The aim of this study was to investigate the in vitro effects of an ethanolic extract of propolis on the growth and adherence of Acanthamoeba castellanii trophozoites and cysts.The effect of propolis with concentrations of 8.0, 6.0, 5.0, 4.0, 3.0, and 2.0 mg/mL on the proliferation of A. castellanii trophozoites, and with a concentration of 62.25, 31.25, 15.62, 7.81, 3.90, 1.95, and 0.97 mg/mL on the proliferation of A. castellanii cysts, at 1, 3, 6, 12, 24, 48, and 72 h were examined in vitro.After 1-72 h, incubation in concentrations between 2.0 and 6.0 mg/mL, its effect was amoebistatic; at concentrations of 8.0 mg/mL and higher, its effect was amoebicidal. After 48 h or longer incubation times at 15.62 mg/mL and at higher concentrations, the propolis extract was cysticidal. At concentrations of 1.97 mg/mL or lower, there was no observable effect at any time point.These findings indicate that ethanolic extract of propolis has amoebicidal, as well as cysticidal, properties for Acanthamoeba trophozoites and cysts. Propolis alone, or in combination with other amoebicidal agents, may be used in clinical practice after further investigations.