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Adult male fathead minnow were exposed for 14 or 28-days under flow-through conditions to undiluted filtered water samples from the rivers Meuse and Rhine in the Netherlands. The experiment included two vessels per treatment each containing 10 fish and samples of five fish were taken after 14 and 28 days. Additional groups were exposed to 17α-ethinylestradiol (EE2) as a reference and untreated drinking water as a negative control. Major endpoints examined included induction of vitellogenin (VTG) synthesis, VTG mRNA activity, hepato- and gonadosomatic indices (HSI and GSI) and gonadal histology. No significant difference was recorded in body weight or mean GSI values between the various treatments. Only exposure to Meuse water resulted in significantly higher HSI means after 14 days. Histological examination showed no apparent effects on gonadal tissue except for eosinophilic blood plasma in fish exposed to Meuse water or EE2. After 14 and 28 days, elevated VTG and VTG mRNA levels were measured in most livers of the fish exposed to Meuse water, but not in the fish exposed to Rhine water. This was confirmed by measuring estrogenic responses in the in vitro ER CALUX® assay. Induction of VTG synthesis proved to be the most sensitive endpoint in the Non Spawning Male Fish Assay for in vivo detection of bio-available estrogenic activity supplementary to a sensitive in vitro assay. The other endpoints examined varied too much and required a higher number of fish or replicates to achieve sufficient power for statistical testing making them less animal friendly. Keywords: Estrogenic activity, Non spawning male fish assay, Fathead minnow, Vitellogenin, River waters, Drinking water

To estimate the impact of accidental oil-spills on cod fisheries a model framework is developed in which a Dynamic Energy Budget (DEB) model is applied to assess mortality caused by petroleum substances in early life stages. In this paper we report on a literature search and DEB analyses, aiming for cod specific DEB-parameters. Furthermore, we explored the relevance of Fathead minnow DEB-parameters as surrogate by comparing LC50 values calculated from DEB-parameters with literature. Cod specific DEB-parameters could not be estimated based on available literature. LC50 values calculated from Fathead minnow DEB-parameters were higher than literature LC50 for early life stages of fish. Applying an extrapolation factor of 50 to the DEB-parameters resulted in LC50 values that were below literature irrespective of life stage. Therefore, we propose to use the last as an estimate for early life stages in cod and recommend relevant experiments with individual petroleum substances on cod.