By: Rachel Schmid Historical Use Of Tea Tree Oil (tto) Small, summer flowering tree native to Australia



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The effect of the essential oil and its components from Melaleuca alternifolia on endospore germination in Bacillus cereus

  • By: Rachel Schmid


Historical Use Of Tea Tree Oil (TTO)

  • Small, summer flowering tree native to Australia

  • First used by Bundjalong Aborigines in New South Wales for skin problems and respiration aliments (Carson and Riley 1993).

  • 1925: distilled oil’s antimicrobial properties published by Penfold and Grant

  • Since then extensive research done on oil’s uses



Uses of the Oil

  • Published evidence of antibacterial, antifungal, antiprotozan, antiviral, and anti-inflammatory properties

  • Also used to treat athlete’s foot, head lice, acne, and other skin irritations

  • Oil readily available for everyday use without a prescription

  • Found in shampoos, skin treatments, etc.



The major components of TTO



Previously Found Active Components

  • terpinen-4-ol thought to be most active ingredient (Carson and Riley 1995)

  • terpinen-4-ol and α-terpineol cause majority of the antibacterial and antifungal action (Carson et al., 2006)

  • α-pinene, linalool, and limonene also shown to have antibacterial properties (Raman et al., 1995)

  • 1,8-cineole thought to play role in allowing active components into cell



The present study

  • TTO has many antimicrobial abilities

  • Can it prevent endospore germination?

  • If so, what component of the oil can do this?



Endospores



Bacillus spp.

  • Using B. cereus as model for B. anthracis

  • 2001 bioterrorism attacks using anthrax spores on mailed envelopes

  • 22 mail workers infected and 5 died from exposure

  • Most infections from anthrax are cutaneous



Methods



Methods

  • B. cereus bacteria placed in LB on shaker for 8 days

  • Heat treatment

  • Spread on LB plate

  • 3-4 3M discs were placed on each plate



Methods

  • B. cereus bacteria placed in LB on shaker for 8 days

  • Heat treatment

  • Spread on LB plate

  • 4 3M discs were placed on each plate

  • Added small amount of TTO or components: terpinen-4-ol, γ-terpinene, α-terpinene, 1,8-cineole, α-pinene, p -cymene, α-terpineol, or limonene



Methods

  • B. cereus bacteria placed in LB on shaker for 8 days

  • Heat treatment

  • Spread on LB plate

  • 4 3M discs were placed on each plate

  • Added small amount of TTO or components: terpinen-4-ol, γ-terpinene, α-terpinene, 1,8-cineole, α-pinene, p -cymene, α-terpineol, or limonene

  • Incubated for 24 hours at 32°C



Methods

  • B. cereus bacteria placed in LB on shaker for 8 days

  • Heat treatment

  • Spread on LB plate

  • 4 3M discs were placed on each plate

  • Added small amount of TTO or components: terpinen-4-ol, γ-terpinene, α-terpinene, 1,8-cineole, α-pinene, p -cymene, α-terpineol, or limonene

  • Incubated for 24 hours at 32°C



Results

  • TTO inhibited endospore germination

  • terpinen-4-ol, α-terpinene, and α-terpineol components active

  • None significantly more active than the others or TTO



Synergisms

  • Two active components:

  • terpinen-4-ol and α-terpineol

  • Combination significantly more effective than either component

  • F = 40.17, df = 2, p < 0.0001



Synergisms

  • Active and inactive:

  • α-terpinene and 1,8-cineole

  • F = 26.24, df = 2, p < 0.0001

  • α-terpinene and p-cymene

  • F = 10.50, df = 2, p = 0.0014



Synergisms

  • Active and inactive:

  • α-terpineol and 1,8-cineole

  • F = 56.43, df = 2, p < 0.0001

  • α-terpineol and γ-terpinene

  • and F = 19.86, df = 2, p < 0.0001



GC/MS

  • The ten most abundant components of the commercial sample of TTO.

  • The relative percentages in the oil as observed by GC/MS.

  • The normal range for α-terpinene is 5-13%.



GC/MS

  • Composition of commercially purchased components that were active or part of a significant synergism



Discussion

  • Terpinen-4-ol is not the only active component, α-terpineol and α-terpinene are just as active

  • Terpenes are shown to cause a loss of membrane integrity and disrupt proton motive force (Sikkema et al. 1995; Cox et al. 1998)



  • These components are not active on their own but contribute to the overall activity of the oil

  • In bacteria, 1,8-cineole has been shown to disrupt the cell membrane to allow active components in (Carson et al. 2006)



Suggested Studies

  • Revise ISO for TTO to contain more α-terpinene

  • Use of TTO in alternative treatments of infectious disease

  • More work with TTO and anthrax endospores in containment labs

  • Clinical trials for prevention/healing of cutaneous infections in places where refrigeration of antibiotics is impossible



Literature Cited

  • • Carson, C. F., K. A. Hammer, and T. V. Riley. 2006. Melaleuca (Tea Tree) Oil: a review of antimicrobial and other medicinal properties. Clinical Microbiology Review 19: 50-62.

  • • Carson, C. F., and T. V. Riley. 1993. Antimicrobial activity of essential oil of Melaleuca alternifolia. Letters in Applied Microbiology 16: 49-55.

  • • Carson, C. F., and T. V. Riley. 1995. Antimicrobial activity of the major components of the essential oil of Melaleuca alternifolia. J. of Applied Bacteriology 78: 264-269.

  • • Cox, S. D., J. E. Gustafson, C. M. Mann, J. L. Markham, Y. C. Liew, R. P. Hartland, H. C. Bell, J. R. Warmington, and S. G. Wyllie. 1998. Tea tree oil causes K+ leakage and inhibits respiration in Escherichia coli. Letters Applied Microbiology 26: 355-358.

  • • Raman, A, U. Weir, and S. F. Bloomfield. 1995. Antimicrobial effects of tea tree oil and its major components on Staphylococcus aureus, Staphylococcus epidermidis, and Propionibacterium acnes. Applied Microbiology 21: 242-245.

  • • Sikkema, J., J. A. De Bont, and B. Poolman. 1995. Mechanisms of membrane toxicity of hydrocarbons. Microbiological Reviews 59: 201–222.



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