Detection and identification of six Cryptospordium species in livestock in Slovakia by amplification of SSU and GP60 genes with the use of PCR analysis.

INTRODUCTION: In this study we examined 200 faecal samples from pigs and calves with suspected cryptosporidiosis were examined by the PCR methods: nested PCR for amplification of SSU region; nested PCR for amplification of GP60 region; and with restriction analysis of DNA (PCR-RFLP). The sequencing identified the following species: Cryptosporidium muris (2), Cryptosporidium andersoni (1), Cryptosporidium bovis (4), Cryptosporidium suis (2), Cryptosporidium scrofarum (10), mixed infection caused by C. scrofarum and C. muris (1),and Cryptosporidium parvum (10) genotype A subtype IIaA17G2R1. RESULTS AND CONCLUSIONS: The findings suggest that livestock can be an important source of zoonotic species or genotypes of Cryptosporidium, which may adversely affect the public health of human populations. This is the first time in our country that the Cryptosporidium species has been identified in livestock in Slovakia. The identification and genotyping of this pathogen in Slovakia, completes the epidemiological situation in Europe for Cryptosporidum species.

Molecular characterization and first report of Cryptosporidium genotypes in human population in the Slovak Republic.

In our study, we examined 91 fecal samples from five different groups of people containing HIV patients, hemodialysis patients, kidney transplant recipients, immunocompetent humans without clinical signs, and humans with suspected cryptosporidiosis. The purpose of our study was to determine species and genotype composition of representatives of Cryptosporidium spp. using PCR analysis of small subunit ribosomal RNA gene and 60-kDa glycoprotein gene and examine their phylogenetic relationship. In HIV-positive/AIDS-infected group of patients and in hemodialysis patients, no presence of Cryptosporidium species was detected. In two kidney transplant recipients, we detected species/genotypes Cryptosporidium parvum IIaA13G1T1R1 (KT355488) and Cryptosporidium hominis IaA11G2R8 (KT355489) and in two immunocompetent patients with clinical symptoms, we identified Cryptosporidium muris and C. hominis IbA10G2T1 (KT355490). In the group of healthy immunocompetent individuals without clinical signs, we identified species/genotype C. hominis IbA11G2 (KT355491) in one sample.