Abstract: Background and Objective: Vernonia mespilifolia is widely used in folk medicine in the Eastern Cape province, South Africa. This study evaluated the toxicity of acetone, aqueous and ethanol extracts of Vernonia mespilifolia using brine shrimp model. Materials and Methods: Different concentrations (0.0625-1 mg mL–1) of the extracts were used to incubate the cysts and nauplii (hatched cysts) of brine shrimp (Artemia salina) to evaluate their effects on the hatching success of the cyst and lethality of the nauplii respectively. The percentage hatching success of cyst and Lethal Concentration (LC50) to kill 50% of the nauplii were recorded. Results: The hatching success was in order: Aqueous extract (48.6%)>acetone extract (38.2%)>ethanol extract (26.8%). The hatching of nauplii was in a concentration dependent fashion, with hatching success decreasing with increase in concentration of extracts. Lethality of extract was based on Meyer’s index of toxicity. Conclusion: All the three extracts showed high levels of toxicity with LC50 <1 mg mL–1 which signify toxicity in a brine shrimp model. In this respect, V. mespilifolia possesses cytotoxic behavior suggesting the presence of potential bioactive chemical component in the plant extract. Further in vivo and cell lines cytotoxicity test is recommended to substantiate these findings.