A step towards eco-friendlier and more efficient microalgal harvesting: Inducing flocculation in the non-naturally flocculating strain Chlorella vulgaris (13-1) without chemical additives

Flocculation is often regarded as a cost-effective and reliable method for microalgal harvesting. However, the traditional method often requires the addition of chemical agents to induce flocculation. This carries certain disadvantages including the chemical contamination of the biomass and the subsequent need to remove the flocculants from the medium. To address these issues, this study aimed to induce flocculation in a naturally non-flocculating strain (Chlorella vulgaris 13-1) without resorting to chemical additives, with the ultimate goal of increasing harvesting efficiency. Scanning electron microscopy showed that Scotelliopsis reticulata UFA-2, a naturally flocculating strain, produces extracellular polymeric substances (EPS) whereas 13-1 does not. As a result, two methods were used to induce flocculation in 13-1: co-cultivation of UFA-2 and 13-1, and insertion of EPS produced by UFA-2 into the growth medium of 13-1. The co-cultivation of 13-1 with UFA-2 significantly increased the flocculation efficiency compared to that of 13-1 alone (30 % higher flocculation efficiency after one hour of settling and 52 % higher after three hours of settling). Alternatively, the insertion of dry tightly-bound (TB) UFA-2 EPS into 13-1 cultures also improved flocculation efficiency (by 19 % compared to the control), while addition of soluble or loosely-bound (LB) EPS was less efficient (less than 1 % and 10 %, respectively). FTIR results showed that the composition of TB-EPS was different to that of LB and soluble EPS. TB-EPS contained higher proportions of proteins and different types of carbohydrates, potentially contributing to its increased efficacy in inducing flocculation in microalgae.