Global environmental monitoring has indicated that the structure and function of some aquatic ecosystems has been significantly altered by human activities. There are many potential causes for these changes; however, one major concern is the increasing release of anthropogenic contaminants into aquatic environments. Although toxicological responses of individual organisms are typically well characterized, few studies have focused on characterizing toxicity at the ecosystem level. In fact, because of their scale and complexity, changes in ecosystem integrity are rarely considered in assessments of risks to ecosystems. This work attempts to move the conversation forward by defining integrity of ecosystems, reviewing current and historical approaches to measuring ecosystem integrity status (e.g., structural and functional measurements), and highlighting methods that could significantly contribute to the field of ecosystem toxicology (e.g., keystone species, environmental energetics, ecotoxicological modeling, and adverse outcome pathways [AOPs]). Through a critical analysis of current and historical methodologies, the present study offers a comprehensive, conceptual framework for the assessment of risks of contaminant exposure for whole ecosystems and proposes steps to facilitate better diagnoses of the integrity of aquatic systems. Environ Toxicol Chem 2019;38:289-301. C 2018 SETAC Environmental Toxicology and Chemistry, 2019;38:289-301-E.M. Maloney C 2018 SETAC wileyonlinelibrary.com/ETC