A general investigation of digenean parasites was carried out from four fish species; Fahka puffer fish (Tetraodon lineatus), Nile tilapia (Oreochromis niloticus), Sharp tooth catfish (Clarias gariepinus) and Bayad (Bagrus bajad) from different localities of the River Nile in Qena and Aswan governorates, Egypt during October 2015 to August 2016. Four adult digeneans were found, belonging to four genera; Astiotrema, Orientocreadium, Glossidium and Acanthostomum. These were Astiotrema impletum 90% (27/ 30), Orientocreadium batrachoides 83.33% (25/ 30), Glossidium pedatum 60% (12/ 20) and Acanthostomum spiniceps 86.66% (26/ 30). Additionally, two larval trematodes (metacercariae) were detected, belonging to two genera; Clinostomum and Euclinostomum. These were Clinostomum complantum 85.57% (60/ 70) and Euclinostomum heterostomum 70% (35/ 50). Morphological and morphometric characters were discussed for each species in details and compared with the previously published data. This study considered Glossidium pedatum, Acanthostomum spiniceps and Euclinostomum heterostomum as new records from the River Nile in Qena Governorate.