Responses to silver nanoparticles and silver nitrate in a battery of biomarkers measured in coelomocytes and in target tissues of Eisenia fetida earthworms.

The current use and development of applications with silver nanoparticles (Ag NPs) could lead to potential inputs of these NPs to soils. Consequently, it is crucial to understand the ecotoxicological risks posed by Ag NPs in the terrestrial compartment. In the present investigation, the effects produced by PVP-PEI coated Ag NPs were assessed in Eisenia fetida earthworms in comparison with the soluble form (AgNO). Earthworms were exposed for 1, 3 and 14 days to a range of sublethal concentrations of Ag (0, 0.05 and 50mg/kg) and at each exposure time, apart from mortality and weight loss of individuals, metallothionein (MT) protein concentration and catalase (CAT) activity were quantified in earthworm tissues. In addition, cellular and molecular level endpoints (cell viability, absolute and relative trophic indices and transcription levels of catalase-cat- and metallothionein-mt-) were measured in coelomocytes extruded from exposed earthworms. Despite the lack of effects in traditional endpoints (mortality and weight loss), Ag NPs and AgNO posed changes at lower levels of biological complexity (biochemical, cellular and molecular levels). Both Ag forms induced similar changes in the metal detoxification mechanism (MT, mt) and in the antioxidant response system (CAT, cat) of E. fetida. In contrast, Ag form dependant cytotoxicity and subpopulation ratio alterations (eleocytes/amoebocytes) were recorded in extruded coelomocytes. Complementarily, the use of coelomocytes to assess molecular level endpoints represented a relevant alternative for development of non-invasive biomarkers.