The U.S. Geological Survey and collaborators from EcoAnalysts, Inc., completed field and laboratory studies during 2016–19 to evaluate the toxicity of metals to freshwater mussels in streams draining the Tri-State Mining District. This project consisted of (1) sampling and analysis of metals in water and sediment, (2) surveys of mussel assemblages at sites with suitable mussel habitat, (3) toxicity tests with juvenile mussels exposed to zinc or to a mixture of metals (zinc, lead, and cadmium) in water, and (4) toxicity tests to evaluate the contributions of metals in sediment and metals in overlying water to toxic effects on mussels. Field sampling at sites in the Spring River and Neosho River and their tributaries demonstrated wide ranges of metal contamination in water and sediment. Zinc was the predominant toxic metal in water, and concentrations of lead and cadmium were much lower. Mussel areal density and species richness were greater at reference sites with low sediment metal concentrations (for example, zinc, 29–141 micrograms per gram) than at test sites that had higher concentrations of sediment zinc (416–3,420 micrograms per gram) as a result of effects of upstream mining activity. Juvenile mussels were highly sensitive to zinc in water in 12-week toxicity tests compared to previous water-only tests, and adding low levels of waterborne lead and cadmium typical of their occurrence in Tri-State Mining District streams produced greater toxicity. Thresholds for mussel toxicity were at or less than waterborne metal concentrations detected in Tri-State Mining District streams, and sites with waterborne metal concentrations exceeding thresholds had decreased mussel density and decreased mussel species richness. The 12-week toxicity tests with juvenile mussels in Tri-State Mining District sediments also demonstrated negative mussel responses with metal exposure. Thresholds for reductions in survival, growth, or biomass were at sediment metal concentrations less than thresholds reported for previous 4-week tests. We documented strong associations between reduced survival in laboratory tests and reduced species richness in community surveys. Attempts to estimate combined toxicity thresholds for metals in sediment and overlying water were not successful. These inconclusive results may be attributable to several factors, including (1) unexpected losses of waterborne metals from solution, (2) differences in sensitivity of different age/size classes of juvenile mussels, (3) disruption of sediment-water equilibria and changes in metal bioavailability, and (4) behavioral or physiological responses allowing juvenile mussels to temporarily reduce or avoid metal exposure. We also observed differences in metal toxicity thresholds between sediment toxicity tests started with different ages/sizes of test organisms. A followup study that combined exposure to Tri-State Mining District sediments with exposures to multiple levels of waterborne metals demonstrated toxic effects of sediments with low metal concentrations; however, some treatments also indicated unexpected reversals of concentration-response trends and reduced toxicity in treatments that had high metal concentrations in overlying water. These unusual responses may reflect development of physiological tolerance to metal toxicity by induction of metal-binding proteins (for example, metallothionein) in response to high metal levels in water.

Results of laboratory and field studies indicated strong associations between metal exposure in Tri-State Mining District streams and toxic effects on juvenile freshwater mussels. Mussel community characteristics corresponded to differences in metal concentrations in sediment and water among Tri-State Mining District sampling sites. Responses of juvenile mussels in 12-week water and sediment exposures were strongly correlated with the status of mussel assemblages in Tri-State Mining District streams. The combined results support the hypothesis that exposure to metals from historical mining activities adversely affects freshwater mussel communities in the Spring River/Neosho River drainage.