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The intensive speciation in this region might be due to the presence of numerous lakes which serve as glacial refugia during the Late Pleistocene, connections and disconnections of river basins, and variability of recolonization routes in the past 10, 14, 15. The most frequently found species in Central Europe are B. cederströmii at least partly overlap, and also some hybrids between these species were detected based on morphological analyses 10, 13. Most Bythotrephes species and forms are concentrated in north and central Europe, which coincides with the region of the maximum Pleistocene glaciation 10. represents the relict populations in the prealpine regions 11, 12. However, the recent detailed morphological revision of this genus clearly distinguished at least seven morphospecies spread throughout Eurasia 10, 11. The most widespread view of recent decades has been that Bythotrephes is a monotypic genus, represented by only a single morphologically highly variable species, B. The taxonomic status of the genus Bythotrephes has been debated since the second half of the XIX century 8, 9, 10. In Europe, Bythotrephes has also invaded lakes and reservoirs in the non-glaciated part of Germany, Belgium, Netherland 5, 6, but generally it is more common in Central Europe 7. In 1970s or 1980s it invaded North American lakes and is currently considered as one of the most invasive zooplankton species 3, that has established in more than 150 lakes in Canada and the United States 4. Its patchy distribution in Central Europe and in prealpine lakes is typical for ice age relicts 1, 2. The spiny waterflea genus Bythotrephes Leydig (Crustacea: Cladocera) is a predatory, cladoceran native to Northern Eurasia, where inhabits mostly oligo- and mesotrophic lakes. Furthermore, this haplotype was the source variant from which most other haplotypes were derived. Nonetheless, we have identified 29 new haplotypes of mtDNA COI, and one which was the same as the haplotype found in North America and Finland. On the other hand, the results of our work could point out that the COI gene is insufficient to evaluate the taxonomic status of Bythotrephes. There were no significant differences between all analyzed sequences, which supports the hypothesis that Bythotrephes is a monotypic genus, with only one highly polymorphic species. For the genetic analysis, we used newly obtained 113 sequences of mtDNA COI gene of the 535-bp length Bythotrephes from Central Europe and sequences downloaded from GenBank. Based on the morphology we identified two species in Central Europe, B. Therefore, we performed a multi-lake survey in Central Europe to give new insight into the taxonomic status of Bythotrephes by combining genetic analysis with traditional morphology-based taxonomy. However, the recent detailed morphological revision of this genus clearly distinguishes at least seven species.
The most widespread view of recent decades has been that Bythotrephes is a monotypic genus, which was support by preliminary molecular data. The taxonomic status of the genus Bythotrephes Leydig (Crustacea: Cladocera) has been debated since the second half of the XIX century.
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