Existing in vivo tests (with the exception of the full lifecycle test) are not adequate for assessing the reproductive effects of endocrine disrupting chemicals (EDCs) on fish, and hence the need for partial life-cycle tests has been recognized internationally. In this paper we describe the development of a short-term (6 week) reproductive performance test for EDCs using pair-breeding fathead minnows (Pimephales promelas). In the test, reproductive performance in paired fish is assessed over two 3 week periods, one with exposure to the test chemical and one without. The test is highly integrative and measures effects of exposure to chemicals on fecundity, gonadosomatic index (GSI), vitellogenin (VTG) induction, and secondary sexual characteristics (fat pad and tubercles in males). In this test, exposure to butyl benzyl phthalate (BBP) at a nominal concentration of 100 µg/L (measured concentration between 69 µg/L and 82 µg/L) had no discernible effects on reproductive performance. In contrast, all reproductive parameters measured were affected by exposure to 4-NP, albeit some (e.g. VTG induction and reduction in the prominence of secondary sexual characteristicslowest effective dose between 0.65 µg/L and 8.1 ( 1 µg/L [measured]) were more sensitive than others (e.g. number of eggs and spawnings, where the lowest effective dose was between 8.1( 1 µg/L and 57.7 ( 3 µg/L [measured]). Concentrations of 4-NP at or above 48 µg/L [measured] inhibited reproduction completely.
Concern about possible adverse effects caused by the inadvertent exposure of humans and wildlife to endocrine active chemicals, has led some countries to develop an in vitro-in vivo screening programme for endocrine effects.In this paper, a previously described estrogen-inducible recombinant yeast strain (Saccharomyces cerevisiae), is used to investigate a number of issues that could potentially lead to the mislabelling of chemicals as endocrine disruptors. The chemicals studied were; 17 -estradiol, dihydrotestosterone, testosterone, estradiol-3-sulfate, 4-nonylphenol, 4-tertoctylphenol, 4-tert-butylphenol, bisphenol-A, methoxychlor, 2,2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane, butyl benzyl phthalate, 4-hydroxytamoxifen, and ICI 182,780. Many of the issues raised will also affect other in vitro assays for endocrine activity, and some will be relevant to the interpretation of data from in vivo assays. These examples illustrate that considerable care and thought must be applied when interpreting results derived from any single assay. Only by using a suite of assays will we minimise the chances of wrongly labelling chemicals as endocrine disruptors.2
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