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Diatom is a unicellular photosynthetic microalga that is found in diverse environments. These are decorated with siliceous cell walls called frustules. Diatoms have long been favoured by grazers such as microscopic protozoa and dinoflagellates. However, grazers typically remain intact in laboratory culturing and feed on diatom in culturing vessels and reducing biomass yield. The isolation and cultivation of diatoms in laboratories hamper diatoms’ diversity and vast industrial potential. Chitosan, a biopolymer, has been widely used with other polyelectrolytes to flocculate various organic and inorganic colloids at acidic pH. Dissolved chitosan (acidic pH) has been used in various natural water samples and wastewater system for dewatering. However, untreated chitosan flakes have never been evaluated in a heterogeneous natural water environment. Since diatoms have silica surfaces, we tested chitosan for diatom separation and optimized chitosan concentration and other parameters to obtain grazer-free diatom starter culture from raw water. We also elucidated the mechanism for chitosan flakes-mediated diatom flocculation through adsorption kinetics and molecular dynamic simulation analysis. The results of this study are statistically optimized and validated, with a significant R2 value of 0.99 for the proposed model.

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