We conducted multiple toxicity tests to (1) evaluate the survival and growth of juvenile mussels (Fatmucket, Lampsilis siliquoidea) in a standard 4-week and a longer-term 12-week exposures following a method refined from the standard method using two common reference toxicants (potassium chloride [KCl] and nickel [Ni]); (2) evaluate reproductive endpoints from reproductive KCl and Ni toxicity tests. Reproductive toxicity tests started with Fatmucket brooding mature glochidia to five concentrations of K and Ni for 6 weeks. Subsamples of glochidia were then removed from the adults to determine (1) the viability of brooded glochidia; (2) the viability of free glochidia in a 24-h additional exposure; and (3) success of glochidia parasitism. In this data release we report individual biological, chemical, and water quality measurements from each toxicity test.