The Microscopic Detection and Identification of Acanthamoeba Spp. In Water Samples Following Enrichment by in Vitro-Cultivation Under Two Distinct Temperature Conditions.

Abstract Acanthamoeba is a genus of free-living amoeba commonly found in environmental sources such as water, soil and air, and can infect humans. There is a significant challenge in the detection and identification of members of this genus in water samples. In order attempt to overcome this challenge, we investigated using in vitro culture, under distinct temperature conditions, to grow and enrich amoeba prior to detection and identification. Aliquots of 150 water samples, collected from Rabat (30 from each river, fountain, sea, public bath and tap water), were individually inoculated into standard culture medium non-nutritive agar and incubated for two weeks at 25 °C and 30 °C under otherwise standard conditions. PCR was used to confirm the presence of Acanthamoeba DNA in positive samples. The findings showed that Acanthamoeba grew more rapidly at 30 °C than 25 °C, allowing improved microscopic detection and identification at the former temperature. This investigation shows clearly that the diagnostic sensitivity of an in vitro based culture system is temperature-dependent.