Abstract: The urinary excretion of estrone sulfate ($\text{E}{1}\text{S}$) by 25 free-roaming feral horses (Equus caballus) was measured by radioimmunoassay applied to extracts of urine-soaked soil. Twelve of 15 mares having $\text{E}{1}\text{S}$ concentrations >1.0 mg/mg creatinine (x = 2.64 +- 1.02 [SD]) produced foals. All 10 mares with $\text{E}{1}\text{S}$ concentrations <1.0 mg/mg creatinine (x = 0.44 +- 0.26) did not foal. Extracting urine from soil and measuring $\text{E}{1}\text{S}$ and creatinine can be used to determine pregnancy in free-roaming feral horses without the stress of capture or immobilization.