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BackgroundSpecies with strict ecological requirements may provide new insights into the forces that shaped the geographic variation of genetic diversity. The Pyrenean desman, Galemys pyrenaicus, is a small semi-aquatic mammal that inhabits clean streams of the northern half of the Iberian Peninsula and is endangered in most of its geographic range, but its genetic structure is currently unknown. While the stringent ecological demands derived from its aquatic habitat might have caused a partition of the genetic diversity among river basins, Pleistocene glaciations would have generated a genetic pattern related to glacial refugia.ResultsTo study the relative importance of historical and ecological factors in the genetic structure of G. pyrenaicus, we used mitochondrial and intronic sequences of specimens covering most of the species range. We show, first, that the Pyrenean desman has very low levels of genetic diversity compared to other mammals. In addition, phylogenetic and dating analyses of the mitochondrial sequences reveal a strong phylogeographic structure of a Middle Pleistocene origin, suggesting that the main lineages arose during periods of glacial isolation. Furthermore, both the spatial distribution of nuclear and mitochondrial diversity and the results of species distribution modeling suggest the existence of a major glacial refugium in the northwestern part of the Iberian Peninsula. Finally, the main mitochondrial lineages show a striking parapatric distribution without any apparent exchange of mitochondrial haplotypes between the lineages that came into secondary contact (although with certain permeability to nuclear genes), indicating incomplete mixing after the post-glacial recolonization. On the other hand, when we analyzed the partition of the genetic diversity among river basins, the Pyrenean desman showed a lower than expected genetic differentiation among main rivers.ConclusionsThe analysis of mitochondrial and intronic markers in G. pyrenaicus showed the predominant effects of Pleistocene glaciations on the genetic structure of this species, while the distribution of the genetic diversity was not greatly influenced by the main river systems. These results and, particularly, the discovery of a marked phylogeographic structure, may have important implications for the conservation of the Pyrenean desman.

Information about the degree of contemporary dispersal is important when trying to understand how populations interchange individuals and identify the specific barriers that prevent these movements. In the case of endangered species, this can represent crucial information when designing appropriate conservation strategies. Here we analyse relatedness between individuals from different localities and use these data to infer whether dispersal occurred in recent generations. We applied this approach to the Pyrenean desman (Galemys pyrenaicus), a semiaquatic and endangered species endemic to the Iberian Peninsula. We studied this species in four primary rivers in the Iberian Range, where two ancient mitochondrial lineages are separated by a strict contact zone, suggesting the existence of complex dispersal patterns. Using next-generation sequencing, we obtained 912 SNPs from each specimen and estimated relatedness values between them. While relatedness networks were dense within each river, we found surprisingly few relationships between individuals from different rivers despite their close proximity in some cases, indicating much lower dispersal between rivers compared to dispersal within a single river. In agreement with this result, the degree of inbreeding was exceedingly high in most individuals. These data show that relatedness information can be crucial to understand the contemporary dispersal patterns and conservation status of specific populations of endangered species.

The European mink Mustela lutreola is one of Europe's most endangered carnivores, with few vulnerable populations remaining. Surprisingly, a recent phylogeny placed a single mink specimen within the polecat (M. putorius, M. eversmannii) group, suggesting a recent speciation and/or the effects of hybridization. The analysis has now been extended to a further 51 mink and polecats. As before, phylogenetic methods failed to resolve the relationships between the species. One haplotype (C11) was found in both species, and predominated in European mink from Spain and eastern Europe. The known M. lutreola fossils are of very young date, so either mink arose recently, or else the situation is confused by hybridization and a biased fossil recovery. The study highlights the dangers of using a single genetic marker in defining Evolutionarily Significant Units (ESUs). Polecats and European mink are clearly distinct in their morphology and ecology, and should still be considered as separate ESUs, but without further data it is difficult to define Management Units. Following the precautionary principle, we recommend that for the moment European mink in eastern Europe (Belarus, Estonia and Russia) and Spain should be managed separately.

Molecular techniques allow non-invasive dietary studies from faeces, providing an invaluable tool to unveil ecological requirements of endangered or elusive species. They contribute to progress on important issues such as genomics, population genetics, dietary studies or reproductive analyses, essential knowledge for conservation biology. Nevertheless, these techniques require general methods to be tailored to the specific research objectives, as well as to substrate- and species-specific constraints. In this pilot study we test a range of available primers to optimise diet analysis from metabarcoding of faeces of a generalist aquatic insectivore, the endangered Pyrenean desman (Galemys pyrenaicus, É. Geoffroy Saint-Hilaire, 1811, Talpidae), as a step to improve the knowledge of the conservation biology of this species. Twenty-four faeces were collected in the field, DNA was extracted from them, and fragments of the standard barcode region (COI) were PCR amplified by using five primer sets (Brandon-Mong, Gillet, Leray, Meusnier and Zeale). PCR outputs were sequenced on the Illumina MiSeq platform, sequences were processed, clustered into OTUs (Operational Taxonomic Units) using UPARSE algorithm and BLASTed against the NCBI database. Although all primer sets successfully amplified their target fragments, they differed considerably in the amounts of sequence reads, rough OTUs, and taxonomically assigned OTUs. Primer sets consistently identified a few abundant prey taxa, probably representing the staple food of the Pyrenean desman. However, they differed in the less common prey groups. Overall, the combination of Gillet and Zeale primer sets were most cost-effective to identify the widest taxonomic range of prey as well as the desman itself, which could be further improved stepwise by adding sequentially the outputs of Leray, Brandon-Mong and Meusnier primers. These results are relevant for the conservation biology of this endangered species as they allow a better characterization of its food and habitat requirements.

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on behalf of the investigators of the PREV-ICTUS Study Abstract-The objective was to assess the stroke risk and prevalence of the cardiovascular risk factors and to analyze their relationship with the specific stroke rates of mortality in each of the autonomic communities of Spain. We conducted a multicenter, cross-sectional study of population Ͼ60 years old in Spanish primary care centers. In all of the subjects, clinical, biochemical, and electrocardiographic data were obtained, and the 10-year stroke risk was calculated using the Framingham score.