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Effects of ZnO NPs on the survival of E. fetida were evaluated in sand and on filter paper 93. Although high toxicity occurred in the filter paper test, it is a less realistic medium and causes greater stress to the organisms 93. In addition, for ZnO, which is photo reactive, the filter paper test would also have added a potentially new mechanism of toxicity through effects of ROS generation from noninternalized ZnO, something that would not occur in the more realistic soil tests. The same authors also determined the effects of ZnO NPs on earthworm reproduction in sand and artificial soil. Reproduction showed a decrease with increasing NPs concentration in both tested media; however, the values were not significantly different from the control 93. The bioaccumulation of ZnO NPs in the isopod P. scaber was dose dependent and was attributable to the dissolution of Zn rather than the accumulation of ZnO particles 41. For the collembolan Folsomia candida, effects of ZnO NPs ( a LC50 of 1,000?mg/kg. The median effective concentration (EC50) for reproduction was 1,964 (95% CI?=?1,635�C2,293), 1,591, and 298 (95% CI?=?181�C415) mg/kg for ZnO NPs, non-nano ZnO, and ZnCl2, respectively 59. When the reproduction EC50 was based on Zn concentration in pore water, it was similar for ZnO NPs and ZnCl2 (10.1 and 16.8?mg/L, respectively), indicating that the toxicity of the NPs can be from dissolved ionic zinc 59. The LC50 for effect on the survival of C. elegans was not significantly different between the Zn forms tested (bulk ZnO, ZnCl2, and ZnO NPs) 42. Similarly, no significant difference in mortality was found between ZnCl2 and ZnO NPs in a buffered K-medium 95. However, when testing in unbuffered medium, higher mortality rates were observed for ZnCl2, which was attributable to a reduced complexation of Zn+ ions with other species in the solution, in comparison with the buffered K-medium 95. Great differences in LC50 were found for C. elegans between these studies. Ma et al. 95 found a mean LC50 of 789?mg Zn/L for ZnO, whereas Wang et al. 42 reported an LC50 of 2.3?mg/L for ZnO, possibly because of differences in media used. For effects on reproduction and avoidance behavior, no differences were found between ZnCl2 and ZnO NPs, in either medium tested (buffered K-medium and unbuffered media) 95.