The detection, and identification of Cryptosporidium species by nested polymerase chain reaction (nested-PCR) and restriction fragment length polymorphism (RFLP) of calves in Shahroud town

Cryptosporidium species belong to Apicomplexa phylum and are opportunistic protozoans that infect the gastrointestinal and respiratory systems of some animals and humans. This study was conducted to investigate the prevalence, detection, and identification of Cryptosporidium species by nested PCR and restriction fragment length polymorphism (RFLP) in calves in Shahroud town.  A total number of 256 calves was collected from fecal samples of pre-weaned calves (≤2 months) in Shahroud town, and positive samples were stained by Ziehl-Neelsen (ZN) staining method. Specific primers were used to investigate the Nested-PCR and sub-species were investigated by RFLP method. The microscopic results showed that 27 samples (10.54 %) of the samples were positive. The results for nested-PCR showed that out of samples, 92.59 % and 7.41 % of samples were positive for C. parvum and C. Andersoi, respectively. The results showed that 25 samples were affected by VSP enzyme in regions of 104 and 628 bp that belong C. parvum bovine, and genotype A gene sub-species. The samples were affected by Ssp I enzyme and the results showed bands in regions of 385 and 448 bp that show C. muris/C. andersoni sub-species. The results for dde enzyme showed bands in regions of 156, 186, and 224 bp that confirm C. muris sub-specie. The different species and sub-species were identified by different methods and can help to control of the cryptosporidiosis.