This research was undertaken to evaluate volatile chemicals which retained mycobactericidal activity in cattle slurry. Mycobacterium bovis, suspended in sterilized cattle slurry, was treated with different concentrations of five volatile chemicals with mycobactericidal activity. Following treatment of the slurry for specified time intervals, the reaction mixture was lyophilized to remove the volatile chemicals, and samples of the reconstituted slurry were used to inoculate flasks of Lowenstein-Jensen medium to determine survival or inactivation of the mycobacteria. Acetone, at a concentration of 22.5%, inactivated M. bovis in less than 24 h. Ammonium hydroxide, at a concentration of 1%, was mycobactericidal after 36 h. Chloroform at a concentration of 0.5%, ethyl alcohol at a concentration of 17.5% and xylene at a concentration of 3% inactivated the mycobacteria within 48 h. Some of the volatile chemicals with mycobactericidal activity are potentially useful at farm level.