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Effects of Tea Tree Oil on Biofilm Formation

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Effects of Tea Tree Oil on Biofilm Formation Effects of Tea Tree Oil on Biofilm Formation RIRDC Publication No. 08/140 RIRDCInnovation for rural Australia Effects of Tea Tree Oil on Biofilm Formation by KA Hammer, CF Carson, T-J Tan & TV Riley September 2008 RIRDC Publication No 08/ 140 RIRDC Project No PRJ-000451 © 2008 Rural Industries Research and Development Corporation. All rights reserved. ISBN 1 74151 729 X ISSN 1440-6845 Effects of Tea Tree Oil on Biofilm Formation Publication No. 08/140 Project No. PRJ-000451 The information contained in this publication is intended for general use to assist public knowledge and discussion and to help improve the development of sustainable regions. You must not rely on any information contained in this publication without taking specialist advice relevant to your particular circumstances. While reasonable care has been taken in preparing this publication to ensure that information is true and correct, the Commonwealth of Australia gives no assurance as to the accuracy of any information in this publication. The Commonwealth of Australia, the Rural Industries Research and Development Corporation (RIRDC), the authors or contributors expressly disclaim, to the maximum extent permitted by law, all responsibility and liability to any person, arising directly or indirectly from any act or omission, or for any consequences of any such act or omission, made in reliance on the contents of this publication, whether or not caused by any negligence on the part of the Commonwealth of Australia, RIRDC, the authors or contributors. The Commonwealth of Australia does not necessarily endorse the views in this publication. This publication is copyright. Apart from any use as permitted under the Copyright Act 1968, all other rights are reserved. However, wide dissemination is encouraged. Requests and inquiries concerning reproduction and rights should be addressed to the RIRDC Publications Manager on phone 02 6271 4165. Researcher Contact Details Katherine A. Hammer Microbiology and Immunology School of Biomedical, Biomolecular and Chemical Sciences The University of Western Australia Phone: (08) 9346 1986 Fax: (08) 9346 2912 Email: [email protected] In submitting this report, the researcher has agreed to RIRDC publishing this material in its edited form. RIRDC Contact Details Rural Industries Research and Development Corporation Level 2, 15 National Circuit BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604 Phone: 02 6271 4100 Fax: 02 6271 4199 Email: [email protected]. Web: http://www.rirdc.gov.au Published in September 2008 by Union Offset ii Foreword The ability of tea tree oil to inhibit and kill a wide range of microorganisms means that it has great potential not only as a topical antimicrobial agent for use in humans but also as a biocide in industrial applications. One of these potential applications is in the prevention of biofilm formation or the remediation of surfaces on which biofilms have formed. This report provides data on the ability of tea tree oil to prevent and disinfect or disrupt biofilm. It provides additional evidence for the antimicrobial activity of the oil and yet another avenue for future product development. Biofilm formation is becoming recognised as a key step in many infections. Prevention of this process as well as destruction of pre-formed biofilm are key strategies in controlling infectious disease. Apart from the key role of biofilm formation in many infectious disease processes, biofilm formation is a major problem in industrial settings such as water reticulation, sewage treatment, food manufacture and any setting where sufficient water and nutrients are available for microorganisms to grow. With careful formulation designed to preserve the antimicrobial activity of tea tree oil, followed by large-scale testing of these products on biofilm formation and biofilm destruction, this may represent another property of tea tree oil that has widespread application in medical and industrial settings. The purpose of this project was to provide initial data characterising the effect of tea tree oil on biofilms which is why RIRDC has invested in this report. The importance of this report is that it provides basic data demonstrating the ability of tea tree oil to inhibit the formation of biofilms and to destroy pre-existing ones. It will be a useful basis for those contemplating investment or formulating products and will help support the marketability of tea tree oil internationally. This project was funded from industry revenue which is matched by funds provided by the Australian Government. This report, an addition to RIRDC’s diverse range of over 1800 research publications, forms part of our Tea Tree Oil R&D program, which aims to investigate the efficacy of tea tree oil and develop new products. Most of our publications are available for viewing, downloading or purchasing online through our website: • downloads at www.rirdc.gov.au/fullreports/index.html • purchases at www.rirdc.gov.au/eshop Peter O’Brien Managing Director Rural Industries Research and Development Corporation iii Acknowledgments This work was supported with an industry contribution and tea tree oil samples from Pat and Paul Bolster, Gelair Pty. Ltd. We are grateful for the technical, financial and institutional support of the Discipline of Microbiology & Immunology, School of Biomedical, Biomolecular and Chemical Sciences at The University of Western Australia and the Division of Microbiology and Infectious Diseases at PathWest Laboratory Medicine WA. Abbreviations ATCC American Type Culture Collection GRAS Generally recognized as safe MCC Minimum cidal concentration MIC Minimum inhibitory concentration NCTC National Collection of Type Cultures nm Nanometres OD Optical density PBS Phosphate buffered saline RT Room temperature SBF Specific biofilm formation TSB Trypticase soy broth TSBG Trypticase soy broth with 0.25% glucose TSBS Trypticase soy broth supplemented with 1% sodium chloride v/v Volume for volume w/v Weight for volume XTT 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide YEPD Yeast extract peptone dextrose iv Contents Foreword.............................................................................................................................................. iii Acknowledgments ............................................................................................................................ iv Abbreviations ..................................................................................................................................... iv Contents ............................................................................................................................................... v List of Tables...................................................................................................................................... vi List of Figures ................................................................................................................................... vii Executive Summary........................................................................................................................ viii Introduction ......................................................................................................................................... 1 Materials and Methods...................................................................................................................... 2 Tea tree oil............................................................................................................................................2 Organisms ............................................................................................................................................2 Determination of minimum inhibitory and cidal concentrations .........................................................2 Biofilm formation.................................................................................................................................3 Destruction of formed biofilm..............................................................................................................3 Quantification of biofilm......................................................................................................................4 Crystal violet staining ......................................................................................................................4 XTT metabolism ..............................................................................................................................4 Regrowth of viable biofilm organisms.............................................................................................4 Standardisation of data and statistical analyses................................................................................5 Results .................................................................................................................................................. 6 C. albicans........................................................................................................................................9 V. harveyi.........................................................................................................................................9 S. maltophilia ...................................................................................................................................9 Destruction of pre-formed biofilm .....................................................................................................25
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