A dike failure at the Tennessee Valley Authority Kingston Fossil Plant in East Tennessee, United States, in December 2008, released approximately 4.1 million m(3) of coal ash into the Emory River. From 2009 through 2012, samples of mayfly nymphs (Hexagenia bilineata) were collected each spring from sites in the Emory, Clinch, and Tennessee Rivers upstream and downstream of the spill. Samples were analyzed for 17 metals. Concentrations of metals were generally highest the first 2 miles downstream of the spill, and then decreased with increasing distance from the spill. Arsenic, B, Ba, Be, Mo, Sb, Se, Sr, and V appeared to have strong ash signatures, whereas Co, Cr, Cu, Ni, and Pb appeared to be associated with ash and other sources. However, the concentrations for most of these contaminants were modest and are unlikely to cause widespread negative ecological effects. Trends in Hg, Cd, and Zn suggested little (Hg) or no (Cd, Zn) association with ash. Temporal trends suggested that concentrations of ash-related contaminants began to subside after 2010, but because of the limited time period of that analysis (4 yr), further monitoring is needed to verify this trend. The present study provides important information on the magnitude of contaminant exposure to aquatic receptors from a major coal ash spill, as well as spatial and temporal trends for transport of the associated contaminants in a large open watershed.