It has long been recognized that thyroid hormone (TH) is essential for normal brain development in both humans and animals, and there is growing evidence that environmental chemicals can disrupt the thyroid system. In the present work, we used a two-hybrid yeast assay to screen for agonistic or antagonistic thyroid receptor (TR) mediated effects in drinking waters. We found no TR agonistic, but TR antagonistic activities in all samples from the drinking water processes. The TR antagonistic activities in organic extracts of water samples were then calibrated regarding to a known TR-inhibitor, NH3, and were expressed as the NH3 equivalents (TEQbio). The observed TEQbio in waters ranged from 180.8+/-24.8 to 280.2+/-48.2 microg/L NH3. To identify the specific compounds responsible for TR disrupting activities, the concentrations of potentially thyroid-disrupting chemicals including organochlorine pesticides (OCPs), phenols, and phthalates in organic extracts were quantitatively determined and their toxic equivalents with respect to NH3 (TEQcal) were estimated from their concentration-dependent relationships, respectively, using the same set of bioassays. Based on the TEQ approach, it was revealed that dibutyl phthalate (DBP) accounted for 53.7+/-8.2% to 105.5+/-16.7% of TEQbio. There was no effective removal of these potential thyroid disrupting substances throughout drinking water treatment processes.