Combination of in vitro bioassays encompassing different mechanisms to determine the endocrine-disrupting effects of river water

In this study, the total toxic effects of river water samples were assessed using a series of cell culture bioassays which encompassed different mechanisms, based on specific modes of action. River water samples were collected from three tributaries of the Youngsan River in the western portion of Korea. We confirmed that Youngsan River water was polluted with a complex mixture of estrogenic and dioxin-like compounds. The total toxic effects of the downstream water samples were found to be higher than that of the upstream water samples. In the upstream water samples, total estrogenic activity was measured to be between 0.005 and 0.049 ng-EEQ/l (17β-estradiol-equivalent concentration) and no CYP1A activity was detected. In the downstream water samples, however, total estrogenic activity was measured to be between 0.021 ng-EEQ/l and 1.918 ng-EEQ/l, and total CYP1A activity was between 0.63 and 3.55 μg-MEQ/l (3-methylcholanthrene-equivalent concentration). When assessed according to a concentration–response curve, downstream water sample extracts exerted dual actions on estrogen receptors, depending on the concentration volume of the samples. The concentration volume range proximal to the original water sample exhibited estrogenic activity, whereas antiestrogenic activity was observed at high concentration volumes (more than 5 times concentration) in the extracts. This study involved a combination of in vitro bioassays, designed to encompass different mechanisms. The bioassays used included the estrogen receptor binding affinity test, E-screen assay, aromatase assay, and EROD assay. These tests provided a great deal of useful information regarding the potency and action modes of estrogenicity and antiestrogenicity inherent in the sampled river water. Although further study is necessary to determine the relationship between toxic responses in in vitro bioassay systems and chronic toxicity in aquatic organisms, our approach is expected to be fairly accurate with regard to the detection of endocrine-disrupting effects in an aquatic environment.