The aging of microplastics in the environment changes their physicochemical properties. While this may affect their toxicity, comparative data on the effects of aged compared to pristine microplastics are scarce. One of those aging processes is the sorption of chemicals, which has mainly been studied for individual pollutants present in marine ecosystems. To investigate how the sorption of a complex mixture of freshwater pollutants affects the toxicity of microplastics, we incubated irregular polystyrene particles (≤63 μm) in either wastewater or ultrapure water. We exposed Daphnia magna to these aged microplastics and their pristine counterparts (80, 400, 2000, and 10,000 particles mL –1 ) over four generations using food limitation as an additional, environmentally realistic stressor. Both particle types affect the survival, reproduction, adult and neonate body lengths, and growth. An exposure to pristine microplastics results in the extinction of the third generation of daphnids. In contrast, wastewater-incubated particles induced a lower mortality. The incubation with wastewater does not change the microplastics’ size, surface charge, and structure. Consistent with the literature, we assume that the adsorption of dissolved organic matter is a key aging process reducing the toxicity of microplastics. Consequently, toxicity testing using pristine microplastics may overestimate the effects of plastic particles in nature.

The ubiquity of microplastics in aquatic ecosystems has raised concerns over their interaction with biota. However, microplastics research on freshwater species, especially mollusks, is still scarce. We, therefore, investigated the factors affecting microplastics ingestion in the freshwater mussel Dreissena polymorpha. Using polystyrene spheres (5, 10, 45, 90 µm), we determined the body burden of microplastics in the mussels in relation to 1) exposure and depuration time, 2) body size, 3) food abundance, and 4) microplastic concentrations. D. polymorpha rapidly ingested microplastics and excreted most particles within 12 h. A few microplastics were retained for up to 1 wk. Smaller individuals had a higher relative body burden of microplastics than larger individuals. The uptake of microplastics was concentration-dependent, whereas an additional food supply (algae) reduced it. We also compared the ingestion of microplastics by D. polymorpha with 2 other freshwater species (Anodonta anatina, Sinanodonta woodiana), highlighting that absolute and relative uptake depends on the species and the size of the mussels. In addition, we determined toxicity of polystyrene fragments (≤63 µm, 6.4-100 000 p mL -1 ) and diatomite (natural particle, 100 000 p mL -1 ) in D. polymorpha after 1, 3, 7, and 42 d of exposure, investigating clearance rate, energy reserves, and oxidative stress. Despite ingesting large quantities, exposure to polystyrene fragments only affected the clearance rate of D. polymorpha. Further, results of the microplastic and diatomite exposure did not differ significantly. Therefore, D. polymorpha is unaffected by or can compensate for polystyrene fragment toxicity even at concentrations above current environmental levels.