A total of 136 samples of tap water were collected from state and municipal schools between March and November 2009. The samples were filtered through cellulose nitrate membranes that were seeded at non-nutrient agar 1.5% containing an overlayer of Escherichia coli suspension. Thirty-one (22.79%) tap water samples investigated were found positive for free-living amoebae (FLA). From these, 13 presented as FLA that seems to belong to the genus Acanthamoeba. All samples of FLA were cloned and identified as belonging to the genus Acanthamoeba by the morphology of cysts and trophozoites and by PCR using genus-specific primers that amplify the ASA.S1 region of 18S rDNA gene. Physiological tests of thermotolerance and osmotolerance were used to evaluate the pathogenicity of the isolates. The sequencing analysis by comparing the sequences submitted to GenBank, showed genotype distribution into groups T2, T2/T6, T6, and T4. In tests of thermotolerance and osmotolerance, 50% of the isolates had a low pathogenic potential. The results indicated the presence of Acanthamoeba in tap water in Rio Grande do Sul, Brazil, revealing its importance and the need for more epidemiological studies to determine their distribution in the environment and its pathogenic potential.
The present study investigated the susceptibility of Acanthamoeba spp. trophozoites to two multipurpose systems for cleaning and maintenance of contact lenses. Three strains of trophozoites from the ATCC (A. castellani T4, A. castellani Neff, and A. polyphaga) and two Acanthamoeba isolates obtained from swimming pools (PT5 and PO1) were placed in monoxenic culture. To test their survival in cleaning solutions for contact lenses, the trophozoites were exposed for 4 and 24 h to two multipurpose solutions (A and B), and were then inoculated into a new monoxenic culture. Amoebic growth on the plates was observed after 72 h of incubation. Trophozoites from all three ATCC strains and one isolate from a swimming pool (PO1) grew in all plates after 4 h of exposure to solutions A and B. After 24 h, the ATCC strains and the PO1 isolate showed growth in most of the plates treated. Only the PT5 isolate showed susceptibility to both solutions over the time intervals tested. The two solutions were not completely effective against most strains and isolates over the time intervals tested. These results are important, since species of Acanthamoeba are widely distributed in the environment and are potential agents of eye pathologies.
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