Thermophilic potentially pathogenic amoebae isolated from natural water bodies in Poland and their molecular characterization

Abstract

The free-living amoebae (FLA) may live in the environment and also within other organisms as parasites and then they are called amphizoic. They are potentially pathogenic for humans and animals and are found in water that is a source of infection. The aim of this study was molecular detection and identification of these FLA in natural water bodies in North-Western Poland to evaluate the risk of the pathogenic amoebae infections. We examined surface water samples collected from 50 sites and first, the tolerance thermic test was performed in order to select thermophilic, potentially pathogenic strains. For molecular identification of FLA, regions of 18S rDNA, 16S rDNA and intergenic spacers were amplified. Acanthamoeba T4 and T16 genotypes of 18S rDNA gene and 18S rDNA of H. vermiformis were detected. We identified two variants of Acanthamoeba T4 genotype, two variants of Acanthamoeba T16 genotype and one variant of H. vermiformis. Identification of the T16 genotype and H. vermiformis in water was for the first time in Poland. Additionally, we made attempts to adapt the RLB method for detection and differentiation of FLA species and strains. PCR seems to be more sensitive than RLB hybridization, though.

The Acanthamoeba spp. in Water Sources from Zanjan Province, Northwest of Iran

“…The food availability for the growth of Acanthamoeba spp and the resistance of the cyst stage of these amoebae to chlorination, ozonation and filtration by biofilm formation, probably creates the potential for high distribution of Acanthamoeba spp in the aquatic environments (25,(37)(38)(39). Thus, this alarmingly frequency of this amoeba in water resources represents a significant hazard for the high risk populations, including the contact lens wearers and immunocompromised individuals [13,21,26].…”

Molecular Characterization of Acanthamoeba spp. Occurring in Water Bodies and Patients in Poland and Redefinition of Polish T16 Genotype

“…Admittedly, the analysis of > 2,000‐bp fragments of the 18S rRNA gene accomplished in this study revealed that these two isolates, and nine other Polish isolates recently sequenced by Adamska et al. () and Leońska‐Duniec et al. () are the most similar to the T20 genotype, but in 2009 they in fact represented a new genotype.…”

“…(), as well as from natural (Adamska et al. ) and artificial (Leońska‐Duniec et al. ) water bodies were analyzed.…”

“…The sequences of Acanthamoeba 18S rRNA gene of length 2,121–2,178 bp were obtained from 24 isolates obtained earlier (Table ) and originated from: 15 natural water bodies (rivers, ponds, and lakes; Adamska et al. ), five artificial water bodies (swimming pools and fountains; Leońska‐Duniec et al. ) in north‐western Poland as well as from four clinical samples (bronchoaspirate fluid and bronchoalveolar lavage; Łanocha et al.…”

“…) and environmental (Adamska et al. ; Leońska‐Duniec et al. ) samples were used to conduct their genotyping and phylogenetic analysis that allow classifying them to correct genotypes.…”