The toxicity of silver nanoparticles to zebrafish embryos increases through sewage treatment processes

Silver nanoparticles (AgNPs) are widely believed to be retained in the sewage sludge during sewage treatment. The AgNPs and their derivatives, however, re-enter the environment with the sludge and via the effluent. AgNP were shown to occur in surface water, while evidence of a potential toxicity of AgNPs in aquatic organisms is growing. This study aims to examine the toxicity of AgNPs to the embryos of the aquatic vertebrate model zebrafish (Danio rerio) before and after sewage treatment plants (STPs) processes. Embryos were treated with AgNP (particle size: \textgreater90 \% \textless20 nm) and AgNO3 in ISO water for 48 h and consequently displayed effects such as delayed development, tail malformations and edema. For AgNP, the embryos were smaller than the controls with conspicuously smaller yolk sacs. The corresponding EC50 values of 48 hours post fertilization (hpf) were determined as 73 μg/l for AgNO3 and 1.1 mg/l for AgNP. Whole-mount immunostainings of primary and secondary motor neurons also revealed secondary neurotoxic effects. A TEM analysis confirmed uptake of the AgNPs, and the distribution within the embryo suggested absorption across the skin. Embryos were also exposed (for 48 h) to effluents of AgNP-spiked model STP with AgNP influent concentrations of 4 and 16 mg/l. These embryos exhibited the same malformations than for AgNO3 and AgNPs, but the embryo toxicity of the sewage treatment effluent was higher (EC50 = 142 μg/l; 48 hpf). On the other hand, control STP effluent spiked with AgNPs afterwards was less toxic (EC50 = 2.9 mg/l; 48 hpf) than AgNPs in ISO water. This observation of an increased fish embryo toxicity of STP effluents with increasing AgNP influent concentrations identifies the accumulation of AgNP in the STP as a potential source of effluent toxicity.