A hazard assessment was conducted based on information derived from two reproduction studies conducted with endangered razorback suckers (Xyrauchen texanus) at three sites near Grand Junction, CO, USA. Selenium contamination of the upper and lower Colorado River basin has been documented in water, sediment, and biota in studies by US Department of the Interior agencies and academia. Concern has been raised that this selenium contamination may be adversely affecting endangered fish in the upper Colorado River basin. The reproduction studies with razorback suckers revealed that adults readily accumulated selenium in various tissues including eggs, and that 4.6 μg/g of selenium in food organisms caused increased mortality of larvae. The selenium hazard assessment protocol resulted in a moderate hazard at the Horsethief site and high hazards at the Adobe Creek and North Pond sites. The selenium hazard assessment was considered conservative because an on-site toxicity test with razorback sucker larvae using 4.6 μg/g selenium in zooplankton caused nearly complete mortality, in spite of the moderate hazard at Horsethief. Using the margin of uncertainty ratio also suggested a high hazard for effects on razorback suckers from selenium exposure. Both assessment approaches suggested that selenium in the upper Colorado River basin adversely affects the reproductive success of razorback suckers.