Nowadays, bioflocculation is considered as a potential technology that could be able to alleviate microalgae dewatering cost regarded as the cornerstone hindrance of their full-scale application. However, most bioflocculation studies reported are laboratory scales. This study examined a pilot-scale and in situ flocculation of freshwater microalgae Desmodesmus brasiliensis by microbial bioflocculant. Biochemical composition of microalgal biomass was analyzed to evaluate the applicability of bioflocculation for microalgae-based biofuel production. The flocculation efficiency >98 % was achieved at both pilot-scale and in situ treatment. Bioflocculation is simple, effective, economic, and environmentally friendly. Even though total proteins recovered from biomass harvested by centrifugation and that harvested by bioflocculation were significantly different, there was no significant difference in total carbohydrates and total lipids recovered from either biomass harvested by centrifugation or biomass harvested by bioflocculation. The results herein presented, doubtlessly demonstrated that the γ-PGA bioflocculant produced by Bacillus licheniformis CGMCC 2876 is applicable for commercial-scale microalgae harvesting. In addition, bioflocculation process cost could greatly be reduced by in situ operation as no investment cost is needed for a separate flocculation tank and mixing device. Furthermore, bioflocculation method developed is a worthy microalgae harvesting method for algal-based biofuel production.