INVESTIGATION OF ACANTHAMOEBA SPP. WITH CULTURE AND MOLECULAR METHODS IN THE ENVIRONMENTAL WATER SAMPLES

<strong>Objective:</strong> Acanthamoeba spp. are free-living amoebae found in a variety of environments, including seawater, lakes, rivers, stagnant waters, swimming pools, bottled waters, ventilation ducts, air conditioning units, sewage systems, soil, and in-hospital dialysis and eye wash units. Although infections caused by Acanthamoeba spp. are infrequent, they are characterised by high mortality and can lead to serious clinical problems. Acanthamoeba causes Acanthamoeba keratitis (AK) in healthy individuals and contact lens users. In immunocompromised individuals, it can lead to granulomatous amoebic encephalitis (GAE) and lung and skin infections. In this study, we investigated the presence of Acanthamoeba species in environmental water samples collected from various cities across Turkey. We employed a range of culture and molecular methods for this analysis. <strong>Material and Methods: </strong>A total of 100 samples were collected from different cities and water sources in Turkey. Water samples were filtered through a 0.45 μm diameter cellulose nitrate membrane filter using a vacuum device, allowing approximately 100 ml to pass in about 30 s. The filtered water samples were cultured on Non-Nutrient Agar E.coli (NNA-E.coli) and Buffered Charcoal Yeast Extract Agar media. Samples that were considered culture-positive were stored at -20 °C for DNA isolation. <strong>Results:</strong> Among the 100 samples, 27 (27%) Acanthamoeba spp. were detected without differentiation by the media. In the NNA-E. coli medium, 22 (22%) Acanthamoeba spp. were produced, while 19 (19%) were produced in the BCYE medium. No significant difference was found between the two media. A total of 25 Acanthamoeba spp. were detected using RT-PCR (25%). The compatibility of the media with RT-PCR was found to be statistically significant (p&lt;0.005) and bidirectional, as calculated by the kappa coefficient. Samples in which Acanthamoeba spp. cysts were detected were evaluated based on morphological criteria. Based on the size and shape of the cysts, it was observed that the majority of the detected samples belonged to groups II and III. <strong>Conclusion: </strong>There are few studies comparing Acanthamoeba growth in different culture media. There is a need to discover newer and more accessible culture media for this amoeba.