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Antimicrobial Agents and Chemotherapy, June 2002, p. 1914-1920, Vol. 46, No. 6
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.6.1914-1920.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Mechanism of Action of Melaleuca alternifolia (Tea Tree) Oil on Staphylococcus aureus Determined by Time-Kill, Lysis, Leakage, and Salt Tolerance Assays and Electron Microscopy

Christine F. Carson,1* Brian J. Mee,1 and Thomas V. Riley1,2

Department of Microbiology, The University of Western Australia, Crawley,1 Division of Microbiology and Infectious Diseases, The Western Australian Centre for Pathology and Medical Research, Nedlands, Western Australia 6009, Australia2

Received 24 April 2001/ Returned for modification 19 August 2001/ Accepted 6 February 2002

The essential oil of Melaleuca alternifolia (tea tree) has broad-spectrum antimicrobial activity. The mechanisms of action of tea tree oil and three of its components, 1,8-cineole, terpinen-4-ol, and {alpha}-terpineol, against Staphylococcus aureus ATCC 9144 were investigated. Treatment with these agents at their MICs and two times their MICs, particularly treatment with terpinen-4-ol and {alpha}-terpineol, reduced the viability of S. aureus. None of the agents caused lysis, as determined by measurement of the optical density at 620 nm, although cells became disproportionately sensitive to subsequent autolysis. Loss of 260-nm-absorbing material occurred after treatment with concentrations equivalent to the MIC, particularly after treatment with 1,8-cineole and {alpha}-terpineol. S. aureus organisms treated with tea tree oil or its components at the MIC or two times the MIC showed a significant loss of tolerance to NaCl. When the agents were tested at one-half the MIC, only 1,8-cineole significantly reduced the tolerance of S. aureus to NaCl. Electron microscopy of terpinen-4-ol-treated cells showed the formation of mesosomes and the loss of cytoplasmic contents. The predisposition to lysis, the loss of 260-nm-absorbing material, the loss of tolerance to NaCl, and the altered morphology seen by electron microscopy all suggest that tea tree oil and its components compromise the cytoplasmic membrane.

* Corresponding author. Mailing address: Department of Microbiology, The University of Western Australia, Crawley, Western Australia 6009, Australia. Phone: 61 8 9346 3288. Fax: 61 8 9346 2912. E-mail: ccarson{at}cyllene.uwa.edu.au.

Antimicrobial Agents and Chemotherapy, June 2002, p. 1914-1920, Vol. 46, No. 6
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.6.1914-1920.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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