We assayed the toxicity of polystyrene nanoparticles (PS-NP, 52 nm) to Daphnia galeata. Survival and reproduction were significantly decreased in individuals exposed to 5 mg/L of PS-NP for 5 days, and embryos showed abnormal development, including a low hatching rate. Using fluorescence confocal microscopy, we recorded the transfer of PS-NP from the external surface of the body to the internal organs, including the thoracic appendices, ovaries, caudal appendices, and brood chamber, as well as PS-NP storage in lipid droplets. Although embryos were exposed to PS-NP in the brood chamber, they did not internalize PS-NP. Exposed D. galeata adults that were not pregnant stored significantly fewer lipid droplets than did the control group, and the lipid droplets that they did store were smaller; meanwhile, there were no significant changes in lipid storage in exposed pregnant individuals. Some embryos showed a high level of lipid storage, a response that occurs when embryos experience an abnormal state, and these embryos showed a very low hatching rate. However, the offspring of exposed adults showed normal survival and lipid storage. This study provides visual evidence that confirms the transfer and effects of PS-NP in Daphnia species, and suggests a relationship between toxicity and lipid storage.

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With increasing concerns about the release of silver nanoparticles (AgNPs) into the environment and the risks they pose to ecological and human health, a number of studies of AgNP toxicity to aquatic organisms have been conducted. USEPA and EU JRC have published risk assessment reports for AgNPs. However, most previous studies have focused on the adverse effects of AgNPs on individual species. Hazardous concentration (HC) of AgNPs for protection of aquatic ecosystems that are based on species sensitivity distributions (SSDs) have not yet been derived because sufficient data have not been available. In this study, we conducted multispecies toxicity tests, including acute assays using eight species from five different taxonomic groups (bacteria, algae, flagellates, crustaceans and fish) and chronic assays using six species from four different taxonomic groups (algae, flagellates, crustaceans and fish). Using the results of these assays, we used a SSD approach to derive an AgNP aquatic HC5 (Hazard concentrations at the 5% species) of 0.614 μg/L. To our knowledge, this is the first report of a proposed HC of AgNPs for the protection of aquatic ecosystems that is based on SSDs and uses chronic toxicity data.

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