An environmentally realistic method to test fullerene (C(60) ) toxicity to the benthic organism Chironomus riparius was created by allowing suspended fullerenes to settle down, making a layer on top of the sediment. To test the hypothesis that higher food concentrations will reduce toxic responses, two food concentrations were tested (0.5 and 0.8% Urtica sp.) in sediment containing fullerene masses of 0.36 to 0.55 mg/cm(2) using a 10-d chronic test. In the 0.5% food level treatments, there were significant differences in all growth-related endpoints compared with controls. Fewer effects were observed for the higher food treatment. Fullerene agglomerates were observed by electron microscopy in the gut, but no absorption into the gut epithelial cells was detected. In the organisms exposed to fullerenes, microvilli were damaged and were significantly shorter. The potential toxicity of fullerene to C. riparius appears to be caused by morphological changes, inhibiting larval growth.
Increasing usage of fullerenes (C60 ) increases their opportunities to be released into the environment. For risk assessment, it is important to understand the environmental fate and ecotoxicological effects of C60 . In the present study, fullerene settling was measured during a 1-yr period with 4 different lake waters and an artificial freshwater, and Daphnia magna immobilization and fullerene accumulation was also measured in each of the lake waters. Depending on the characteristics of the lake waters, fullerenes either exhibited extended water stability or settled rapidly; in all waters, there was a fraction that remained stable after 1 yr. Water stability was affected by the quality and molecular size distribution of dissolved natural organic matter (DNOM). Increasing DNOM molecular sizes with high aromatic content enhanced water stability. Immobilization of D. magna was generally quite low (under 20%) and highly variable after 24 h and 48 h at initial fullerene concentrations up to 10 mg/L. Substantial settling occurred during the time period for acute toxicity assays (i.e., 48 h), which should be anticipated when conducting toxicity assays. There were no significant differences in the quantity of accumulated fullerenes among the different lake waters at fullerene concentrations of 0.5 mg/L, but there were differences at 2 mg/L.
A key component of understanding the potential environmental risks of fullerenes (C60) is their potential effects on benthic invertebrates. Using the sediment dwelling invertebrate Chironomus riparius we explored the effects of acute (12 h and 24 h) and chronic (10 d, 15 d, and 28 d) exposures of sediment associated fullerenes. The aims of this study were to assess the impact of exposure to C60 in the sediment top layer ((0.025, 0.18 and 0.48) C60 mg/cm2) on larval growth, oxidative stress and emergence rates and to quantify larval body burdens in similarly exposed organisms. Oxidative stress localization was observed in the tissues next to the microvilli and exoskeleton through a method for identifying oxidative stress reactions generated by reactive oxygen species. Rapid intake of fullerenes was shown in acute experiments, whereas body residues decreased after chronic exposure. Transmission electron microscopy analysis revealed oxidative damage and structural changes in cells located between the lipid droplets and next to the microvilli layer in fullerene exposed samples. Fullerene associated sediments also caused changes in the emergence rate of males and females, suggesting that the cellular interactions described above or other effects from the fullerenes may influence reproduction rates.
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