Morphological and molecular characterization of Paramphistomum epiclitum (Platyhelminthes: Paramphistominae) infecting rumen of Capra hircus in Chandigarh (India)

The morphological and molecular characterization of an amphistomid trematode, Paramphistomum epiclitum Fischoeder (1901) collected from Capra hircus (goat) in Chandigarh using Gower's carmine stain was studied and the nucleotide sequences of 28S ribosomal RNA and internal transcribed spacer 2 (ITS2) gene markers were investigated. A total of 10 rumen of C. hircus were examined and cut open to check the infection with trematodes. The worms collected were morphologically identified as P. epiclitum after evaluating different morphological characters like paramphistomum type pharynx, oesophageal length, acetabulum diameter and having epiclitum type of genital atrium. The BLAST analysis of 28S gene of the study specimen P. epiclitum (KY786333) showed maximum homogeneity of 99% with P. epiclitum (JX628580) and Orthocoelium streptocoelium (JX678277). Only 3 gaps were found between the aligned sequences of the study species and P. epiclitum (JX628580) and Orthocoelium streptocoelium (JX678277). Phylogenetic tree revealed close relatedness of the study specimen of Paramphistomum with Orthocoelium streptocoelium (JX678277) infecting B. indicus from India with high bootstrap value, thus showed it to be a sister taxa as both of them belonged to the same family and have the same host and locality. The BLAST analysis of ITS2 gene of the present species P. epiclitum (KY786334) showed maximum homogeneity of 97% with P. epiclitum (JF834888), Calicophoron daubneyi (KP201674), C. microbothrium (KP639632), C. calicophorum (GU133057), C. microbothrioides (AB056570); P. epiclitum (KF564870); P. leydeni (KJ995526). Phylogenetic tree also revealed close relatedness of the study specimen of Paramphistomum with P. epiclitum (KF564870, JF834888) infecting Bubalus bubalis and Bos indicus from India. The molecular results revealed the ITS2 region as effective for the study of the interspecific variations and the 28S gene was unsuitable as this marker showed mixing with the members at sub family level