Prevalence and first molecular identification of Sarcocystis species in cattle and water buffaloes in India

The importance of Sarcocystis hominis in causing zoonotic infections is well known. Recently, S. hominis like cysts have been reported from water buffalo in China. Previous studies indicate prevalence of Sarcocystis species in bovine populations in India but molecular evidence is required for proper species differentiation. We examined two hundred and ninety six cardiac tissue samples of Indian water buffaloes and cattle from northern and western parts of the country. Tissues were examined for Sarcocystis using intact cyst isolation method, pepsin acid digestion method and Sarcocystis 18S rRNA PCR. The combination of primers was used for 18S rRNA PCR amplification followed by sequencing. Twenty five representative samples were sent for sequencing and 19 readable sequences were obtained for phylogenetic analysis. Overall, the Sarcocystis cysts/zoites were recorded in 44% (95% CI 38-49%), 58% (95% CI 53-64%) and 68% (95% CI 63-73%) from both cattle and buffalo samples using intact cyst isolation, pepsin-HCl digestion method and conventional PCR, respectively. The results indicate that pepsin-HCl digestion method and conventional PCR are more sensitive than intact cyst isolation for detection of Sarcocystis species in tissue samples. The prevalence of Sarcocystis species was high in buffalo as compared to cattle intermediate hosts. Phylogenetic analysis indicated that more than one Sarcocystis species are circulating in cattle and water buffaloes in India. The results further indicate that experimental transmission studies are required to re-confirm the identities and host ranges of the Sarcocystis species in cattle and water buffaloes in India.