This study focused on the monitoring and assessment of aquatic ecosystem functioning based on the abiotic and biotic features of Lake Nasser in the post-flood and pre-flood periods as the last ones before water storing behind the Grand Ethiopian Renaissance Dam. The physicochemical parameters, distribution and structure of biotic communities, including bacteria, phytoplankton, zooplankton, macrophytes, epiphytes and fish were analyzed at 15 sites. The values of most parameters (primarily temperature, total suspended solids, pH, orthophosphates, carbonates, sulphates and some cations) were higher in the pre-flood period, whereas higher values of conductivity, total dissolved solids, total solids, nitrites, nitrates, chemical oxygen demand, bicarbonates and chlorophyll a were recorded in the post-flood period. Cyanobacteria-dominated phytoplankton and total coliform bacteria were more abundant in the pre-flood period, and especially at sites 10–15, whereas higher abundances of zooplankton were recorded in the post-flood period. All these changes in parameters were significantly differentiated, and only the concentrations of dissolved oxygen, biochemical oxygen demand, ammonium and silicates were at similar level in both periods. The study also indicated that environmental factors had affected the macrophyte distribution and the adaptation of the invasive species Myriophyllum spicatum to thrive under different environmental factors in the lake. Besides, macrophytes provided the habitat to other aquatic organisms, especially epiphytes, and also helped maintaining the good water quality, nutrient cycling and stabilizing rivers’ and lakes’ banks. Generally, these conditions were rather favorable for biological processes and fish production.