A four-hour yeast bioassay for the direct measure of estrogenic activity in wastewater without sample extraction, concentration, or sterilization |
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Authors: | Heather A. Balsiger Roberto de la Torre Wen-Yee Lee Marc B. Cox |
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Affiliation: | a Department of Biological Sciences, University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, United States b Environmental Science and Engineering Ph.D. Program, University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, United States c Department of Chemistry, University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, United States d Border Biomedical Research Center, University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, United States |
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Abstract: | The assay described here represents an improved yeast bioassay that provides a rapid yet sensitive screening method for EDCs with very little hands-on time and without the need for sample preparation. Traditional receptor-mediated reporter assays in yeast were performed twelve to twenty four hours after ligand addition, used colorimetric substrates, and, in many cases, required high, non-physiological concentrations of ligand. With the advent of new chemiluminescent substrates a ligand-induced signal can be detected within thirty minutes using high picomolar to low nanomolar concentrations of estrogen. As a result of the sensitivity (EC50 for estradiol is ∼ 0.7 nM) and the very short assay time (2-4 h) environmental water samples can typically be assayed directly without sterilization, extraction, and concentration. Thus, these assays represent rapid and sensitive approaches for determining the presence of contaminants in environmental samples. As proof of principle, we directly assayed wastewater influent and effluent taken from a wastewater treatment plant in the El Paso, TX area for the presence of estrogenic activity. The data obtained in the four-hour yeast bioassay directly correlated with GC-mass spectrometry analysis of these same water samples. |
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Keywords: | Effluent Environmental estrogens Estrogenic activity Wastewater Yeast bioassay Yeast estrogen screen |
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