Early ecotoxic effects of ZnO nanoparticle chronic exposure in Mytilus galloprovincialis revealed by transcription of apoptosis and antioxidant-related genes

Recently, China became one of the largest nanomaterial markets in the world. The wide use of ZnO nanoparticles in a number of products implies an increasing release in marine environment and consequently the evaluation of the potential effects upon marine organisms largely cultured in China for commercial purposes, such as invertebrate bivalves is a current need. To this aim, survival, bioaccumulation, and transcription pattern of key genes, p53, PDRP, SOD, CAT, and GST, involved in DNA damage/repair and antioxidation, in Mytilus galloprovincialis digestive gland, exposed to ZnO NPs (<100 nm) and ZnO bulk (150-200 nm) for 4 weeks, were evaluated. ZnSO₄ was also assessed to appraise the role of zinc ions. Starting from 72 h, increasing mortality values along the exposure time were observed for all ZnO compounds. The highest difference was evident after 28 d when NPs resulted three times more toxic than bulk, (LC₅₀) = 0.78 mg Zn/L (confidence limits: 0.64, 1.00) and 2.62 mg Zn/L (confidence limits: 1.00, 4.00), respectively. For ZnSO₄ the (LC₅₀) was always the lowest reaching the minimum value at 28 d 0.25 mg Zn/L (confidence limits: 0.10-0.40). Digestive gland showed higher uptake rate of ionic Zn respect to ZnO NPs and bulk during the first three days of exposure. In particular at the end of the exposure time (28 d) at 1 mg Zn/L the rank of Zn uptake rate was Zinc ion > ZnO NPs > ZnO bulk. The relative expression of investigated genes evidenced that distinct actions of apoptosis and antioxidation occurred in M. galloprovincialis exposed to ZnO NPs with a peculiar pattern dependent on exposure time and concentration. Application of the qRT-PCR technique revealed evidence of sensitivity to the nanomaterial since the first time of exposure.