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+.