"Nanosize effect" in the metal-handling strategy of the bivalve Scrobicularia plana exposed to CuO nanoparticles and copper ions in whole-sediment toxicity tests

Highlights: • CuO NPs exert a specific toxic effect compared to their bulk counterpart. • Different subcellular metal handling strategies depending on the metal form • Delayed toxic effect in clams exposed to sediment-associated CuO NPs To date, the occurrence, fate and toxicity of metal-based NPs in the environment is under investigated. Their unique physicochemical, biological and optical properties, responsible for their advantageous application, make them intrinsically different from their bulk counterpart, raising the issue of their potential toxic specificity or "nanosize effect". The aim of this study was to investigate copper bioaccumulation, subcellular distribution and toxic effect in the marine benthic species Scrobicularia plana exposed to two forms of sediment-associated copper, as nanoparticles (CuO NPs) and as soluble ions (CuCl₂). Results showed that the exposure to different copper forms activated specific organism's metal handling strategies. Clams bioaccumulated soluble copper at higher concentrations than those exposed to sediment spiked with CuO NPs. Moreover, CuO NPs exposure elicited a stronger detoxification response mediated by a prompt mobilization of CuO NPs to metal-containing granules as well as a delayed induction of MT-like proteins, which conversely, sequestered soluble copper since the beginning of the exposure at levels significantly different from the control. Eventually, exposure to high concentrations of either copper form led to the same acute toxic effect (100% mortality) but the outcome was delayed in bivalves exposed to CuO NPs suggesting that the mechanisms underlying toxicity were copper form-specific. Indeed, while most of soluble copper was associated to the mitochondrial fraction suggesting an impairment of the ATP synthesis capacity at mitochondrial level, CuO NPs toxicity was most likely caused by the oxidative stress mediated by their bioaccumulation in the enzymatic and mitochondrial metabolically available fractions.