Molecular identification of Pseudodiscus collinsi from wild Indian elephant (Elephas maximus indicus) based on ITS-2

Domestic and wild animals can contract amphistomosis, a disease caused by digenetic trematodes belonging to the superfamily Paramphistomoidea. The importance of these flukes is underestimated worldwide due to their ubiquity and abundance among hosts. Pseudodiscus collinsi is a member of the family Paramphistomatidae that infects the colon of equines and elephants. In the present study, the flukes were recovered from the colon of a dead wild Indian elephant (Elephas maximus indicus). The flukes were stained using acetyl alum carmine and morphologically identified as P. collinsi based on the presence of oral pouches and position of the testes. The polymerase chain reaction (PCR) for amplification of the internal transcribed spacer 2 (ITS-2) region and sequence analysis were performed. The phylogenetic analysis using the Maximum Likelihood (ML) method, based on the Kimura 2-parameter model, revealed the separation of P. collinsi (elephant) as a distinct species from the other amphistomes of different hosts. This is the first molecular marker of P. collinsi to be presented.