Effects of Cr(VI) on Ca²⁺-ATPase activity in the earthworm Eisenia andrei

The effect of Cr(VI) as a soil contaminant on the edaphic worm Eisenia andrei was studied by evaluating the activity of Ca²⁺-ATPase in the intestinal mucosa. In eukaryotes, Ca²⁺-ATPase is a key mediator of cell signaling although comparatively little is known about its activity in earthworms. Size and anatomical constraints (i.e. small and complex) led us to develop and optimize a cyto-biochemical method to measure Ca²⁺-ATPase activity in earthworms. The principal site of enzyme activity was found to be the post clitellar intestinal tract; immunohistochemistry then identified plasma membrane Ca²⁺-ATPase (PMCA ATPase) in the apical area of the intestinal epithelium. Earthworms exposed for 28days to OECD soil contaminated with 1, 2, and 15mg/Kg Cr(VI) demonstrated about 70% inhibition of Ca²⁺-ATPase activity at the low Cr (VI) concentration (the half of the Italian law limit for residential areas), rising to approximately 84% inhibition at the highest concentration. Reduced enzyme activity was accompanied by decreased enzyme content and reduced lysosomal membrane stability (LMS), which is a well established early warning biomarker of stress. These data demonstrate the potential utility of Ca²⁺-ATPase activity as a sensitive parameter with which to detect environmental stress in earthworms.