Cross-species comparison of fluoxetine metabolism with fish liver microsomes

Pharmaceuticals discharged in municipal wastewater are of emerging concern because of their potential for inducing biological effects in aquatic organisms. Selective serotonin reuptake inhibitors (SSRIs), pharmaceuticals prescribed to treat chronic depression, have been detected in receiving and wastewaters. Fluoxetine is a highly prescribed model SSRI used to assess impacts of antidepressants on aquatic organisms. In this study, in vitro hepaticfluoxetine metabolism was determined in several model fish species: rainbow trout, goldfish, zebrafish and killifish. Incubation of fluoxetine with hepatic microsomes from trout pre-treated with carbamazepine showed a time-dependant loss of fluoxetine, concomitant with an increase in norfluoxetine, the major mammalian demethylated metabolite. However, fluoxetine was not well metabolized in reactions with hepatic microsomes from untreated fish. Fluoxetine loss was greater than norfluoxetine production, indicating that norfluoxetine is not the predominant fluoxetine biotransformation product in fish. Furthermore, norfluoxetine was often undetected, possibly indicating that fluoxetine demethylation is a minor metabolic pathway in fish. Inter-species differences in fluoxetine metabolism were not evident because of high intra-species variability, although killifish appeared to have the highest hepatic metabolic capacity for fluoxetine. Fluoxetine metabolism in mammals is catalyzed by cytochrome P450 (CYP) enzymes. Trout were exposed to knownCYP inducers, carbamazepine and 3-methylcholanthrene, to assess potential induction of hepatic fluoxetine metabolism. Microsomes from 3-methylcholanthrene treated fish did not induce detectable changes in fluoxetine concentrations in vitro, indicating that fish CYP1s are not involved in fluoxetine metabolism; the CYPs involved are still unclear. Identification of metabolites other than norfluoxetine warrants further investigation.