Acute toxicity of dichlorvos to Aphanius iberus (Cuvier & Valenciennes, 1846) and its anti-cholinesterase effects on this species

This study evaluates the toxic effects of the organophosphate pesticide (OP) dichlorvos to the endangered Iberian toothcarp (Aphanius iberus). To this end, the lethal toxicity of dichlorvos based on 96 h-LC50 bioassays was determined in saline water (50 g/L), and in vivo effects of dichlorvos on cholinesterase (ChE) activity were investigated in adult female and male specimens. The 96 h-LC50 value determined by probit analysis was 3.17 mg/L (95% confidence limits: 1.34–3.97). The characterisation of the ChE using different substrates and specific inhibitors was also carried out in head and muscle tissues. Acetylthiocholine was the substrate preferred by both head and muscle ChE in males and females. Eserine sulphate and BW284C51 significantly inhibited both head and muscle enzyme activity at low concentrations (μM range), and iso-OMPA had no significant effect. These results indicate that in the head and muscle the predominant ChE form is acetylcholinesterase (AChE) for both sexes. The kinetic parameters for ChE activity (Km and Vmax) were similar in both sexes. The 96 h-LC50 value obtained for adult specimens of Iberian toothcarp was 3.17 mg/L. ChE activity in head and body tissues of both sexes was significantly inhibited in all concentrations tested (0.5, 1, 2 and 4 mg/L) after “in vivo” dichlorvos exposure. However, Iberian toothcarp was able to tolerate high concentrations of dichlorvos, and resist high levels of brain and muscle ChE inhibition without mortality. Both ChE inhibition and recovery followed a similar time-course pattern in response to sub-lethal exposure to dichlorvos (1 mg/L), and the enzyme activity did not return to control levels after 96 h in clean water. The results of this study show that ChE activity is a good biomarker of exposure to OP in the Iberian toothcarp adults.