Rumen flukes are widely distributed digenetic trematodes belong to the superfamily Paramphistomoidea which hit domestic and wild ruminants and cause a parasitic disease known as paramphistomiasis or paramphistomosis. The economic importance of paramphistomes is globally underestimated because of their proliferation and abundance among ruminants. Substitutional approaches to improving species identification are required. The paramphistomes scrutinised during this present study were adult flukes. Our five samples include four samples of paramphistomes from cattle and only one from sheep were identified according to the ITS-2 sequencing as followers of the genera Paramphistomum, Calicophoron, and Gastrothylax. The DNA of each worm was isolated, and the ITS-2 rDNA was amplified, purified, and sequenced. Then, the sequences were uploaded to GenBank, which allocated them the following accession numbers: OK216189, OK216190, OK216191, OK216192, and OK216193. MEGA X has been used to execute the phylogenetic analysis on these sequences. This result reveals that ITS-2 is an excellent molecular marker, and can be beneficially used for paramphistomes identification to determine the affinity of samples among the different genera of the superfamily Paramphistomoidea. The current investigation will help build a scope to make the identification of paramphistomes very straightforward and, eventually, the command of this underappreciated cosmopolitan digenean group.