Prevalence of free-living amoebae in five rivers associated with high human activity in Tehran province, Iran

Free-living amoebae (FLA) are ubiquitous protozoa capable of enduring harsh environmental conditions. These microorganisms are commonly found in water, soil, and air and can be transmitted to humans in areas with high human activity. This study aimed to investigate the prevalence of FLA and their associated genotypes/species in five rivers in Tehran province, Iran. A total of 60 water samples were collected from the Jajrud, Kan, Farhzad, Darakeh, and Shadchay rivers. Samples were subjected to filtration and cultivation onto non-nutrient agar. The genera/species of FLA were characterized based on the amplification and sequencing of the specific genetic fragments. Microscopic analysis suggested the presence of trophozoites and cysts of FLA in 18/60 (30%) of samples, of which <i>Acanthamoeba</i> spp., <i>Vermamoeba</i> sp., and Vahlkampfiidae were identified. Sequence analysis showed the presence of the genotypes T11, T4, T3, and T5 in five, five, four, and one isolates, respectively. The molecular analysis of the T4 genotype showed gene flow between the current isolates with previously described sequences. The findings suggest a clear association between environmental and clinical isolates of <i>Acanthamoeba</i> spp. Therefore, scheduled monitoring of environmental waters, particularly in regions with high human activities, is highly recommended.