Effects of biochar and thermally treated biochar on Eisenia fetida survival, growth, lysosomal membrane stability and oxidative stress.

Despite its known positive impacts when added to soil, the negative effects of biochar on earthworms are not fully understood. Here, we investigated the toxicity of nine biochars of three feedstock origins, animal (cow dung), plant (corncob) and microorganism (sewage sludge), produced at three pyrolysis temperatures (350 °C, 550 °C and 750 °C) on earthworms. Vermitoxicity was first assessed using acute toxicity test, neutral red retention time (NRRT) assay and oxidative stress response assay. Furthermore, we evaluated whether the thermal treatment of biochars could reduce their vermitoxicity using an acute toxicity assay. We found that, according to LC and earthworm weight loss, cow dung biochar was more toxic than corncob or sewage sludge biochar; thus, production feedstock is apparently important to biochar vermitoxicity. Furthermore, NRRTs indicated cow dung biochar disrupted lysosomal membrane stability in earthworm coelomocytes, providing further evidence for the toxicity of this biochar to earthworms. Disturbed antioxidant enzyme activities and elevated malondialdehyde content showed that earthworm suffer oxidative stress, also implying a potential vermitoxicity. However, thermal treatment of cow dung biochar substantially improved its LC and decreased earthworm weight loss, implying that the PAHs in this biochar might be damage factors and that heating could reduce the potential toxicity of biochar. Besides, NRRT assay was first used to evaluate the effects of biochar on earthworms and clear dose-effect relationships indicated that NRRT assay might be a useful tool for assessing the potential negative effects of biochar. Overall, given the different effects of various biochars, including toxicity, reported here, our findings will help improve understanding of biochar vermitoxicity mechanisms, serve to improve biochar ecological risk assessments and provide a reference for the proper application of biochar amendments.