In order to estimate acute-to-chronic toxicity ratios (ACRs) relevant to a coldwater stream community, we exposed rainbow trout (Oncorhynchus mykiss) to cadmium (Cd), lead (Pb), and zinc (Zn) in 96-h acute and 60+ day early-life stage (ELS) exposures. We also tested the acute and sublethal responses of a mayfly (Baetis tricaudatus) and a midge (Chironomus dilutus, formerly C. tentans) with Pb. We examine the statistical interpretation of test endpoints and the acute-to-chronic ratio concept. Increasing the number of control replicates by 2 to 3x decreased the minimum detectable differences by almost half. Pb ACR estimates mostly increased with increasing acute resistance of the organisms (rainbow trout ACRs ?? mayfly < Chironomus). The choice of test endpoint and statistical analysis influenced ACR estimates by up to a factor of four. When calculated using the geometric means of the no- and lowest-observed effect concentrations, ACRs with rainbow trout and Cd were 0.6 and 0.95; Zn about 1.0; and for Pb 3.3 and 11. The comparable Pb ACRs for the mayfly and Chironomus were 5.2 and 51 respectively. Our rainbow trout ACRs with Pb were about 5-20x lower than earlier reports with salmonids. We suggest discounting previous ACR results that used larger and older fish in their acute tests. ?? 2007 GovernmentEmployee: U.S. Geological Survey.