In recent years, biochars have gained increasing interest in mitigating climate changes and revitalizing contaminated or drained soil. Studies determining their impact on the ecosystem, especially on soil invertebrates, however, are still scarce and the neurotoxic potential of biochars has never been evaluated before. Using the model organism Caenorhabditis elegans we determined the neurotoxic effect of biochar produced from rice straw by pyrolysis at 500 °C at concentrations ranging from 0 to 2000 mg C∙L-1. Biochar had a hormetic effect on locomotion behavior. Furthermore, high concentrations impaired defecation as well as the recognition and response to a chemical attractant. None of the potential toxic chemicals in the biochar had sufficient high concentrations to explain the detected neurotoxic effect. Using electron paramagnetic resonance (EPR) spectroscopy, we detected free radicals in the biochar. Detrimental reaction of free radicals with biotic macromolecules can induce oxidative stress responses and are a potential reason for the evaluated neurotoxic effect of biochar. Overall, we were able to prove that biochars have the potential to act as weak neurotoxins to soil organisms and effects of persistent free radicals should be investigated further.

As an effective soil amendment, biochars require a comprehensive ecological evaluation before they can be widely used in agriculture because endogenous contaminants, such as environmentally persistent free radicals (EPFRs), certainly pose an ecological risk to soil invertebrates. In this study, Caenorhabditis elegans (C. elegans) was used as a model organism to investigate the neurotoxicity of two rice straw biochars pyrolyzed at 500 and 700 °C. After 24 h exposure to unwashed biochar, washed biochar, and leaching fluids (supernatants), the neurobehavioral parameters of C. elegans were determined in a liquid toxicity test. The results showed that the washed 700 °C biochar particles significantly impaired locomotion and prolonged the defecation interval at a biochar concentration of 4 g·well⁻¹, while the unwashed biochar and supernatants caused no apparent impairment. Supporting this, electron paramagnetic resonance (EPR) results showed that the intensity of EPFRs in unwashed 700 °C biochar was stronger than that of the corresponding washed particles. This indicates that, in the liquid test, the EPR signal alone is not indicative of particle toxicity. The accessibility and activity of the EPFRs should be considered. Dissolved organic matter (DOM) was observed in the leaching fluids. The neurotoxic activity of the washed biochar was alleviated after the re-addition of leaching fluids to the washed biochar, suggesting that the dissolved organic materials modulate the reactivity of the EPFRs in the liquid phase. This study suggests that the leaching process may increase the risk of biochar when used in the field environment.