Genotyping of free-living acanthamoeba species in enviroment

Acanthamoeba species are free-living protozoan parasites that diffuse the enviroment and can be found in tap, mineral waters, contact lens solutions and soil, dust, air, swimming pools and thermal waters. Acanthamoeba species lead to Granulomatous Amebic Encephalitis (GAE), a chronic nervous system disaeses of immunocompromised humans and animals and amoebic keratitis. Also species of Acanthamoeba cause infections of the lungs, skin, bones, adrenal gland, and sinuses. Acanthamoeba species identified of more than 24 species have been found based on morphological criteria. Currently, moleculer techniques, sequencing of 18S rRNA genes, are being used to the species phylogeny of Acanthamoeba. Based on sequence differences 16 genotypes (T1-T16) and of these genotype T4 predominates among enviromental sources. In this study 50 water samples and 50 soil samples were taken from Adana, Afyon, Kütahya, Niğde and Mersin cities of Turkey. After using microscopy and cultivation methods 18S rRNA gene as PCR target for Acanthamoeba species and all of samples were examined by PCR method. The sensitivity and specificity of both culture and PCR methods were 100% in water and soil samples. Twenty one (42%) of the 50 water samples were found positive, 29 (58%) water samples were found negative, 23 (46%) soil samples were found positive and 27 (54%) soil samples were found negative according to PCR method. In this investigation, 100% Acanthamoeba castellanii (T4) species were isolated from water samples in Adana, Afyon, Niğde and Mersin cities while 90% Acanthamoeba castellanii and 10% Acanthamoeba griffini (T3) were isolated from water samples in Kütahya city. However, 100 % Acanthamoeba castellanii (T4) were isolated from soil samples in Afyon, Niğde and Mersin cities. Also 67% Acanthamoeba castellanii (T4), 33% Acanthamoeba griffini isolated from soil samples in Adana, while 80% Acanthamoeba castellanii (T4) and 20% Acanthamoeba griffini (T3) were isolated from soil samples in Kütahya. As a result of the study, it has also been shown that Acanthamoeba castellanii dominant species among of Acanthamoeba species in Turkey. KEYWORDS: A.castellanii, A. griffini, water samples, soil samples, Turkey