Cu, Ni, and Zn effects on basic physiological and stress parameters of Raphidocelis subcapitata algae

The submitted work observed Cu, Ni, and Zn effects on selected physiological and stress parameters of the alga Raphidocelis (Pseudokirchneriella) subcapitata. In 96-h experiments, EC50 values for algal specific growth rates (SGR) inhibition in Cu, Ni, and Zn presence were estimated as 0.15, 0.50, and 0.20 mg l^-1. In addition to growth inhibition, the effect of metals at various concentrations on algal SGR was also monitored. While these experiments confirmed approximately the same toxicity of Zn and Cu on SGR, Ni toxicity on this parameter was observed as the lowest. In terms of the effect of metals on the level of selected photosynthetic pigments, chlorophyll a, chlorophyll b, and carotenoids, the following inhibition orders can be established: Zn > Cu > Ni, Ni > Cu > Zn, and Ni > Cu ≥ Zn, respectively. As a novelty of our research, we included monitoring and evaluation of the intensity of stress, which was the response of algal cells to the presence of Cu, Ni, and Zn, and its correlation with respect to production factors and metal accumulation in algal cells. As stress factors, thiol (-SH) group and TBARS (thiobarbituric acid reactive substances) as significant indicators of lipid level peroxidation were determined. The content of -SH groups depended on the concentration of metal, and its level was the most stimulated by Zn, less by Cu and Ni. The TBARS content was 2 to 5 times higher in Cu than in Zn or Ni presence. In the presence of Zn and Ni, TBARS content reached approximately the same levels. For this parameter, the following rank order can be arranged: Cu >> Ni ≥ Zn. While Cu and Ni accumulation in R. subcapitata was confirmed, Zn accumulation was not determined or was below the detectable limit. Regression analyses revealed significant positive correlation between Cu accumulation and TBARS while carotenoids as possible antioxidants confirmed with TBARS mostly negative correlations.