Molecular, biochemical and behavioral responses of Daphnia magna under long-term exposure to polystyrene nanoplastics.

The presence and potential toxicity of nanoplastics (NPs) in aquatic ecosystems is an issue of growing concern. Although many studies have investigated the adverse effects of short-term exposure to high concentrations of NPs to aquatic organisms, the information on the consequences caused by the administration of low NPs concentrations over long-term exposure is limited. The present study aimed at investigating the effects induced by a long-term exposure (21-days) to two sub-lethal concentrations of polystyrene nanoplastics (PS-NPs; 0.05 and 0.5 µg/mL) on Daphnia magna. A multi-level approach was performed to assess potential sub-individual (i.e., molecular and biochemical) and individual (i.e., behavioural) adverse effects. At molecular level, the modulation of the expression of genes involved in antioxidant defence, response to stressful conditions and specific physiological pathways was investigated. Oxidative stress (i.e., the amount of pro-oxidants, the activity of antioxidant and detoxifying enzymes and lipid peroxidation) and energetic (i.e., protein, carbohydrate, lipid and total caloric content) biomarkers were applied to assess effects at the biochemical level, while swimming activity was measured to monitor changes in individual behavior. Although the 21-days exposure to PS-NPs induced a slight modulation of gene involved in oxidative stress response, biochemical analyses showed that D. magna individuals did not experience an oxidative stress condition. Significant changes in energy reserves of individuals exposed for 21 days to both the PS-NPs concentrations were observed, but no alterations of swimming activity occurred. Our results highlighted that the exposure to low concentrations of PS-NPs could pose a limited risk to D. magna individuals and suggested the importance of a multi-level approach to assess the risks of NPs on aquatic organisms.