Assessment of the potential cyto-genotoxicity of the nonsteroidal anti-inflammatory drug (NSAID) diclofenac on the zebra mussel (Dreissena polymorpha)

A battery of eight biomarkers was used on the freshwater bivalve Dreissena polymorpha in order to evaluate potential sub-lethal effects of the nonsteroidal anti-inflammatory drug diclofenac (DCF; 2-[(2,6-dichlorophenyl)amino]phenylacetic acid). By an in vivo approach, mussels were exposed for 96 h to increasing concentrations (0.3, 1, and 2 nM) of DCF perfectly comparable with current surface water levels. We determined the single cell gel electrophoresis assay, the apoptotic frequency (DNA Diffusion assay), the micronucleus test (MN test), and the lysosomal membrane stability (Neutral Red Retention Assay) in mussel hemocytes. Moreover, the activity of catalase, superoxide dismutase, glutathione peroxidase, and the phase II detoxifying enzyme glutathione S-transferase was measured in the cytosolic fraction extracted from a pool of entire bivalves to reveal possible alterations of the oxidative status of exposed specimens. The biomarker battery pointed out a negligible cyto- and genotoxicity on zebra mussel hemocytes since only a slight decrease of lysosomal membrane stability from baseline levels was measured at the end of exposures at the highest concentration (2 nM). In addition, environmental concentrations of DCF seem to have a negligible effect on the activities of antioxidant and detoxifying enzymes.