Because of their insolubility in water, nanoparticles have a limitation concerning toxicity experiments. The present study demonstrated a plant agar test for homogeneous exposure of nanoparticles to plant species. The effect of Cu nanoparticles on the growth of a plant seedling was studied, and bioaccumulation of nanoparticles was investigated. All tests were conducted in plant agar media to prevent precipitation of water‐insoluble nanoparticles in test units. The plant species were Phaseolus radiatus (mung bean) and Triticum aestivum (wheat). Growth inhibition of a seedling exposed to different concentrations of Cu nanoparticles was examined. Copper nanoparticles were toxic to both plants and also were bioavailable. The 2‐d median effective concentrations for P. radiatus and T. aestivum exposed to Cu nanoparticles were 335 (95% confidence level, 251–447) and 570 (450–722) mg/L, respectively. Phaseolus radiatus was more sensitive than T. aestivum to Cu nanoparticles. A cupric ion released from Cu nanoparticles had negligible effects in the concentration ranges of the present study, and the apparent toxicity clearly resulted from Cu nanoparticles. Bioaccumulation increased with increasing concentration of Cu nanoparticles, and agglomeration of particles was observed in the cells using transmission‐electron microscopy–energy‐dispersive spectroscopy. The present study demonstrated that the plant agar test was a good protocol for testing the phytotoxicity of nanoparticles, which are hardly water soluble.

This study investigated the trophic transfer, individual impact, and embryonic uptake of fluorescent nano-sized polystyrene plastics (nanoplastics) through direct exposure in a freshwater ecosystem, with a food chain containing four species. The alga Chlamydomonas reinhardtii, water flea Daphnia magna, secondary-consumer fish Oryzias sinensis, and end-consumer fish Zacco temminckii were used as test species. In the trophic transfer test, algae were exposed to 50 mg/L nanoplastics, defined as plastic particles <100 nm in diameter; higher trophic level organisms were exposed through their diet. In the direct exposure test, each species was directly exposed to nanoplastics. Microscopic analysis confirmed that the nanoplastics adhered to the surface of the primary producer and were present in the digestive organs of the higher trophic level species. Nanoplastics also negatively affected fish activity, as measured by distance traveled and area covered, and induced histopathological changes in the livers of fish that were directly exposed. Additionally, nanoplastics penetrated the embryo walls and were present in the yolk sac of hatched juveniles. These observations clearly show that nanoplastics are easily transferred through food chain, albeit because of high experimental dosages. Nevertheless, the results strongly point to the potential health risks of nanoplastic exposure.