Contemporary Clinical Trials 32 (2011) 802808

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Contemporary Clinical Trials

j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / c o n c l i n t r i a l

Comparative evaluation of efcacy of 0.2% Chlorhexidine, Listerine and Tulsi

extract mouth rinses on salivary Streptococcus mutans count of high school
childrenRCT
Pooja Agarwal a,, L. Nagesh b
a
b

Sumandeep Vidyapeeth, K.M.Shah Dental College and Hospital, Piparia, Vadodara, India
Bapuji Dental College and Hospital, Davangere, Karnataka, India

a r t i c l e

i n f o

Article history:
Received 8 February 2011
Received in revised form 27 May 2011
Accepted 28 June 2011
Available online 6 July 2011
Keywords:
Chlorhexidine
Listerine
Tulsi
Latin square design
Streptococcus mutans

a b s t r a c t
Objective: To assess and compare the effect of 0.2% Chlorhexidine mouth rinse, Listerine mouth
rinse and 4% Tulsi extract mouth rinse on salivary Streptococcus mutans level.
Methods: The present study is an experimental study of cross over type, employing Latin square
design. 45 school children aged 1415 years were divided into three groups/blocks. The
baseline unstimulated saliva samples were obtained from each group and assessed for
Streptococcus mutans counts. The study was divided in to three phases, each phase lasted for
8 days separated by a washout period of 15 days in between them. Groups A, B and C were
treated with 0.2% Chlorhexidine, Listerine and 4% Tulsi extract mouth rinses respectively in the
phase I. The study subjects were instructed to use the assigned mouth rinse twice daily for
1 min for 7 days. On day 8th the subjects were instructed to use the mouth rinse only once in
the morning. The follow up unstimulated saliva samples were collected 1 h after the use of the
assigned mouth rinse and assessed for salivary Streptococcus mutans counts. After phase I,
mouth rinses were crossed over as dictated by the Latin square design in phase II and III.
Results: All the three mouth rinses have individually shown a statistically significant reduction
in the salivary Streptococcus mutans counts. When the three mouth rinses were compared the
difference did not reach statistical signicance.
Conclusion: Tulsi has stood the test and is as effective as Chlorhexidine and Listerine in reducing
the salivary S. mutans levels.
2011 Elsevier Inc. All rights reserved.

1. Introduction
Dental caries is the most common microbial disease
affecting humans. It is a chronic disease of complex etiology
and is the predominant cause of tooth loss in children and
young adults. In the recent years, the prevalence of dental caries
in most western countries has steadily declined; the most
common reason for the decline of dental caries is attributed to

Corresponding author at: Sumandeep Vidyapeeth, K.M. Shah Dental

College and Hospital, Waghodia, Piparia Vadodara, Gujarat India. Tel.: + 91
9727585950.
E-mail address: agarwaldrpooja@yahoo.com (P. Agarwal).
1551-7144/$ see front matter 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.cct.2011.06.007

the widespread use of uorides. However it has shown

increasing trend in developing countries including India [1].
Research in the eld of caries prevention has been focusing on
ways for reducing or totally eradicating cariogenic ora from oral
cavity. Studies have shown that caries can be prevented by
regular tooth brushing and ossing [2]. However, most of the
studies have shown it is difcult to eliminate Streptococcus
mutans from pits, ssures and approximal surfaces by mechanical
means alone. For effective caries control, these methods should
be combined with the chemoprophylactic agents. These agents
are generally delivered as mouth rinses, toothpastes, gels, and
varnishes. Antiseptic mouth rinse represents one form of attack
on oral bacteria. Therapeutic rinsing was especially popular
among the Europeans, and persisted until the early 18th century.

P. Agarwal, L. Nagesh / Contemporary Clinical Trials 32 (2011) 802808

The incorporation of broad spectrum antimicrobial mouth rinses

as adjuncts to patient's oral hygiene regimens has assumed
greater importance with the recognition that most individuals
are unable to maintain adequate oral hygiene [3,4]. A variety of
chemotherapeutic agents have been examined for their ability to
control oral microorganisms.
The bisbiguanide chlorhexidine has been studied extensively for over 20 years and is currently the most potent
chemotherapeutic agent against Mutans streptococci [5,6]. It
has bactericidal activity against both gram positive and gram
negative bacteria. Its effect against Mutans streptococci is
greater than against Streptococcus sanguis and lactobacilli [7].
Studies have demonstrated that Chlorhexidine has an
antimicrobial activity against Streptococcus mutans [8,9].
Listerine mouth rinse is a product which is about 115 years
old. It is a broad spectrum antiseptic mouth rinse, whose active
ingredients are the four essential oils namely thymol, menthol,
eucalyptol and methylsalicylate. It is effective against a wide
range of Gram-negative and Gram-positive micro-organisms,
commonly implicated in dental caries. Its antimicrobial activity
has been tested against plaque and salivary Streptococcus mutans
and it is found to be effective [10]. Finding healing powers in
Plants is an ancient idea. Tulsi, scientically known as Ocimum
sanctum, is a time tested premier medicinal herb. It is a plant of
Indian origin, worshipped by Hindus and used in Ayurvedic
medicine since ancient times. It is bestowed with enormous
antimicrobial property. Literature review reveals that no studies
have been conducted to assess the antimicrobial activity of Tulsi
against Streptococcus mutans. As studies related to herbal mouth
rinses are lacking and as clinicians, we confront piquant
situation when patient seek our opinion about such herbal
mouth rinses, research in this area is necessary to generate the
required evidence. Hence the current study was conducted with
an aim to assess and compare the effect of 0.2% Chlorhexidine
mouth rinse, Listerine mouth rinse and 4% Tulsi extract mouth
rinse individually on salivary Streptococcus mutans level on
1415 year old school children of Davangere city.
2. Materials and methods
2.1. Study design and population
The present study is an experimental, in vivo, cross over
study (within subjects) employing Latin square design
conducted among selected school children aged 1415 years
in Davangere city, India. The study was conducted for a period
of 3 months from January 2009 to March 2009. The study
protocol was approved by the Ethical review board of Bapuji
Dental College and Hospital, Davangere. Permission to
conduct the study was obtained from the Deputy Director of
Public Instruction (DDPI) and school authorities. Davangere
city was arbitrarily divided into 4 zones (North east, North
west, South east and South west) utilizing the municipal block
divisions which have been done for administrative purpose.
From each zone 2 schools were selected randomly by
employing lottery method, from the selected 8 schools three
schools were selected randomly by employing lottery method.
The three selected schools were randomly assigned codes as
schools A, B and C. List of all the students of 1415 years age
group in each of the selected school was made from school
records. Only those children who had at least one cavitated

803

active carious lesion(clinical selection criteria) and a salivary

Streptococcus mutans count of equal to or more than 10 5 CFU
per ml of saliva (microbiological selection criteria) were
included in the study. Dental caries was examined using the
Nyvads criteria [11]. Examination for caries was done in the
school premises in the available natural light using sterilized
plane mouth mirror and CPI probe. Subjects giving a history of
hypersensitivity to any product used in the study, suffering
from diseases which might affect the salivary ow, unable to
comply with the study appointment schedules, having a
history of antibiotic therapy in the previous 1 month till the
start of the study, having any systemic diseases and who were
already using any mouth rinses were excluded from the study.
2.2. Sample size
The total sample size was forty-ve (15 subjects from each
school). The sample size was calculated for error xed at
b 5% (p b 0.05) and xed at 20%, expected mean difference
2.281 and standard deviation 2.563. Based on the above
calculation the minimum sample required in each group was
12 subjects. In the current study the sample size was 15
subjects in each group to compensate for any attrition up to
20%. The selected fteen children from each school were
treated as one group/block. Written informed consent was
obtained from selected children and from their parents before
the beginning of examination by discussing with them the
purpose of the study.
2.3. Baseline saliva collection and microbial analysis
All subjects undergoing investigations were given clear
instructions to refrain from eating for 1 h before collection of
saliva. 1 ml of unstimulated whole saliva was collected by asking
the children to let saliva collect in the oor of the mouth without
swallowing it for at least 1 min, and then to expectorate into the
sterile test tube. The test tubes were then immediately
transported to the oral pathology laboratory, where it was
processed immediately. The saliva sample was homogenized
manually by stirring using a stirrer. Hundred microliters of saliva
was diluted with 1 ml of normal saline (1:10 dilution). Using an
inoculation loop (2 mm inner diameter) 5 l of the 1:10 dilution
sample was streaked on Mitis Salivarius Bacitracin agar (MSB), a
selective medium for Streptococcus mutans. The MSB agar plates
were incubated for 48 h at 37 C in an atmosphere of 95%
Nitrogen and 5% carbon dioxide. After 48 h of incubation period
Streptococcus mutans appeared on the culture plate as small,
rough, raised and adherent colonies. Those colonies which were
atypical were further conrmed by mannitol and sorbitol test.
Colonies so identied were counted using an electronic colony
counter (Deep Vision Company, India).
2.4. Blinding and calibration
All the microbiological procedures were performed by the
investigator, who was blinded with respect to culture plates
of different groups throughout the entire course of the study.
The oral pathology faculty and the investigator underwent a
calibration session. The aim of this session was to train the
investigator in counting and recording the Streptococcus
mutans counts as colony forming units. The kappa coefcient

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P. Agarwal, L. Nagesh / Contemporary Clinical Trials 32 (2011) 802808

value for inter examiner and intra examiner variability was

0.86 and 0.89.
2.5. Random allocation
Three mouth rinses namely, 0.2% Chlorhexidine mouth
rinse((ICPA Health Products Ltd), Listerine mouth rinse
(Johnson and Johnson) and 4% Tulsi extract mouth rinse
(self prepared) constituted three interventions.
As the present study was a crossover study employing
Latin square design; each group/block was subjected to all
three interventions in a phased manner (block randomization) as shown in the schematic diagram (Fig. 1). Random
allocation of mouth rinses using the lottery method is done at
phase I only, further in the next two phases that is phase II
and phase III allocation was automatically determined as it
was a Latin square design. The respective mouth rinse as
assigned was distributed to all 15 selected high school
children from each school. Children were instructed to rinse
for 7 days, twice daily, morning after breakfast and night before
going to bed, with 10 ml (undiluted) of the assigned mouth
rinse for 1 min and then expectorate the rinse. A measuring cup
was provided to the children to dispense 10 ml of the assigned
mouth rinse. They were instructed not to eat or drink anything
for a minimum of half an hour after rinsing. On day 8th subjects
were instructed to rinse once in the morning after breakfast.
One milliliter of unstimulated saliva samples were collected 1 h
after the subjects had rinsed with the assigned mouth rinse. The
collected saliva was subjected to microbiological procedure as
explained earlier during baseline salivary Streptococcus mutans
CFU/ml count estimation. The use of mouth rinse was followed
by a washout period of 15 days during which children were
asked to stop using the assigned mouth rinse. After the washout
period of 15 days the selected subjects were assigned to the
next mouth rinse in a phased manner (Fig. 1). During the entire
course of the study the subjects were instructed to stick to their
routine oral hygiene habits.
2.6. Compliance
A checklist was provided to each subject to assess
compliance. They were instructed to mark a tick () after

CHX 0.2% Chlorhexidine mouth rinse

using the assigned mouth rinse; if they had missed using the
rinse on any day, they were asked to mark a cross (X) and
mention the reason for not using the mouth rinse. A
compliance questionnaire was also given to the subjects to
assess if they experienced any adverse effects with any of the
rinse. Mouth rinsing was supervised by the examiner in the
morning by visiting the schools every alternate day. Parents
were instructed to supervise the rinsing at night times.
2.7. Statistical analysis
For intragroup and intergroup comparison of salivary
Streptococcus mutans counts at baseline and on day 8th paired
t test and one way ANOVA were used respectively. Chi square
test was used to assess the difference in the compliance when
the subjects used the three mouth rinses.
3. Results
In a cross over trial, analysis of the data can be done by the
following two methods:
1) By comparing the observations done on each group
individually at different phases of the trial.
or
2) By compiling, summarizing and comparing the observations done on the total sample with respect to each
intervention (each mouth rinse).
In the current study both the analysis have been done i.e.
I) Phase I, Phase II and Phase III of group A are compared,
similarly Phase I, II, III of group B and group C are
compared (Tables 1, 2, 3).
II) The overall results for 0.2% Chlorhexidine, Listerine and 4%
Tulsi extract mouth rinses for the total sample are
compared (Table 4).
Individually each of the mouth rinse signicantly reduced
the salivary Streptococcus mutans count which was reected
in the difference between pretest and post test counts. When
all the three mouth rinses were compared for their ability to
reduce salivary Streptococcus mutans count although there

TUL 4% Tulsi extract mouth rinse

LIST Listerine mouth rinse

Phase I
Group A

Group B

Group C

B
a
s
e
l
i
n
e

CHX

LIST

TUL

Washout Period Phase II Washout period Phase III

P
o
s
t
t
e
s
t

B
a
s
e
l
i
n
e

CHX

LIST

TUL

P
o
s
t
t
e
s
t

B
a
s
e
l
i
n
e

CHX

LIST

TUL

P
o
s
t
t
e
s
t

Fig. 1. Latin square design depicting the order of phases in group A, B and C and the period of baseline saliva collection at the beginning of every phase and post test saliva
collection at the end of every phase followed by a washout period. CHX0.2% Chlorhexidine mouth rinse TUL4% Tulsi extract mouth rinse. LISTListerine mouth rinse.

P. Agarwal, L. Nagesh / Contemporary Clinical Trials 32 (2011) 802808

was a numerical difference statistically it was found to be not

signicant (Table 4).
3.1. Compliance
All subjects in the present study used the mouth rinse as
per the instructions given by the investigator. Few subjects
did not use the 0.2% Chlorhexidine, Listerine and 4% Tulsi
extract mouth rinses for all the 8 days assigned. Most subjects
said that they had not used the mouth rinse on some
particular day as they were out of station on that day and
hence could not use the mouth rinse on that particular day,
while few other subjects reported that they had forgotten to
use the mouth rinse on some day. However over all this
difference was not statistically signicant with any of the
mouth rinses (Tables 5 and 6).
4. Discussion
The present study is an Active control trial of a cross over
design (within subjects) of Latin square type. The advantage
of a within subjects design is that the subjects themselves act
as their own controls and hence the expected subject variance
is minimized to least i.e. all subject related factors are literally
identical across treatments; because of the reduced subject
variance, within subject design is more powerful. The order in
which the interventions are instituted to the subjects in a
cross over trial is said to induce some errors which are named
under order effects. Latin square design is a carefully
constructed cross over trial design which eliminates the
order effects hence it can control and eliminate such an error.
The study employed block randomization method i.e. 15
children from each school were considered as a block/group
for random allocation of the mouth rinse. This method was
employed in order to blind the subjects with respect to the
different mouth rinses used. As the commercial mouth rinses
tested were of different color, taste, and avor, the only way
to blind the subjects and prevent cross contamination of
product information was by resorting to block randomization.
The particular age group of 1415 years was chosen in the
present study as children of this age are easily accessible, the
compliance can be better expected and 15 years is an age
group recommended as representative age group by basic
oral health survey. The study aimed to test the different
mouth rinses in real life situation without exercising control
on subject's routine oral hygiene procedures and dietary
habits. The results obtained with each mouth rinse are
ascribed to the whole mouth rinse and not to the active
ingredient alone, as the study design does not incorporate

805

suitable negative controls. The aim of the study is to compare

the relative efcacy of selected mouth rinses in reducing the
salivary Streptococcus mutans count and there is no scope for
attributing the obtained results exclusively to the active
ingredients. In the present study Tulsi extract was used in
the form of mouth rinse and investigated for its efcacy on
Streptococcus mutans. Exploration of the available literature
has revealed that no studies have been conducted to assess
the effect of Tulsi on Streptococcus mutans. The Tulsi extract
was prepared by the investigator at 4% concentration. This
particular concentration was chosen as it produced the
maximum zone of inhibition against Streptococcus mutans
among the 15 different concentrations that were investigated
in the pilot study [12]. In The current study Subjects were
instructed to use the assigned mouth rinse undiluted, twice
daily as per the instructions given by the manufacturers (ICPA
Health products Ltd, Johnson and Johnson). Most manufacturers recommend the use of 10 to 20 ml of mouth rinse for 30
to 60 s. In the present study a volume of 10 ml and rinsing
time of 1 min was used in order to standardize the use of
mouth rinse with respect to the volume used and the rinsing
time. The washout period utilized was 15 days based on
previous studies [10,1315]. Taking into account the aforementioned data a washout period of 15 days was used in the
current study. This period was sufcient to reduce the
carryover effect of the intervention as the salivary Streptococcus mutans counts returned to baseline levels within 15 days
hence eliminating the possibility of any carryover effect.
0.2% Chlorhexidine when used as a mouth rinse was found
to be effective in reducing the salivary Streptococcus mutans
counts to a statistically signicant level (p b 0.001). Similar
results were obtained in previous studies [8,9,16]. However in
few studies the concentration of Chlorhexidine mouth rinse
used was 0.12% [9,10]. In the current study Chlorhexidine
mouth rinse was used at 0.2% concentration. However studies
have also used 0.2% Chlorhexidine and have found a
reduction in the salivary Streptococcus mutans [17].
Listerine mouth rinse was effective in reducing the
salivary Streptococcus mutans counts to a statistically significant level (p b 0.001). Similar results were obtained in
previous studies [10,17]. However in the study done by Fine
et al. the subjects were asked to use 20 ml of the rinse twice
daily for 11 days. In our study the subjects were asked to use
10 ml of Listerine mouth rinse twice daily for 7 days and once
on day 8th after which the post test salivary samples were
collected and analyzed. The current study used relatively less
volume (10 ml) of Listerine mouth rinse for a shorter period
(8 days) but still demonstrated a signicant reduction in
salivary Streptococcus mutans counts. This is a clear evidence

Table 1
The mean baseline, mean post test and mean difference between baseline and post test values (mean reduction) of salivary Streptococcus mutans in Group A at
different phases of the trial expressed as 105 CFU/ml.
Group A

Mean baseline

Mean post-test

t-Value

Signicance level p

Mean reduction (baselinepost-test)

PHASE I (CHX)
PHASE II (LIST)
PHASE III (TUL)
ANOVA p level

16.3 5.9
16.4 5.0
16.8 5.1
F = 0.03
p = 0.97 (NS)

6.3 3.7
9.3 5.6
11.1 3.8

6.45
5.26
4.50

b 0.001 (HS)
b 0.001 (HS)
b 0.001 (HS)

10.0 6.0
7.1 5.3
5.7 4.9
F = 2.43
p = 0.10 (NS)

HShighly signicant, NSnot signicant.

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P. Agarwal, L. Nagesh / Contemporary Clinical Trials 32 (2011) 802808

Table 2
The mean baseline, mean post test and mean difference between baseline and post test values (mean reduction) of salivary Streptococcus mutans in Group B at
different phases of the trial expressed as 105 CFU/ml.
Group B

Mean baseline

Mean post-test

t-Value

Signicance level p

Mean reduction (baselinepost-test)

PHASE I (LIST)
PHASE II (TUL)
PHASE III (CHX)
ANOVA p level

17.3 5.8
16.5 3.6
18.0 5.1
F = 0.38
p = 0.68 (NS)

7.6 3.3
9.5 4.4
8.7 3.1

8.87
5.44
6.74

b0.001 (HS)
b0.001 (HS)
b0.001 (HS)

9.7 4.2
6.9 4.9
9.3 5.4
F = 1.44
p = 0.25 (NS)

*HShighly signicant, NSnot signicant.

to suggest that Listerine is a potent antibacterial agent even

when used in smaller volume for less number of days. An in
vitro study has also demonstrated the antimicrobial effect of
Listerine against Streptococcus mutans [18]. The bactericidal
efcacy of essential oil containing mouth rinse used in this
study has been long recognized and represents the primary
mechanism by which it exerts its clinical effects.
The results of 4% Tulsi extract mouth rinse could not be
compared with other studies as no studies have been
reported in literature which has tried to assess the effect of
Tulsi on Streptococcus mutans. However studies have been
reported which suggest that Tulsi possess antifertility,
anticancer, antifungal, antidiabetic, antiemetic and antimicrobial property [19]. Eugenol (1-hydroxy-2-methoxy-4allylbenzene), the active constituent present in Tulsi (Ocimum sanctum) has been found to be largely responsible for
the therapeutic potentials of Tulsi. In the current study we
have made an attempt to assess the antimicrobial effect of 4%
Tulsi extract against Streptococcus mutans. This makes the
present study unique as hitherto no similar studies have been
reported in literature. As Tulsi has been found to be effective
against Streptococcus mutans the use of it may be promoted
further among Indians. As literature is totally devoid of any ill
effects of Tulsi on human beings one can condently
recommend the use of Tulsi as a mouth rinse specially for
rural population which has an easy access to Tulsi and perhaps
highly cost effective. Further research is required to elucidate
and understand the action of Tulsi on oral microbiota.
All the three mouth rinses have antibacterial effect on
salivary Streptococcus mutans. The results of these comparisons
could not be compared with other studies as no studies have
been reported in literature which has tried to compare 0.2%
Chlorhexidine, Listerine and 4% Tulsi extract on salivary
Streptococcus mutans counts together in one single study.
However studies have been conducted which have tried to
assess the effect of 0.2% Chlorhexidine and Listerine individually

on salivary Streptococcus mutans and have found a signicant

reduction in the count as mentioned above. Usually this kind of
results are interpreted as negative results and the immediate
question asked is whether the power of the study is sufcient
enough to show the difference in outcomes in case it exists. In
the current study the sample size was determined by xing the
power of the study at 80% hence the study cannot fail to show a
difference in case it really existed. Hence it can be purported
that all the three mouth rinses namely 0.2% Chlorhexidine,
Listerine and 4% Tulsi extract are showing similar effect on
salivary Streptococcus mutans.
With respect to adverse effects, most subjects reported an
alteration of taste sensation with 0.2% Chlorhexidine and a
bitter taste with 4% Tulsi extract mouth rinse. Alteration in
taste sensation with the use of Chlorhexidine is a common
side effect of Chlorhexidine. In a previous study it was
hypothesized that the probable reason for the alteration in
taste sensation is that Chlorhexidine binds to a specic
sodium receptor molecule in the taste bud which is uniquely
different from receptors for sweet, bitter and sour stimuli[20].
The bitter taste with Tulsi extract mouth rinse could be
perhaps due to the concentration at which Tulsi extract was
prepared i.e. at 4%, this particular concentration was chosen
as it showed the maximum antimicrobial activity against
Streptococcus mutans among the 15 different concentrations
investigated in the pilot study. With respect to Listerine
mouth rinse an initial burning sensation was experienced by
few subjects. This could be attributed to eucalyptol, which is
an essential oil present in Listerine mouth rinse.
4.1. Limitations
The results of Tulsi extract mouth rinse could not be
compared with other studies as no studies have been reported
in literature which has tried to assess the antimicrobial effect
of 4% Tulsi extract against Salivary Streptococcus mutans.

Table 3
The mean baseline, mean post test and mean difference between baseline and post test values (mean reduction) of salivary Streptococcus mutans in Group C at
different phases of the trial expressed as 105 CFU/ml.
Group C

Mean baseline

Mean post-test

t-Value

Signicance level p

Mean reduction (baselinepost-test)

PHASE I (TUL)
PHASE II (CHX)
PHASE III (LIST)
ANOVA p level

13.0 5.3
12.1 5.3
13.9 4.5
F = 0.46
p = 0.67 (NS)

5.8 3.1
7.0 2.4
9.5 3.5

6.64
5.12
7.95

b0.001 (HS)
b0.001 (HS)
b0.001 (HS)

7.2 4.2
5.1 3.9
4.4 2.2
F = 2.46
p = 0.10 (NS)

HShighly signicant, NSnot signicant.

P. Agarwal, L. Nagesh / Contemporary Clinical Trials 32 (2011) 802808

807

Table 4
The mean baseline, mean post test , difference between baseline and post test values (mean reduction) and the percentage reduction of salivary Streptococcus
mutans in the total sample with respect to 0.2% Chlorhexidine, Listerine and 4% Tulsi extract mouth rinses expressed as 105 CFU/ml.
Mouth rinses

Mean baseline

Mean post-test

t-Value

Signicance level p

Mean reduction (%)

0.2% Chlorhexidine
Listerine
4% Tulsi Extract
ANOVA p level

15.5 5.9
15.9 5.2
15.4 4.9
F = 0.09
p = 0.92 (NS)

7.3 3.2
8.7 4.2
8.8 4.3
F = 2.07
p = 0.13 (NS)

9.95
10.45
9.57

b 0.001 (HS)
b0.001 (HS)
b0.001 (HS)

8.2 5.5 (53%)

7.1 4.6 (45%)
6.6 4.6 (43%)
F = 1.19
p = 0.31 (NS)

HShighly signicant, NSnot signicant.

Table 5
Distribution of study subjects based on the use of mouth rinses for all the 8 days.

Yes
No
Total

CHX

LIST

TUL

38 (84%)
7 (16%)
45 (100%)

34 (76%)
11 (24%)
45 (100%)

39 (87%)
6 (13%)
45 (100%)

2 = 2.13
p = 0.34 (NS)

NSnot signicant.

Table 6
Distribution of study subjects based on the side effects experienced during the use of the mouth rinses.

Yes
No
Total

CHX

LIST

TUL

8 (18%)
37 (82%)
45 (100%)

14 (31%)
31 (69%)
45 (100%)

9 (20%)
36 (80%)
45 (100%)

2 = 2.60
p = 0.27 (NS)

NSnot signicant.

Further studies are required to assess the effect of Tulsi on

Streptococcus mutans.

5. Conclusion
Tulsi has stood the test by demonstrating similar antimicrobial effect on salivary Streptococcus mutans when compared with the bench mark control (0.2% Chlorhexidine) and
positive control (Listerine). This is an encouraging result
which clearly favors promotion of Tulsi as a mouth rinse
among rural communities especially belonging to low
socioeconomic strata as Tulsi is easily accessible. However as
this is the rst attempt to assess the effect of Tulsi on
Streptococcus mutans further long term studies need to be
performed to investigate the effect of Tulsi on dental caries
before it can be condently recommended .Researchers need
to have a fresh look in the area of Green medicine as there may
be many such potential herbs which may possess signicant
antimicrobial property but require scientic evidence, thus to
generate the required evidence research in the area of Green
medicine is deemed necessary.

Acknowledgements
We duly acknowledge the cooperation of Bapuji Pharmacy
College, Davangere in preparation of the Tulsi extract mouth
rinse.

References
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global increase in dental caries. A pending public health crisis. Am J Dent
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