The present study proposes a sustainable circular bioeconomy approach for municipal wastewater (MWW) treatment, employing a sub-pilot two-stage sequential microalgal co-cultivation system (TSSCS), and evaluates the impact of hydraulic retention time (HRT) on MWW treatment efficiency, as well as biomass and biomolecule productivity. In the first stage of TSSCS, 2 ratios of Tetraselmis indica and one ratio of Picochlorum sp. (2TS:1PC) were used for the treatment of 75% raw MWW + 25% ASN-III, while in the second stage, 2PC:1TS was employed for the treatment of 50% 2TS:1PC effluent+ 50% ASN-III. At the end of the first and second stages of TSSCS, nutrient removal efficiencies were as follows: total nitrate (TN) was <80% and 90% on HRT-10 and HRT-12; total phosphate (TP) was <85% and 95% on HRT-16, and chemical oxygen demand (COD) was <75% and <85% on HRT-14 and HRT-10, respectively. In the TSSCS, 2TS:1PC (S1) and 2PC:1TS (S2) showed maximum biomass productivity of 2.65 and 2.78g/L on /L on HRT-14 and HRT-12, respectively. Additionally, biomass of 2TS:1PC (S1) and 2PC:1TS (S2) contains lipid content (38.67 and 40.67%), β-carotene (4.09 and 3.00mg/g), and astaxanthin (0.69 and 0.31mg/g), respectively.
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