Oral Microbiology Immunology 2003: 18: 389±392

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Susceptibility of oral bacteria to K. A. Hammer1, L. Dry1, M. Johnson2,

E. M. Michalak1, C. F. Carson1,
T. V. Riley1,2

Melaleuca alternifolia (tea tree) oil

1
Discipline of Microbiology, School of
Biomedical and Chemical Sciences, The
University of Western Australia, Crawley,
Western Australia, Australia, 2Division of

in vitro Microbiology and Infectious Diseases, Western

Australian Centre for Pathology and Medical
Research, Queen Elizabeth II Medical Centre,
Nedlands, Western Australia, Australia

Hammer KA, Dry L, Johnson M, Michalak EM, Carson CF, Riley TV. Susceptibility of
oral bacteria to Melaleuca alternifolia (tea tree) oil in vitro.
Oral Microbiol Immunol 2003: 18: 389±392. ß Blackwell Munksgaard, 2003.

The in vitro activity of Melaleuca alternifolia (tea tree) oil against 161 isolates of oral
bacteria from 15 genera was determined. Minimum inhibitory concentrations (MIC) and
minimum bactericidal concentrations (MBC) ranged from 0.003 to 2.0% (v/v).
MIC90 values were 1.0% (v/v) for Actinomyces spp., Lactobacillus spp., Streptococcus Key words: tea tree oil; Melaleuca alternifo-
mitis and Streptococcus sanguis, and 0.1% (v/v) for Prevotella spp. Isolates of lia; minimum inhibitory concentration; time
Porphyromonas, Prevotella and Veillonella had the lowest MICs and MBCs, and kill
isolates of Streptococcus, Fusobacterium and Lactobacillus had the highest. Time
Katherine A. Hammer, Discipline of
kill studies with Streptococcus mutans and Lactobacillus rhamnosus showed that Microbiology (M502), School of Biomedical
treatment with 0.5% tea tree oil caused decreases in viability of >3 log colony and Chemical Sciences, The University of
forming units/ml after only 30 s, and viable organisms were not detected after 5 min. Western Australia, 35 Stirling Hwy, Crawley,
These studies indicate that a range of oral bacteria are susceptible to tea tree oil, Western Australia, 6009, Australia
Tel.: ‡61 8 9346 4730; fax: ‡61 8 9346 2912;
suggesting that tea tree oil may be of use in oral healthcare products and in the e-mail: khammer@cyllene.uwa.edu.au
maintenance of oral hygiene. Accepted for publication June 27, 2003

The essential oil of Melaleuca alternifolia,
also known as tea tree oil, has been used
medicinally in Australia for more than 80
years (5). The tree itself has been used
therapeutically for even longer, being
one of the plants used in traditional med-
icine by the Bundjalung aborigines of
northern New South Wales (5). The essen-
tial oil is obtained by steam distillation and
contains approximately 100 components,
which are mostly monoterpenes (4). Com-
mercial oils must comply with the percen-
tage composition ranges for components
stipulated in the International Standard
4730 for `Oil of Melaleuca, terpinen-4-ol
type' (12). Tea tree oil, and several of
the components, has broad-spectrum
antimicrobial (5) and anti-in¯ammatory
(3, 14) activity in vitro. These properties
have formed the basis of its use in the
treatment of a range of super®cial com-
plaints such as cuts, insect bites, boils, acne
and tinea (2, 5). Furthermore, data from
recent clinical studies indicate that super-
®cial infections or conditions caused by
bacteria (2), fungi (13, 22) and viruses (6)
respond clinically to treatment with tea tree
oil. Anecdotal and scienti®c evidence also
suggest that tea tree oil may be useful in the
maintenance of oral hygiene and the pre-
vention of dental disease (9, 19, 23). How-
ever, the in vitro activity of tea tree oil
against the organisms found in the oral
cavity, or the potential uses of tea tree
oil in the oral cavity have not been studied
extensively. Therefore, the purpose of this
study was to investigate the in vitro sus-
ceptibility of a wide range of oral bacteria
to tea tree oil.
Material and methods
Tea tree oil
Tea tree oil (batch 97/1) was kindly
donated by Australian Plantations Pty
Ltd., Wyrallah, NSW. The oil composition
was determined by gas-chromatography
mass spectrometry performed by the Wol-
longbar Agricultural Institute, Wollong-
bar, NSW, as described previously (11).
Levels of terpinen-4-ol were 41.5% and
levels of 1,8-cineole were 2.1%, in com-
pliance with the International Standard
4730 (12).
Bacterial isolates
A collection of 161 bacterial isolates
was obtained from the culture collections
of the Discipline of Microbiology at The
University of Western Australia (n ˆ 9),
the Division of Microbiology and Infec-
tious Diseases at the Western Australian
Centre for Pathology and Medical Research
(n ˆ 123) and from the oral cavities of
volunteers (n ˆ 29) (Tables 1 and 2).
Reference isolates were Escherichia coli
NCTC 10418, Veillonella parvula NCTC
11810, Actinobacillus actinomycetemco-
390 Hammer et al.
Table 1. In vitro susceptibility of viridans streptococci (n ˆ 78) to tea tree oil Streptococcus spp. and A. actinomycetem-
MIC (% v/v) MBC (% v/v) comitans, which were incubated for 24 h in
Organism (n) Range 901 Range 901 air plus 5% CO2. After incubation, micro-
titre trays were subcultured by mixing the
S. bovis (1) 0.5 1
S. constellatus (8) 0.25±1 0.25±1 contents of each well, removing 10 ml
S. gordonii (2) 0.5 0.5±1 aliquots and spot inoculating onto pre-
S. intermedius (6) 0.12±2 0.25±2 dried Rogosa agar or blood agar. MICs
S. mitis (11) 0.25±1 1 0.25±1 1 were determined from subcultures as the
S. mutans (2) 0.25±2 0.25±2 lowest concentration resulting in the main-
S. oralis (5) 0.25±1 0.25±1
S. parasanguis (3) 0.25±0.5 0.25±0.5
tenance or reduction of the inoculum, and
S. salivarius (2) 0.25 0.25 the MBC was determined as the concen-
S. sanguis (19) 0.25±1 1 0.25±2 2 tration resulting in the death of 99.9% of
S. sobrinus (1) 1 2 the inoculum. The reference isolate E. coli
Streptococcus spp. (18) 0.25±1 1 0.25±2 2 was included in all assays as a control.
1
Percentage tea tree oil inhibitory or bactericidal to 90% of isolates. Assays were repeated at least twice for
each isolate and modal MIC and MBC
values were selected.
Table 2. In vitro susceptibility of oral bacteria (n ˆ 83) to tea tree oil Isolates of Porphyromonas endodonta-
MIC (% v/v) MBC (% v/v) lis, Prevotella intermedia and one isolate
Organism (n) Range 901 Range 901 each of Actinomyces viscosus and Clostri-
dium glycolicum did not produce suf®cient
Actinobacillus actinomycetemcomitans (1) 0.06 0.06
Actinomyces spp. (13) 0.1±1 1 0.1±2 1 growth in the microdilution format and
Branhamella sp. (1) 0.06 0.06 were tested by broth macrodilution. Dilu-
Capnocytophaga sp. (3) 0.03±0.06 0.03±0.06 tions of tea tree oil were prepared in 1 ml
Clostridium glycolicum (1) 0.05 0.1 volumes of BHIB and, after inoculation,
Eikenella corrodens (5) 0.03±0.06 0.03±0.06 ®nal concentrations of tea tree oil ranged
Fusobacterium spp. (5) 0.25±2 0.25±2
Lactobacillus spp. (18) 0.03±2 1 0.06±2 2
from 0.5±0.001% (v/v). Controls were pre-
Neisseria sp. (1) 0.25 0.25 pared with no tea tree oil. Macrodilutions
Peptostreptococcus asaccharolyticus (3) 0.25±0.5 0.5±1 were pre-reduced for approximately 60 min,
Porphyromonas endodontalis (8) 0.025±0.1 0.025±0.1 inoculated with 1 ml volumes of inocula
Prevotella intermedia (15) 0.003±0.1 0.1 0.003±0.1 0.1 prepared as described above, then incu-
Prevotella spp. (3) 0.016±0.06 0.016±0.06
bated for 48 h. MICs and MBCs were
Stomatococcus sp. (1) 0.5 0.5
Veillonella spp. (5) 0.016±1 0.03±1 determined by subculturing as described
1 above.
Percentage tea tree oil inhibitory or bactericidal to 90% of isolates.

Time kill assays

mitans ATCC 43718 and Lactobacillus
rhamnosus NCTC 10302. Oral bacteria
were isolated by rubbing a sterile cotton-
tipped swab over the teeth, gums and
tongue and then placing the swab into a
glass Bijou bottle containing 1 ml of phos-
phate buffered saline. The bottle was
mixed thoroughly using a vortex mixer
and the resulting bacterial suspension
was inoculated onto a range of selective
and non-selective culture media. Media
were incubated anaerobically at 358C for
3±10 days and pure cultures obtained. Iso-
lates were identi®ed using biochemical
pro®les determined with API 32A strips,
antibiotic susceptibilities determined using
Microring AN discs (Medical Wire and
Equipment Co. (Bath) Ltd., Wiltshire,
England) and other standard microbiolo-
gical methods (20).
In vitro susceptibility assays
Inocula for susceptibility tests were pre-
pared by growing organisms for 24±48 h
on Rogosa agar (Oxoid Ltd., Basingstoke,
England) for Lactobacillus spp. or blood
agar for the remaining organisms and sus-
pending colonies in sterile distilled water.
All isolates were grown anaerobically,
except for Streptococcus spp. and A. acti-
nomycetemcomitans, which were grown in
air plus 5% CO2. Suspensions were
adjusted to the turbidity of a 0.5 McFarland
standard using a nephelometer and diluted
as necessary to result in ®nal inocula con-
centrations of approximately 5  105 col-
ony forming units (cfu)/ml, as con®rmed
by viable counts.
For the microdilution assay, doubling
dilutions of tea tree oil ranging from 4 to
0.004% (v/v) were prepared in 100 ml
volumes in a 96-well microtiter tray in
the relevant growth medium. Todd Hewitt
Broth (Oxoid) was used for Streptococcus
spp., de Mann, Rogosa and Sharpe (MRS)
broth (Oxoid) was used for Lactobacillus
spp. and Brain Heart Infusion Broth
(BHIB) (Oxoid) was used for all other
organisms. A ®nal concentration of
0.001% (v/v) Tween 80 was included to
enhance tea tree oil solubility. After inocu-
lation, tests were incubated for 24 h under
anaerobic conditions except for tests with
A clinical isolate of Streptococcus mutans
and L. rhamnosus NCTC 10302 were used
in time kill studies. Inocula were prepared
by growing each isolate on the appropriate
solid media for 48±72 h, suspending
growth in 0.85% saline and adjusting to
0.5 McFarland to result in ®nal inocula
concentrations of approximately 107 cfu/
ml. Treatments containing tea tree oil ran-
ging from 4±0.12% (v/v) were prepared in
1 ml volumes of double-strength BHIB (S.
mutans) or MRS (L. rhamnosus) with
0.002% Tween 80. Treatments were pre-
reduced for at least 30 min before 1 ml
volumes of prepared inocula were added.
Samples were removed at 30 s, 5 and
10 min for viable counts. Samples were
diluted 10-fold in 0.85% saline and 10
ml aliquots were spot inoculated in dupli-
cate onto pre-dried blood agar or Rogosa
agar. Inoculation, sampling and viable
counting was performed in the anaerobic
chamber. Agar plates were incubated anae-
robically for 48±72 h and viable counts
were calculated from each replicate 10 ml
spot having one or more colony. The limit

of detection was 1  103 cfu/ml. Assays
were repeated at least twice for each tea
tree oil concentration and the mean, stan-
dard deviation and standard error of the
viable count data were calculated. Data
were compared using a Student's t-test,
two-tailed, two-sample assuming unequal
variance. P-values of <0.05 were consid-
ered signi®cant.
Results
Results of in vitro susceptibility assays are
shown in Tables 1 and 2. MICs and MBCs
were similar for all Streptococcus species,
and MICs ranged from 0.12 to 2% and
MBCs ranged from 0.5 to 2%. The MIC90
for all streptococci was 1% and the MBC90
was 2%. Some of the non-streptococcal
bacteria were considerably more suscepti-
ble to tea tree oil, with MICs and MBCs as
low as 0.016% for isolates of Prevotella
and Veillonella. Both the MIC90 and
MBC90 for all non-streptococcal bacteria
was 1%. For reference isolates, MIC/MBC
values were 0.06/0.06% for A. actinomy-
cetemcomitans, 0.25/0.5% for L. rhamno-
sus, 0.016/0.03% for V. parvula and 0.25/
0.25% for E. coli.
Time kill data are shown in Fig. 1. Sig-
ni®cant decreases in the viability of
S. mutans occurred after 30 s treatment
108
107
Viable count (cfu/ml)
106
105
104
103
10 2
0 2 4 6
Time (min)
108
Viable count (cfu/ml)
107
106
105
104
103
102
0 2 4 6
Time (min)
Fig. 1. Time kill curves for S. mutans (A) and L.
rhamnosus (B). Cells were treated with 0.5%
(~), 0.25% (&), 0.12% (*) and 0% (^) tea
tree oil and viable counts were determined after
30 s, 5 min and 10 min. Mean  standard error.
For S. mutans, the MIC and MBC was 2% and
for L. rhamnosus the MIC was 0.25% and the
MBC was 0.5%.
with 4, 2, 1 and 0.5% tea tree oil. Numbers
of viable organisms recovered from the 2
and 4% treatments after 30 s were 3.2 
103 and 3.3  103 cfu/ml, respectively (data
not shown in Fig. 1). Viable counts of S.
mutans after treatment with 0.25% tea tree
oil differed signi®cantly from controls at
30 s, 5 and 10 min, whereas viable counts
of cells treated with 0.12% differed sig-
ni®cantly from controls at 5 and 10 min
only. For L. rhamnosus, numbers of viable
cells were below detectable limits after
30 s treatment with 1% tea tree oil. Treat-
ment with 0.5% tea tree oil reduced the
numbers of viable organisms by >3 log
within 30 s and, at 5 min, viable organisms
were no longer detectable. Treatment with
0.25% tea tree oil resulted in a modest
reduction in viability, which was signi®-
cant at 5 and 10 min, whereas treatment
with 0.12% had no signi®cant effects.
Discussion
The tea tree oil susceptibility data obtained
in the present study are similar to those
from previous studies of oral bacteria,
which found MIC ranges of 0.03±1.25%
(15) and 0.11 >0.6% (19) for nine and six
isolates, respectively. Data were also simi-
lar to the MIC range of 0.02±0.08% found
for 12 isolates of oral bacteria by Walsh &
Longstaff (23), although these MICs were
of Melasol, a solution containing 40% tea
tree oil and 13% isopropyl alcohol (23). In
addition, a range of anaerobic and micro-
aerophilic vaginal bacteria, similar to those
found in the oral cavity, have been shown
to be susceptible to tea tree oil, with MICs
ranging from 0.03±2% (10). Although data
from the current and previous studies were
A generally similar, notably different was the
MIC of 1.25% for P. intermedia obtained
8 10
by Kulik et al. (15) which was consider-
ably higher than the MICs determined for
this organism in the present study and,
conversely, the values obtained in the cur-
rent study for Fusobacterium spp. were
considerably higher than those found by
previous authors (19, 23). Factors that may
have contributed to these divergent results
include differences between the types and
B numbers of bacterial isolates tested in each
of the studies, and the methods used,
8 10
including the criteria for determining
MICs. In particular, where possible, multi-
ple isolates from each species were tested
in the current study, whereas previous
studies often tested only single isolates.
Time kill studies demonstrated rapid
killing of both S. mutans and L. rhamnosus
after 30 s treatment with as little as 0.5%
tea tree oil, suggesting that tea tree oil may
Oral bacteria and tea tree oil 391
be an effective active ingredient in anti-
septic mouthrinses. Time kill studies of
30 s have been recommended for evaluat-
ing mouthrinses in vitro (24) and many
mouthrinses are recommended by manu-
facturers to be used for 30 s twice daily,
with volumes of approximately 20 ml (8,
17). However, the signi®cant bactericidal
effects seen with 0.5% tea tree oil in vitro
may not necessarily re¯ect what may be
occurring in the oral cavity when rinsing
with a tea tree oil mouthwash solution, for
several reasons. Firstly, in vitro and in vivo
®ndings may not directly correlate, as
found previously for another mouthrinse
product, which gave promising in vitro
results but did not perform well in vivo
(17). Secondly, the presence of exogenous
protein in the oral cavity may adversely
affect the activity of tea tree oil (11) and it
is therefore important to repeat the time-
kill studies in the presence of protein to
examine this effect (24). Ultimately, clin-
ical trials are required to determine in vivo
ef®cacy and investigate optimal tea tree oil
concentrations.
Traditional or alternative oral hygiene
measures, such as mastic chewing gum and
traditional chewing sticks, have been the
subject of recent studies (7, 21). The use of
mastic chewing gum was shown to signi®-
cantly reduce bacterial numbers in saliva,
plaque index and gingival index in 20
volunteers, compared to placebo gum (21).
Similarly, use of a eucalyptus-extract gum
signi®cantly reduced the plaque index in
15volunteerswhencomparedtocontrolgum
(18). It has been suggested that the effec-
tiveness of these traditional oral hygiene
tools may be due, in part, to the antimicro-
bial plant compounds found within these
traditional medicaments (7, 18). Further-
more, it has been suggested for some time
that tea tree oil may be an effective agent
for both the treatment and prevention of
oral infections or conditions, although
scienti®c reports are few. In 1937, Penfold
& Morrison reported that oral conditions
such as thrush, aphthous stomatitis, mouth
ulcers, gingivitis and pyorrhoea all
responded favorably to treatment with
tea tree oil (16). More recently, the effect
of mouthwashes containing tea tree oil
(approximately 0.34%) or 0.1% chlorhex-
idine on plaque formation was compared to
placebo in eight volunteers (1). However,
the plaque index and plaque vitality after
use of the tea tree oil mouthwash did not
differ from placebo mouthwash on any day,
whereas the chlorhexidine mouthwash
group differed signi®cantly from placebo
on all days (1). Another study compared
tea tree oil (0.2%), garlic (2.5% solution)
392 Hammer et al.
and chlorhexidine (0.12%) by determining
levels of S. mutans and other oral micro-
organisms in saliva samples after the use of
each mouthwash (9). All products pro-
duced signi®cant immediate reductions
in viable counts and reductions were main-
tained for 2 weeks following the cessation
of mouthwash use, but only for the tea tree
oil and garlic mouthwashes, not chlorhex-
idine (9). Tea tree oil mouthwash has also
been evaluated for the treatment of oral
candidiasis in two different studies with
overall clinical response rates of 67% (13)
and 60% (22). These clinical data illustrate
that tea tree oil can be used effectively in
the oral cavity for yeast infections, how-
ever, whether tea tree oil can reduce or
prevent the formation of microbial plaque
has not yet been established. A further
rationale for the oral use of tea tree oil is
that it has anti-in¯ammatory effects (3, 14)
and may therefore be helpful in the treat-
ment or prevention of gingivitis. Many oral
hygiene products may reduce gingival
in¯ammation indirectly by reducing pla-
que, whereas tea tree oil has the potential to
reduce both gingivitis and plaque mass
simultaneously.
Safety issues associated with the use of
tea tree oil in the oral cavity need to be
addressed. Adverse effects such as a burn-
ing sensation (9, 13, 22), stinging (22) and
unpleasant taste (9) have all been noted
previously by patients using tea tree oil
mouthwashes. However, in two of these
studies the tea tree oil solution used was
alcohol-based, and it is possible that the
alcohol may have caused the burning,
rather than the tea tree oil. Also, the mild
to moderate burning sensation was noted
largely during the ®rst week of therapy and
gradually decreased thereafter (13, 22),
which may be due to accommodation.
Patients using Listerine1 (which contains
26.9% alcohol and 0.26% essential oils)
have reported very similar reactions of an
initial burning sensation and bitter taste,
both of which decreased over subsequent
days (24). The possibility of reactions to
tea tree oil within the oral cavity or sys-
temic toxicity from ingestion of products
requires further research.
In conclusion, in vitro susceptibility data
from this study show that the bacteria
found in the mouth are susceptible to,
and rapidly killed by, tea tree oil. As such,
tea tree oil, incorporated into appropriate
oral hygiene products such as mouthrinses,
dentifrices and dental gels or irrigators,
may be suitable for use in the oral cavity.
Acknowledgments
This work was supported by grants UWA-
58A and UWA-55A from the Rural Indus-
tries Research and Development Corpora-
tion, and Australian Bodycare Pty. Ltd.,
Vissenbjerg, Denmark.
References
1. Arweiler NB, Donos N, Netuschil L, Reich
E. Clinical and antibacterial effect of tea
tree oil ± a pilot study. Clin Oral Invest
2000: 4: 70±73.
2. Bassett IB, Pannowitz DL, Barnetson RStC.
A comparative study of tea-tree oil versus
benzoylperoxide in the treatment of acne.
Med J Aust 1990: 153: 455±458.
3. Brand C, Ferrante A, Prager RH, Riley
TV, Carson CF, Finlay-Jones JJ, et al. The
water soluble components of the essential
oil of Melaleuca alternifolia (tea tree oil),
suppress the production of superoxide by
human monocytes, but not neutrophils,
activated in vitro. In¯amm Res 2001: 50:
213±219.
4. Brophy JJ, Davies NW, Southwell IA, Stiff
IA, Williams LR. Gas chromatographic
quality control for oil of Melaleuca terpi-
nen-4-ol type (Australian tea tree). J Agric
Food Chem 1989: 37: 1330±1335.
5. Carson CF, Riley TV. Antimicrobial acti-
vity of the essential oil of Melaleuca alter-
nifolia. Lett Appl Microbiol 1993: 16:
49±55.
6. Carson CF, Ashton L, Dry L, Smith DW,
Riley TV. Melaleuca alternifolia (tea tree)
oil gel (6%) for the treatment of recurrent
herpes labialis. J Antimicrob Chemother
2001: 48: 450±451.
7. Darout IA, Albandar JM, Skaug N. Period-
ontal status of adult Sudanese habitual
users of miswak chewing sticks or tooth-
brushes. Acta Odontol Scand 2000: 58:
25±30.
8. Fine DH, Furgang D, Barnett ML, Drew C,
Steinberg L, Charles CH, et al. Effect of an
essential oil-containing antiseptic mou-
thrinse on plaque and salivary Streptococ-
cus mutans levels. J Clin Periodontol 2000:
27: 157±161.
9. Groppo FC, Ramacciato JC, Simoes RP,
Florio FM, Sartoratto A. Antimicrobial
activity of garlic, tea tree oil, and chlorhex-
idine against oral microorganisms. Int Dent
J 2002: 52: 433±437.
10. Hammer KA, Carson CF, Riley TV. In vitro
susceptibilities of lactobacilli and organ-
isms associated with bacterial vaginosis
to Melaleuca alternifolia (tea tree) oil.
Antimicrob Agents Chemother 1999: 43:
196.
11. Hammer KA, Carson CF, Riley TV. In¯u-
ence of organic matter, cations and surfac-
tants on the antimicrobial activity of
Melaleuca alternifolia (tea tree) oil in vitro.
J Appl Microbiol 1999: 86: 446±452.
12. International Organization or Standardisa-
tion. ISO 4730:1996: oil of Melaleuca,
terpinen-4-ol type (tea tree oil). Geneva:
International Organization or Standardisa-
tion, 1996.
13. Jandourek A, Vaishampayan JK, Vazquez
JA. Ef®cacy of Melaleuca oral solution for
the treatment of ¯uconazole refractory oral
candidiasis in AIDS patients. AIDS 1998:
12: 1033±1037.
14. Koh KJ, Pearce AL, Marshman G, Finlay-
Jones JJ, Hart PH. Tea tree oil reduces
histamine-induced skin in¯ammation. Br J
Dermatol 2002: 147: 1212±1217.
15. Kulik E, Lenkeit K, Meyer J. Antimicrobial
activity of tea tree oil (Melaleuca alterni-
folia) against oral microorganisms. Acta
Med Dent Helv 2000: 5: 125±130.
16. Penfold AR, Morrison FR. Some notes on
the essential oil of Melaleuca alternifolia.
Australas J Pharm 1937: 18: 274±275.
17. Pitten FA, Kramer A. Antimicrobial ef®-
cacy of antiseptic mouthrinse solutions. Eur
J Clin Pharmacol 1999: 55: 95±100.
18. Sato S, Yoshinuma N, Ito K, Tokumoto T,
Takiguchi T, Suzuki Y, et al. The inhibitory
effect of furoran and eucalyptus extract-
containing chewing gum on plaque forma-
tion. J Oral Sci 1998: 40: 115±117.
19. Shapiro S, Meier A, Guggenheim B. The
antimicrobial activity of essential oils and
essential oil components towards oral bac-
teria. Oral Microbiol Immunol 1994: 9:
202±208.
20. Summanen P, Baron EJ, Citron DM, Strong
C, Wexler HM, Finegold SM. Wadsworth
anaerobic bacteriology manual, 3rd edn.
Belmont: Star Publishing Company, 1993.
21. Takahashi K, Fukuzawa M, Motohira H,
Ochiai K, Nishikawa H, Miyata T. A pilot
study on antiplaque effects of mastic chew-
ing gum in the oral cavity. J Periodontol
2003: 40: 501±505.
22. Vazquez JA, Zawawi AA. Ef®cacy of
alcohol-based and alcohol-free Melaleuca
oral solution for the treatment of ¯ucona-
zole-refractory oral candidiasis in patients
with AIDS. AIDS 2002: 12: 1033±1037.
23. Walsh LJ, Longstaff J. The antimicro-
bial effects of an essential oil on selected
oral pathogens. Periodontology 1987: 8:
11±15.
24. Wu CD, Savitt ED. Evaluation of the safety
and ef®cacy of over-the-counter oral
hygiene products for the reduction and con-
trol of plaque and gingivitis. Periodontol
2000 2002: 28: 91±105.