Influences and mechanisms of nanoparticles on pentachloronitrobenzene accumulation by earthworms

Pesticides and nanoparticles may coexist in soil; however, influences of nanoparticles on accumulation of pesticides in terrestrial organisms are still unclear. This study aims to investigate the influences and mechanisms of metal oxide nanoparticles (nano ZnO and nano CuO) on accumulation of pentachloronitrobenzene (PCNB) in earthworms and their combined toxicity. The earthworms were cultivated in the soil spiked with nanoparticles (10, 50, 250 mg/kg) and PCNB (100 μg/kg) for 21 days. The concentrations of PCNB in earthworms in binary exposure treatments (PCNB + ZnO and PCNB + CuO) reached 2.47 and 3.13 times of that in individual PCNB exposure treatment, indicating that nanoparticles facilitated the accumulation of PCNB in earthworms. The contents of reactive oxygen species (ROS) in earthworms in treatments PCNB + ZnO 250 and PCNB + CuO 250 reached 379 and 316 fluorescence intensity/mg Protein, respectively, which were significantly higher than that in control group (183 fluorescence intensity/mg protein), indicating that nanoparticles would cause oxidative stress to earthworms. Earthworm coelomocytes were extracted from healthy earthworms and cultivated in culture media in cytotoxicity tests. Changes of intracellular ROS contents and cell viability suggested that PCNB and nanoparticles caused serious oxidative damage to earthworm coelomocytes, thus leading to the damage of cell membrane and cell death. In in vivo tests, changes of biomarkers (ROS and malondialdehyde) demonstrated that these pollutants injured the earthworms. Increased accumulation of PCNB in binary exposure treatments was due to the damage of body cavity caused by nanoparticles. This study provides a novel hypothesis for nanoparticles facilitating organic pollutants entering terrestrial organisms and determines whether nanoparticles would bring about greater environmental risks of other pollutants.