Suitability of lysosomal membrane stability in Eisenia fetida as biomarker of soil copper contamination

Accumulated metals in soils negatively affect dwelling organisms. Earthworms, which are widespread and perform various essential functions, are able to accumulate metals that can damage the coelomic cells. The aim of this work was to evaluate the effect on Eisenia fetida lysosomal membrane stability both during and after copper exposure, and finally to link this to internal concentrations. E. fetida specimens were exposed to a reference soil and two Cu-spiked soils (35 and 350 mg kg−1 d.w.) for 14 days (uptake period) and then transferred into the reference soil for other 18 days (elimination period). After 3 days of uptake, internal Cu concentrations increased and were higher in the specimens exposed to soils spiked with 350 mg Cu kg−1 d.w. After 2 days of elimination, a strong decrease of internal Cu concentrations was always observed. The lysosomal membrane stability, measured as neutral red retention-times, was approximately 50 min for the earthworms exposed to the reference soils, whereas it decreased, at the end of the uptake period, to 21 and 13 min, respectively, for the organisms exposed to soils spiked with 35 and 350 mg Cu kg−1 d.w. A full recovery of the lysosomal membrane stability was reached after 14 and 18 days of the elimination period, respectively, for the organisms exposed to soils spiked with 35 and 350 mg Cu kg−1 d.w. The neutral-red assay would seem a good biomarker since the lysosomal membrane stability of E. fetida appeared to respond rapidly and strongly to soil copper contamination.