Protozoa of the genus Acanthamoeba are organisms that can be generally found in the environment. The focus of this study is the detection of the presence of Acanthamoeba in different water sources and samples taken from airconditioning units. The identification of Acanthamoeba isolates was based on the morphology of cysts and trophozoites as well as PCR amplification with a genus specific primer pair JDP1 and JDP2. Growth characteristics and temperature tolerance were monitored. The pathogenic potential was tested in vitro on Vero cell cultures. Genotype identification was based on the sequencing of the GTSA.B1 PCR amplimer of 18S ribosomal DNA. The data obtained revealed that the isolates belong to T3 and T4 genotypes. One T3 and one T4 isolate contain a group I intron. The 933 base pair intron found in a genotype T4 isolate is considerably larger compared to formerly described introns of Acanthamoeba griffini (genotype T3) and A. lenticulata (genotype T5). This is the first report detailing the environmental distribution of the Acanthamoeba genotypes in the region of Bratislava, Slovakia.

Acanthamoeba keratitis (AK) is a relatively rare disease worldwide. Over the past 10 years, five cases of AK were reported in Slovakia. Four preserved Slovak strains and one strain from the Czech Republic isolated from corneal scrapes of patients with AK are characterised in this study. Genotype identification of isolates is based on sequences of the PCR amplimer GTSA.B1 amplified from 18S ribosomal DNA. A strain isolated from the first patient in 1999 was classified as a rare sequence type T15. This is just the second report in which genotype T15 has been associated with AK. The other three Slovak strains were identified as belonging to the most common genotype T4. The only strain originating from the Czech Republic was classified as sporadically appearing sequence type T3. All isolates were also studied for their temperature tolerance and growth characteristics. The cythopatic effect was tested in vitro on Vero cell cultures.

SUMMARYEnteroviruses (EVs) infections occur worldwide. Although, infections by these viruses are often asymptomatic and go unnoticed, they can be shed in stool for several weeks. The EVs are associated with sporadic outbreaks and a wide range of clinical symptoms, occasionally accompanied with fatal consequences. Presently in the Slovak Republic (SR) recreational waters are tested only for bacterial indicators. Our aim was to monitor EVs in recreational waters. Water samples were collected during the years 2012-2014 from different recreational natural lakes in Central and West regions of SR. The samples were concentrated by centrifugation using the two-phase separation method recommended by the World Health Organization (WHO) used for EVs surveillance in the treated sewage waste water. Each of the two phases collected from the samples was analysed by polymerase chain reaction for detection of EVs and primary sequencing was done. Our study demonstrated presence of EVs in three localities consecutively for three years, indicating a probability of constant local source of faecal contamination. This is the first monitoring report on the occurrence of EVs in the natural recreational waters in SR.