The toxicity of purified microcystin-LR (MC-LR) and algal material collected during the winter and summer seasons (2005/2006) from the Hartebeespoort dam, South Africa, was investigated using the enzyme-linked immunosorbent assay (ELISA), mouse bioassay, catfish primary hepatocytes (in vitro assay) and protein phosphatase inhibition (PPi) assays. Microcystis aeruginosa, known producer of microcystins, was the dominant cyanobacteria present in the water samples. Exceptionally high cell numbers per millilitre were observed, especially with the summer samples (1.442×108 cells/ml), indicating a severe algal bloom in the dam. The toxin concentration as detected by ELISA and PPi assay in the winter and summer extracts was at least 1000 times more than the provisional guideline value (1 μg/l) set by the World Health Organization (WHO) for MC-LR in drinking water. Hepatotoxic effects and death of mice were observed after dosing with the summer extracts, while no hepatotoxic effects were observed with winter extracts. The EC50 values obtained after exposure of the catfish primary hepatocytes for 72 h to MC-LR, winter and summer extracts was about 0.091, 0.053 and 0.014 mg/l, respectively. Similar toxicity results were obtained when the mouse bioassay and primary hepatocytes were used.