Effects of polystyrene microplastics on copper toxicity to the protozoan Euglena gracilis: emphasis on different evaluation methods, photosynthesis, and metal accumulation.

Microplastics (MPs) released into aquatic environment interact with other pollutants that already exist in water, potentially altering their toxicity, which poses a new problem for aquatic ecosystems. In the present study, we first evaluated the effects of polystyrene MPs (mPS) on copper (Cu) toxicity to the protozoan Euglena gracilis using three methods based on 96-h acute toxicity, orthogonal test and 12-d sub-acute toxicity data. Thereafter, the 12-d sub-acute exposure was employed to investigate protozoan growth, photosynthetic parameters and pigments, soluble protein, total antioxidant capacity and trace metal accumulation in E. gracilis after exposure to either 1.5 mg/L of Cu, 75-nm mPS (1 and 5 mg/L) or a combination therein, with the objective to understand the underlined mechanisms. The results show that the concentration and exposure time are key factors influencing the effects of the mPS on Cu toxicity. A mPS concentration of 5 mg/L caused significantly more dissipation energy, which is used for photosynthesis and thus decreased photosynthetic efficiency, but this effect weakened after 12 d of exposure. Exposure to Cu alone resulted in significantly high Cu accumulation in the cells and inhibited uptake of manganese and zinc. The presence of mPS did not influence the effects of Cu on trace metal accumulation. Our result suggests that application of multiple methods and indices could provide more information for a comprehensive understanding of the effects of mPS on toxicity of other pollutants. In addition, long-term exposure seems necessary for evaluating mPS toxicity.