Microbiological

Communication

Biosci. Biotech. Res. Comm. 10(2): 225-232 (2017)

Evaluation of antibacterial effect of

Eucalyptus

and

ginger on dentinal tubules

Enterococcus faecalis

in the

presence of smear layer

Samira Shahsiah,

* Eskandar Moghimipour,

Nooshin Shamsizadeh

and Azardokht Khosravi

Department of Endodontics, Faculty of Dentistry, Ahvaz Jundishapur University of Medical Sciences,

Ahvaz, Iran

Medicinal Plant Research Center, Faculty of Pharmacy, Ahvaz Jundishapur University of Medical Sciences,

Ahvaz, Iran

Dentist

Professor of Clinical Microbiology, Ahvaz Jundishapur Univesity of Medical Sciences, Ahvaz, Iran

ABSTRACT

Enterococcus faecalis is a resistant microorganism and plays an important role in root canal treatment failure. One of

the most critical steps in root canal treatment is chemical cleaning of the root canal with endodontic irrigation solu-

tions. The most commonly used solution is Sodium hypochlorite which in spite of its broad antimicrobial activity, has

many disadvantages. Therefore, this study was designed to evaluate the antibacterial effect of eucalyptus and ginger

on dentinal tubules Enterococcus faecalis in the presence of smear layer. In this study dentin cylinders from 80 single

rooted human teeth were prepared and contaminated with Enterococcus faecalis with smear layer created in each

dentin disk. Experimental groups were as follows: Group 1: Sodium hypochlorite 5.25%, Group 2: ginger extract,

Group 3: eucalyptus extract, Group 4: Normal saline (positive control), Group 5: sterile cylinders. Then dentin chips

were collected from three different depths of dentin including 0.1,0.2 and 0.3 millimeters. Optical density comparison

of different experimental groups showed that there is a signi cant difference between groups in 0.1 and 0.2 millim-

eters depth (p<0.01), however no signi cant difference in 0.3 millimeters depth was obtained. Antimicrobial ef cacy

of eucalyptus and Sodium hypochlorite in the presence of smear layer showed no signi cant differences, while this

effect was weaker with ginger compared to Sodium hypochlorite.

KEY WORDS:

ENTEROCOCCUS FAECALIS

, SMEAR LAYER, EUCALYPTUS, GINGER

225

ARTICLE INFORMATION:

*Corresponding Author: shahsiahs@gmail.com

Received 31

March, 2017

Accepted after revision 29

June, 2017

BBRC Print ISSN: 0974-6455

Online ISSN: 2321-4007 CODEN: USA BBRCBA

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Online Contents Available at: http//www.bbrc.in/

226 EVALUATION OF ANTIBACTERIAL EFFECT OF

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Samira Shahsiah et al.

INTRODUCTION

Today different in canal detergents are used. The most

popular one is sodiumhypochlorite (homemade bleach).

Main advantage of this material is solving necrotic tis-

sue and its anti-microbial properties against wide range

of micro-organisms such as Enterococcus faecalis. This

material has some disadvantages too which include

unpleasant smell and taste, toxicity during contact with

pre radicular tissues, removing cloth’s color, and capacity

of metal corrosion (Torabinejad and Walton 2003; Gulve

and Gulve 2010). Studies indicated that sodiumhypochlo-

rite has very less ability for removing smear layer. In addi-

tion, overusing this material causes reduction of dentinal

elasticity strength (Hargreaves 2011).

We selected herbals for this research because of fol-

lowing reasons: Eucalyptus (which out of them Euca-

lyptus camaldulensis is most popular in Iran) and ginger

are found in Iran region, there is no need to import this

medicine from other countries, providing their effective

concentrate has less price than chemical medicine, in

compare to chemical medicine, these are more adap-

tive to natural environment of the body and has less

unfavorable effect. No research has so far made about

investigating this material as a detergent in canal at the

presence of dentine and smear layer so there is need

to research in this case. Historically, numerous combi-

nations of aqueous solution are recommended as root

canal detergents; including ineffective sodium chloride

(saline) and so toxic and allergist biocids such as formal-

dehyde (Zehnder 2006).

There are different factors including numbers and

diameter of dentinal tubules, scleroticdentin, and also

smear layer can affect dentine permeability and can

avoid penetration of anti-microbial materials to dentinal

tubules. During preparation of canal walls, some parts of

dentinal wall which has contact with tools are covered

by one smear layer (Hargreaves and et al, 2011; McCom-

bEand et al, 1976). Smear layer is amorf, granular and

disordered that is made from minerals such as dentianls

and organic materials including pulp remnants, odonto-

blastic excrescence, suliva, and blood cells (Kakehashi

and et al, 1965; Gwinnett 1984). (Figure 1).

Maekawa et al performed a research in 2013 in Brazil.

They investigated effect of ginger and propolis1 concen-

trate on some microorganisms in canal and endotoxin.

According to the results, researchers recommended that

propolis and ginger concentrate in combination with

calciumhydroxide can be used as a medicine in alterna-

tive canal although there is need more studies on bio-

compatibility, chemical interactions of these materials

and their clinical effects to be made so their use in endo-

dontic treatment is approved. In another study of Martos

et al in 2012 in Brazil, they considered anti-microbial

activities of chloroform solution, eucalyptus oil essence,

and orange oil essence lonely and also with Cetrimide

in 0.05%, 0.1%, 0.2%, 0.3% densities. These research-

ers observed that bio lm reduction in orange oil and

eucalyptus oil in 2 minutes indicated no signi cant dif-

ference; however, bio lm removal percent by orange oil

in 5 minutes was more than eucalyptus oil.

Shari an et al (2011) in Tehran performed a research

in order to investigating anti-microbial properties of lime

on Enterococcus faecalis in dentinal tubules in presence

of smear layer on 140 dentinal cylinderprovided from

central incisor of cow. These researchers concluded that

FIGURE 1. Root dentine section which is covered with smear layer resulted

from canal working tools

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS EVALUATION OF ANTIBACTERIAL EFFECT OF

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AND GINGER 227

Samira Shahsiah et al.

at the presence of smear layer, anti-microbial properties

of lime was less than NaOCl while there was no differ-

ence between anti-microbial properties of lime skin and

NaOCl. Abdul Rahman et al in 2010 studied anti-micro-

bial and bio chemical properties of food  avors in meat

products. In this experiment, Disk diffusion method was

used and it was indicated that cinnamon is the most

effective  avor against all under studied micro-organ-

isms except micrococcusluteus. In addition, ginger had

anti-microbial activity against Enterococcus faecalis.

Gulve et al.,( 2010 )from India, managed study for

comparing invitro anti-microbial properties of ginger

concentrate 2% sodiumhypochlorite against Enterococ-

cus faecalis on 18 growth medium by Agar diffusion.

These two researchers observed that ginger concentrate

signi cantly avoid bacteria’s growth in compare to

sodiumhypochlorite. This study aimed at  nding better

alternative for sodium hypochlorite as a relative ideal

root canal detergent. Thus, by aim of determining anti-

microbial properties of eucalyptus concentrate - ginger

concentrate- sodiumhypochlorite, and 5.25%– normal

saline against Enterococcus faecalis at presence of smear

layer in 0.1, 0.2 and 0.3 millimeter depth of dentine

and comparing anti-microbial properties of eucalyptus

concentrate – ginger concentrate- sodiumhypochlorite

and 5.25% –normal saline in 0.1, 0.2 and 0.3 millimeter

depth of dentine at presence of smear layer.At the fol-

lowing we will discuss materials and methods of col-

lecting data, and collecting samples. Then in section 3

we will explain research results and statistical tests, in

addition to results of the test and  nally we explained

the conclusion.

MATERIALS AND METHODS

In this study, in order to determining of eucalyptus con-

centrate – ginger concentrate - 5.25% sodium hypochlo-

rite- normal saline against enterococcus faecalis in 0.1,

0.2 and 0.3 milliliter depth from dentine at the presence

of smear layer.We selected 80 human teeth which were

withdrawn due to different diseases including periodon-

tal, orthodontia, dental corruption were selected from

therapeutical centers in Ahvaz. These single root teeth

have maxilar anseizure and mandible premolars. After

withdrawn of these teeth, tissues connected to outer

surface of root were separated with one court. Then in

order to disinfection of teeth surface, they put in 2.5%

sodium hypochlorite, and they were kept in steal normal

saline at the room temperature. Eucalyptus leaves, 70%

ethanol, dried rhizome of ginger, stilled water, stand-

ard strain of Enterococcus faecalis , growth medium BHI

Broth (Merck-Germany), growth medium of BHI Agar

(Merck-Germany), laboratory tube, microbial cultivation

plate, cotton, gloves, 15 milliliter falcon tube (Maxell-

China), sampler (Trasnferpette-Germany), head sampler,

5.25% sodium hypochlorite (Golrang-Iran), laboratory

Handpiece (NSK-Japan), coal disk, medin broach (Mani-

Japan), number 4Gates-Gliddendrills(Mani- Japan), nail

polish, aluminum foil, self-cure acryl (Acro Pars-Iran),

EDTA17% (Cerkamed-Poland), needlegauge 23.

In order to provide plant concentrate, maceration

method was used (Martos j, et al, 2013). For providing

detergent, dried concentrate obtained from freezer drier

were solve into stilled water slowly until a homogenous

solution. Mass concentration for ginger solution was

calculated equaled to gr/ml0.51and for eucalyptus con-

centrate solution equled to gr/ml0.57. then these deter-

gents were transferred to sanitary-therapeutic center in

Golestan Hospital of Ahvaz and then they were sterile ed

with X-ray (Shalaby E. A, 2011). Standard ndard strain

of 29212 ATCC

Enterococcus faecalis bacteria was pro-

vided from Pastor Institute of Iran in Tehran. Brain heart

infusion broth steal Growth medium (Merck-Germany)

was prepared and added into Lyophilizated in order to

solve it completely. Then it was cultivating on BHI Agar

growth medium (Merck-Germany) and they were incu-

bated in aerobiccondition in C°37 for 24 hours. Then

by biochemical tests for bacteria identi cation (smear

provision, PYR test, Bile esculin, Catalaz) we assured

from its purity. So 24 hours colonies provided fro,m

pure enterococcus faecalis. For providing bacteria sus-

pension, bacteria clonies were taken from solid growth

medium and solved in a tube with sterile BHI Broth and

then we obtained a suspension with turbidityMc farland

(cfu/ml 3×

Samples withdrawn from autoclave were opened

under laminar air ow hood and they were all inoculated

by 20-10 micro litter bacteria suspension of enterococcus

faecalis with strain 1 of Mc Farland (cfu/ml 3× except

negative control group. Then cylinders were covered

with a lid made from sterile aluminum paper and growth

medium with bacteria were added into each cylinder

every two days by sample (Trasnferpette-Germany) and

trough sterile head sampler. In negative control group,

sterile BHI Broth growth medium were added into teeth.

This was repeated 4 weeks for providing maximum pen-

etration of bacteria into dentinal tubules (Shabahang S

and Torabinejad M, 2003; Ahangari Z, et al, 2008) and

the samples were incubated in aerobiccondition and in

When inoculation ended, I order to make sure of

micro-organisms penetration into dentinal tubules, 10

samples were randomly selected out of samples. Then

acrylic layer were removed from lumenof these sam-

ples and patterned by low speed and Gates Gilden

(Mani-Japan) number 5 (o.1 milliliter depth), Gates

American type culture collection

228 EVALUATION OF ANTIBACTERIAL EFFECT OF

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Samira Shahsiah et al.

Gilden (Mani-Japan) number 6 (0.2 milliliter depth) and

pisormer (Mani-Japan) number 6 (0.3 milliliter depth)

from dentine.

Provided dentine chips from different dentinal sec-

tions were placed separately in 15 milliliter falcon tubes

(Maxell-China) with 3 milliliter sterile BHI Broth and

were incubated 48 hours in 37 C° temperature. Then, out

of 30 mentioned tube some samples were cultivated on

plates with BHI Agar and were incubated in aerobic con-

dition and in 37

C temperature. After incubation end,

micro-organisms identi cation test in order to be sure

about that bacteria are single strain and its penetration

in 3 depths.

In other samples, smear layer was created by sterile

hedstroms number 50 (Mani-Japan). In a way that hed-

strom number 50 were withdrawn three times and by

push and pull movement on canal wall. In order to avoid

pollution of environment implying to Luman cylinders

each hedstrom were changed after one time use.

In next step, samples in group 1,2,3 and 4 were

washed separately by 5 milliliter sterile syringe (Homa-

Sorang-Iran) and 23 needles with gauge and from neg-

ative control group, patterning was made in 3 depth.

Washing syringe were changed for each sample after

using them once in order to minimize environmental

pollution. Group washing was made as follow:

Group 1: 2 milliliter Naocl 5.25% in each sample

for 20 minutes.

Group 2: 2 milliliter ginger concentrate with mg/

ml127.5 density in each sample for 20 minutes.

Group 3: 2 milliliter eucalyptus concentrate with

mg/ml17.8 density in each sample for 20 minutes.

Group 4 (positive control): 2 milliliter sterile nor-

mal saline in each sample for 20 minutes.

Group 5 (negative control): washing was not made

in this group. In order to be con dant about lack

of cylinder pollution penetration into another

bacteria and test correction was patterned except

Enterococcus faecalis.

When acryliclayer washing, cylinders were remove

from lumen of then dentine sampling was performed

by low speed and Gates Gilden (Mani-Japan) number 5

drills (o.1 milliliter depth ), Gates Gilden (Mani-Japan)

number 6 drills (0.2 milliliter depth) and pisormer (Mani-

Japan) number 6 (0.3 milliliter depth) from dentine. In a

way that 15 milliliter falcon tubes (Maxell-China) with

3 milliliter sterile BHI Broth was prepared and dentine

chips obtained from shaving were transferred into these

tubes in each step from dentine cylinders.

It should be mentioned that tubes were incubated 48

hours in 37 C° temperature. Then, out of 30 mentioned

tube some samples were cultivated on plates with BHI

Agar and were incubated in aerobic condition and in

C temperature. After incubation end, micro-organ-

isms identi cation test in order to be sure about that

bacteria are single strain and its penetration in 3 depths.

In order to measure strain from microbial growth, 2.5

milliliter was extracted from BHI Broth in tubes with

dentinal chips removed from incubator and its optical

density was studied by Spectrophotometer (WPA-bio-

wave2-USA) 540 nano-meter in wave length. Optical

density of groups under experiment and positive control

groups were compared together.

RESULTS AND DISCUSSION

Optical density related to experimental groups in three

depths (0.1, 0.2, 0.3) milliliter from dentine was meas-

ured by Spectrophotometer (Biochrom WPA - USA).

Mean and standard deviation of OD were mustered for

different depths in table 1.

Comparing experimental groups considering OD rate

in 3 measured dentinal depths was performed based on

statistical analysis of One-way ANOVA and SPSS soft-

ware.

According to table 2, and One-way ANOVA in 0.1

millimeter of dentine there was signi cant difference

between under studied groups (p<0.01). in addition, in

0.2 millimeter of dentine there was signi cant differ-

ence between under studied groups (p<0.01). But in 0.3

millimeter of dentine there was no signi cant difference

between under studied groups.

Shortly, comparing different groups with consider-

ing OD in each depth by one-way ANOVA analysis indi-

cated that there is signi cant difference between groups

Table 1. Mean ±standards deviation of optical density of provided  lings from three dentinal depth under study

Mean (±standards deviation)

0.3 millimeter depth

Mean (±standards deviation)

0.2 millimeter depth

Mean (±standards deviation)

0.1 millimeter depth

Detergent solutions

(0.08±)0.23 (0.11±)0.22 (0.04±)0.016 Sodium hypochlorite 1- Sodium hypochlorite

(0.25±)0.34 (0.21±)0.34 (0.07±)0.24 2- ginger concentrate

(0.21±)0.26 (0.04±)0.21 (0.04±)0.18 3- eucalyptus concentrate

(0.08±)0.40 (0.07±)0.46 (0.07±)0.41

4- Normal saline

(positive control)

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS EVALUATION OF ANTIBACTERIAL EFFECT OF

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AND GINGER 229

Samira Shahsiah et al.

DIAGRAM 1. Mean of optical density of provided  lings from three dentinal depth under

study

Table 2. comparing OD between groups under study in 0.1 millimeter depth from dentine

Depth Sum of Squares df Mean Square F Sig.

0.1 Between Groups .308 4 .077 28.084 .000

Within Groups .178 65 .003

Total .486 69

0.2 Between Groups .376 4 .094 5.005 .001

Within Groups 1.220 65 .019

Total 1.596 69

0.3 Between Groups .219 4 .055 1.606 .183

Within Groups 2.214 65 .034

Total 2.433 69

Table 3. comparing pairwise OD of under study groups in 0.1 milliliter depth from dentine

Depth

(I)

group

(J)

group

Mean Difference

(I-J)

Std. Error Sig.

95% Con dence Interval

Lower Bound Upper Bound

0.1 1

2 -.085650* .016549 .000 -.11870 -.05260

3 -.018150 .016549 .277 -.05120 .01490

4 -.251650* .026166 .000 -.30391 -.19939

1 .085650* .016549 .000 .05260 .11870

3 .067500* .016549 .000 .03445 .10055

4 -.166000* .026166 .000 -.21826 -.11374

1 .018150 .016549 .277 -.01490 .05120

2 -.067500* .016549 .000 -.10055 -.03445

4 -.233500* .026166 .000 -.28576 -.18124

1 .251650* .026166 .000 .19939 .30391

2 .166000* .026166 .000 .11374 .21826

3 .233500* .026166 .000 .18124 .28576

Samira Shahsiah et al.

230 EVALUATION OF ANTIBACTERIAL EFFECT OF

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AND GINGER BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

under study in 0.1 and 0.2 milliliter depth from dentine

(p<0.01). but this difference in 0.3 depth is not signi -

cant. According to post hoc test of Fisher’s least signi -

cant differences

(LSD) with 95% con dence we performed comparing

groups as pairwise in each depth. Group numbering is

as follow:

Group number 1: 5.25% sodiumhypochlorite

Group number 2: ginger concentrate

Group number 3: eucalyptus concentrate

Group number 4: normal saline (positive control)

According to post hoc test and table 3 in 0.1 mil-

liliter depth of dentine, anti-microbial effect of 5.25%

sodium hypochlorite was signi cantly better that ginger

concentrate and eucalyptus concentrate (p<0.01). While

there was no signi cant difference between 5.25%

sodium hypochlorite and eucalyptus concentrate.

In addition comparing ginger concentrate and euca-

lyptus concentrate indicated that in 0.1 milliliter depth

of dentine, there is no signi cant difference between

these two concentrate anti- microbial effect (p<0.01).

and eucalyptus concentrate had better anti- microbial

effect on enterococcus faecalis in dentinal tubules at the

presence of smear layer.

In 0.1 milliliter depth of dentine, anti- microbial

effect of5.25% sodium hypochlorite, ginger concentrate

and eucalyptus concentrate was signi cantly better than

normal saline (p<0.01).

According to post hoc test and table 4 in in 0.2 mil-

liliter depth of dentine, 5.25% sodium hypochlorite was

signi cantly better that ginger concentrate and normal

saline (p<0.02). While there was no signi cant differ-

ence between 5.25% sodium hypochlorite and eucalyp-

tus concentrate.

In addition comparing ginger concentrate and euca-

lyptus concentrate indicated that in 0.2 milliliter depth

of anti- microbial effect dentine there is no signi cant

difference (p<0.01). and eucalyptus concentrate has bet-

ter anti- microbial effect on enterococcus faecalis in

dentinal tubules at the presence of smear layer.

In 02 milliliter depth of dentine, anti- microbial effect

of 5.25% sodium hypochlorite and eucalyptus concen-

trate was signi cantly better that normal saline (p<0.01)

while there was no signi cant difference between anti-

microbial effect of ginger concentrate and normal saline.

According to post hoc test and table 5 in 0.3 milliliter

depth of dentine, there is no signi cant difference in

anti-microbial effect of 5.25% sodium hypochlorite, gin-

ger concentrate , eucalyptus concentrate , and normal

saline was signi cantly better that ginger concentrate

and normal saline (p<0.02). While there was no signi -

cant difference between 5.25% sodium hypochlorite and

eucalyptus concentrate.

3-1 results of growth and lack of growth after applying

canal detergents

In this study, in addition to measure optical density of

all environments under experiment, some samples were

cultivated as what is described in methodology. Growth

and lack of growth of bacteria on growth medium was

reported in each dentinal cylinder. Results of samples

cultivation was reported in table 8.

Table 8 indicates that:

In group 1(5.25% sodiumhypochlorite) 30% canals

had alive batteries.

In group 2( ginger concentrate ) 45% canals had

alive batteries.

In group 3( eucalyptus concentrate ) 20% canals

had alive batteries.

Table 4. comparing pairwise OD of under study groups in 0.2 milliliter depth from dentine

Depth

(I)

group

(J)

group

Mean Difference

(I-J)

Std. Error Sig.

95% Con dence Interval

Lower Bound Upper Bound

0.2 1

2 -.111750* .043327 .012 -.19828 -.02522

3 .013000 .043327 .765 -.07353 .09953

4 -.234400* .068507 .001 -.37122 -.09758

1 .111750* .043327 .012 .02522 .19828

3 .124750* .043327 .005 .03822 .21128

4 -.122650 .068507 .078 -.25947 .01417

1 -.013000 .043327 .765 -.09953 .07353

2 -.124750* .043327 .005 -.21128 -.03822

4 -.247400* .068507 .001 -.38422 -.11058

1 .234400* .068507 .001 .09758 .37122

2 .122650 .068507 .078 -.01417 .25947

3 .247400* .068507 .001 .11058 .38422

Samira Shahsiah et al.

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS EVALUATION OF ANTIBACTERIAL EFFECT OF

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AND GINGER 231

In group 4( normal saline (positive control ) 100%

canals had alive batteries.

In group 5( negative control ) no canals had alive

batteries.

Group number 1: 5.25% sodiumhypochlorite

Group number 2: ginger concentrate

Group number 3: eucalyptus concentrate

Group number 4: normal saline (positive control)

Many studies have been made so far on anti-micro-

bial properties of plant (Karlovic´ et al, 2000; Hammer

et al, 1999; Rahman et al, 2010; Adeniyi et al, 2006;

Shalaby et al, 2011; Dulger and Gonaz 2004) and there

are many studies that considered anti-microbial prop-

erties of plant as detergents (Gulve and Gulve 2010;

Shari an et al, 2011; Maekawa et al 1013). It is clear

that this topic is very important. Consequently, in many

researches, no groups under study were able to remove

bacteria of canal at the presence of smear layer. Nor-

mal saline groups indicated 100% growth. This material

has no inhibition properties against Enterococcus faeca-

lisand its effect on existing micro-organisms in canal

happens through mechanical act. In sterile dentinal cyl-

inder (negative control), no growth was observed which

indicates that there is no pollution in environment and

applied tools so accuracy of performance was approved.

Anti- microbial effect of Eucalyptus camaldulensis con-

centrate on enterococcus faecalis (ATCC29212) in cylin-

ders provided from human teeth at the presence of smear

layer, with 5.25% sodium hypochlorite has no signi cant

difference. In addition, anti- microbial effect of Zingiber

of cinale on Enterococcus faecalis (ATCC29212) in cyl-

inders provided from human teeth at the presence of

smear layer is weaker than 5.25% sodium hypochlorite.

Table 5. comparing pairwise OD of under study groups in 0.3 milliliter depth from dentine

Depth

(I)

group

(J)

group

Mean Difference

(I-J)

Std. Error Sig.

95% Con dence Interval

Lower Bound Upper Bound

0.3 1

2 -.114500 .058369 .054 -.23107 .00207

3 -.036700 .058369 .532 -.15327 .07987

4 -.176650 .092289 .060 -.36096 .00766

5 -.017050 .092289 .854 -.20136 .16726

1 .114500 .058369 .054 -.00207 .23107

3 .077800 .058369 .187 -.03877 .19437

4 -.062150 .092289 .503 -.24646 .12216

5 .097450 .092289 .295 -.08686 .28176

1 .036700 .058369 .532 -.07987 .15327

2 -.077800 .058369 .187 -.19437 .03877

4 -.139950 .092289 .134 -.32426 .04436

5 .019650 .092289 .832 -.16466 .20396

1 .176650 .092289 .060 -.00766 .36096

2 .062150 .092289 .503 -.12216 .24646

3 .139950 .092289 .134 -.04436 .32426

5 .159600 .116737 .176 -.07354 .39274

Table 6. numbers of samples with bacterial growth

Lack of growth growth numbers of samples Group

14 6 20 5.25% sodium hypochlorite

11 9 20 Gin

16 4 20 eucalyptus concentrate

0 5 5 Normal saline (positive control)

505

Dentinal cylinder (negative

control)

Samira Shahsiah et al.

232 EVALUATION OF ANTIBACTERIAL EFFECT OF

EUCALYPTUS

AND GINGER BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

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