Pharmaceuticals in the aquatic environment: β-blockers as a case study

This thesis was submitted for the degree of Doctor of Philosophy and awarded by Brunel University, 20/09/2008.The presence of many human pharmaceuticals in the aquatic environment is now a worldwide concern and yet little is known of the chronic effects that these bioactive substances may be having on aquatic organisms. This study used mammalian pharmacodynamics to predict the mode of action of the 13-blocker, propranolol, on fish, in order to identify chronic effects in fathead minnows. β-blockers target β1- and β2-adrenergic receptors in humans and hence these receptors were characterised in the fathead minnow. It was found that fish possess β1- and β2-ARs that are structurally very similar to their mammalian counterparts. Further, the distributions of these two β-ARs in various organs of the fathead minnow were similar to those in mammals. Pair-breeding assays were conducted, in which fathead minnows were exposed to various concentrations of propranolol. To discover whether β-ARs had been up or down regulated by propranolol, molecular analysis was conducted on different tissues of the exposed fish using real-time polymerase-chain reactions (RT-PCR). Exposure of fathead minnows to propranolol caused acute toxicity at 10 mg/L. Propranolol caused a statistically significant decrease in reproduction at 1.0 mg/L, dose-related decreases in male weight, condition index and fatpad weight, and a dose-related increase in female GSI. Molecular analysis of βl- and β2-AR expression levels revealed a dose-related decrease in β2-AR expression in fathead liver and heart. LOEC and NOEC values were 0.1 mg/L and 0.01 mg/L, respectively. Propranolol plasma concentrations in fish exposed to water concentrations of 0.1 and 1.0 mg/L were greater than the human therapeutic concentration and hence these data very strongly support the fish plasma model proposed by Huggett et al. (2001).European Union (as part of the ERAPharm consortium