The aim of the present study was to investigate the effects of the synthetic progestin levonorgestrel (LNG) on the reproductive endocrine system of a teleost fish, the roach (Rutilus rutilus). Pubertal roach were exposed for 28 days in a flow-through system to four concentrations of LNG (3, 31, 312, and 3124 ng/l). Both males and females treated with 3124 ng/l LNG exhibited the upregulated levels of vitellogenin and oestrogen receptor 1 mRNA in the liver. At the same concentration, LNG caused a significant upregulation of the mRNA expression of the gene encoding luteinising hormone β-subunit (lhβ) and the suppression of the mRNA expression of the gene encoding follicle-stimulating hormone β-subunit (fshβ) in the pituitary of both male and female roach. A lower LNG concentration (312 ng/l) suppressed mRNA expression of fshβ in males only. Females treated with 3124 ng/l LNG exhibited significantly lower plasma 11-ketotestosterone (11-KT) and oestradiol (E2) concentrations, whereas their testosterone (T) level was higher compared with the control. Females exposed to 312 ng/l LNG presented significantly lower plasma E2 concentrations. Males exposed to ≥31 ng/l LNG exhibited significantly reduced 11-KT levels. As determined through a histological analysis, the ovaries of females were not affected by LNG exposure, whereas the testes of males exposed to 31 and 312 ng/l LNG exhibited a significantly higher percentage of spermatogonia B compared with the control. The results of the present study demonstrate that LNG disrupts the reproductive system of pubertal roach by affecting the pituitary gonadotropin expression and the sex steroid levels. This disruption was determined to occur in males after exposure to an environmentally relevant concentration (31 ng/l). Moreover, the highest tested concentration of LNG (3124 ng/l) exerted an oestrogenic effect on fish of both sexes.

Department of Ecophysiology and Aquaculture Leibniz Institute of Freshwater Ecology and Inland Fisheries Mueggelseedamm 310 D 12587 Berlin Germany

Department of Ecophysiology and Aquaculture Leibniz Institute of Freshwater Ecology and Inland Fisheries Mueggelseedamm 310 D 12587 Berlin Germany; Department Ecological Impact Research and Ecotoxicology Technische Universität Berlin Ernst Reuter Platz 1 10587 Berlin Germany

Department of Ecophysiology and Aquaculture Leibniz Institute of Freshwater Ecology and Inland Fisheries Mueggelseedamm 310 D 12587 Berlin Germany; Department of Endocrinology Humboldt University Berlin Invalidenstrasse 42 D 10099 Berlin Germany

Research Institute of Fish Culture and Hydrobiology South Bohemian Research Centre of Aquaculture and Biodiversity of Hydrocenoses Faculty of Fisheries and Protection of Waters University of South Bohemia in Ceske Budejovice Zatisi 728 2 CZ 38925 Vodnany Czech Republic

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