In-vivo and in-vitro tests to assess toxic mechanisms of nano ZnO to earthworms

Abstract Increasing applications of engineered nanomaterials lead to the release of nanoparticles into various environmental media, especially soil. However, the environmental behavior of nano ZnO in soil and the toxic mechanism to terrestrial invertebrates were not fully understood. In this study, the concentrations of nano ZnO in earthworms ( Eisenia fetida ) were measured to assess its bioaccumulation. The ratio of nano ZnO in earthworms to soil in 250 mg/kg treatment group was lower than that in 10 mg/kg treatment group as the earthworms would not take up too much nano ZnO to protect themselves from the damage. Combination of in-vivo and in-vitro tests was adapted to investigate the toxic mechanism of nano ZnO to earthworms. In in-vivo test, biomarkers including ROS, SOD, and MDA suggested that the toxic effects of nano ZnO to earthworms were caused by the oxidative stress. To further elucidate its toxic mechanism, in-vitro toxicity test was carried out by employing earthworm coelomocytes. The biomarkers, intracellular ROS, extracellular LDH, and cell viability showed concentration-dependent manner with nano ZnO in the culture media, demonstrating that in-vitro toxicity test could be utilized to reveal the toxic mechanism of nano ZnO to earthworms or other organisms.