The Athabasca River basin, located in Alberta, Canada, covers 157, 000 km(2) and holds significant cultural and economic importance. Recent research assessed changes in several water quality and quantity parameters that have changed both spatially (along the river continuum) and temporally (pre-development and present day) in the Athabasca River Basin. In particular, parameters such as salinity and dissolved sulphate have changed significantly across the Athabasca River mainstem over the past five decades. Further laboratory testing has linked concentrations of these parameters to changes in fathead minnow reproduction. Research is required to determine whether these changes observed in the laboratory can be applied to actual in-river conditions. The objectives of the present study were to twofold: assess changes in fathead minnow response metrics (i.e., condition, liver and gonad size, egg production, and gill histology) associated with increasing concentrations of salinity and dissolved sulphate and determine whether sublethal effect thresholds established in laboratory experiments correspond to actual in-river concentrations using water from the mouth and headwaters of the Athabasca River. Three dose-response experiments (NaCl, SO4, and water sampled from the mouth of the Athabasca River) were conducted at Jasper National Park, Alberta, Canada. Significant increases in mean eggs per female per day occurred at the 50% treatment for the mouth experiment and thresholds previously developed in the laboratory were verified.