The relationship between the North American bumble bee species Bombus moderatus Cresson and the European and Asian species in the subgenus Bombus sensu stricto has long been in question. Bombus moderatus has either been regarded as a distinct species or has been synonymized with B. lucorum (L.). We surveyed 10 Bombus s.str. species at 26 enzyme loci, using horizontal and vertical starch-gel electrophoresis. We found that B. moderatus differs from B. lucorum at three loci but is identical in these and all other loci surveyed with B. cryptarum (Fabricius) and B. magnus Vogt. The present distribution of B. moderatus, together with the observation that its closest relatives are the European B. cryptarum and B. magnus and the Asian B. hypocrita Pérez, suggests that differentiation occurred after dispersal to North America via Beringia.