Aims The study established the inactivation kinetic parameters of an Acanthamoeba cyst isolate subjected to heating and chlorination. Methods and Results A strain of Acanthamoeba was isolated and purified from an area surrounding a pilot food plant. Mature cysts (14 days) were subjected to heat inactivation studies at 71, 76, 81, 86 and 91 degrees C; and chlorination at 100, 200, 300, 400 and 500 ppm. The decimal reduction times (D-values) at 71, 76, 81, 86 and 91 degrees C were 18 center dot 31, 9 center dot 26, 7 center dot 35, 4 center dot 52 and 1 center dot 81 min respectively. The calculated thermal resistance constant (z-value) was 21 center dot 32 degrees C (R-2 = 0 center dot 96-0 center dot 97). The D-value in 100, 200, 300, 400 and 500 ppm chlorine-treated water were 47 center dot 17, 25 center dot 06, 24 center dot 51, 23 center dot 70 and 18 center dot 55 min respectively. The chlorine resistance constant (z-value) was 1179 ppm chlorine (R-2 = 0 center dot 65-0 center dot 74). Conclusions Results demonstrated high resistance of the isolated Acanthamoeba cysts towards the common methods applied in ensuring food and food processing environment sanitation. Significance and Impact of the Study The resistance parameters of the test organisms established in this study may be used in the establishment of Sanitation Standard Operating Procedures (SSOPs), which are often based on inactivation of bacteria. These SSOPs could render better protection to food and food processing environments.