Background: Free-living amoebas are very abundant in nature, especially in water and soil. Some species of amoebas cause serious and sometimes fatal infections such a keratitis in people with normal and suppressed immune systems. Based on the prevalence of amoebic keratitis in Iran, it is important to consider the free-living amoeba. Objectives: The current study aimed to determine the presence of Acanthamoeba and Hartmannella spp. in soil samples of Sari, North of Iran. Patients and Methods: A total of 96 soil samples from three areas of Sari were collected to be examined for soil contamination with Acanthamoeba and Hartmannella spp. Soil samples were investigated for the presence of these parasites by Sheather's flotation technique and Gimsa staining method. The identification of Acanthamoeba and Hartmannella spp. at the genus level in this study was based on distinctive features of double walled cysts. Results: Of the 96 studied samples, from dierent environmental locations in Sari, 38 (39.6%) Acanthamoeba and 5 (5.2%) Hartman- nella species were identified. Conclusions: The results of the present study revealed that soil resources of these areas were contaminated with opportunistic amoebas such as Acanthamoeba and Hartmannella spp. and this may lead to severe diseases in high-risk people, such as immuno- compromised patients.