Identification of pig-specific Cryptosporidium species in mixed infections using Illumina sequencing technology

Nowadays molecular methods are widely used in epidemiological studies of Cryptosporidium infections in humans and animals. However to gain better understanding of parasite species or genotypes, especially when mixed infections are noticed, highly sensitive tools with adequate resolution power need to be employed. In this article, we report an application of the next generation sequencing method (NGS) for detection and characterisation of Cryptosporidium species concurrently present in pig faeces. A mixture of Cryptosporidium DNA obtained from two faecal samples was amplified at the 18 SSU rRNA gene locus and the resulting amplicons were subsequently used for MiSeq sequencing. Although initial molecular analyses indicated the possible presence of another Cryptosporidium species other than Cryptosporidium scrofarum and Cryptosporidium suis, deep sequencing only confirmed the presence of pig-specific Cryptosporidium.