Are steroids really the cause for fish feminization? A mini-review of in vitro and in vivo guided TIEs

Feminization of fish has been reported throughout the world in freshwater and marine systems. While the population impacts are conflictive, enough negative effects warrant additional research into causation. In order to ascertain the identities of specific feminizing agents, variants of toxicity identification evaluations (TIEs) have been employed. The majority of these evaluations have utilized in vitro estrogen receptor-based cell-lines to identify chromatographic fractions that possess biological activity from predominately wastewater derived from municipal treatment facilities and have concluded that synthetic and natural estrogens are the primary cause for feminization of fish. This paper will focus on three aquatic systems impacted by wastewater originating from purely domestic, and industrial/domestic secondary treatment systems. Wastewater and sediment extracts were evaluated by in vitro and in vivo biological responses in a TIE fractionation design. While in vitro responses tended to mirror in vivo responses in purely domestic wastewater systems, in vitro responses tended to severely underestimate in vivo estrogenic activity when normalized to estradiol equivalents in more complex systems. TIE fractionation schemes using in vivo biological responses failed to indicate any relationship to steroids in either wastewater or sediment extractions. These data consistently support the view that mechanisms other than direct ER binding and activation by toxicants may be important in the feminization of fish particularly residing in habitats that receive complex wastewater or agricultural effluents.