations that can include international waters.This study investigated the phylogeography and genetic structure among 17 flatback turtle rookeries across their range by sequencing an 810 bp portion of the mitochondrial DNA in 889 samples and genotyping 10 microsatellite loci in 598 samples.There was low phylogenetic divergence among haplotypes and evidence of recent population expansion, likely in the late Pleistocene. A predominant haplotype was found across all rookeries, but other haplotype groups were regionally specific.In general, there was agreement in patterns of genetic differentiation in the mitochondrial DNA and microsatellite data, and in some pairwise comparisons a higher mutation rate of microsatellites provided stronger evidence of differentiation.These results suggest natal philopatry operates in the choice of breeding locations for males as well as females.Evidence of genetic connectivity among neighbouring rookeries led to the identification of seven genetic stocks. Geographic boundaries of rookeries used by genetic stocks varied widely (160–1,300 km), highlighting a need for field studies to better understand movement patterns.Hierarchical analysis of molecular variance identified significant genetic differentiation based upon genetic stock, nesting phenology (summer vs. winter nesters), and a west–east discontinuity across Torres Strait. A pattern of isolation by distance was identified, which was most strongly observed in the microsatellite data.In combination with tagging and telemetry studies, these results will allow better quantification of stock‐specific threats along migratory routes and in foraging habitats. Implications of climate change will be stock specific and may depend upon the extent of genetic connectivity between neighbouring stocks. [ABSTRACT FROM AUTHOR]

The Upper Rhine Valley, a "hotspot of biodiversity" in Germany, has been treated with the biocide Bacillus thuringiensis var. israelensis (Bti) for mosquito control for decades. Previous studies discovered Bti nontarget effects in terms of severe chironomid abundance reductions. In this study, we investigated the impact of Bti on species level and addressed the community composition of the nontarget family Chironomidae by use of community metabarcoding. Chironomid emergence data were collected in three mosquito‐control relevant wetland types in the Upper Rhine Valley. For all three sites the chironomid species composition, based on operational taxonomic units (OTUs), was different to varying degrees in the Bti‐treated samples versus control samples, ranging from a significant 63% OTU reduction to an OTU replacement. We assumed that predatory chironomids are less prone to Bti than filter feeders, as the latter feed on floating particles leading to direct ingestion of Bti. However, a comparable percentage of predators and filter feeders (63% and 65%, respectively) was reduced in the Bti samples, suggesting that the feeding strategy is not the main driver for Bti sensitivity in chironomids. Finally, our data was compared to a three‐year‐old data set, indicating possible chironomid community recovery due to species recolonization a few years after the last Bti application. Considering the currently discussed worldwide insect decline we recommend a rethinking of the usage of the biocide Bti, and to prevent its ongoing application especially in nature protection reserves to enhance ecological resilience and to prevent boosting the current biodiversity loss. [ABSTRACT FROM AUTHOR]

The American rusty crayfish, Faxonius rusticus , is an invasive species in parts of North America where it displaces native crayfish species. In Europe, various invasive North American crayfish species are carriers of the crayfish plague disease agent Aphanomyces astaci that causes devastating population declines of European crayfish. Little is known about either the presence or the influence of A. astaci in North America where it originates. Here, we attempted to isolate A. astaci from North American F. rusticus for the first time and compare it to the strains detected in Europe. We tested the infection status of 84 F. rusticus samples from 10 different locations in the midwestern United States that are outside of the F. rusticus native distribution range. We used quantitative real-time PCR (qPCR) to assess the infection status in each individual, and we determined the mitochondrial haplotypes and multilocus microsatellite genotypes when it was possible. We detected A. astaci DNA in 9 individuals from 4 out of 10 locations. Analyses of the axenic culture isolates and the crayfish tissue samples by sequence analyses, haplotyping, and genotyping confirmed the results and revealed a novel A. astaci microsatellite genotype. Our results clearly identify F. rusticus as a host of A. astaci in North America. The threat of these strains to endangered crayfish species in North America remains unknown, but the potential of A. astaci infections should be considered when developing and implementing invasive species management plans for conservation purposes. [ABSTRACT FROM AUTHOR]

The biocide Bacillus thuringiensis var. israelensis (Bti) is widely applied for mosquito control in temporary wetlands of the German Upper Rhine Valley. Even though Bti is considered environmentally friendly, several studies have shown non-target effects on chironomids, a key food resource in wetland ecosystems. Chironomids have been proposed as important indicators for monitoring freshwater ecosystems, however, morphological determination is very challenging. In this study, we investigated the effectiveness of metabarcoding for chironomid diversity assessment and tested the retrieved chironomid operational taxonomic units (OTUs) for possible changes in relative abundance and species diversity in relation to mosquito control actions in four temporary wetlands. Three of these wetlands were, for the first year after 20 years of Bti treatment, partly left Bti-untreated in a split field design, and one wetland has never been treated with Bti. Our metabarcoding approach detected 54 chironomid OTUs across all study sites, of which almost 70% could be identified to species level comparisons against the BOLD database. We showed that metabarcoding increased chironomid species determination by 70%. However, we found only minor significant effects of Bti on the chironomid community composition, even though Bti reduced chironomid emergence by 65%. This could be due to a time lag of chironomid recolonization, since the study year was the first year of Bti intermittence after about 20 years of Bti application in the study area. Subsequent studies will have to address if and how the chironomid community composition will recover further in the now Bti-untreated temporary wetlands to assess effects of Bti. [ABSTRACT FROM AUTHOR]

Several North American crayfish species have so far been identified as carriers of the crayfish plague agent Aphanomyces astaci. The pathogen is responsible for the declines of thousands of European crayfish populations. Currently, one of the introduction pathways of North American crayfish species is the aquarium trade which may sometimes be followed by intentional release or unintentional escape of the pet species into the wild. We investigated 85 samples of North American and New Guinean species, available through the aquarium trade, for their possible infection with A. astaci. Crayfish plague infection was examined by applying real-time PCR. Besides morphological identification, we sequenced a fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene for crayfish species confirmation. Additionally, sequence analysis of nuclear DNA was conducted to identify the A. astaci lineage of moderate to highly infected crayfish. A total of 11 of the 85 analyzed crayfish individuals tested positive for an A. astaci infection, of which nine species are for the first time identified as carriers of A. astaci. No new genetic lineages of A. astaci were identified. The results confirm that, due to the positive carrier status of tested crayfish, the aquarium trade in Europe can facilitate the spread of A. astaci and can thus be a significant threat to the indigenous crayfish and the environment. [ABSTRACT FROM AUTHOR]

We generated RNA-seq data to assemble the transcriptome of the noble crayfish ( Astacus astacus ) from four combined tissues (abdominal muscle, hepatopancreas, ovaries, green glands). A total of 194 million read pairs with a length of 100 bp were generated. The transcriptome was assembled de novo using Trinity software, producing 158,649 non-redundant transcripts. Lowly expressed transcripts were filtered out leaving 45,415 transcripts of which 14,559 were found to contain open reading frames with predicted gene function. The Transrate software revealed that 91% of the total reads were realigned to the assembly. Furthermore, BUSCO analysis indicated that our assembly is 64% complete. A total of 13,770 transcripts were assigned at least one GO term. This first de novo transcriptome assembly is an important foundation for future genomic research on the noble crayfish and adds to the general knowledge and further characterization of transcriptomes of non-model organisms. [ABSTRACT FROM AUTHOR]

Reconstructing the phylogeographic history of a species can aid in defining areas of conservation priority. For freshwater species, historical river structure plays a significant role in explaining genetic differentiation and population structure. However, human-induced translocations can erase the natural genetic structure, especially for species of commercial interest such as the noble crayfish ( Astacus astacus)., Our aim was to reconstruct the current genetic structure of the endangered noble crayfish in central Europe to identify refugial areas that are hotspots of genetic diversity. We analysed a fragment of the mitochondrial cytochrome oxidase subunit I, and the 16 S r RNA from 540 noble crayfish specimens from 156 sampling sites distributed around five European sea basins. Additionally, we conducted a microsatellite analysis of 289 individuals from 22 sites., Both mitochondrial and nuclear markers revealed genetically relatively homogenous populations in central Europe that had been influenced by anthropogenic translocations. However, some areas (Eider catchment in northern Germany and Rhineland-Palatinate in south-western Germany) show a distinct genetic structure with endemic haplotypes and private alleles indicating (i) that these areas were refugia for A. astacus in central Europe and (ii) that these populations have not been subject to anthropogenic translocations. Further, we found the highest genetic diversity in the Black Sea basin and particularly high differentiation between populations from the western Balkans and the remaining Black Sea populations. The split between Western Balkan and the remaining European populations is estimated to have occurred approximately 700 k years before present, whereas remaining differentiations occurred within the last 450 k years. Using migration modelling, we detected that the North Sea basin and the Baltic Sea basin were colonised independently via different colonisation paths from the eastern Black Sea basin, while the western Balkans did not contribute to this colonisation., Our results suggest the existence of at least two refugial areas in south-eastern Europe. To conserve maximum genetic diversity, conservation priorities for noble crayfish should focus on the south-eastern European genetic hotspots and on populations in central Europe that hold an autochthonous genetic structure (e.g. Langsee in the Eider catchment area). We further propose that each river catchment should form a separate management unit to reduce anthropogenic genetic homogenisation. [ABSTRACT FROM AUTHOR]

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Non-invasive DNA sampling is an important tool in amphibian conservation. Buccal swabs are nowadays replacing the wounding toe-clipping method. Skin and cloaca swabbing are even less invasive and easier to handle than buccal swabbing, but could result in contaminations of genetic material. Therefore, we test if external skin and cloaca swabs are as reliable as buccal swabs for genetic analysis of amphibians. We analysed eight microsatellite loci for the common frog ( Rana temporaria, Linnaeus 1758) and compared genotyping results for buccal, skin and cloaca swabs regarding allelic dropouts and false alleles. Furthermore, we compared two DNA extraction methods regarding efficiency and cost. DNA quality and quantity (amplification success, genotyping error rate, in nanogram per microlitre) were comparable among DNA sources and extraction methods. However, skin and cloaca samples exhibited high degrees of contamination with foreign individuals, which was due to sample collection during mating season. Here, we established a simple low budget procedure to receive DNA of amphibians avoiding stressful buccal swabbing or harmful toe clipping. However, the possibility of contaminations of external swabs has to be considered. [ABSTRACT FROM AUTHOR]

Summary 1. We investigated the Pleistocene and Holocene history of the rare mayfly Ameletus inopinatus EATON 1887 (Ephemeroptera: Siphlonuridae) in Europe. We used A. inopinatus as a model species to explore the phylogeography of montane, cold-tolerant aquatic insects with arctic-alpine distributions. 2. Using species distribution models, we developed hypotheses about the species demographic history in Central Europe and the recolonisation history of Fennoscandia. We tested these hypotheses using mitochondrial cytochrome oxidase I (mtCOI) sequence data and compared our genetic results with previously generated microsatellite data to explore genetic diversity distributions of A. inopinatus. 3. We observed old lineages, deep splits and almost complete lineage sorting of mtCOI sequences among mountain ranges. These results support a periglacial survival, i.e. persistence at the periphery of Pleistocene glaciers in Central Europe. 4. There was strong differentiation between the Fennoscandian and all other populations, indicating that Fennoscandia was recolonised from a refugium not accounted for in our sampling. High degrees of population genetic structure within the northern samples suggest that Fennoscandia was recolonised by more than one lineage. However, this structure was not apparent in previously published microsatellite data, consistent with secondary contact without sexual incompatibility or with sex-biased dispersal. 5. Our demographic analyses indicate that (i) the separation of northern and Central European lineages occurred during the early Pleistocene; (ii) Central European populations have persisted independently throughout the Pleistocene and (iii) the species extended its range about 150 000 years ago. [ABSTRACT FROM AUTHOR]

We describe the isolation of 11 polymorphic trinucleotide microsatellite loci from the stonefly Arcynopteryx compacta. Loci were highly variable with 3 to 14 alleles (mean = 6.45). Observed heterozygosity ranged from 0 to 0.867. Seven loci showed significant deviation from Hardy–Weinberg equilibrium across both populations. There was no evidence for null alleles, and thus, Hardy–Weinberg departures could have resulted from genetic structure between populations or subpopulations. No linkage between loci was found. The 11 loci should prove highly informative for population genetic studies. [ABSTRACT FROM AUTHOR]

We describe the isolation of ten polymorphic microsatellite loci from the mayfly Ameletus inopinatus. Loci had di- or trinucleotide repeat motifs and were highly variable with three to 17 alleles (mean = 7.15). Observed heterozygosity ranged from 0.143 to 0.905. One locus (Ami_202) showed significant deviation from Hardy–Weinberg equilibrium in one population, but no evidence for null alleles. One locus (Ami_73) was significantly linked with three other loci. The remaining nine loci should prove highly informative for population genetic studies. [ABSTRACT FROM AUTHOR]

• We used telemetry to track common toads in an agricultural landscape. • Toads are using vineyards as habitats during their post-breeding migration. • Comparing used and available habitats shows that toads tend to avoid vineyards. • Up to 24% of the breeding population are exposed to pesticides per day. • A heterogeneous cultural landscape is vital for the conservation of amphibian populations. Two important drivers of the global amphibian decline are habitat destruction due to an intensification of farming and a related increase of pesticide applications. Recent studies have shown that there might be an underestimated risk of pesticides on terrestrial amphibians. However, there are too few data on the terrestrial habitat use of amphibians in agricultural landscapes to estimate the exposure risk. To fill this knowledge gap, we used telemetry to investigate the post-breeding migration of 51 common toads (Bufo bufo) from a breeding pond in a vineyard-dominated landscape in Southern Palatinate (Germany). We expected most toads to migrate to the nearby Palatinate Forest as a terrestrial habitat. However, only four individuals reached the forest, suggesting that a part of the population is inhabiting the agricultural landscape over large parts of the year. Individuals were also found directly in the vineyards (15% of all relocations), but 23% less often than expected from a random choice and therefore tend to avoid vineyards as terrestrial habitat. To estimate a possible spatial–temporal overlap of toad migration and pesticide application, we combined telemetry data with information about pesticide applications from local wine growers. Seven individuals had a high probability (>75%) of being directly exposed to a pesticide application. Taking spray drift and the half-life values of applied pesticides into account, the number of toads potentially exposed raised to 15 individuals. We estimated that, on a single day up to 24% of the whole breeding population came in contact with pesticides, resulting in a high overall exposure risk. Pesticides can have negative effects on amphibians, and toads try to avoid vineyards as habitats. Therefore, we conclude that a heterogeneous cultural landscape, with buffer strips around ponds, uncultivated patches and migration corridors, might be the best management measure for sustaining amphibians in the agricultural landscape. [ABSTRACT FROM AUTHOR]