Incomplete knowledge of biodiversity remains a stumbling block for conservation planning and even occurs within globally important Biodiversity Hotspots (BH). Although technical advances have boosted the power of molecular biodiversity assessments, the link between DNA sequences and species and the analytics to discriminate entities remain crucial. Here, we present an analysis of the first DNA barcode library for the freshwater fish fauna of the Mediterranean BH (526 spp.), with virtually complete species coverage (498 spp., 98% extant species). In order to build an identification system supporting conservation, we compared species determination by taxonomists to multiple clustering analyses of DNA barcodes for 3165 specimens. The congruence of barcode clusters with morphological determination was strongly dependent on the method of cluster delineation, but was highest with the general mixed Yule-coalescent (GMYC) model-based approach (83% of all species recovered as GMYC entity). Overall, genetic morphological discontinuities suggest the existence of up to 64 previously unrecognized candidate species. We found reduced identification accuracy when using the entire DNA-barcode database, compared with analyses on databases for individual river catchments. This scale effect has important implications for barcoding assessments and suggests that fairly simple identification pipelines provide sufficient resolution in local applications. We calculated Evolutionarily Distinct and Globally Endangered scores in order to identify candidate species for conservation priority and argue that the evolutionary content of barcode data can be used to detect priority species for future IUCN assessments. We show that large-scale barcoding inventories of complex biotas are feasible and contribute directly to the evaluation of conservation priorities.

Distribution records (historical, contemporary)

Data are provided on the distribution, abundance, early development and biology of the endangered Greek endemic species Valencia letourneuxi Sauvage 1880, along with a record of its occurrence at new localities. V. letourneuxi is a small-bodied and short-lived insectivorous species, exhibiting cryptic colouration and sexual dimorphism. It matures in the first year of life, reproduces serially in late spring and summer, and deposits spherical eggs, around 2 mm, on aquatic plants. Most morphometric characters show size-specific trends, which complicate comparisons among populations or with other species. Of specific systematic importance is the relative position of the anal and dorsal fins, which remains almost unaltered throughout development, and allows safe distinction from A. fasciatus. The species was found mostly in deep areas with clean and slow running water, usually associated with freshwater springs. Rich submerged vegetation is the prominent ecological feature of all sites in which the species was found. Using as criteria of rarity the limited geographic distribution, the confinement of the species in few localities of each aquatic system and the low local densities, V. letourneuxi can be characterised as a "restricted and locally rare species". The restricted distribution, coupled with the narrow ecological requirements, makes the species vulnerable to extinction. Its disappearance from at least four aquatic systems and the serious population decline in a number of other systems seems to be connected with habitat loss or degradation caused by human activities. The prospects of conservation are discussed.