A sub-individual multilevel approach for an integrative assessment of CuO nanoparticle effects on Corbicula fluminea.

Because they are widely used, copper oxide nanoparticles (CuO NPs) are likely to enter the aquatic environment and then reach the sediment. We have examined the effect of CuO NPs in the freshwater endobenthic bivalve Corbicula fluminea. Some previous studies have investigated effects at biochemical and physiological levels, but molecular endpoints are still poorly studied despite they are sensitive in early detection of NPs effect. In the present study, we have investigated short-term effects (96 h) of CuO NP (12, 30 nm; 0, 20 and 100 µg/L) using molecular endpoints as well as more conventional biochemical and physiological markers. The expression of antioxidant (CuZnSOD, MnSOD, Cat, Se-GPx, Trxr) and antitoxic (GST-Pi, HSP70, MT, Pgp, MRP1) related genes was measured at the mRNA level while antioxidant (SOD, TAC) and antitoxic (GST, ACP) defenses, energetic reserves and metabolism (ETS, Tri, LDH), and cellular damages (LPO) were assessed using a biochemical approach. The filtration rate measured at 96 h provided information at the physiological scale. Gene expression and filtration rate were responsive to CuO NPs but the effects differed according to the NP size. The results suggest that defense mechanisms may have been set up following 30 nm-NP exposure. The response to 12 nm-NP was lower but still showed that exposure to 12 nm-NP led to activation of cellular elimination mechanisms. The lowering of the filtration rate may have protected the organisms from the contamination. However, this raised the question of further repercussions on organism biology. Together, the results (i) indicate that CuO NP may exert effects at different levels even after a short-term exposure and (ii) point out the precocity of molecular response.

Corbicula fluminea gene expression modulated by CeO2 nanomaterials and salinity.

Cerium dioxide nanomaterials (CeO2 NMs) are used in different fields and incorporated in daily products. Several studies highlighted their effects on organism physiology, although molecular studies remain scarce. NM behavior is strongly dependent on the environment but few data are available using complex exposure media, raising the question of its environmental impacts. The aim of the present work was to assess the toxic potential of three CeO2 NMs in Corbicula fluminea at a molecular level by RT-qPCR under a more realistic scenario of exposure, in a multistress context at two different salinities (1.5 and 15 psu). C. fluminea was exposed for 28 days to pulses of the three selected NMs (reference, manufactured, and aged manufactured). In bivalves, the gills and digestive gland are two key organs used for ecotoxicological studies. The expression change of 12 genes was measured in control organisms after 28 days in both organs, allowing us to clearly separate the responses for both organs and salinities. As gills come in contact with the environment first, we monitored gene the expression at intermediate time points (7, 14, and 21 days) for this organ in order to highlight clams responses to NM and salinity. Two genes (Se-GPx, MnSOD) had a salinity-dependent level of expression. HSP70, Se-GPx, and Trxr mRNAs presented significant changes in their expressions in the presence of NM. This study was completed using an integrated statistical approach. The exposed organisms differed more from control at field salinity than those exposed to hyper-saline conditions. At 15 psu, salinity pressure seems to cause the first molecular impact. At 1.5 psu, gene expression patterns allowed the effect of each NM to separate clearly. These results confirmed the usefulness of gene expression studies. Moreover, we highlighted the necessity to assess the environmental toxicity of the different forms of manufactured NM.

Genotoxicity and physiological effects of CeO2 NPs on a freshwater bivalve (Corbicula fluminea).

The rapid development of nanotechnology and the increased use of nanomaterials in products used in everyday life have raised the question of the potential release of nanoparticles into the aquatic environment. Their fate and effects in natural ecosystems are not currently well understood but harmful effects of nanoparticles have been demonstrated at low concentrations on some freshwater and marine species. Cerium dioxide nanoparticles (CeO2 NPs) are produced in large quantities and used in products in many different fields, such as automotives or optics. Because of their widespread use in daily products, CeO2 NPs are included in the OECD priority list of manufactured nanomaterials for human and environmental assessment. Indeed some studies have been conducted to assay various enzymatic biomarkers, which showed the CeO2 NPs potential to modify anti-oxidative defenses and cellular membrane stability. Nevertheless, only a few studies were performed on their genotoxic potential. The aim of this work was to evaluate the genotoxic and physiological effects of CeO2 NPs on a widespread freshwater bivalve Corbicula fluminea by using comet assay and a multi-enzymatic biomarker approach. Exposure to two CeO2 NP concentrations during a short term experiment (6 days) was set up. The first one (10 µg/L) was chosen in order to work with low but measurable concentrations whereas the second one was ten times higher (100 µg CeO2 NPs/L). DNA damage was significantly more pronounced compared with control for both concentrations tested as early as two days of exposure and seemed to increase with time. Some enzymatic biomarkers of anti-oxidative defenses (total antioxidant capacity, catalase activity), anti-toxic mechanisms (glutathione-S-transferase activity, caspase-3 activity) or metabolism (lactate dehydrogenase activity) tended to increase after 6 days of exposure but only the induction of caspase pathway and DNA damages appeared significant for exposed organisms. In this study, time and concentration effects of CeO2 NPs were highlighted by coupling genotoxic and cellular biomarker assessments.