Tetracycline (TC) antibiotics are one of the class of drugs widely used in clinical practice but also constitute a significant environmental concern. However, the adverse effects of TC on non-target organisms have not been well studied. The aim of this study was to examine the influence of exposure to high levels of TC on thalli of lichens to determine the impact on (1) physiological parameters including integrity of cell membranes, photosynthetic efficiency and viability, (2) oxidative stress response such as membrane lipid peroxidation, and (3) enzymatic antioxidant activities as catalase (CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR). Data demonstrated that exposure to tetracycline did not markedly affect the lichen membrane damage as indicated by no change in conductivity. This antibiotic diminished the potential photosystem II efficiency (FV/FM) indicating enhanced susceptibility as evidenced by lower chlorophyll fluorescence and chlorophyll content. The viability of lichens exposed to high concentrations of tetracycline was significantly reduced. The concentrations of thiobarbituric acid reactive substances were markedly elevated with increasing concentrations of antibiotics. At higher TC concentrations, 500 mg/L SOD activity was significantly elevated. In the case of CAT, APX and GR, TC at higher concentrations significantly decreased these enzymic activities. The findings of this study contribute to the knowledge that TC antibiotics exert adverse ecotoxicological effects on lichens at high concentrations and provided a better understanding of the mechanisms underlying toxicity. Data also indicates that lichens may serve as an effective biomonitoring species for TC antibiotic exposure. Keywords: tetracycline; lichen; viability; chlorophyll fluorescence; oxidative stress; antioxidant enzyme.