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The genus  Acanthamoeba  is a free-living amoeba widely distributed in various aquatic environments. It is an etiologic cause of amoebic encephalitis and keratitis particularly for immunocompromised individuals. The purpose of the present study was to investigate  Acanthamoeba  species prevalence in household and hospital potable water in Beni-Suef governorate, Egypt, and to employ sequencing methods to identify positive  Acanthamoeba  species isolates and their potential health risks. Sixty tap water samples (30 household and 30 governmental and private hospital settings) collected from Beni-Suef governorate, Egypt were filtered, cultured on non-nutrient agar, identified by morphotyping keys after staining with Giemsa stain and then confirmed by PCR using  Acanthamoeba  specific primers. Twenty positive samples were successfully genetically characterized and phylogenetically analyzed to identify  Acanthamoeba  species. The total detection rate for  Acanthamoeba  was 48/60 (80%);  Acanthamoeba  contamination in water collected from domestic houses was higher than in hospitals; 27/30 (90%) versus 21/30 (70%) with statistical significant value ( P  value = 0.05). Sequencing of 20 positive isolates revealed  Acanthamoeba  T4 in 65% and T2 in 35%. To our knowledge, this is the first research that documents the occurrence and phylogeny of  Acanthamoeba  species in Beni-Suef, Egypt. The presence of a higher percentage of  Acanthamoeba  species in tap water, in particular T4, highlights the potential health hazards for immunocompromised individuals and emphasizes the urgent need for the implementation of effective filtration and disinfection measures.

Molecular characterization and phylogenetic analysis of Acanthamoeba isolates in tap water of Beni-Suef, Egypt