A broth micro-dilution method was used to examine the susceptibility of Escherichia coli (n = 110) and Staphylococcus aureus (n = 105) to the essential oil of Melaleuca alternifolia (tea tree oil). The detergent Tween 80 was used successfully to enhance the solubility of tea tree oil in the test medium. The MIC90 of tea tree oil for E. coli was 0.25% while for S. aureus it was 0.50%.