Owing to the modification of testing methods of disinfectants or antiseptics, variations of bacteria according to characteristics of regions and resistance changes of bacteria, it is necessary that the bactericidal activities of disinfectants or antiscptics should be reevaluated nowadays. This study was carried out to reevaluate in the vitro bactericidal activity of Chlorhexidine gluconate solution. The results of experiment were summarized as follows. 1. For Chlorhexidine gluconate solution, minimal inhibitory concentrations of total bacteria taken from sewage water and Legionella bozemanii were $2.0 imes 10^{-3}$%, $1.0 imes 10^{-2}$%, respectively and were comparatively high. Minimal inhibitory concentration of Shigella flexneri was $1.6 imes 10^{-4}$%, and was comparatively low. 2. For total bacteria taken from sewage water, it was killed within 15 minute in 0.1% Chlorhexidine gluconate solution when number of cells was $1.6 imes 10^7$/ml. 3. For 0.0125% Chlorhexidine gluconate solution, decimal reduction times of Ps. aeruginosa, S. typhi, E. Coli were 45 sec, 25 sec, 18 sec repectively. For 1%, 0.125% Chlorhexidine gluconate solution, decimal reduction times of Legionella bozemanii were 10 sec, 45 sec respectively. 4. There was significant difference in the bactericidal activity of Chlorhexidine gluconate solution according to temperattire. Phenol coefficient of Chlorhexidine gluconate solution as using Staph. aureus was 100 and comparatively higher than that of other disinfectants. In comparison with other disinfectants, Legionella bozemanii was killed within 5 minutes in 0.02% KMnO$_4$ and 0.125% Chlorhexidine giuconate solution but was not killed within 3 minutes in 1% 0-cresol, 1% Phenol.