Attempts to better understand causal factors affecting estrogenicity in municipal wastewater have primarily focused on analytical evaluation of specific chemical estrogens and the use of estrogen receptor (ER) based in vitro assays. To compare analytical,in vitro, and in vivo assays for estrogenicity, wastewater from four New York and one Texas municipal wastewater facilities was evaluated for estrogenic activity using the yeast estrogen screen assay (YES) and an in vivo fish vitellogenin (VTG) assay. Estrogenic activity, as measured by the YES assay, was observed in methanol and/or methylene chloride eluents from C18 extracts in two of the New York treatment facilities and the Texas facility. Estradiol equivalents for the YES assay data ranged from

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