Biochars are used as amendments to improve soil quality, but their effects on edaphic organisms such as earthworms remain controversial. This study aimed to assess the effects of adding a poultry manure-derived biochar into a contaminated technosol on trace element (TE) (i.e. As, Cd, Cu, Pb, and Zn) bioavailability for two earthworm species, Aporrectodea icterica and Aporrectodea longa. Three components of the bioavailability concept were determined using a pot experiment: (1) total soil TE (potentially reactive) and TE concentrations in the soil pore water (environmental availability), (2) TE concentrations in depurated whole earthworm bodies (environmental bioavailability) and (3) ecophysiological and biochemical effects on earthworms (toxicological bioavailability). Biochar addition increased TE concentrations in the soil pore water respectively from 1.8, 2.7, 9.4, 0.7 and 959 to 6, 6.2, 19.3, 6.9, and 3003 µg L−1 for As, Cd, Cu, Pb and Zn. Biochar addition did not influence TE environmental bioavailability for earthworms, except a decreased As concentration (32.5 to 15.2 µg g−1) in A. icterica. This suggests an inter-specific variability in As homeostasis in the Aporrectodea genus. In line with this internal As decrease, the Glutathione-S-transferase (GST) activity decreased by 42% and protein and lipid contents slightly increased (14 and 25%, respectively) in A. icterica tissues. The body weight of both earthworm species decreased for the biochar-amended soil. Environmental TE availability depended on both the biochar addition and the earthworm activity in the contaminated soil, while environmental and toxicological bioavailabilities resulted from the earthworm species, the targeted TE and biochar supply to the soil.Read less <