Abstract: The species of liver fluke of the genus Fasciola (phylum platyhelminthes, order Digenea, Family Fasciolidae) are obligatory parasites that inhabit the large biliary ducts of herbivore animals as well as man. Reports on the species of Fasciola present in the Nile Delta, Egypt, appear controversial. In the current study a precise identification of Fasciola isolates from cattle in Qena province, Upper Egypt was done based on examination of the second Internal Transcribed Spacer (ITS2) and the mitochondrial cytochrome oxidase subunit I (COI). Amplification, sequencing and phylogenetic examination revealed that the collected Fasciola isolates represent only one species which is Fasciola hepatica.