Morphological and Molecular Identification of Acanthamoeba Spp from Surface Waters in Birjand, Iran, During 2014-2015

Background & Aims of the Study: Free-living amoebae (FLA) are opportunistic and ubiquitous protozoa that are widely found in various environmental sources. They are known to cause serious human infections including a fatal encephalitis, a blinding keratitis, and pneumonia. So, due to their medical importance, the identification of free living amoeba in water resources, as a source of human infection, is necessary. The objective of this study was to isolate the Acanthamoebaspp from the surface waters of Birjand, Iran, during 2014-2015 by Morphological and molecular method. Materials and Methods:  In a cross-sectional study, 50 samples were collected from different localities of Birjand city including the surface waters, pools and fountains in parks,squares and water stations from the October 2014 to the January 2015.Each sample was filtered through a nitrocellulose membrane filters and cultured on non-nutrient agar (NNA) with Escherichia coli suspension and incubated for 1 week to 2 months at room temperature.The plates were examined by the microscopy to morphologically identify Acanthamoeba species. Following DNA extraction, PCR specific primers was used to confirm the identification morphologically. Results:  Out of 50 water samples, 19 (38%) were positive for Acanthamoebatrophozoites and cysts according to the morphological criteria. In addition, Acanthamoebaspp was identified by PCR method, using genus specific primers pairs in 15 (78.9%) cases of positive cultures, showing anearly 500bp band. Conclusion: According to the prevalent of Acanthamoebaspp in the surface stagnant waters of Birjand, more attention to the potential role of such waters in transmission of infection by the regional clinicians and health practitioners is necessary.