PCR-RFLP at 18S rRNA gene for identification of Sarcocystis species and their prevalence in cattle of Andhra Pradesh

Abstract

In the present study, the species specific determination of Sarcocystis cysts isolated from 137 naturally infected and slaughtered cattle from different regions of Chittoor district, Andhra Pradesh, India was achieved by PCR-RFLP. Diagnosis of sarcocysts in cattle was based on macroscopic examination and pepsin digestion method. Genomic DNA was extracted individually from bradyzoites of all 137 sarcocystosis positive cattle and all the isolates were characterized at 18S rRNA. Digestion of PCR amplicons (900 bp) with restriction endonuclease revealed 3 different electromorphs which were referred to Sarcocystis cruzi (513 and 343 bp), S. hirsuta (525, 241 and 141 bp) and S. fusiformis (532 and 335 bp). Sarcocystis cruzi (93.43%) was significantly more prevalent in comparison with the S. hirsuta (4.38%) and S. fusiformis (2.19%). Infection of cattle with S. hominis was not observed in the study area. Occurrence of S. fusiformis in cattle supports that Sarcocystis species of buffaloes are not strictly intermediate host specific however can infect cattle.