The mutagenic activity of XAD-2 adsorbates and water extracts recovered from nine locations of the Kumho River was tested on S. typhimurium TA98 strain to identify the source of the mutagenicity. A sampling site, receiving effluents from the textile industrial complex located in Daegu City, showed extraordinarily high mutagenic activity, especially in the presence of S9 mixture, at all sampling time in both XAD-2 adsorbates and dichloromethane extracts. This indicated the existence of the frame-shift mutagens in the Kumho River, same type of mutagens detected in previous studies by other researchers in the Nakdong River into which the Kumho River discharges. The fractionation study showed that the mutagenic chemicals in the river water are mid-polar. Furthermore, mean tail length obtained by single cell gel electrophoresis assay (Comet assay) showed consistent dose-dependent DNA damage, indicating that the chemicals in the river water not only act as frame-shift mutagens but also break human lymphocytes DNA strain. Chemical identification of the mutagens should be required.