The aim of this study was to evaluate the recovery and survival of four species of unionid mussles [pimpleback, Quadrula pustulosa pustulosa (I. Lea, 1831); spike, Elliptio dilatata (Rafinesque, 1820); Higgins eye, Lampsilis higginsii (I. Lea, 1857); and pocketbook, Lampsilis cardium (Rafinesque, 1820)] that were experimentally relocated to in situ refugia in the St Croix River of Minnesota and Wisconsin, USA. In 1996, 150 mussels of each of the first three species (450 total) were relocated to three 5 × 5 m study grids (Site A), one near Lakeland, Minnesota, which served as a source-site control, and two in the experimental refuge 48 km upstream, near Franconia, Minnesota. In a second relocation in 1997, L. cardium was substituted for L. higginsii and 150 mussels of this and each of the other two species (450 total), were relocated to two study grids (Site B). The source site control was near Sunrise, Minnesota and the experimental refuge was 14 km downstream near Almelund, Minnesota. Mussel recovery, survival and substratum characteristics were evaluated annually at Site A for 2 years and for 3 years at Site B. Mean annual recovery of all three species ranged from 90 to 100% at Site A, and from 34 to 70% at site B. The mean annual survival of recaptured mussels ranged from 85 to 100% at Site A, and from 88 to 100% at Site B. The textural characteristics of the substratum differed significantly between the control and the two refuge locations at the beginning of the study, but did not differ from this initial status among subsequent years at Site A. At Site B, there was a significant shift in textural characteristics from large to smaller fractions over the four years. The relatively high survival of mussels during this study demonstrates the importance of proper handling and transport protocols when relocating mussels and the selection of suitable relocation habitat with stable substratum. When established correctly, in situ refugia may be a viable tool for preserving unionid mussels.