Abstract
The use of silver nanoparticles (AgNPs) in agriculture and many consumer products has led to a significant release of Ag in the environment. Although Ag toxicity in terrestrial organisms has been studied extensively, very little is known about the accumulation capacity and coping mechanisms of organisms in Ag-contaminated soil. In this context, we exposed Eisenia fetida earthworms to artificial OECD soil spiked with a range of concentrations of Ag (AgNPs or AgNO3). The main aims were to (1) identify the location and form of accumulation of Ag in the exposed earthworms and (2) better understand the physiological mechanisms involved in Ag detoxification. The results showed that similar doses of AgNPs or AgNO3 did not have the same effect on E. fetida survival. The two forms of Ag added to soil exhibited substantial differences in speciation at the end of exposure, but the Ag speciation and content of Ag in earthworms were similar, suggesting that biotransformation of Ag occurred. Finally, 3D images of intact earthworms obtained by X-ray micro-computed tomography revealed that Ag accumulated preferentially in the chloragogen tissue, coelomocytes, and nephridial epithelium. Thus, E. fetida bioaccumulates Ag, but a regulation mechanism limits its impact in a very efficient manner. The location of Ag in the organism, the competition between Ag and Cu, and the speciation of internal Ag suggest a link between Ag and the thiol-rich proteins that are widely present in these tissues, most probably metallothioneins, which are key proteins in the sequestration and detoxification of metals.
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Acknowledgments
The authors wish to thank Dominique Dubois, Olivier Proux, Géraldine Sarret, Ana Elena Pradas Del Real, Kerstin Hund-Rinke and Régine Leroux for their help and fruitful discussions.
Funding
This study was funded mainly by the ANSES in the ETNA2 project context, by a grant of the University of Lille and the SMRE doctoral school and by a public grant overseen by the French National Research Agency (ANR) as part of the French platform Nano-ID (EQUIPEX project ANR-10-EQPX-39-01).
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Supplementary 1
Scheme of the experimental design. “Control” corresponds to the microcosm without any inputs. “AgNPs” corresponds to silver nanoparticles. “Dis” corresponds to dispersant. “AgNO3” corresponds to silver nitrate. The concentrations C1, C2, C3 and C4 correspond to 33 (± 16), 71 (± 8), 117 (± 11) and 277 (± 24) mg kg-1 of Ag (dry matter) (the mean concentrations of AgNO3 and AgNPs were not significantly different). The volumes D1, D2, D3 and D4 correspond to the volumes of dispersant added to the microcosms. Dispersant was added in the same amount as in the corresponding AgNPs microcosms, that is, 1.599, 2.666, 5.331 and 10.662 mL. (PPTX 65.4 kb)
Supplementary 2
Metal contents in earthworms (mean in mg kg-1). The results were obtained by ICP analysis. “Control” corresponds to the microcosm without silver addition. “AgNPs” corresponds to silver nanoparticles. “Dis” corresponds to dispersant. “AgNO3” corresponds to silver nitrate. The concentrations C1, C2, C3 and C4 correspond to 33 (± 20), 71 (± 10), 117 (± 15) and 277 (± 45) mg kg-1 (dry matter) (the mean concentrations of AgNO3 and AgNPs were not significantly different). Stars (*) indicate significant differences between the condition with Ag and the associated control without Ag. Standard deviations are in parentheses. (DOCX 13.7 kb)
Supplementary 3
3D imaging of an earthworm by micro-CT. (top) Selection of FOV for high-resolution micro-CT scan. (bottom) 3D image analysis procedure for isolating Ag accumulation areas (normalization and thresholding step). (PPTX 692 kb)
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Courtois, P., Rorat, A., Lemiere, S. et al. Accumulation, speciation and localization of silver nanoparticles in the earthworm Eisenia fetida. Environ Sci Pollut Res 28, 3756–3765 (2021). https://doi.org/10.1007/s11356-020-08548-z
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DOI: https://doi.org/10.1007/s11356-020-08548-z