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The zebra mussel, Dreissena polymorpha is an aquatic nuisance invasive species originally native to the Ponto-Caspian region where it is found in lakes and delta areas of large rivers draining into the Black and Caspian seas. The dispersal of D. polymorpha began at the end of the 18th century, at a time when shipping trade become increasingly important and many canals were built for linking different navigable river systems in Europe. Over the past 200 years, zebra mussels spread to most of the lakes, rivers and waterways in Europe by a combination of natural and anthropogenic dispersal mechanisms. D. polymorpha invaded Spain around 2001, being found for the first time in the Riba-roja reservoir at the lower part of the Ebro River, North-East Spain. The relatively late invasion of Spain was most likely caused by the presence of the Pyrenees, which isolated the Iberian Peninsula from the rest of the European continent, and acted as a barrier to the dispersal of D. polymorpha. In recent studies, molecular genetic methods have successfully been used to determine phylo-geographic relationships, which may reflect invasion corridors and can help retrace source populations. Zebra mussels from populations in Great Britain, The Netherlands, Belgium, France, Germany, Spain, Italy, Romania and North America were analyzed using PCR based amplified fragment length polymorphism (AFLP)-fingerprinting to determine the source population of D. polymorpha in Spain. The phylogenetic analyses and pair-wise genetic distances revealed that the recent invasion of zebra mussels in Spain is most likely from France.