Morpho-molecular detection and characterization of rumen fluke in cattle at northern area of Bangladesh

Rumen fluke infection is a common disease of ruminants caused by different species of paramphistomes and causes heavy economic loss worldwide. This study was conducted to morphological identification, molecular detection and characterization with genotyping of rumen fluke in cattle of Bangladesh. Adult fluke specimens were collected from rumen, reticulum as well as bile duct of slaughtered cattle then washed extensively with physiological saline and flukes were identified morphologically as paramphistomes depending on family and genus specific structural characteristics for molecular study. In our research, paramphistomes were identified through morphological and molecular analysis, subsequently confirmed through sequence analysis and the sequences were submitted to GenBank and assigned as accession numbers of OP751135. PCR was standardized with extracted DNA using published primers and annealing temperatures amplifying ITS2 ribosomal gene sequences with 520bp amplicon sizes. The results of sequence alignment indicated that, the study sequences shared 98-100% similarity with sequences of different species of paramphistomes from India, China and other countries and the collected isolate was considered as paramphistomes. The collected fluke was also confirmed as paramphistomes on the basis of ITS2 gene partial fragment with 53% G+C and 47% A+T composition. Phylogenetic tree reconstruction demonstrated that they all clustered together with high homogeneity within their respective groups. PCR method, based on ITS2 gene sequences, proved as specific and essential approach for detection, characterization and classification of various rumen fluke like. The findings from baseline molecular research could serve as cornerstone for future applied molecular research and control of this group of trematodes.