The first records of Niphargus hrabei and N. potamophilus in Ukraine and Bulgaria significantly enlarge the ranges of these species

Niphargus comprises hundreds of narrowly-endemic West Palaearctic subterranean taxa. However, a few exceptional species inhabit surface waters and have remarkably large ranges. Herein, based on morphological and molecular analyses, we provide important new records for two of these species. N. potamophilus, previously known from the eastern Azov Sea lowlands, is reported for the first time from Ukraine and Bulgaria from localities adjacent to the Black Sea. These findings expand its range westward by more than 1000 km along the coastline. From Bulgaria, we also report for the first time N. hrabei, a species previously known to occur along the middle and lower Danube lowlands and in isolated populations at the foothills of the Northern Caucasus. Our new record thus extends its range southwards by more than 150 km. Both species contained unique haplotypes at all of the sampled localities. These were, nevertheless, not very divergent from more distant populations, emphasizing their good dispersal ability. Ecologically, the sampling localities were generally characterized by stagnant to low running water, dense vegetation, and muddy substrate. Overall, our results bring important insights, shedding more light on the biogeography and ecology of Niphargus

Molecular markers and SEM imaging reveal pseudocryptic diversity within the Ponto-Caspian low-profile amphipod invader Dikerogammarus bispinosus

The Ponto-Caspian amphipod Dikerogammarus bispinosus was originally described from the Black Sea basin. Its recent discovery in the Caspian Sea basin was puzzling because it was unknown whether it was an invasive or an overlooked native species in this area. Here, we examined specimens collected from both the Black and Caspian Sea basins by means of molecular species delimitation based on nuclear (28S) and mitochondrial (COI) DNA sequences, as well as scanning electron microscopy (SEM). Our analyses reveal that D. bispinosus comprises three evolutionary independent lineages that are molecularly and morphologically distinct. One lineage occurs throughout rivers in the Black Sea basin, while the other two inhabit the Caspian Sea and were found in sympatry, further reinforcing that they are distinct species. Our time-calibrated phylogeny indicates that these lineages split during the Late Miocene-Pliocene, a period corresponding with the separation of the Black and Caspian basins via the Caucasus mountain uplift. SEM imaging revealed morphological differences with respect to setal patterns on the gnathopod propodi among all three lineages. Therefore, our results clearly indicate not only that D. bispinosus is native in the Caspian region, but that it has been overlooked for a long time. Additional populations covering the entire range of this species complex need to be further studied in order to gain a more complete picture of its evolutionary history and resolve its taxonomy