Tests for potentially pathogenic amoebae were carried out in order to determine the presence of free-living amoebae of genus Acanthamoeba in soil and beach sand sources related to human environments in Tenerife, Canary Islands, Spain. Acanthamoeba identification was based on the morphology of cyst and trophozoite forms and PCR amplification with a genus specific primer pair. The pathogenical potential of Acanthamoeba isolates was characterized by temperature and osmotolerance assays and PCR reactions with two primer pairs related to Acanthamoeba pathogenesis. The results demonstrate the presence of potential pathogenic strain in both sources. Thus, some of the amoebae found in these habitats could act as opportunistic pathogens and may present a risk to human health.