Differential toxicity of the UV-filters BP-3 and BP-4 in Chlamydomonas reinhardtii: A flow cytometric approach

Abstract

Due to the concern about the negative effects of exposure to sunlight, UV-filters are being introduced in all kind of cosmetic formulas. Wastewater treatment plants are not able to remove and/or degrade them; consequently they find their way into rivers, lakes and oceans. These chemicals are acquiring a concerning status due to their increasingly common use and the potential risk for the environment. Benzophenone-3 (BP-3) and Benzophenone-4 (BP-4) are broad-spectrum UV-filters used for the same purpose in personal care products, insecticides and plastic bags; however, after 96 h of exposure to several concentrations of these UV-filters, the growth of C. reinhardtii was more affected by BP-3 than by BP-4, being the 96 h-EC₅₀ for growth 5 mg L^-1 and 38 mg L^-1, respectively. Based on these values Chlamydomonas reinhardtii cultures were exposed during 24 h to 2.5, 5 and 10 mg L^-1 of BP-3 and 19, 38 and 76 mg L^-1 of BP-4. A cytometric panel was carried out to evaluate the effect of sublethal concentrations of these UV-filters, thus several cytotoxicity biomarkers were analysed, including chlorophyll a fluorescence, viability, metabolic activity, oxidative stress, cytoplasmic and mitochondrial membrane potentials, and intracellular pH. BP-3 and BP-4 affect C.reinhardtii cells in a different way, showing differences for three of the examined parameters. Chlorophyll a fluorescence and mitochondrial membrane potential showed a significant increase (p < 0.05) in BP-3 and a significant decrease in BP-4, whereas viability only decreased significantly in the highest concentrations of BP-3. Regarding to the other parameters analysed, a similar pattern of cytotoxicity was observed. Growth rate, vital population and metabolic activity (esterase activity) and intracellular pH decreased significantly and cytoplasmic membrane potential and ROS levels increased significantly in cultures exposed to both pollutants.

Keywords: Cytotoxicity; Emerging pollutants; Flow cytometry; Microalga; UV-filters.