Morphological and molecular characterization of Gastrothylax crumenifer (Platyhelminthes: Gastrothylacidae) infecting rumen of Capra hircus in Chandigarh (India)

Morphological and molecular characterization of the amphistomid trematode, Gastrothylax crumenifer (Creplin, 1847) Poirier 1883 collected from Capra hircus goat in Chandigarh was carried out using Gower's carmine stain and by investigations on the nucleotide sequences of 28S and internal transcribed spacer 2 (ITS2). A total of 10 guts of C. hircus were examined for infection with trematodes. All the worms collected were morphologically identified as G. crumenifer by comparing the arrangement of median sagittal sections of adult amphistomes. The BLAST analysis of 28S gene of the study specimen G. crumenifer (KY786331) included 100 hits and showed maximum homogeneity of 99% with G. crumenifer as only 7 gaps in the case of G. crumenifer (JX518971, JX518960, JX518959, JX518970); 8 gaps in the case of G. crumenifer (JX518969) and 18 gaps in case of G. crumenifer (KJ559529). Phylogenetic tree revealed close relatedness of the present specimens with G. crumenifer infecting Bos indicus and Bos frontalis from India with 0.01 and 0.0 (nil) evolutionary divergence, respectively. The BLAST analysis of ITS2 gene of the study species G. crumenifer (KY786332) included 100 hits and showed maximum homogeneity of 97% to 99% with G. crumenifer as only 1 gap in the case of G. crumenifer (HM159382) and 5 gaps in the case of G. crumenifer (KU530204) were found between them. Phylogenetic tree revealed close relatedness of the present specimens of Gastrothylax with G. crumenifer infecting Bos indicus and Capra hircus from India with 0.01 and 0.0 (nil) evolutionary divergence, respectively. The molecular results revealed ITS2 as the better gene marker as it formed clear monophyly of the present species with G. crumenifer but in the case of 28S, the tree was somewhat more towards the polyphyly of Gastrothylacidae. Thus on the basis of morphological and molecular data based on 28S and ITS2, the study species was identified as G. crumenifer