Integrative phylogenetic, phylogeographic and morphological characterisation of the Unio crassus species complex reveals cryptic diversity with important conservation implications.

The global decline of freshwater mussels and their crucial ecological services highlight the need to understand their phylogeny, phylogeography and patterns of genetic diversity to guide conservation efforts. Such knowledge is urgently needed for Unio crassus, a highly imperilled species originally widespread throughout Europe and southwest Asia. Recent studies have resurrected several species from synonymy based on mitochondrial data, revealing U. crassus to be a complex of cryptic species. To address long-standing taxonomic uncertainties hindering effective conservation, we integrate morphometric, phylogenetic, and phylogeographic analyses to examine species diversity within the U. crassus complex across its entire range. Phylogenetic analyses were performed using cytochrome c oxidase subunit I (815 specimens from 182 populations) and, for selected specimens, whole mitogenome sequences and Anchored Hybrid Enrichment (AHE) data on âˆ¼ 600 nuclear loci. Mito-nuclear discordance was detected, consistent with mitochondrial DNA gene flow between some species during the Pliocene and Pleistocene. Fossil-calibrated phylogenies based on AHE data support a Mediterranean origin for the U. crassus complex in the Early Miocene. The results of our integrative approach support 12 species in the group: the previously recognised Unio bruguierianus, Unio carneus, Unio crassus, Unio damascensis, Unio ionicus, Unio sesirmensis, and Unio tumidiformis, and the reinstatement of five nominal taxa: Unio desectusstat. rev., Unio gontieriistat. rev., Unio mardinensisstat. rev., Unio nanusstat. rev., and Unio vicariusstat. rev. Morphometric analyses of shell contours reveal important morphospace overlaps among these species, highlighting cryptic, but geographically structured, diversity. The distribution, taxonomy, phylogeography, and conservation of each species are succinctly described.

First results on the genetic diversity of the invasive signal crayfish Pacifastacus leniusculus (Dana, 1852) in Europe using novel microsatellite loci.

The introduction of non-native crayfish in aquatic ecosystems is very common due to human activities (e.g. aquaculture, recreational and commercial fisheries). The signal crayfish, Pacifastacus leniusculus (Dana, 1852), is one of the most widespread invasive species in Europe. Although several important ecological and economic impacts of this species have been reported, its European population genetic characterisation has never been undertaken using nuclear markers. Thus, the aim of this study was to develop and characterise new microsatellite markers for signal crayfish that can be useful in future studies in its invaded range, since only five are available so far. In total, 93 individuals from four geographically distinct European populations (Portugal, Great Britain, Finland and Sweden) were scored for the new markers and for those previously described, with the Bayesian analysis revealing a clear distinction among populations. These markers are suitable for future studies of the population genetic structure of this important invasive species, by increasing information about the possible pathways of introduction and dispersal, and by giving insights about the most important vectors of introduction.

Impacts of climate change and land-use scenarios on Margaritifera margaritifera, an environmental indicator and endangered species.

In this study, we assess the impacts of future climate and land-use in the Beça River (northern Portugal) under different scenarios and how this will translate into the conservation status of the endangered pearl mussel Margaritifera margaritifera (Linnaeus, 1758). This species is currently present in several stretches of the Beça River that still hold adequate ecological conditions. However, the species is threatened by projected declines in precipitation for the 21st century, with implication on the river flows and water depths that might decrease below the species requisites. This situation could be especially critical during summer conditions since the ecological flows may not be assured and several river stretches may be converted into stagnant isolated pools. The habitat connectivity will also be affected with reverberating effects on the mobility of Salmo trutta, the host of M. margaritifera, with consequences in the reproduction and recruitment of pearl mussels. In addition, human-related threats mostly associated with the presence of dams and an predicted increases in wildfires in the future. While the presence of dams may decrease even further the connectivity and river flow, with wildfires the major threat will be related to the wash out of burned areas during storms, eventually causing the disappearance of the mussels, especially the juveniles. In view of future climate and land-use change scenarios, conservation strategies are proposed, including the negotiation of ecological flows with the dam promoters, the replanting of riparian vegetation along the water course and the reintroduction of native tree species throughout the catchment.

Isolation, purification and characterization of glycosaminoglycans in the fluids of the mollusc Anodonta cygnea.

The isolation, purification and characterization of Anodonta cygnea haemolymph, and extrapallial fluid glycosaminoglycans (Gags), which have high calcium affinity, were carried out in order to better understand the process of nacreous shell biomineralization. Our results show the existence of two different Gags with similar contents in the two fluids, throughout the year, but with significant seasonal variations for both. In the winter months, we identified by electrophoresis, only one kind of Gag chain (hyaluronic acid) while two different Gags (hyaluronic acid and heparan sulphate-like) from both fluids of A. cygnea were found in summer months. Quantification showed the total Gag fractions of both fluids in average, at their highest concentration (79.8 mg/L) in the highest calcification summer period. In contrast, the period of the year with the lowest concentration of total Gags (54.6 mg/L) occurred during the winter when calcification is reduced. This significant decrease between seasons is correlated mainly with the sulphated fraction, being 37.1 mg/L in the summer while only 9.2 mg/L in the winter haemolymph. The present data suggest that a heparan sulphate-like Gag has a relevant role in the biomineralization mechanisms acting as the calcium carbonate nucleator in the shell.