The present work was planned to isolate Free living amoebae from differentenvironmental sources in Egypt e.g. water, soil, and air. Identification of FLA wasbased on morphological differences detected by light microscope. FLA wereidentified morphologically on the basis of trophozoites, cysts, locomotion,flagellation test. Twenty five samples were obtained and examined; fifteen watersamples (five swimming pool, five River Nile and five tap water samples), fivesoil and five air samples. All samples were cultivated on NNA plates enrichedwith E.coli and incubated at 37ºC for 2- 7 days. Free living amoebae were isolatedfrom seventeen samples out of twenty five samples (68%); five swimming pool,five River Nile, three tap water, one air and three soil samples. Acanthamoebawas isolated from thirteen samples out of the seventeen positive samples (76.5%)while Naegleria was isolated from four samples (23.5%). Naegleria was positivefor flagellation test. Their trophozoites showed rounded smooth lobopodia.Naegleria cysts were single walled. On the other hand Acanthamoeba wasnegative for flagellation test, trophozoites showed spiky acanthapodia and their cysts were having double wrinkled walls. The effect of water chlorination onamoebic growth was studied. Naegleria was found to be sensitive to chlorine level of 0.6-0.7 μg/ml. This ideal level of chlorination of swimming pools makes themnegative for Naegleria. Chemotherapeutic agents including azithromycin andneurazine were tried in vitro against FLA. Azithromycin was found to be effectiveagainst FLA at MIC of 10 μg/ml. Neurazine was effective against N.fowleri atMIC of 10μg/ml while it was effective against Acanthamoba at MIC of 100μg/ml.