The in vitro activity of Melaleuca alternifolia (tea tree) oil against dermatophytes (n = 106) and filamentous fungi (n = 78) was determined. Tea tree oil MICs for all fungi ranged from 0.004% to 0.25% and minimum fungicidal concentrations (MFCs) ranged from <0.03% to 8.0%. Time-kill experiments with 1-4 x MFC demonstrated that three of the four test organisms were still detected after 8 h of treatment, but not after 24 h. Comparison of the susceptibility to tea tree oil of germinated and non-germinated Aspergillus niger conidia showed germinated conidia to be more susceptible than non-germinated conidia. These data demonstrate that tea tree oil has both inhibitory and fungicidal activity.