In this study, three different sizes of cerium oxide (CeO₂) nanoparticles were synthesized and exposed to Caenorhabditis elegans to investigate the potential harmful effect of CeO₂ nanoparticles on the environment. The effects of the CeO₂ nanoparticles on C. elegans were assessed at multiple levels, such as with respect to stress response gene expression, growth, reproduction and mortality. Moreover, to test the ecotoxicological relevance of CeO₂-induced gene expression. The overall results suggest that CeO₂ nanoparticles may provoke ecotoxicity in C. elegans especially with respect to gene expression, reproduction and survival, which can com-prise an important contribution to knowledge on the ecotoxicity of CeO₂ nanoparticles, about which little data are available. This is particularly valuable in the biomarker research on ecotoxicology, as ecological relevance is a crucial criterion for the applicability of the biomarker in field biomonitoring and ecological risk assessment.