New insights into the genetic diversity of the stone crayfish: taxonomic and conservation implications.

BACKGROUND: Austropotamobius torrentium is a freshwater crayfish species native to central and south-eastern Europe, with an intricate evolutionary history and the highest genetic diversity recorded in the northern-central Dinarides (NCD). Its populations are facing declines, both in number and size across its entire range. By extanding current knowledge on the genetic diversity of this species, we aim to assist conservation programmes. Multigene phylogenetic analyses were performed using different divergence time estimates based on mitochondrial and, for the first time, nuclear DNA markers on the largest data set analysed so far. In order to reassess taxonomic relationships within this species we applied several species delimitation methods and studied the meristic characters with the intention of finding features that would clearly separate stone crayfish belonging to different phylogroups. RESULTS: Our results confirmed the existence of high genetic diversity within A. torrentium, maintained in divergent phylogroups which have their own evolutionary dynamics. A new phylogroup in the Kordun region belonging to NCD has also been discovered. Due to the incongruence between implemented species delimitation approaches and the lack of any morphological characters conserved within lineages, we are of the opinion that phylogroups recovered on mitochondrial and nuclear DNA are cryptic subspecies and distinct evolutionary significant units. CONCLUSIONS: Geographically and genetically isolated phylogroups represent the evolutionary legacy of A. torrentium and are highly relevant for conservation due to their evolutionary distinctiveness and restricted distribution.

Potamophylaxhumoinsapiens sp. n. (Trichoptera, Limnephilidae), a new species from the Sharr Mountains, Republic of Kosovo.

Background: The Sharr Mountains are one of the most important hotspots of terrestrial and freshwater biodiversity in the Balkan Peninsula, with many endemic and rare species. The caddisfly studies in this area increased during the past years, although insufficiently investigated areas still remain. New information: In this paper, we describe a new species, Potamophylaxhumoinsapiens sp. n. from the Sharr Mountains in the Republic of Kosovo, which is morphologically closest to Potamophylaxidliri Ibrahimi, Bilalli & Kucinic, 2022 and Potamophylaxjuliani Kumanski, 1999. The males of the new species differ from all known species of the Potamophylaxwinneguthi Species Group by their uniquely-shaped parameres, which are long, bulbous in their basal half and thin in the remaining length, with a bunch of very thin and long, hair-like spines, grouped uniformly at the apex. The new species further differs from its most similar congeners by its very wide distance between the dorsal and ventral edges of the apical part of inferior appendages in lateral view. The new species was found at three localities from 1416 to 1505 m a.s.l.Similar to the other species of the Potamophylaxwinneguthi Species Group, which have very narrow distribution areas, we posit that Potamophylaxhumoinsapiens sp. n. is a microendemic of the Sharr Mountains. The new species is the second known caddisfly species occurring only in the Kosovan part of the Sharr Mountains.

Frequencies of erythrocyte nuclear abnormalities and of leucocytes in the fish Barbus peloponnesius correlate with a pollution gradient in the River Bregalnica (Macedonia).

Integrated chemical and biomarker approaches were performed to estimate if there is ongoing toxicity in the River Bregalnica, namely connected with the presence of metals. The study was performed in water, sediment, and barbel (Barbus peloponnesius), collected in two seasons, from two suspected polluted and one reference zones. The water analyses revealed higher mean values in polluted sites for most of the examined physicochemical parameters. Metal concentrations (Zn, Cu, Cd, Mn, Pb, and Fe) in water were more or less constant, whereas in sediment, they were higher at the two polluted locations. Condition factor (CF), as a general health indicator, revealed better overall condition in barbel from the reference site. In general, blood parameters revealed higher values in the polluted localities. Irrespective of sex and/or season, the frequency of micronuclei (MN) and vacuolated nuclei (VN) were with higher rates in polluted sites. Similarly, the frequencies of the leucocytes (Le), binuclei (BN), and irregularly shaped nuclei (ISN) were also significantly increased in the polluted localities, but they seemed prone to be influenced by sex and/or season. However, strong positive correlations between blood biomarkers and most water physicochemical parameters and metal in sediment were estimated. Our data support that the River Bregalnica's lower course receives significant genotoxic pollution, likely via metal industry effluents, agricultural runoff, and domestic sewage, and reinforced the utility of MN and other nuclear abnormalities as sensitive and suitable biomarkers for genotoxicity when used in monitoring studies.