Adverse impacts on the health of some fish populations, such as skewed sex distributions, have been noted in surface waters and in laboratory experiments with relatively low concentrations (above 25 ng/L) of natural estrogen (17 beta-estradiol--E2). Sources of E2 to surface and ground waters can include avian, human, and mammalian waste products. The Ozark Plateau Aquifer (OPA) is a karstic basin that receives a significant portion of its water through losing reaches of rivers. Thus, there is a direct connection between surface water and ground water. The OPA was targeted for an E2 study to assess the potential for adverse health effects to aquatic organisms living in the system. Eight springs, which drain the aquifer, were sampled quarterly. The concentrations of E2 in the OPA ranged from 13 to 80 ng/L. For any one sampling event, the concentrations of E2 at the spring waters were statistically similar; however, the concentrations of E2 at all springs varied throughout the year. At Maramec Spring, one of the larger springs, the E2 concentration, was correlated with discharge. Based on the correlation between discharge and E2 concentration, aquatic organisms living in the plateau or in its discharged waters, including the threatened southern cavefish T. subterraneus, are exposed to concentration of E2 above 25 ng/L approximately 60% of the time. This implies that organisms living in karst basins throughout the OPA are likely exposed to E2 concentrations that may adversely impact their reproductive success for a significant portion of each year.