Since 2011 the Joint Oil Sands Monitoring (JOSM) program has been conducted in the lower Athabasca River by the Governments of Canada and Alberta to assess the freshwater health in areas associated with oil sands development. The majority of the benthic invertebrate assemblage of the Athabasca River and its tributaries are Chironomidae larvae. Assessments of such benthic assemblages are made difficult because the identification of Chironomidae larvae is costly and time consuming. To facilitate this identification process, we aimed to develop a simple taxonomic key for Chironomidae larvae of this region. This taxonomic reference and identification key makes use of the known taxonomic details on these Chironomidae species. Moreover, we provide details on their geographical distribution, ecology, habitats, environmental tolerance values for species, and trait-based morphological characters. Our main goal was to make this information readily available to both non-specialists and specialists so that biomonitoring programs can more readily utilise these organisms in biomonitoring.