The estrogen receptor agonist fate of hexane extracts from various locations and phases (liquid and solid) within one pilot-scale and two full-scale wastewater treatment facilities were examined by use of the receptor-binding yeast estrogen screen (YES assay). Estrogenic activity was found in samples that contained a high concentration of biological solids and was particularly high in the suspended solid fraction from biosolids treatment facilities. Mass balances revealed that the estrogenic activity associated with the processed biosolids constituted between 5 and 10% of the influent estrogenic activity, while the treated liquid effluent prior to disinfection contained between 26 and 43%. Overall, this suggests that between 51 and 67% of the estrogenic activity contained in the influent wastewater was either biodegraded during the wastewater or biosolids treatment processes or was unavailable to the extraction/detection procedure. In both aerobic and anaerobic digestion, mass balances revealed an increase in estrogenic activity as treatment progressed and biosolids destruction occurred. The estrogenic activity associated with the solid phase decreased during mesophilic aerobic digestion. A correlation was observed between the estrogenicity of mixed liquor suspended solids and aerobic sludge age and suggests that wastewater treatment facilities can be designed and operated to enhance the sorption and removal of estrogenic compounds from the liquid phase.

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