Monogenea is a class constituting the ectoparasitic flukes mainly on skin and gills of fish, with greater diversity of species worldwide. In the present study, four different species of monogenean parasites were recorded; they were isolated from gills of four freshwater fish species captured from locations along the River Nile, Qena governorate, Egypt. These were Cichlidogyrus aegypticus from Oreochromis niloticus (93.33, 28/30); Protoancylodiscoides malapteruri from Malapterurus electricus (90%, 18/ 20). Quadricanthus bagrae from Bagrus bajad (30%, 15/ 50), and Ancyrocephalus sp. from Mormyrus kannume (75%, 30/ 40). The recovered parasites were described by light microscopy, where the different sclerotized parts of the haptor and copulatory organs were used as key of identification of each species. C. aegypticus can be differentiated by the presence of haptor with two pairs of strongly developed anchors, two long projections of the complex bar and a narrow copulatory tube with a wide basal funnel. P. malapteruri was characterized by the presence of broad haptor with two large dorsal and two small ventral hamuli. Ancyrocephalus sp. was characterized by a haptor which was not clearly demarcated from the anterior body with ventral and dorsal anchors similar in shape. Quadriacanthus bagrae was discriminated by possessing a complex bar with long projection at the center and two long ends, the dorsal and ventral anchors were without distinct roots. The present study represented new locality records for these species off the River Nile, Qena governorate, Egypt.