Lopes da Silva, TeresaSilva, ThiagoFrança, Bruna ThomazinhoRibeiro, BelinaReis, Alberto2025-09-172025-09-172025-07Lopes da Silva, T., Silva, T. A., França, B. T., Ribeiro, B., & Reis, A. (2025). Monitoring C. vulgaris Cultivations Grown on Winery Wastewater Using Flow Cytometry. In: Fermentation, 2025, vol. 11 (8), article 442. https://doi.org/10.3390/fermentation11080442http://hdl.handle.net/10400.9/6059ABSTRACT: Winery wastewater (WWW), if released untreated, poses a serious environmental threat due to its high organic load. In this study, Chlorella vulgaris was cultivated in diluted WWW to assess its suitability as a culture medium. Two outdoor cultivation systems-a 270 L raceway and a 40 L bubble column-were operated over 33 days using synthetic medium (control) and WWW. A flow cytometry (FC) protocol was implemented to monitor key physiological parameters in near-real time, including cell concentration, membrane integrity, chlorophyll content, cell size, and internal complexity. At the end of cultivation, the bubble column yielded the highest cell concentrations: 2.85 x 106 cells/mL (control) and 2.30 x 106 cells/mL (WWW), though with lower proportions of intact cells (25% and 31%, respectively). Raceway cultures showed lower cell concentrations: 1.64 x 106 (control) and 1.54 x 106 cells/mL (WWW), but higher membrane integrity (76% and 36% for control and WWW cultures, respectively). On average, cells grown in the bubble column had a 22% larger radius than those in the raceway, favouring sedimentation. Heterotrophic cells were more abundant in WWW cultures, due to the presence of organic carbon, indicating its potential for use as animal feed. This study demonstrates that FC is a powerful, real-time tool for monitoring microalgae physiology and optimising cultivation in complex effluents like WWW.engChlorella VulgarisWinery wastewaterFlow cytometryjournal article10.3390/fermentation110804422311-5637