In 2021, the Nile River between Aswan and Sebaiyia (Egypt) was subjected to seasonal analyses of major biological parameters and the surrounding environmental conditions, such as temperature (ºC), conductivity (EC), pH, dissolved oxygen (DO), nitrite (NO₂), nitrate (NO₃), orthophosphate (PO₄), carbonate (CO₃), organic matter (O.M.), and chlorophyll-a. 32 taxa from 16 groups of macrobenthic invertebrates were identified; these included a crustacean, one Hirudineas, three oligochaetes, ten chironomid larvae, and fourteen molluscs. Furthermore, three unidentified species that belong to adult Corixidae, Ephemeroptera, and Zygoptera nymphs were discovered. Throughout the study period, the numerous groups of the Nile zoobenthos were represented by the combined contributions of molluscs, chironomid larvae, and oligochaetes. With 39.8 GFW m^-2, the average yearly number of macrobenthic invertebrates was 4560 ind.m^-2. According to the findings of the canonical corresponding analysis (CCA), many ecological parameters that were examined appeared to play a significant role in affecting the biomass and abundance of zoobenthos. Furthermore, the abundant macrophyte population at the western sites may be the cause of the biota differences between the east and west banks. Furthermore, the effects of industrial pollution were not directly present in those sample locations. Most significantly, the knowledge gathered from this study will be useful for managing the Nile, especially following the construction of Ethiopia's Renaissance Dam.