Toxicity tests with Vibrio fischeri, Daphnia magna, and Lemna minor were used to evaluate acute and chronic toxicity of cork-boiling wastewaters and correlation analysis was performed with physicochemical parameters. Acute toxicity values (EC(50)) ranged from 2.3% to 24.2% in the Microtox test and from 4.4% to 29.5% in the Daphnia test. According to these values, 78% of the samples were classified as acutely toxic to the most sensitive species (V. fischeri). Significant correlation was obtained between EC(50) and chemical oxygen demand, biochemical oxygen demand after 5 days, total organic carbon, tannin and lignin and total solids. No significant correlation was found between the two acute tests. The D. magna chronic test showed significant differences between all tested concentrations and the control, with an EC(50) of 0.32%. An EC(50) value of 26.0% was calculated for L. minor. For the analysis of cork-boiling wastewater toxicity and routine biomonitoring, the use of the Microtox test is suggested, to afford greater environmental protection.