Inhibition of egg production in zebrafish by fluoxetine and municipal effluents: A mechanistic evaluation

This study explored the impact of nominal concentrations of ethinylestradiol (EE2; 10 ng/L), fluoxetine (FLU; 0.32, 3.2, 32 mu g/L), and 1-50% treated municipal effluent on reproduction and liver function in sexually mature female zebrafish over a 7-day period. Compared with the control groups, FLU (32 mu g/L) and 50% effluent treatment significantly reduced the average eggs spawned by approximately 4.5 and 2 fold, respectively. FLU also decreased ovarian levels of 17 beta-estradiol (E-2) without affecting the gonadosomatic indices of the fish. The expression of ovarian genes involved in the production of prostaglandins, steroid biosynthesis, and gonadotropin receptors were quantified to determine a potential mechanism underlying the reduced egg production in FLU and effluent exposed fish. Quantitative real-time PCR analysis determined that ovarian aromatase, follicle stimulating hormone receptor (FSHr), and luteinizing hormone receptor (LHr) gene expression were significantly reduced by FLU suggesting that disruptions to the synthesis of ovarian steroids and the actions of gonadotropins may underlie the negative influence of FLU on ovarian E-2 and spawning levels. Potential mechanisms involved in the modest effects of the effluent on reproduction remain unknown, but the elevated levels of total ammonia and nitrite in the 50% effluent treatment groups compared with the other treatments should not be discounted. Liver expression of CYP3A65 was significantly induced by all treatments of effluent, while EE2 caused a reduction in the expression levels of CYP1A1, PXR, and CYP3A65. The results of the present study suggest that SSRI may disrupt reproductive functioning at concentrations greater than those found in receiving environments; yet, more research is warranted into to the possible effects of low levels of synthetic estrogens on liver function in exposed fish, particularly the PXR-CYP3A pathway. (C) 2009 Elsevier B.V. All rights reserved.