Aim Population fragmentation represents a leitmotif of conservation biology, but the impact of population reconnection is less well studied. The recent recolonization of large carnivores in Europe is a good model for studying this phenomenon. We aim to show novel data regarding distribution and population genetic structure of the grey wolf in Central Europe, a region considered a frequent crossroad and contact zone of different phylogeographic lineages, in a biogeographic context. Location Western Carpathians, Central Europe. Methods In concordance with the presumption of a highly mobile mammal, individual‐based Bayesian clustering and a posteriori definition of populations were used. Integrating the frameworks of landscape genetics and biogeography enabled the identification of transitions in population architecture. These patterns could be ascribed to isolating factors based on historical knowledge about species demography. Results Genetic differentiation mirrors population isolation and recognized environmental clusters, suggesting ecotypic variation. The east–west split in the Western Carpathians likely represents the signature of range fragmentation during bottlenecks in the 20th century. Mitochondrial variability is more depleted than nuclear variability, indicating founder‐flush demography. Microsatellites show finer‐scale differentiation in the Carpathians compared to the European plain, corresponding to topographic heterogeneity. Long‐range dispersal of a Carpathian wolf (ca. 300 km), the establishment of enclaves originated from the lowland population and admixture with mountain wolves were ascertained, indicating a population fraction producing large‐scale gene flow. Main conclusion Carpathian wolves are characterized by periods of population and range decline due to eradication, facilitating refugial role of alpine habitats and peripatric effects, followed by expansions and fusions probably caused by forest transition, population adaptation and efforts in conservation management. New occurrence and hybridization events predict further contacts between formerly isolated populations, with potential opposing effects of heterosis and outbreeding depression. Population recovery might be hindered due to isolation by environment and anthropogenic impacts.

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Voltammetric behavior of the genotoxic environmental pollutant 2‐aminofluoren‐9‐one (2‐AFN) was investigated using direct current voltammetry (DCV) and differential pulse voltammetry (DPV) at a glassy carbon electrode (GCE) in both negative and positive potential regions. For the determination of 2‐AFN based on the cathodic reduction of the carbonyl group, optimum conditions were found in a methanol–BrittonRobinson (BR) buffer pH 4.0 (1 : 9, v/v) medium, with the limits of quantification (LQs) of 0.4 and 0.2 µmol L−1 for DCV and DPV, respectively. For the determination of 2‐AFN based on the anodic oxidation of the amino group, optimum conditions were found in a mixture of methanol–BR buffer pH 8.0 (1 : 9, v/v), with the LQs of 0.8 and 0.6 µmol L−1 for DCV and DPV, respectively. The practical applicability of the newly developed voltammetric methods was verified on the direct determination of 2‐AFN in model samples of drinking and river water. Moreover, the interaction between 2‐AFN and double‐stranded DNA (dsDNA) was investigated by DPV (performed at the bare GCE when both dsDNA and 2‐AFN were present in the measured solution) and square‐wave voltammetry (SWV) (carried out at the dsDNA/GCE biosensor after its incubation in the solutions of 2‐AFN for various times and at various concentrations of 2‐AFN) to characterize damaging effects of the test substance on the dsDNA structure in vitro. The intercalation of 2‐AFN between the dsDNA base pairs was the predominant supramolecular interaction observed.

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