With the advancement in nanotechnology, particularly the use of TiO2 nanoparticles (NPs), there is a need to study their release into the environment and assess the related risk in an environmentally relevant contamination scenario. In the present study, the transfer and toxicity of TiO2 NPs in microcosms mimicking terrestrial and aquatic ecosystems were evaluated. The contaminated soil was prepared by spiking natural soils, with these then used as the basis for all exposure systems including preparation of soil leachates for amphibian exposure. Results demonstrated significant reductions in bacterial (-45%) and archaeal (-36%) nitrifier abundance; significant translocation of Ti to M. truncatula leaves (+422%); significant reductions in plant height (-17%), number of leaves (-29%), and aboveground biomass (-53%); nonsignificant Ti uptake in snail foot and viscera, and excretion in feces; and genotoxicity to X. laevis larvae (+119% micronuclei). Our study highlights a possible risk of engineered TiO2 NPs in the environment in terms of trophic transfer and toxicity in both terrestrial and aquatic environments.