BACKGROUND: Cryptosporidium felis, a host-specific protozoan with possible zoonotic potential, is a common parasite of cats (Felis catus). To date, there have been few studies on the molecular subtyping of C. felis worldwide, and no have been conducted in Central Europe. The aim of this study was to analyse the prevalence and genetic variability of C. felis in domestic and stray cats in Central Europe, particularly in the Czech Republic, Poland and Slovakia. METHODS: Faecal samples were collected from domestic and stray cats. The presence of Cryptosporidium spp. was analysed by light microscopy with aniline-carbol-methyl violet staining and polymerase chain reaction (PCR)/sequencing of the small subunit rRNA gene (18S rDNA). All PCR-positive samples were further subtyped using PCR and sequencing of the 60 kDa glycoprotein gene (gp60). A chi-square test and odds ratio (OR) analysis were used to compare infection rates and assess the risk of C. felis infection between domestic and stray cats. RESULTS: A total of 711 faecal samples were collected - 350 from domestic cats and 361 from stray cats. The overall infection rate of Cryptosporidium spp. was 4.5% (32/711), with stray cats being significantly more frequently infected (6.7%) than domestic cats (2.3%). Oocysts of Cryptosporidium spp. were not detected microscopically in any of the samples. There were no significant differences between the infection rates in the three countries. All isolates were identified as C. felis, and analysis of the gp60 gene revealed five different subtypes, all belonging to the XIXa subtype family of C. felis. These subtypes formed five well-supported phylogenetic clusters, none of which had been previously reported worldwide. Only one subtype was found in domestic cats, whereas all five subtypes were found in stray cats. The subtypes identified in stray cats showed a clear geographical distribution in the study region. CONCLUSIONS: The results of this study extend our knowledge of the genetic variability of C. felis and indicate a possible geographical distribution of the detected subtypes. The significance of the observed genetic variability in terms of geographical distribution, host specificity and zoonotic potential remains unclear and requires further investigation.

Department of Biology and Medical Parasitology Wroclaw Medical University Wroclaw Poland

Faculty of Agriculture and Technology University of South Bohemia in České Budějovice České Budějovice Czech Republic

Faculty of Forestry Technical University in Zvolen Zvolen Slovakia

Institute of Parasitology Biology Centre of the Academy of Sciences of the Czech Republic 5 V 1 Branišovská 31 370 05 České Budějovice Czech Republic

National Agricultural and Food Centre Lužianky Slovakia

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