Formation of agglomerates and their rapid sedimentation during aquatic ecotoxicity testing of nanoparticles is a major issue with a crucial influence on the risk assessment of nanomaterials. The present work is aimed at developing and testing a new approach based on the periodic replacement of liquid media during an ecotoxicological experiment which enabled the efficient monitoring of exposure conditions. A verified mathematical model predicted the frequencies of media exchanges which checked for formation of agglomerates from silver nanoparticles AgNP with 50 nm average size of the original colloid. In the model experiments, embryos of common carp Cyprinus carpio were exposed repeatedly for 6 h to AgNPs (5-50 μm Ag L(-1)) either under semistatic conditions (exchange of media after 6 h) or in variants with frequent media exchanges (varying from 20 to 300 min depending on the AgNP colloid concentration and the desired maximum agglomerate size of 200 or 400 nm). In contrast to other studies, where dissolved free metals are usually responsible for toxic effects, our 144-h experiments demonstrated the importance of AgNP agglomerates in the adverse effects of nanosilver. Direct adsorption of agglomerates on fish embryos locally increased Ag concentrations which resulted in pronounced toxicity particularly in variants with larger 400 nm agglomerates. The present study demonstrates the suitability of the novel methodology in controlling the conditions during aquatic nanomaterial toxicity testing. It further provided insights into the mechanisms underlying the effects of AgNP, which rank on a global scale among the most widely used nanomaterials.
Acute toxicity of zinc oxide nanoparticle (ZnO-NP, mean particle size diameter of 10 nm) powder and water-soluble salt of zinc (ZnCl) to annelid Enchytraeus crypticus was tested using an agar-based nutrient-enriched medium with the addition of kaolin and humic acids (HA). Adults of the E. crypticus were cultivated in pure agar and in three types of modified exposure media containing different proportions of model soil constituents. Potworms were exposed to zinc in both forms (1-1000 mg kg of agar) for 96 h. In experiments with ZnCl, toxicity of zinc was the highest in pure agar followed by agar with HA and agar with kaolin and HA and the lowest toxicity was observed in agar with kaolin. The corresponding LC values were 13.2, 28.8, 39.4, and 75.4 mg kg respectively. In contrast, zinc in the form of ZnO-NPs was most toxic in the presence of HA followed by pure agar, agar with kaolin, and kaolin with HA. In this case, LC values were 15.8, 43.5, 111, and 122 mg kg respectively. Scanning electron microscopy revealed that the smallest agglomerates occurred in the presence of kaolin, where ZnO-NPs were sealed in a kaolin shell. This effect reduced the bioavailability and toxicity of the NPs. In contrast, larger agglomerates were observed in the presence of HA but a larger amount of zinc was dispersed in the volume of agar.
Chloroacetanilide herbicides are widely used in the agricultural sector throughout the world. Because of their poor biodegradability, high water solubility, and long persistence, chloroacetanilide herbicides have a high potential to contaminate water, and conventional water treatment processes do not ensure sufficient removal. Therefore, heterogeneous photocatalysis using TiO2/UV-A was investigated for the degradation of alachlor, acetochlor, and metolachlor from water. Two commercially available TiO2 (P25 and AV-01) were used as photocatalysts. Different experimental setups were also tested. In addition, the toxicity of single herbicides and mixtures of their photocatalytic degradation products to the freshwater alga Chlorella kessleri was investigated via a growth inhibition test. The maximum removal efficiency for alachlor, acetochlor, and metolachlor was 97.5%, 93.1%, and 98.2%, respectively. No significant differences in the removal efficiency of chloroacetanilide herbicides were observed for the photocatalysts used. Although the concentrations of all herbicides during photocatalysis decreased, the toxicity of the resulting mixtures of degradation products increased or remained the same, indicating the formation of toxic degradation products.
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