This survey provides information on sediment toxicity and structural characteristics of the macroinvertebrates and benthic phytoplankton at 10 locations in the Cache la Poudre River after long-term exposure to coal tar sediment. The application of the Australian river bioassessment system (AUSRIVAS) as well as a biotest battery was used to evaluate the river ‘health’ condition. Coal tar is a dense nonaqueous-phase liquid of significant environmental concern due to its toxicity and persistence in the subsurface. Organisms like Selenastrum capricornutum, Daphnia magna and Chironomus tentans, representing different complexities in the biosphere, were selected as test systems for ecotoxicological studies. The results obtained in this study indicate that a biotest battery, macroinvertebrate and benthic phytoplankton communities are in principle suitable biological tools for evaluation of toxic oil and coal-derived substances in long-term contaminated river sediment.