Vermitoxicity of aged biochar and exploring potential damage factors.

Although biochar is a promising soil amendment, its characteristics change owing to its aging in soil. Studies have shown that some aged biochar is hazardous to plants and soil microbiota. Earthworms are well-known soil ecosystem engineers; nevertheless, the toxic effects of aged biochar on them (vermitoxicity) are yet unknown, and it is necessary to explore the potential risk factors. Here, a series of soil culture experiments were conducted to systematically examine the vermitoxicity of aged biochar at various levels utilizing the earthworm Eisenia fetida and corncob biochar.. Acute toxicity bioassays were also used to evaluate several potential harm factors utilizing modified aged biochar/leaching solutions. The findings showed that both fresh and aged biochar might have adverse effects on earthworms, and that aged biochar was more toxic than fresh biochar with LCs reduced to 6.89%. Specifically, aged biochar caused earthworm death, growth inhibition with a maximum of 36.6%, and avoidance with 100% avoidance at the application rates of 2% at the individual-behavioral level. At the cellular and physiological-biochemical levels, aged biochar damaged coelomocyte lysosomal membrane stability, disrupted antioxidant enzyme activities, and improved the malondialdehyde (MDA) content in earthworms. Heat-treated and pH-modified aged biochar exhibited less acute toxicity on earthworms than aged biochar, whereas aqueous and acetone extracts showed weak vermitoxicity. As a result, earthworms may be harmed by volatile organic compounds (VOCs), an improper pH, and aqueous and acetone extracts. Additionally, the range of neural red retention times (NRRTs) was reviewed as ∼20-70 min mostly. This study, as far as we know, is the first to evaluate the vermitoxicity of aged biochar and its potential damage factors. The results may enhance our understanding of ecological toxicity of biochar, particularly over the long term, and lead to the development of application standards for biochar amendments to the soil.