Isolation and genotyping of Acanthamoeba species and Vahlkampfiidae in the harsh environmental conditions in the centre of Iran.

Different species of free-living amoeba (FLA) have been abundantly isolated in harsh environmental conditions such as hot springs and brackish water. The present study aimed to isolate, genotype, and evaluate the pathogenicity of FLAs in Qom Roud, a large river, in the centre of Iran. About 500 mL of water samples (n = 30) were collected from each sampling site and were investigated for the presence of FLAs using morphological and molecular characters. Genotype identification was performed using DNA sequencing and a phylogenetic tree was constructed with the MEGA X software. The pathogenic potential of all positive isolates was evaluated using the tolerance ability test. Morphological and molecular analysis indicated that 14 (46.66%) and two (6.66%) water samples were positive for Acanthamoeba species and Vahlkampfiidae, respectively. According to sequence analysis, Acanthamoeba isolates related to the T4 genotype and Vahlkampfiidae sequences were similar to Naegleria philippinensis. In the next step, thermo- and osmotolerance tests indicated four Acanthamoeba strains are extremely pathogenic. Our data showed the presence of potentially pathogenic Acanthamoeba T4 genotype and N. philippinensis in the super harsh Qom Roud. Contamination of water with virulent T4 genotype of Acanthamoeba may pose risk factors for contact lens users, children, and immunocompromised people.