Objective: The main objective of this study was to investigate the effect of brief exposure to subtherapeutic concentrations of chlorhexidine gluconate on germ tube formation of Candida albicans isolates obtained from smokers, diabetics, asthmatics using steroid inhalers and healthy individuals. Materials and Methods: Forty isolates of C. albicans were used in this study. All these isolates were quantified for germ tube formation without exposure to the drug and were used as the control group for data analysis. Isolates were also exposed to three subtherapeutic concentrations of chlorhexidine gluconate (0.00125, 0.0025 and 0.005%) for 30 min (limited exposure); the antiseptic was then removed and germ tube formation of these isolates was quantified microscopically following incubation in a germ tube-inducing medium. Results: Compared with the unexposed controls, brief exposure to all concentrations of chlorhexidine gluconate suppressed the ability of the C. albicans isolates to form germ tubes in increasing order by 13.72% (p < 0.001 to p = 0.02), 46.16% (p < 0.001) and 72.46% (p <0.001). Conclusions: These findings show that brief exposure to subtherapeutic concentrations of chlorhexidine gluconate may modulate germ tube formation of C. albicans isolates, thereby suppressing their pathogenicity, and further elucidate the pharmacodynamic mechanisms by which chlorhexidine gluconate may operate in vivo.