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The role of the genus Acanthamoeba in the process of maintaining, multiplying and transmitting human pathogens has not been well recognized. The aim of our study was to evaluate the association of potentially pathogenic Acanthamoeba spp. with bacteria in strains recently isolated from Lake Malta near the city of Poznań, Poland. Most of the isolated amoebae belonged to the species A. castellanii and A. rhysodes. The majority of the isolates were pathogenic for mice and amoebae could be recovered from their brains and lungs. In those cases when amoebae could not be recovered from the autopsy material histopathological analysis showed changes in tissues indicating bacterial infection. We found that approximately 50% of the isolates were associated with endosymbiotic or endocytobiotic bacteria, e.g., Proteus sp., Micrococcus sp., Chryseobacterium meningosepticum, Clostridium perfringens, Escherichia coli, Salmonella sp., Legionella pneumophila and other species. The presence of bacteria residing inside free-living amoebae possess a great challenge in terms of disease control and sanitation of contaminated water sources.
Acanthamoeba are widespread free-living amoebae which may cause granulomatous amoebic encephalitis (GAE), keratitis, skin ulcerations and disseminated tissue infection. An important diagnostic and prognostic factor for the treatment of infection is a quick and correct diagnosis of amoebae strains. The aim of our study was to develop a rapid method for detection and identification of pathogenic Acanthamoeba spp. strains from diagnostic material collected from water. In this study we analysed five amplification-based genetic markers (Aca 16S, Ac6/210, GP, JDP, Nelson) used for identification of pathogenic Acanthamoeba spp. strains isolated in water sources in Poland, Iceland and Sweden. Our results demonstrated the presence of pathogenic Acanthamoeba strains in tap water. PCR assay appeared to be a more rapid and sensitive method to detect the presence of amoebae than the limited conventional techniques. Based on our observations, we can confirm that the use of four out of five genetic markers (Aca 16S, Ac 6/210, JDP, GP, Nelson) may be helpful in identification of Acanthamoeba spp. strains, but only one Aca 16S primer pair is a highly specific marker that distinguishes between pathogenic strains of Acanthamoeba and other free-living amoeba families.
The present study investigated the susceptibility of Acanthamoeba spp. trophozoites to two multipurpose systems for cleaning and maintenance of contact lenses. Three strains of trophozoites from the ATCC (A. castellani T4, A. castellani Neff, and A. polyphaga) and two Acanthamoeba isolates obtained from swimming pools (PT5 and PO1) were placed in monoxenic culture. To test their survival in cleaning solutions for contact lenses, the trophozoites were exposed for 4 and 24 h to two multipurpose solutions (A and B), and were then inoculated into a new monoxenic culture. Amoebic growth on the plates was observed after 72 h of incubation. Trophozoites from all three ATCC strains and one isolate from a swimming pool (PO1) grew in all plates after 4 h of exposure to solutions A and B. After 24 h, the ATCC strains and the PO1 isolate showed growth in most of the plates treated. Only the PT5 isolate showed susceptibility to both solutions over the time intervals tested. The two solutions were not completely effective against most strains and isolates over the time intervals tested. These results are important, since species of Acanthamoeba are widely distributed in the environment and are potential agents of eye pathologies.
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