Many herpetofauna species have been introduced outside of their native range. MtDNA barcoding is regularly used to determine the provenance of such populations. The alpine newt has been introduced across the Netherlands, the United Kingdom and Ireland. However, geographical mtDNA structure across the natural range of the alpine newt is still incompletely understood and certain regions are severely undersampled. We collect mtDNA sequence data of over seven hundred individuals, from both the native and the introduced range. The main new insights from our extended mtDNA phylogeography are that 1) haplotypes from Spain do not form a reciprocally monophyletic clade, but are nested inside the mtDNA clade that covers western and eastern Europe; and 2) haplotypes from the northwest Balkans form a monophyletic clade together with those from the Southern Carpathians and Apuseni Mountains. We also home in on the regions where the distinct mtDNA clades meet in nature. We show that four out of the seven distinct mtDNA clades that comprise the alpine newt are implicated in the introductions in the Netherlands, United Kingdom and Ireland. In several introduced localities, two distinct mtDNA clades co-occur. As these mtDNA clades presumably represent cryptic species, we urge that the extent of genetic admixture between them is assessed from genome-wide nuclear DNA markers. We mobilized a large number of citizen scientists in this project to support the collection of DNA samples by skin swabbing and underscore the effectiveness of this sampling technique for mtDNA barcoding.

Studies on the impact of agricultural practices on plant diversity provide important information for policy makers and the conservation of the environment. The aim of the present work was to evaluate wild plant diversity across the agroecosystems in two contrasting regions of Bulgaria; Pazardzhik-Plovdiv (representing agroecosystems in the lowlands) and Western Stara Planina (the Balkan Mountains, representing agroecosystems in the foothills of the mountains). This study conducted a two-year assessment of plant diversity in different types of agricultural and forest ecosystems, representing more than 30 land use types. Plant diversity, measured by species number, was affected by the land use type only in Pazardzhik-Plovdiv region. More pronounced was the effect of the groups of land use types on the diversity, measured by the mean species number per scoring plot. Climatic conditions, measured by 19 bioclimatic variables, were the most important factor affecting plant species diversity. Six bioclimatic variables had a significant effect on the plant diversity, and the effect was more pronounced when the analysis considered pooled data of the two regions. The highest plant diversity was found on grazing land with sparse tree cover, while the lowest one was in the land use types representing annual crops or fallow. The study also established a database on weed species, relevant to agriculture. A number of common weeds were found in the Pazardzhik-Plovdiv region, while the most frequent species in the Western Stara Planina region were indigenous ones. Overall, the natural flora of Western Stara Planina was more conserved; eleven orchid species with conservation significance were found in the pastures and meadows in that region. The present study is the first attempt in Bulgaria to characterize the plant diversity across diverse agroecosystems representing many different land use types and environmental conditions. The results can contribute to nature conservation, biodiversity, and the sustainable use of plant resources.

Snakes rely heavily on chemical cues when foraging, searching for mates, etc. Snakes’ sex attractiveness pheromones comprise mainly heavy, semi-volatile compounds such as ketones. Here we investigated the composition of skin secretions of adult Vipera ammodytes (Linnaeus, 1758) individuals. The samples were analyzed by gas chromatography/mass spectrometry and the identification of the compounds was performed using commercial mass spectral libraries and retention times. The relative concentrations of all detected compounds were tested for significant differences between (1) male vs. female live individuals, (2) shed skin vs. live individuals, and (3) pre-reproductive vs. reproductive live individuals. We detected fifty-nine compounds of which six were ketones. Two ketones (2-pentacosanone and 2-heptacosanone) were present in many of the samples and thus may have an important role in the V. ammodytes chemical communication. We did not find significant differences between the relative concentrations of the compounds between male and female individuals (only three compounds are exceptions). Significant differences were found between extracts from shed skins and live individuals and between live pre-reproductive individuals and live reproductive individuals. The results of the study suggest that chemical communication in V. ammodytes involves less compounds in comparison to the known literature data for other species.