Combined effects of goethite nanoparticles with metallic contaminants and an organophosphorus pesticide on Eisenia andrei

The effects of mixtures of nanoparticles (NPs) and other chemicals have been poorly studied in terrestrial invertebrates. In this study, we investigated the effects of binary mixtures of goethite (α-FeOOH) NPs and metallic (Cd and Pb) or organic (chlorpyrifos, CPF) contaminants in Eisenia andrei earthworms. We used the filter paper contact test to evaluate (i) the uptake of NPs in organisms exposed to the mixtures of NPs+Metals and NPs+CPF and (ii) the potential effects of the mixture of NPs+CPF on the CPF-induced inhibition of the biomarker enzymes acetylcholinesterase (AChE) and carboxylesterases (CES). We used the artificial soil test to deepen the study on joint effects of NPs+CPF. All compounds were applied separately and in binary mixtures. In the single exposure treatment, Fe levels decreased significantly in organisms exposed to NPs on filter paper, suggesting systemic effects aimed at eliminating Fe incorporated through NPs. Conversely, earthworms exposed to binary mixtures showed Fe levels similar (NPs+Metals) to or higher (NPs+CPF) than controls. The earthworms single exposed to NPs presented no changes in AChE and CES activities. In the artificial soil test, the only treatment that showed AChE inhibition after 72 h was single CPF exposure, while no significant changes were observed in CES activity. However, after 7-day exposure in artificial soil or 72-h exposure on filter paper, the mixture of NPs+CPF induced a similar degree of AChE and CES inhibition as single CPF exposure. All these suggested that NPs did not produce neurotoxic effects, and that the inhibition of the enzymes’ activities in all cases was due to the presence of the pesticide. On the other hand, the differences in the pattern of Fe accumulation in the earthworms indicate that the presence of other contaminants in the exposure media can modify the uptake and/or the excretion of Fe and evidence the interactions that may be found in binary mixtures of metal oxide NPs and other pre-existing contaminants in the soil ecosystem.