Neutral red retention by lysosomes from earthworm (Lumbricus rubellus) coelomocytes: A simple biomarker of exposure to soil copper

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Abstract
A simple subcellular histochemical staining technique employing the lysosomal probe neutral red has been developed for use with the epiendogeic earthworm Lumbricus rubellus Hoffmeister. Coelomocytes extracted from the coelomic cavity of earthworms into an isotonic earthworm Ringer solution were allowed to adhere to a microscope slide for 30 s before the application of a neutral red dye (80 mg·ml−1). This red dye was rapidly accumulated within the lysosomes. Observation of the loss of this dye from these lysosomes into the surrounding cytosol has enabled the quantification of the degree of lysosomal damage caused to earthworms with exposure to an increasing range of soil copper concentrations, in both laboratory and mesocosm studies. This simple in vitro biomarker has potential for the rapid assessment of the toxic effects to earthworms from soils contaminated with heavy metals and metalloids.

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