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

Background & Aims of the Study: Free-living amoebae (FLA) are opportunistic andubiquitous protozoa that are widely found in various environmental sources. They areknown to cause serious human infections including a fatal encephalitis, a blinding keratitis,and pneumonia. So, due to their medical importance, the identification of free livingamoeba in water resources, as a source of human infection, is necessary. The objective ofthis 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 Birjandcity including the surface waters, pools and fountains in parks,squares and water stationsfrom the October 2014 to the January 2015.Each sample was filtered through anitrocellulose membrane filters and cultured on non-nutrient agar (NNA) with Escherichiacoli suspension and incubated for 1 week to 2 months at room temperature.The plates wereexamined by the microscopy to morphologically identify Acanthamoeba species. FollowingDNA extraction, PCR specific primers was used to confirm the identificationmorphologically.Results:Out of 50 water samples, 19 (38%) were positive for Acanthamoebatrophozoites and cystsaccording to the morphological criteria. In addition, Acanthamoebaspp was identified byPCR 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 stagnantwaters of Birjand, more attention to the potential role of such waters in transmission ofinfection by the regional clinicians and health practitioners is necessary.

Morphological and Molecular Identification of AcanthamoebaSpp from Surface Waters in Birjand, Iran, During 2014-2015 Keywords:

Acanthamoeba , Water , PCR , Birjand , SurfaceWater , Microbiology , Iran