Ozone is widely used to disinfect drinking water and wastewater due to its strong biocidal oxidizing properties. Recently, it was reported that hydroxyl radicals ( ⅐ OH), resulting from ozone decomposition, play a significant role in microbial inactivation when Bacillus subtilis endospores were used as the test microorganisms in pH controlled distilled water. However, it is not yet known how natural organic matter (NOM), which is ubiquitous in sources of drinking water, affects this process of disinfection by ozone-initiated radical reactions. Two types of water matrix were considered for this study. One is water containing humic acid, which is commercially available. The other is water from the Han River. This study reported that hydroxyl radicals, initiated by the ozone chain reaction, were significantly effective at B. subtilis endospore inactivation in water containing NOM, as well as in pH-controlled distilled water. The type of NOM and the pH have a considerable effect on the percentage of disinfection by hydroxyl radicals, which ranged from 20 to 50%. In addition, the theoretical C T value of hydroxyl radicals for 2-log B. subtilis removal was estimated to be about 2.4 ؋ 10 4 times smaller than that of ozone, assuming that there is no synergistic activity between ozone and hydroxyl radicals.Ozone is widely used to disinfect drinking water and wastewater due to its strong biocidal oxidizing properties. Many studies on ozone disinfection have focused on the effects of ozone on various microorganisms and the factors affecting the inactivation of these microorganisms such as pH, temperature, contact time, ozone dose, and ozone demand (1,6,9,19). Among the factors, the role of pH in inactivating microorganisms is not well understood and is somewhat controversial (2,5,12,19,25). No pH requirement for inactivation of microroganisms or fast inactivation by low pH has been reported without much discussion of the exact duration of exposure to ozone or the coexistence of hydroxyl radicals. This is partly due to the limited approaches used which considered mainly the biocidal activity of molecular ozone (5, 11, 12) rather than the indirect effect of hydroxyl radicals resulting from ozone decomposition. On the other hand, several studies did emphasize the importance of hydroxyl radicals in microorganism inactivation (1, 3). Recently, Cho et al. (2) reported that under certain experimental conditions, the presence of hydroxyl radicals produced by ozone disinfection plays a significant role in the inactivation of Bacillus subtilis endospores in pH-controlled ozone-demand-free distilled water. In their study, the observed C T values for achieving a 2-log inactivation were 40% lower at pH 8.2 than at pH 5.6 when time-dependent ozone decay as a function of the reaction time was considered in relation to the pH difference. However, in the presence of excess hydroxyl radical scavengers (t-butanol), where it is difficult for hydroxyl radical to exist, all the observed C T values achieved parity, within a 10% margin of error, ...