Emerging contaminants are at the forefront of modern water quality research. Steroid sex hormones are among the most potent of these compounds. Spatial variations of steroid hormones were evaluated in three watersheds in rural Wyoming. Hormone concentrations were evaluated in the context of proximate upstream land uses and position in the watershed. Hormone contamination was found to be a localized phenomenon as hormones were not found to be transported downstream. Hormone concentrations were correlated with water quality parameters. Water temperature and E.Coli concentrations were found to provide statistically significant correlations with hormone concentration. Testosterone was the most frequently occurring sex hormone within the study area. The highest concentrations of hormones were associated with the Wastewater Treatment Plant and urban land uses. Hormones were also found to be associated with rangeland and irrigated crop land uses.