Molecular and phylogenetic analyses of cryptosporidium spp from dairy cattle in Egypt

ABSTRACT

Cryptosporidium infections in cattle result in considerable economic loss as well as a significant source for zoonotic infection. This study was devoted to spot analysis on Cryptosporidium parasites derived from dairy cattle at Kafr El Sheikh Province, Egypt, using morphological and molecular tools. Fecal specimens from neonatal calves as well as adult cattle were screened for Cryptosporidium oocysts using modified Ziehi-Neelsen technique. Oocysts in positive samples were concentrated using sugar floatation technique. Small size oocysts from calves and large size ones from adults were identified microscopically with measures correspond to those of C parvum and C. muris-C. andersoni, respectively. DNA was extracted from 8 positive specimens; 4 harbored small and 4 had large oocysts and used for PCR amplification, RFLP analysis and nucleotide sequence. RFLP analysis revealed 2 types of banding patterns coinciding with those of Cryptosporidium parvum and C. muris-C. andersoni. Sequence analysis of amplified fragments of small subunit of ribosomal RNA [SSU rRNA] and heat shock protein [HSP70] genes were used as markers for species identification and genotyping of the parasite. Homology search and phylogenetic analysis of the generated sequences versus those deposited in Genbank revealed that small oocysts were belong to C. parvum genotype II, whereas large ones were belong to C. andersoni