The objective of the present study was to evaluate the nutrient removal efficiency and the physiological responses in terms of growth, biochemical composition and photosynthetic activity of the autochthonous freshwater algal strain Desmodesmus communis. Microalgae were grown in a primary municipal effluent under different hydraulic retention times (HRTs) and in a two-phases process using both primary and secondary wastewater effluents. Semi-continuous cultures were operated for 7 day at 5-, 3- and 1.5-day HRT and the different dilution rate showed a greater influence on the biomass composition and nutrient removal efficiency. Removal of N-NH3 and P-PO4 was over 99 % and the highest accumulation of polysaccharides (57.2 wt.%) was obtained at high HRT (5 day); the maximum content of proteins (26.9 wt.%) was achieved at 1.5-day HRT, even if, under this condition, a clear inefficiency in terms of ammonia removal was observed. Moreover the accumulation of N-NH3 occurring at 1.5-day HRT caused the decrease of the photosynthetic response in terms of efficiency of light capture (alpha) and relative electron transport rate (rETR), both parameters extracted from the rapid light curves (RLC) measurements. No significant differences were observed for the total fatty acids (TFAs), with a content of 2-3.5 wt.% for each HRT condition. On the other hand, in the two-phases process, when a nutrient deprivation condition was induced by diluting the culture with the secondary wastewater effluent, the algal cells accumulated TFAs, achieving a maximum content of 9.7 wt.% and a great increment in terms of biomass (1.64 +/- 0.02 g L-1) due to the ability of this algal strain to accumulate intracellular N. The wide and accurate investigation of the different aspects related to the whole process represents a relevant point of novelty in this research field and suggests the operational conditions for the start-up of an open pond system for wastewater treatment and biomass production for further applications.