Molecular analysis reveals multiple native and alien Phoxinus species (Leusciscidae) in the Netherlands and Belgium

Fresh waters are among the most endangered ecosystems, one of the problems being the lack of data on biodiversity. In the center of the missing knowledge are cryptic species, two (or more) species classified as a single one due to their (seemingly) indistinguishable morphology. Lack of research and stabilizing selection are reflected in the cryptic diversity of the genus Phoxinus (Leusciscidae), the studies of which have intensified over the last two decades and reveal undetected taxonomic complexity. Moreover, some of the Phoxinus lineages act as invasive species, while others are endangered by their alien counterparts. Minnows have been intentionally (as food for predatory fish species) or unintentionally (with other fries) stocked causing hybridisation zones in Norway, Portugal, Spain, France, Italy, Germany and Austria. Given that genetic identity and lineage assignment of Phoxinus from Belgium and the Netherlands have not been researched, the goal of the study was to examine available samples from known localities in the area in order to infer- whether they are native or not. For this purpose, the barcoding region cytochrome oxidase I , another mitochondrial gene cytochrome b , a nuclear recombination activating gene 1 and a combination of these markers from a wider neighboring region were analyzed. The study found four different Phoxinus species/lineages occurring in Belgium and the Netherlands: P. phoxinus , P. csikii, P. septimaniae and genetic lineage 11 (possibly P. cf. morella ). While the first seem to be native, the other three were probably introduced.