The environmental behavior and the potential toxicity of copper nanoparticles (nano-Cu) in water are major concerns for assessing their environmental safety. The present study was undertaken to characterize the properties of nano-Cu in E3 medium, such as size changes, solubility, zeta-potential and pH, and to test the toxicity of nano-Cu suspension to zebrafish embryos. Dynamic light scattering and solubility experiments showed that three components coexisted in the nano-Cu exposure system, including small nano-Cu aggregates still suspended in E3 medium, large nano-Cu aggregates deposited on the container bottom and dissolved copper species (Cu(dis)). Both the zeta-potential of nano-Cu particles in E3 medium and the pH of the nano-Cu suspension showed no change during a 24 hour period. It is found that nano-Cu retarded the hatching of zebrafish embryos and caused morphological malformation of the larvae, and high concentrations (>0.1 mg/L) of nano-Cu even killed the gastrula-stage zebrafish embryos. Cu2+ ions were used to study the toxicity caused by nano-Cu dissolution. The embryo toxicity of nano-Cu at 0.01 and 0.05 mg/L showed no significant difference from Cu2+ at the corresponding concentrations (0.006 and 0.03 mg/L), but 0.1 mg/L nano-Cu had a greater toxicity than 0.06 mg/L Cu2+.
To investigate the toxicological effects of nanomaterials, experimental studies on the absorption and accumulation in organisms are of broad interest. In the present study, Caenorhabditis elegans (C. elegans) was used as a ''model'' organism to investigate the bioaccumulation and toxicological effects of engineered copper nanoparticles with a scanning technique of microbeam synchrotron radiation X-ray fluorescence (m-SRXRF). The adult hermaphrodite is anatomically simple with 959 somatic cells and 1 mm in length. The mapping results of the whole organism indicate that the exposure to copper nanoparticles can result in an obvious elevation of Cu and K levels, and a change of bio-distribution of Cu in nematodes. Accumulation of Cu occurs in the head and at a location 1/3 of the way up the body from the tail compared to the un-exposed control. In contrast, a higher amount of Cu was detected in other portion of worm body, especially in its excretory cells and intestine when exposed to Cu 2+ . The results compared well with total Cu levels in nematodes, which were 4.10 AE 0.54, 12.32 AE 0.49 and 5.22 AE 0.63 mg g À1 dry weight for the PBS, Cu 2+ and Cu nanoparticle groups, respectively, measured by ICP-MS. The nondestructive and multi-elemental m-SRXRF provides an important tool for mapping the elemental distribution in the whole body of a single tiny nematode at lower levels.
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