The Chinese mitten crab (Eriocheir sinensis) is an invasive organism of concern, with established non-native populations in Europe and California, USA. The species is thought to pose a risk to other North American waterways, including the Great Lakes and St. Lawrence Seaway. Since 1965, there have been sixteen confirmed adult E. sinensis caught in the North American Great Lakes or adjoining waterways. Analysis of their mitochondrial DNA sequence variation for part of the cytochrome c oxi-dase subunit I gene discerned three haplotypes among seven individuals (caught between 1973 and 2005), identical to common haplotypes in Europe. Analysis of mitochondrial haplotype frequencies and shipping patterns suggests that E. sinensis has been introduced to the Great Lakes from Europe, although we are unable to preclude native Asian populations as putative sources. The species is catadromous, migrating between salt and fresh water to complete its life cycle. This trait makes it unlikely that E. sinensis will establish a breeding population in the Great Lakes proper, which are separated from saltwater by a considerable distance and significant instream barriers such as waterfalls and navigation locks. However, the recent discovery of two confirmed mitten crabs in the St. Lawrence River, which could be more readily colonized, underscores the risk posed by the repeated introduction of this species into the Great Lakes and St. Lawrence Seaway.