Microplastics have been found in freshwater systems, and in turn have been detected in freshwater bivalves. However, there is limited research that defines the toxicity of bicroplastics to native freshwater bivalves that have long been imperiled in North America. Our objective was to determine whether a suite of pristine microplastics has an adverse effect on two early life stages of unionid freshwater mussels. Glochidia of Lampsilis fasciola (a Canadian species at risk) and Lampsilis siliquoidea (widespread across Canada) were individually exposed to spheres of polystyrene (6 and 90 μm), polyethylene (28, 90, and 1000 μm), and cellulose acetate (1000 μm), as well as fibers of polyethylene terephthalate (60 μm). After 24 h, there was no significant decrease in glochidia viability in either species. Juvenile L. siliquoidea mussels were also exposed to spheres of polystyrene (6 and 90 μm) and polyethylene (28 μm), and fibers of polyethylene terephthalate (60 μm) in individual 28-day subchronic tests followed by a 7-day depuration period. Burial was assessed weekly, and ingestion of each microplastic was compared in nondepurated and depurated mussels. There was no sustained effect on juvenile burial with any microplastic tested. Ingestion of microplastics was concentration dependent, and depuration occurred for all particles and size ranges tested. The results suggest that pristine microplastics were not acutely toxic to the early life stages of these freshwater mussels, but that the energetic costs associated with particle uptake and depuration, which were not measured in our study, may have an impact on fitness that warrants further investigation. In addition, testing with other shapes and polymers of microplastics typically detected in the environment is recommended. Environ Toxicol Chem 2024;43:2557-2568. © 2024 The Author(s). Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.