Biochar increases pesticide-detoxifying carboxylesterases along earthworm burrows

Herein, we examined whether synergistic effects of earthworms (Lumbricus terrestris) and pine needle-derived biochar result in biochar-coated burrows with enhanced carboxylesterase (CE) activity (a pesticide-detoxifying enzyme). Biochar was placed at the top of soil columns at two doses (2.5 and 5% w/w dry mass), with an additional biochar-free treatment as control. Carboxylesterase and dehydrogenase activities were measured in the burrow walls sampled at three depths (0-4, 8-12, and 18-22 cm). Biochar was recovered from these samples to confirm its vertical transport and enzymatic activation. We tested whether biochar protected CE activity against desiccation stress of burrow wall samples. Likewise, the role of earthworm mucus in stabilizing CE onto biochar surface was also investigated by measuring the enzyme activity in fresh biochar particles previously incubated in the presence of earthworm mucus and purified esterase. Finally, we checked for the sensitivity of biochar-bound CE activity against selected organophosphorus pesticides. The main results were: i) co-application of earthworms and biochar caused a significant increase of CE activity in the first 12 cm of the soil column, ii) recovered biochar particles displayed CE activity which was significantly higher in the particles collected from the bottom of columns, iii) soil desiccation decreased the enzymatic activity, although such effect depended on biochar treatment and column depth (22-77% inhibition). Nevertheless, CE activity still was higher in the samples from the 5%-biochar treatment than activity in control and 2.5%-biochar treatments, iv) earthworm mucus favored the retention of CE onto the biochar surface, and v) the activity of biochar-bound CE was sensitive to inhibition by chlorpyrifos-oxon. These results suggest that the joint application of L. terrestris and biochar may be a suitable vermiremediation strategy to inactivate OP pesticides.