The phylogenetic relationships and taxonomy of the spirlins in the genus Alburnoides are examined by comparative sequencing analysis of mitochondrial and nuclear markers. Molecular analyses revealed 17 Eurasian lineages divided into two main clades, termed the Ponto-Caspian and European in accordance with the lineage distribution. The indel diagnostics of β-actin and S7 markers and translation of cyt b to the amino acid chain were evaluated as a reliable identifying tool for most of the recognised lineages. Lineage richness is closely connected with the existence of known glacial refugia in most cases. The underestimation of species richness in the genus Alburnoides is confirmed: the genetic analyses support the validity of 11 morphologically accepted species; apart from them, four phylogenetic lineages requiring descriptions as separate species were revealed. The distribution area of the nominotypical species A. bipunctatus s. stricto is newly defined. Two diverging phylogenetic lineages, A. ohridanus, and A. prespensis complex, were observed in the Southeast Adriatic Freshwater Ecoregion, confirmed as a hotspot of endemic biodiversity. A. ohridanus demonstrates high divergence from the A. prespensis complex, represented by three similar mitochondrial lineages with the same nuclear haplotypes and sympatric occurrence. The range restricted endemism was confirmed for at least seven species. The Albanian river systems, as well as the wider Ponto-Caspian basin exhibit complications among definite species delineations and gaps in understanding of microevolutionary processes; these areas require further investigations.

Agricultural University of Tirana Tirana Albania

Faculty of Science Charles University Viničná 7 12844 Praha Czech Republic

Hellenic Centre for Marine Research Institute of Marine Biological Resources and Inland Waters 46 7 Km Athens Sounio Ave 19013 Anavissos Attiki Greece

Institute of Vertebrate Biology of the Academy of Sciences of the Czech Republic v v i Květná 8 60365 Brno Czech Republic

National Museum of Natural History Bulgarian Academy of Sciences Tsar Osvoboditel Blvd 1 1000 Sofia Bulgaria

University of Agriculture in Kraków Department of Ichthyobiology and Fisheries T Spiczakowa 6 30 199 Kraków Poland

University of Prešov Department of Ecology 17 Novembra 1 08116 Prešov Slovakia

Zoological Museum Bolshaya Nikitskaya 6 125009 Moscow Russia

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