Different Acanthamoeba species are amphizoic organisms distributed in wide range of habitats in natural and man-made environments; they are also detected on surfaces of equipment and accessories in health facilities. Some strains of the amoebae are causative agents of the vision-threatening human disease Acanthamoeba keratitis, mainly reported in contact lens wearers. An exceptional high resistance of Acanthamoeba trophozoites and particularly cysts to chemicals, disinfectants and drugs is believed as influencing difficulty resulting in unsuccessful therapeutic management. As Acanthamoeba keratitis is the serious medical problem worldwide, different chemicals with possible activity against environmental and clinical Acanthamoeba strains are tested. In our study, selected disinfectants used in health care settings and laboratories were tested and their efficacy against the corneal strains Acanthamoeba castellanii and A. polyphaga , and environmental A. castellanii Neff strain was assessed. Comparative assessment of results of the assays show that, apart from amoebistatic effects, the disinfectants indicated expected cysticidal efficacy.