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Mechanisms of Resistance

Effects of Melaleuca alternifolia (Tea Tree) Essential Oil and the Major Monoterpene Component Terpinen-4-ol on the Development of Single- and Multistep Antibiotic Resistance and Antimicrobial Susceptibility

Katherine A. Hammer, Christine F. Carson, Thomas V. Riley
Katherine A. Hammer
Discipline of Microbiology and Immunology, School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia, Crawley, Western Australia, 6009, Australia
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Christine F. Carson
Discipline of Microbiology and Immunology, School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia, Crawley, Western Australia, 6009, Australia
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Thomas V. Riley
Discipline of Microbiology and Immunology, School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia, Crawley, Western Australia, 6009, AustraliaDivision of Microbiology and Infectious Diseases, PathWest Laboratory Medicine WA, Queen Elizabeth II Medical Centre, Nedlands, Western Australia, 6009, Australia
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DOI: 10.1128/AAC.05741-11
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Tables

  • Table 1

    Frequencies of single-step antibiotic-resistant mutants occurring in the presence and absence of tea tree oila

    Organism and no. of isolatesAntibioticFold increase in MICFrequency of mutants in:P valueb
    Control culture with:TTO culture with:
    Antibiotic alone (treatment A)Antibiotic + TTO (treatment B)Antibiotic alone (treatment C)Antibiotic + TTO (treatment D)
    S. aureus
        10RIF88.9 × 10−85.9 × 10−81.1 × 10−77.2 × 10−80.0065c
        10MUP87.3 × 10−82.3 × 10−89.9 × 10−83.5 × 10−80.0002d
        7VAN22.4 × 10−71.2 × 10−73.4 × 10−75.9 × 10−70.1268
        4CIP28.1 × 10−85.9 × 10−88.6 × 10−83.7 × 10−80.6725
    E. coli
        10RIF83.7 × 10−82.9 × 10−84.3 × 10−83.8 × 10−80.2083
        9KAN23.3 × 10−62.0 × 10−72.6 × 10−63.1 × 10−7<0.0001e
    • ↵a Values are the geometric means from 4 to 10 isolates. MUP, mupirocin; RIF, rifampin; VAN, vancomycin; CIP, ciprofloxacin; KAN, kanamycin; TTO, tea tree oil.

    • ↵b P values were obtained by repeated-measure one-way ANOVA.

    • ↵c Significant differences exist between treatments B and C (P < 0.01) (Bonferroni post test).

    • ↵d Significant differences exist between treatments A and B (P < 0.01), B and C (P < 0.001), and C and D (P < 0.05).

    • ↵e Significant differences exist between treatments A and B (P < 0.001), A and D (P < 0.01), B and C (P < 0.001), and C and D (P < 0.01).

  • Table 2

    S. aureus MICs of antibiotics (μg/ml) alone, antibiotics with or without tea tree oil (0.062%) or terpinen-4-ol (0.031%), and tea tree oil or terpinen-4-ol without antibiotic, determined by serial subculturea

    Agent (no. of isolates) and parameterTreatmentMIC on day:
    123456
    CIP (10)
        MedianAlone0.511222
    With TTO0.2511222
    With T4ol0.250.51222
        GMAlone0.80.81.11.51.62.3
    With TTO0.251.01.11.62.53.5
    With T4ol0.20.61.01.51.72.3
    MUP (11)
        MedianAlone0.120.250.5111
    With TTO0.120.120.50.50.51
    With T4ol0.120.120.5142
        GMAlone0.10.30.50.91.21.5
    With TTO0.10.20.50.60.51.4
    With T4ol0.10.20.50.91.93.1
    VAN (12)
        MedianAlone124844
    With TTO144844
    With T4ol144844
        GMAlone1.02.74.57.64.03.8
    With TTO1.15.05.06.33.84.3
    With T4ol0.94.04.09.53.65.0
    Tea tree oil (18)
        MedianAlone0.50.51112
        GMAlone0.50.71.00.91.01.7
    Terpinen-4-ol (18)
        MedianAlone0.250.250.250.50.51
        GMAlone0.20.30.30.60.60.8
    • ↵a GM, geometric means. Boldface type indicates that the MIC is more than double the baseline (day 1) value. Single underlining indicates that values differed significantly from antibiotic alone on that day. Double underlining indicates significant differences between tea tree oil and terpinen-4-ol treatments on that day.

  • Table 3

    Summary of E. coli MICs of antibiotics (μg/ml) alone, antibiotics with tea tree oil (0.062%) or terpinen-4-ol (0.031%), and tea tree oil or terpinen-4-ol without antibiotics, determined by serial subculturea

    Agent (no. of isolates) and parameterTreatmentMIC on day:
    123456
    CIP (12)
        MedianAlone0.0080.0160.0160.030.030.12
    With TTO0.0080.0160.030.060.060.12
    With T4ol0.0080.0160.060.060.120.12
        GMAlone0.0100.0250.0340.0700.0700.140
    With TTO0.0070.0140.0290.0450.0640.122
    With T4ol0.0090.0180.0480.0780.1000.147
    KAN (11)
        MedianAlone832166464128
    With TTO41632323264
    +With T4ol41632323264
        GMAlone6.745.320.260.450.8107.6
    With TTO4.812.733.938.142.747.9
    With T4ol2.515.128.533.932.053.8
    AMP (10)
        MedianAlone24881616
    With TTO24881616
    With T4ol24881632
        GMAlone2.14.98.010.611.316.0
    With TTO2.14.68.69.812.112.1
    With T4ol2.14.911.311.317.124.3
    Tea tree oil (18)
        MedianAlone0.511111
        GMAlone0.650.730.961.001.120.96
    Terpinen-4-ol (18)
        MedianAlone0.120.250.250.250.250.25
        GMAlone0.130.170.220.290.250.26
    • ↵a GM, geometric means. Boldface type indicates that the MIC is more than double the baseline (day 1) value. Single underlining indicates that values differed significantly from antibiotic alone on that day. Double underlining indicates significant differences between tea tree oil and terpinen-4ol treatments on that day.

  • Table 4

    MICs of antibiotics (μg/ml), biocides (μg/ml), and tea tree oil and terpinen-4-ol (%, vol/vol) for three Staphylococcus strains serially subcultured with terpinen-4-ol or tea tree oila

    AgentS. aureus NCTC 6571S. aureus ATCC 25923S. epidermidis ATCC 12228
    Passage no.MICPassage no.MICPassage no.MIC
    ControlWith 0.1% T4olControlWith 0.1% T4olWith 0.1% TTOControlWith 0.2% T4ol
    AMX190.120.06180.120.120.121410.5
    220.250.25200.50.251710.5
    CIP170.250.06160.120.120.25140.250.25
    190.120.06180.250.060.12170.50.25
    220.120.06200.250.06
    GEN1712160.50.250.5140.120.5
    1911180.250.120.12170.250.25
    2212200.250.25
    TET170.120.06160.120.120.25140.50.5
    190.25<0.03180.250.120.251710.5
    220.120.06200.250.12
    VAN1711162221444
    1910.5181121744
    220.50.252011
    Benzalkonium170.511620.521421
    Chloride19111820.511721
    22112020.5
    Triclosan190.060.03180.060.030.03170.030.06
    220.120.12200.250.12
    Tea tree oil170.50.25160.120.250.25140.50.5
    1910.5180.50.50.5170.50.5
    220.250.25200.50.5
    Terpinen-4-ol170.250.5160.250.50.5140.50.5
    190.50.5180.50.50.25170.50.5
    220.120.25200.120.25
    • ↵a Boldface type indicates a difference in MIC of 4-fold or more for passaged and nonpassaged strains.

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Effects of Melaleuca alternifolia (Tea Tree) Essential Oil and the Major Monoterpene Component Terpinen-4-ol on the Development of Single- and Multistep Antibiotic Resistance and Antimicrobial Susceptibility
Katherine A. Hammer, Christine F. Carson, Thomas V. Riley
Antimicrobial Agents and Chemotherapy Jan 2012, 56 (2) 909-915; DOI: 10.1128/AAC.05741-11

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Effects of Melaleuca alternifolia (Tea Tree) Essential Oil and the Major Monoterpene Component Terpinen-4-ol on the Development of Single- and Multistep Antibiotic Resistance and Antimicrobial Susceptibility
Katherine A. Hammer, Christine F. Carson, Thomas V. Riley
Antimicrobial Agents and Chemotherapy Jan 2012, 56 (2) 909-915; DOI: 10.1128/AAC.05741-11
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