Indian Journal of Multidisciplinary Research 01 ISSN - 0973 - 2225

Ind. J. Multi. Res. 2008. Vol. 4 (4) : 507 - 516

IN VITRO ANTIBACTERIAL ACTIVITIES OF PLANT ESSENTIAL

OILS AGAINST ORAL BACTERIA
KOTESWARA RAO P.*, VARAPRASAD BOBBARALA**, D. ARYAMITHRA**, P.
SUBHASHINI DEVI* AND T. RAGHAVA RAO*
*Department of Biochemistry, Andhra University, Visakhapatnam, A.P-530003, India.
**For U Biosciences, A/4A, Park lane Residency, East point colony, Visakhapatnam,
A. P-530 017, India., Email: koti_au@yahoo.co.in
(Received 25 July 2008, Accepted 23 October 2008)

SUMMARY

Essential oils, namely Cinnamon, Clove, Lavender, Rose, Eucalyptus, Lemon,

Menthol, Tea tree oil, Mustered and Rosemary oils were screened for potential antibacterial
activity against medicinally important bacterial strains, Which are present in dental plaques,
namely Streptococcus mutans (MTCC 890), Streptococcus mutans (MTCC 497),
Streptococcus salivarius, Streptococcus mitis, Lactobacillus fermentum, Streptococcus
anginosus, Streptococcus gordonii, Lactobacillus acidophilus, and Staphylococcus
aureus. The antibacterial activity was determined by agar well diffusion method. Out of ten
essential oils tested, nine oils showed antibacterial activity against one or more strains.
Cinnamon oil, Clove oil, Lavender oil, Rose oil, Eucalyptus oil, Lemon oil, Menthol, Tea tree
oil and Rosemary oils exhibited significant inhibitory effect. Cinnamon oil showed promising
inhibitory activity even at low concentration; where as Mustered oil did not shown activity
against the tested bacteria. Streptococcus mutans (MTCC 890) was the most resistant
strain, while the most susceptible bacterial strains were Streptococcus anginosus and
Lactobacillus fermentum. Cinnamon and Tea tree oil were showed strong activity against
all the tested bacterial strains. Hence these can be used to discover bio active natural products,
which are reduce the oral bacteria serve as leads in the development of new pharmaceuticals
that address unmet therapeutic needs in oral medicine. Majority of the oils showed antibacterial
activity against the tested strains. However Cinnamon oil, Tea tree oil, Clove oil and Lemon
oils were found to be inhibiting gram-positive bacteria. Cinnamon oil can be a good source of
antibacterial agents.
Key words: Antibacterial activity, Plant essential oils, Oral bacteria

INTRODUCTION

Essential oils are aromatic oily liquids obtained from plant materials (flowers, buds,
seeds, leaves, twigs, bark, herbs, wood, fruits and roots). Essential oils are complex mixers
IND. J. MULTI. RES. Volume - 4 ( No. 4). 2008 507
comprising many single compounds. Essential oils have been shown to possess antibacterial,
antifungal, antiviral insecticidal and antioxidant properties (Burt, 2004 and Kordali et al.
2005) essential oils are a rich source of biologically active compounds. There has been an
increased interest in looking at antimicrobial properties of extracts from aromatic plants
particularly essential oils (Milhau et al. 1997). Chemically they are derived from terpenes
and their oxygenated compounds. Each of these constituents contributes to the beneficial or
adverse effects. Therefore, it is reasonable to expect a variety of plant compounds in these
oils with specific as well as general antimicrobial activity and antibiotic potential (Darokar
et al. 1998). Essential oils such as Cinnamon, Clove, Eucalyptus, Lavender, Lemon, Menthol,
Tea tree, Mustard, Rosemary and Rose oil have been traditionally used by people for various
purposes in different parts of the world. A few recent studies have demonstrated antimicrobial
activity against selected oral pathogens from natural sources Cinnamon, Clove and Rosemary
oils had shown antibacterial and antifungal activity (Ouattara et al. 1997). In spite of all the
information available on the nine oils selected for this study, we were not able to find antibacterial
activity for all those oils. Hence this study was undertaken with the intention of finding out
the efficacy of these essential oils as antimicrobial agents for commercial purposes.

Dental plaque, a film of microorganisms on the tooth surface, plays an important

part in the development of caries and periodontal diseases (Marsh 1992). Mutans streptococci
can colonize the tooth surface and initiate plaque formation by their ability to synthesize
extracellular polysaccharides from sucrose, mainly water-insoluble glucan, using
glucosyltransferase (Gibbons and Van Houte 1975; Hamada and Slade 1980; Jacquelin et
al. 1995). De novo synthesis of water-insoluble glucan is essential for the adherence of
Streptococcus mutans and other oral microorganisms to the tooth surface, forming a barrier
that prevents the diffusion of acids produced by the bacteria. Many antimicrobial compounds
from plants and fungi (Shouji, et al. 2000)] which inhibit oral bacteria, particularly S. mutans,
and control plaque and thus prevent caries have been investigated (Morgan et al. 2001;
Wongkham et al. 2001; Koo et al. 2000; Ooshima et al. 2000; Jagtap and Karkera 1999;
Matsumoto et al. 1999; Park et al. 1998; Sato et al. 1996; Didry, Dubreuil, and Pinkas 1994;
Murio and Kubo 1993; Kubo, Himejima, and Muroi 1992; Kubo 1992; Chen, Lin, and Namba
1989). Use of plant-based alternatives for oral health has been successfully promoted.

This study evaluated the in vitro antibacterial activity of essential oils against some
oral micro organisms, Streptococcus mutans, Streptococcus salivarius, Streptococcus
mitis, Lactobacillus fermentum, Streptococcus anginosus, Streptococcus mutans,
Streptococcus gordonii, Lactobacillus acidophilus and Staphylococcus aureus. Thus,
oils would be a useful compound for development of antibacterial agents against oral pathogens
and has great potential for use in food additives and mouthwash for preventing and treating
dental caries.

IND. J. MULTI. RES. Volume - 4 ( No. 4). 2008 508

 Table 1: List of selected essential oils and their properties
S. BOTANICAL NAME
COMMON NAME PROPERTIES
No (FAMILY)
Cinnamomum zeylanicum Carminative, Stomachic,
1 Cinnamon oil
(Lauraceae) Astringent and Antiseptic.

Eugenia caryophyllus Dental analgesic, Carminative,

2 Clove oil
(Myrtaceae) Stimulant and Antiseptic.

Counter-irritant, Antiseptic,
3 Eucalyptus oil Eucalyptus globulus (Myrtaceae)
Expectorant, Cough reliever.

4 Lavender oil Lavandula angustifolia (Labiatae) Stimulant and Flavouring agent.

Carminative, Stimulant,
5 Lemon oil Citrus limon (Rutaceae)
perfuming and flavouring agent.

Antibacterial, Stimulant
6 Menthol Mentha arvensis (Lamiaceae)
and Flavoring agent.
Perfuming and
7 Rose oil Rosa damascena (Rosaceae)
Antibacterial agent.
Carminative, Stimulant
8 Rosemary oil Rosmarinus officinalis (Labiatae)
and flavouring agent.
Melaleuca alternifolia Chicken pox, Flu,
9 Tea tree oil
(Myrtaceae) Migraine, Urethritis.

MATERIALS AND METHODS

Nine essential oils (Table 1) were obtained from Tegraj & Co (P) Ltd, India. These
oils were selected based on literature survey and their use in traditional medicine. Quality of
the oils was ascertained to be more than 98% pure. The tested microorganisms of
Streptococcus mutans (MTCC 890), Streptococcus salivarius (MTCC 1938), Streptococcus
mitis (MTCC 2696), Lactobacillus fermentum (MTCC 903), Streptococcus anginosus
(MTCC 1929), Streptococcus mutans (MTCC 497), Streptococcus gordonii (MTCC 2695),
Lactobacillus acidophilus (MTCC 447) and Staphylococcus aureus (MTCC 96) were
purchased from The Microbial Type Culture Collection & Gene Bank (MTCC), Institute of
Microbial Technology Sector39-A, Chandigarh, India. Microorganisms were inoculated into
brain heart infusion broth and grown at 37oC for 24hours. The 24 and 48hr cultures were
used in the determination of microbial growth inhibition of oils.
IND. J. MULTI. RES. Volume - 4 ( No. 4). 2008 509
 The readymade Brain Heart Infusion Agar (52g) was suspended in distilled water
(1000ml) and heated up to boiling until it dissolved completely. The medium and the petri
dishes were autoclaved at pressure of 15 lb/inc for 20 min. Agar well diffusion bioassay was
employed for testing antibacterial activity of essential oils following the standard procedure
(Linday 1962). The medium was poured into petri dishes under aseptic conditions in a laminar
flow chamber. When the medium in the plates solidified, 0.5ml of 24h old culture of test
organism was inoculated. After inoculation, cups were scooped out with 6mm sterile cork
borer and the lids of the dishes were replaced. The tested essential oil at 50µl was introduced
into each well and the plates were incubated at 37oC for 24h. Microbial growth was determined
by measuring the diameter of zone of inhibition. For each bacterial strain the experiment was
done two times and the mean values are presented.

RESULTS AND DISCUSSION

Figure 1. Antibacterial activity of some medicinal plant oils against

Streptococcus mutans MTCC
25
890
Antibacterial activity against S. mutans
Zone of Inhibition(mm)

The antibacterial activity of oils of the different

20 plants against Streptococcus mutans
is shown in Figure 1. From which Cinnamon oil, Clove 15 oil, Lavender oil, Rose oil, Eucalyptus

oil, Lemon oil, Menthol, Tea tree oil and Rosemary oils10exhibited significant activity. Maximum
5
Antibacterial activity against S. salivarius

25 0
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Oils
zone of inhibition

Figure 2. Antibacterial activity of some medicinal plant oils against

Streptococcus salivarius MTCC 1938.
IND. J. MULTI. RES. Volume - 4 ( No. 4). 2008 510
activity was observed in Cinnamon oil followed by Lemon oil, Tea tree oil, Clove oil, Rosemary
oil, Eucalyptus oil and Menthol where as minimum antibacterial activity was recorded in
Menthol other oils demonstrated no activity against S. mutans.

All oils exhibited significant activity against Streptococcus salivarius in which figure
2 represents Cinnamon oil, Clove oil, Lavender oil, Rose oil, Eucalyptus oil, Lemon oil, Menthol,
Tea tree oil and Rosemary oils exhibited significant activity. Maximum activity was showed
in Cinnamon oil followed by Lemon oil, Tea tree oil, Clove oil, Rosemary oil, eucalyptus and
Menthol where as minimum antibacterial activity was observed in Lavender oil.
Antibacterial activity against S.mitis
25
Zone of Inhibition(mm)

20

15

10

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Oils zone of inhibition

Figure 3. Antibacterial activity of some medicinal plant oils against

Streptococcus mitis MTCC 2696.
Apart from the results obtained Cinnamon oil, Menthol Clove oil, Lavender oil, Rose
oil, Eucalyptus oil, Lemon oil, Menthol, Tea tree oil and Rosemary oils
Antibacterial exhibited
activity significant
against L. fermemtum
activity. Maximum activity was shown by Cinnamon 25 oil, followed by Lemon oil, Tea tree oil,
Clove oil, Rosemary oil, and eucalyptus. Minimum antibacterial activity was reported in
Z one of In hib itio n(m m )

20
Lavender oil and Eucalyptus oil. 15

10

0
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Oils zone of inhibition

Figure 4. Antibacterial activity of some medicinal plant oils against

Lactobacillus fermemtum MTCC 1938.
IND. J. MULTI. RES. Volume - 4 ( No. 4). 2008 511
 The antibacterial activity of oils of the different plants against Lactobacillus
fermemtum is shown in figure 4. All oils exhibited significant activity against Streptococcus
mutans in this figure. Maximum activity was recorded in Cinnamon oil followed by Lemon
oil, Tea tree oil, Clove oil, Rosemary oil, Eucalyptus oil, Menthol and Rose oil. Minimum
antibacterial activity was observed in Menthol and Rose oil.
Antibacterial activity against S.anginosus

Z o ne of Inhibition(m m ) 25

20

15

10

0
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Oils zone of inhibition

Figure 5. Antibacterial activity of some medicinal plant oils against

Streptococcus anginosus MTCC 1929.

The antibacterial activity of oils of the different plants against Streptococcus anginosus
is shown in Figure 5. From thee oils tested eight oils namely Cinnamon oil, Clove oil, Lavender
oil, Eucalyptus oil, Lemon oil, Menthol, Tea tree oil and Rosemary oils exhibited significant
activity. Maximum activity was shown by Cinnamon oil, followed by Rosemary oil, Lemon
antibacterial activity against s. aureus
oil, Tea tree oil, Clove oil, eucalyptus and Menthol. Menthol shows only minimum antibacterial
activity. Rose oil has no activity against Streptococcus 25 anginosus.
zone of inhibition(m m)

20

15

10

0
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oils zone of inhibition

Figure 6. Antibacterial activity of some medicinal plant oils against

Staphyloccus aureus MTCC 96.
IND. J. MULTI. RES. Volume - 4 ( No. 4). 2008 512
 The antibacterial activity of oils of the different plants against Streptococcus mutans
is shown in Figure 6. From the above observations Cinnamon oil, Clove oil, Eucalyptus oil,
Lemon oil, Menthol, Tea tree oil and Rosemary oils exhibited significant activity. Maximum
activity was shown by Rosemary oil and Tea tree oil. Minimum antibacterial activity was
shown by Eucalyptus oil. Other oils demonstrated no activity against S. aureus.
Antibacterial activity against l. acdophilua
25

20
Z o ne of inhibition(m m )

15

10

0
se

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oils zone of inhibition

Figure 7. Antibacterial activity of some medicinal plant oils against

Lactobacillus acidophilus MTCC 447.
All oils exhibited significant activity against Lactobacillus acidophilus. Cinnamon
oil shows maximum activity against L. acidophilus and Lavender oil showed minimum
antibacterial activity.
Antibacterial activity againsty s. mutans

25

20
Z on e o f in hib itio n(m m )

15

10

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Oils zone of inhibition

Figure 8.Antibacterial activity of some medicinal plant oils against

Streptococcus mutans MTCC 497.
Figure 8 represents Cinnamon oil, Clove oil, Lavender oil, Eucalyptus oil, Lemon oil,
Menthol, Tea tree oil and Rosemary oils exhibited significant activity. Maximum activity was
observed in Cinnamon oil followed by Rosemary oil, Lemon oil, Tea tree oil, Clove oil,
eucalyptus and Menthol. Menthol shows minimum antibacterial activity and Rose oils
demonstrated no activity against Streptococcus anginosus.
IND. J. MULTI. RES. Volume - 4 ( No. 4). 2008 513
 Figure 9. Antibacterial activity of some medicinal plant oils against
Streptococcus gordonii MTCC 2695.
Apart from screening Cinnamon oil, Clove oil, Lavender oil, Eucalyptus oil, Lemon
oil, Menthol, Tea tree oil and Rosemary oils exhibited significant activity. Maximum activity
was shown by Cinnamon oil, followed by Tea tree oil, Lemon oil, Rosemary oil, Clove oil,
Eucalyptus oil, Lavender oil and Menthol. Minimum antibacterial activity was shown by
Menthol. Rose oils demonstrated no activity against Streptococcus gordonii. Thus, Cinnamon
oil would be useful oil for development of antibacterial agents against oral pathogens and has
great potential for use in food additives and mouthwash for preventing and treating dental
caries. Very little work has been done on the possible medicinal applications of these Cinnamon
oil and hence extensive investigation is needed to exploit their therapeutic utility to combat
human oral diseases diseases.

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