While the Strategic Roadmap focuses primarily on human health, ICCVAM also recognizes the need to implement non-animal approaches for ecotoxicity testing. Development and implementation of alternative approaches to reduce and replace animal use for this testing will involve four key steps: (1) defining testing needs, (2) identifying the available alternative approaches, (3) developing defined approaches to testing and assessment, and (4) addressing both scientific and non-scientific (including regulatory) challenges. Accordingly, in 2018 ICCVAM established an ecotoxicology workgroup to provide expertise in identifying and evaluating alternative approaches to identify ecological and environmental hazards using in vitro and/or in silico methods. The workgroup is sponsored by DoD, DOI, and EPA and has representatives from seven ICCVAM agencies. Members are currently compiling a summary of ecotoxicity tests that require the use of animals, emerging technologies for ecotoxicity and environmental safety, and the utility of those technologies in regulatory testing. This summary, planned for publication in 2020, will help advance the development and evaluation of defined approaches for screening, testing, and assessment of relevant endpoints.