Landscape Genetics of the Yellow-Bellied Toad (Bombina variegata) in the Northern Weser Hills of Germany

Anthropogenic influences such as deforestation, increased infrastructure, and general urbanization has led to a continuous loss in biodiversity. Amphibians are especially affected by these landscape changes. This study focuses on the population genetics of the endangered yellow-bellied toad (Bombina variegata) in the northern Weser Hills of Germany. Additionally, a landscape genetic analysis was conducted to evaluate the impact of eight different landscape elements on the genetic connectivity of the subpopulations in this area. Multiple individuals from 15 study sites were genotyped using 10 highly polymorphic species-specific microsatellites. Four genetic clusters were detected, with only two of them having considerable genetic exchange. The average genetic differentiation between populations was moderate (global FST = 0.1). The analyzed landscape elements showed significant correlations with the migration rates and genetic distances between populations. Overall, anthropogenic structures had the greatest negative impact on gene flow, whereas wetlands, grasslands, and forests imposed minimal barriers in the landscape. The most remarkable finding was the positive impact of the underpasses of the motorway A2. This element seems to be the reason why some study sites on either site of the A2 showed little genetic distance even though their habitat has been separated by a strong dispersal barrier.

Effects of Desiccation on Metamorphic Climax in Bombina variegata: Changes in Levels and Patterns of Oxidative Stress Parameters

In this paper, we examined how the oxidative status (antioxidant system and oxidative damage) of Bombina variegata larvae changed during the metamorphic climax (Gosner stages: 42—beginning, 44—middle and 46—end) and compared the patterns and levels of oxidative stress parameters between individuals developing under constant water availability (control) and those developing under decreasing water availability (desiccation group). Our results revealed that larvae developing under decreasing water availability exhibited increased oxidative damage in the middle and end stages. This was followed by lower levels of glutathione in stages 44 and 46, as well as lower values of catalase, glutathione peroxidase, glutathione S-transferase and sulfhydryl groups in stage 46 (all in relation to control animals). Comparison between stages 42, 44 and 46 within treatments showed that individuals in the last stage demonstrated the highest intensities of lipid oxidative damage in both the control and desiccation groups. As for the parameters of the antioxidant system, control individuals displayed greater variety in response to changes induced by metamorphic climax than individuals exposed to desiccation treatment. The overall decrease in water availability during development led to increased oxidative stress and modifications in the pattern of AOS response to changes induced by metamorphic climax in larvae of B. variegata.

Conservation genetics of the yellow-bellied toad (Bombina variegata): population structure, genetic diversity and landscape effects in an endangered amphibian

Revealing patterns of genetic diversity and barriers for gene flow are key points for successful conservation in endangered species. Methods based on molecular markers are also often used to delineate conservation units such as evolutionary significant units and management units. Here we combine phylo-geographic analyses (based on mtDNA) with population and landscape genetic analyses (based on microsatellites) for the endangered yellow-bellied toad Bombina variegata over a wide distribution range in Germany. Our analyses show that two genetic clusters are present in the study area, a northern and a southern/central one, but that these clusters are not deeply divergent. The genetic data suggest high fragmentation among toad occurrences and consequently low genetic diversity. Genetic diversity and genetic connectivity showed a negative relationship with road densities and urban areas surrounding toad occurrences, indicating that these landscape features act as barriers to gene flow. To preserve a maximum of genetic diversity, we recommend considering both genetic clusters as management units, and to increase gene flow among toad occurrences with the aim of restoring and protecting functional meta-populations within each of the clusters. Several isolated populations with especially low genetic diversity and signs of inbreeding need particular short-term conservation attention to avoid extinction. We also recommend to allow natural gene flow between both clusters but not to use individuals from one cluster for translocation or reintroduction into the other. Our results underscore the utility of molecular tools for species conservation, highlight outcomes of habitat fragmentation onto the genetic structure of an endangered amphibian and reveal particularly threatened populations in need for urgent conservation efforts.

Conservation genetics of yellow-bellied toads (Bombina variegata): a matter of geographical scale and isolation

Amphibian populations world-wide are threatened by declines and extinctions mainly due to habitat loss and fragmentation. Habitat fragmentation threatens the yellow-bellied toad Bombina variegata in the northern and western regions of its distribution where it is strictly protected. We studied the genetic structure and diversity of populations at three geographical scales using microsatellite loci to detect potential threats for population persistence. At the local scale, we sampled four neighbouring localities at 1–2.6 km distance to detect effects of short-term (decades) fragmentation on connectivity. At the regional scale, five additional localities in the mountains of the Westerwald (Rhineland-Palatinate, Germany) were studied at up to 50.1 km distance to analyse genetic diversity and population structure. At the continental scale, we included data from regions in the northern distribution with fragmented populations (Hesse and Lower Saxony, Germany) and more continuous populations in the South (Alsace, France; Geneva, Switzerland; Trentino, Italy) to evaluate variation of genetic diversity. At the local scale, short-term fragmentation caused significant genetic differentiation between breeding assemblages only 1.4 km apart from each other. At the regional scale, we found notable genetic distance among localities. At the continental scale, we identified Alsace, Trentino and Geneva in the South as regions with low genetic structuring and high allelic richness, and the northern remaining regions in Germany as deeply structured with reduced allelic richness. We suggest that reduced genetic diversity and habitat fragmentation in northern regions makes these populations particularly vulnerable to decline. In conclusion, informed conservation management of B. variegata should focus on measures maintaining or improving connectivity among neighbouring populations.

Bombina variegata (Linnaeus, 1758) in rain pools in the Carpathian Mountains (Slovakia)

Rain pools (pluviotelmata or puddles) are common, small and non-permanent aquatic ecosystems. Data on the use of rain pools by amphibian taxa in the Carpathian Mountains, especially in Slovakia, are rare. In this study we describe the occurrence of Bombina variegata (Linnaeus, 1758), which may use these small aquatic ecosystems as part of their complex life cycle or where they spend their larval period. The most important hydroperiodic factors affecting the vertebrate fauna were the volume and area of the rain pools, the presence (respectively the absence) of shade, water temperature, biotope structure and the presence of several chemical compounds (NO− 3, GH, KH, pH). This study confirms that micro-habitats, such as rain pools, may play an important role for suitable reproduction sites of B. variegata, as stated by several authors from the different countries and regions of its large distributional range. At the same time, data on the presence or absence of different taxa with important conservation status in the monitored habitat aid in the detection of species.

Carry-over effects of the larval environment on the post-metamorphic performance of Bombina variegata (Amphibia, Anura)

Metamorphs of the yellow-bellied toad, Bombina variegata, vary widely in size at metamorphosis in the field. We performed a replicated outdoor mesocosm study to simulate the environmental factor combinations in permanent and ephemeral breeding sites and to quantify their effects on tadpole development (duration of the larval period, metamorph size and body condition). Looking for potential carry-over effects of the larval environment, we quantified locomotor performance of all metamorph phenotypes originating from the mesocosms immediately after metamorphosis under controlled conditions. In contrast to the prediction of life-history theory, tadpoles were unable to adjust developmental rate to water availability, but metamorphs originating from the ephemeral pond treatment were smaller and had a lower body condition than those from the permanent pond treatment. Size-dependent carry-over effects included the length of the first jump following tactile stimulation, burst performance (total length of spontaneous jumps) and endurance (total distance covered in 10 forced jumps). A size-independent effect of larval environment was the prolonged locomotor effort to escape (5.7 consecutive jumps following initial stimulus) of metamorphs from the ephemeral pond treatment compared to same-sized ones (3.7 jumps) from the permanent pond treatment. Thus, we demonstrate that carry-over effects of larval environment on metamorph phenotype and behaviour cause a considerable variation in fitness in the early terrestrial stage of B. variegata. Informed conservation management of endangered populations in the northern range should therefore include the provision of small permanent breeding ponds promoting larger and fitter metamorphs.