Steroid hormones are naturally synthesized by both humans and animals and are released into the environment. Significant levels of steroid hormones have been detected in sewage effluent around the world. The potential problem is that these hormones may interfere with the normal function of the endocrine systems, thus affecting reproduction and development in wildlife. Due to the major shortage of water in Western Cape, South Africa there is a great need to recycle water by either direct or indirect methods. The treated sewage effluent-natural surface water mixture found in the Kuils and Eerste Rivers is used directly for irrigation of agricultural areas. Sewage effluents were collected from four sites (Jonkershoek, Belville, Zandvliet, and Macassar) and subjected to C(18) solid phase extraction. Commercially available rapid ELISA kits were validated for the quantification of estrogens in these sewage effluent samples. Analysis of estrone, estradiol, and estriol levels showed a significant difference between the control site (Jonkershoek) and sewage effluent from the three sewage treatment works. Steroid hormone concentrations detected in these sewage effluents were similar to reports from Britain, Italy, Germany, Canada, and Netherlands.