Relationships between metal compartmentalization and biomarkers in earthworms exposed to field-contaminated soils

Partitioning tissue metal concentration into subcellular compartments relecting 'toxicologically available'pools may provide good descriptors of the toxicological efects of metals on organisms. Here we investigatedthe relationships between internal compartmentalization of Cd, Pb and Zn and biomarker responses in a model soil organism: the earthworm. The aim of this study was to identify metal fractions reflecting the toxic pressure in an endogeic, naturally occurring earthworm species (Aporrectodea caliginosa) exposed torealistic field-contaminated soils.After a 21 days exposure experiment to 31 field contaminated soils, Cd, Pb and Zn concentrationsin earthworms and in three subcellular fractions (cytosol, debris and granules) were quantified. Differentbiomarkers were measured: the expression of a metallothionein gene (mt), the activity of catalase (CAT) andof glutathione-s-transferase (GST), and the protein, lipid and glycogen reserves. Biomarkers were furthercombined in an integrated biomarker index (IBR).The subcellular fractionation provided better predictors of biomarkers than the total internal contentshence supporting its use when assessing toxicological bioavailability of metals to earthworms. The mostsoluble internal pools of metals were not always the best predictors of biomarker responses. metallothioneinexpression responded to increasing concentrations of Cd in the insoluble fraction (debris + granules). Proteinand glycogen contents were also mainly related to Cd and Pb in the insoluble fraction. On the other hand,GST activity was better explained by Pb in the cytosolic fraction. CAT activity and lipid contents variationswere not related to metal subcellular distribution. The IBR was best explained by both soluble and insolublefractions of Cd and Pb.This study further extends the scope of mt expression as a robust and specific biomarker in an ecologicallyrepresentative earthworm species exposed to field-contaminated soils. The genetic lineage of the individuals,assessed by DNA barcoding with cytochrome c oxidase subunit I, did not inuence mt expression