Effectivity Test of Black Cumin Extract

(Nigella Sativa) on Growth of Staphylococcus Aureus Isolates

Odontogenic Infection Pus

Rahmi Syaflida

, Ahyar Riza

, Abdullah Oes

, Ricky Rianto

Department of Oral and Maxillofacial Surgery, Faculty of Dentistry Universitas Sumatera Utara, Universitas 9 street

Padang Bulan, Medan, Indonesia

Keywords: Odontogenic infection, Staphylococcus aureus, Black cumin, Inhibition zone

Abstract : The majority of infection that manifest in the oral cavity are odontogenic. Based on research, Staphylococcus

aureus is the most dominant bacteria. Antibiotic administration is one of the treatment planning, but

inappropriate antibiotics administration results in resistance. In recent years, many research showed resistance

of antibiotics. Based on this, a new alternative from another material is needed to overcome this problem like

black cumin. This study aims to determine the effect of black cumin extract (Nigella sativa) concentration on

the growth of Staphylococcus aureus.This is a laboratory experimental study with a “post-test with control

group” design. Black cumin (Nigella sativa) extract was prepared at concentrations of 50%,75%,and 100%

and controlled with aquadest. The first step of the trial was bacteria rejuvenation. Furthermore the

antimicrobial test were tested with disc diffusion technique, and the data were analyzed with Kruskkal

Wallis.The result of inhibition zone measurement at 50% concentration (6.08 ± 0.48 cm), 75% concentration

(6.65±1.07 cm), at 100% concentration (9.47 ± 0.88 cm), and there was no inhibition zone of control group.

The average percentage at 50% concentration (56.95 ± 4.24%), 75% concentration (65.55 ± 9,11%), and

100% (92.09 ± 5,87%).

1 INTRODUCTION

The majority of infection that manifest in the oral

cavity are odontogenic. The infections caused by both

aerobic and anaerobic bacteria comprised about 60%

of all odontogenic infections.

(Fragiskos, 2007) Based

on research, Staphylococcus aureus is the most

dominant bacteria.

(Kohli, 2009) Bacterial virulence

can move bacteria freely in all directions. Antibiotic

administration can be used to overcome this, but

inappropriate antibiotic administration results in

resistance.

(Syed, 2011) A research of Benardo and

Ueno showed almost all Staphylococcus aureus

strains were resistant to amoxicilin and amphicilin in

Brazil.

(Foday, 2010) Based on this, a new alternative

from another material is needed to overcome this

problem. Black cumin (Nigella sativa) is cultivated in

many countries in the world like Middle Eastern,

Mediteranian region, India, and Saudi Arabia. Black

cumin grows at 20-90 cm tall. The flowers are white,

yellow, pink, with 5-10 petals. Black cumin has 4

important active ingredients, they are thymoquinone,

tanin, thymol, and dithymoquinone. This ingredients

have antibacterial effect.

An experiment of Wasim

Raja showed about antibacterial effect of black cumin

extract to gram positive bacteria like Enterobacter

showed the average measurement of inhibition zone

at 100% concentration is 11.15 cm.

(Raja, 2016)

2 MATERIAL AND METHOD

This study was an experimental study with post-test

with control group design. The research was conduct

in the Laboratory of Microbiology Faculty of

Mathematics and Natural Science Sumatera Utara

University, Medan, Indonesia. The sampling

calculation in this study use Federer formula, in

which sample in this study was 28 petri dishes.

Sample was take from Laboratory of Microbiology

Faculty of Health Sumatera Utara University.

Maceration technique is used to produce black cumin

(Nigella sativa) extract. Production black cumin

extract is carried out using 96% ethanol solution.

Black cumin extract was prepared at concentrations

of 50%,75%,and 100%. Odontogenic infection pus

904

Syaﬂida, R., Riza, A., Oes, A. and Rianto, R.

Effectivity Test of Black Cumin Extract (Nigella Sativa) on Growth of Staphylococcus Aureus Isolates Odontogenic Infection Pus.

DOI: 10.5220/0010099909040907

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

904-907

ISBN: 978-989-758-449-7

has been previously isolated from Laboratory of

Microbiology Faculty of Health Sumatera Utara

University and from the result of isolation

Staphylococcus aureus is dominant bacteria.

Antimicrobial test started from bacteria rejuvenation.

The bacteria rejuvenation tarted by streaking 1-3 ose

bacteria in Nutrient Agar then incubate for 24 hours

in incubator.

Antimicrobial test was performed on the media

under the following procedures. First step is taken 3

ose from rejuvenated bacteria then put the bacteria in

test tube that contain 5-10 milimeter aquadest, and

vortex. After the vortex process, the absorbance of

suspension was measured with spectrophotometry at

wavelenght of 600 nm until the absorbance value of

0,5 is obtained. Dip the sterile cotton bud into

bacterial suspension, then streak on the surface of

Mueller Hinton Agar. Take the disc paper that has

been soaked in each concetration for 1 hour then

placed on the part that has been marked, then incubate

the petri dish in incubator for 24 hours on 37

Table 1: The result of inhibition zone percentage

Group Concentration Contr

50% 75% 100%

I 57.42 % 46.21% 98.03% 0%

II 57.42% 72.82% 98.97% 0%

III 56.02% 66.29% 88.23% 0%

IV 63.02% 66.76% 94.30% 0%

V 60.22% 66.76% 94.77% 0%

VI 55.08% 66.29% 86.36% 0%

VII 49.48% 73.76% 84.03% 0%

The inhibition zone known based on the

measurement of the clear zone diameter around disc

paper. Measurement performed with caliper.

Measurements were carried out 3 times by measuring

the largest diameter of the inhibitory zone at each

concentration. Data obtained from this experiment

was put into a table. Data processing was done with

computer and analyzed with SPSS using Kruskall-

Wallis test.

3 RESULT

The result of inhibiton zone measurement at 50%

concentration are 6.15 cm, 6.15 cm, 6 cm, 6.75 cm,

6.45 cm, 5.90 cm, and 5.30 cm. Then at 75%

concentration are 4.95 cm, 7.80 cm, 7.10 cm, 7.15 cm,

7.15 cm, 7.10 cm, and 7.90 cm. The biggest

concentration in this study is 100%. The result of the

inhibition zone measurement are 10.50 cm, 10.60 cm,

9.45 cm, 10.1 cm, 10.15 cm, 9.25 cm, and 9 cm, and

there was no inhibition zone of control group. (Table

The result of inhibition zone percentage at 50%

concentration are 57.42%, 57.42%, 56.02%, 63.02%,

60.22%, 55.08%, and 49.48%. Then at 75%

concentration are 46.21%, 72.82%, 66.29%, 66.76%,

66.76%, 66.29%, and 73.76%. The inhibition zone

percentage at 100% concentration are 98.03%,

98.97%, 88.23%, 94.30%, 94.77%, 86.36%, and

84.03%. (Table 2)

The result of average measurement showed at

table 3, the smallest inhibition zone at 50%

concentration (6.08 ± 0.48 cm) and the largest

inhibition zone was found at 100% concentration

(9.47 ± 0.88 cm) and there was no inhibition zone of

control group. (Figure 1,2,3,4) The average

percentage at 50% concentration (56.95 ± 4.24%),

75% concentration (65.55 ± 9,11%), and 100% (92.09

± 5,87%). (Table 3)

Table 2: The result of the inhibition zone measurement

Table 3: The average measurement and percentage of

inhibition zone.

Concentration Inhibition

Zone

Percentage

of Inhibition

Zone

value

Mean±SD

(cm)

Mean±SD

(%)

50% 6.08±0.48 56.95±4.24 0.000

75% 6.65±1.07 65.55±9.11

100% 9.47±0.88 92.09±5.87

Control 0 0

Group Concentration Contr

50% 75% 100%

I 6.15 cm 4.95 cm 10.05

0 cm

II 6.15 cm 7.80 cm 10.60

0 cm

III 6.00 cm 7.10 cm 9.45 cm 0 cm

IV 6.75 cm 7.15 cm 10.10

0 cm

V 6.45 cm 7.15 cm 10.15

0 cm

VI 5.90 cm 7.10 cm 9.25 cm 0 cm

VII 5.30 cm 7.90 cm 9.00 cm 0 cm

Effectivity Test of Black Cumin Extract (Nigella Sativa) on Growth of Staphylococcus Aureus Isolates Odontogenic Infection Pus

905

Figure 1: Petri dish at 50% concentration.

Figure 2: Petri dish at 75% concentration.

Figure 3: Petri dish at 100% concentration.

Figure 4: Petri dish for control.

The result of kruskall wallis statistical test was

significant (p=0.000; p< 0.05). This means black

cumin extract can inhibit Staphylocococcus aureus

growth. Beside that, there is an increase inhibition

zone value as the concentration value increase.

Graphic 1: The average measurement and percentage of

inhibition zone.

4 DISCUSSION

The inhibition zone is a clear area that appears on

media in petri dish after the disc has placed. This clear

area indicates an inhibition of the growth of

microorganism by antimicrobial agents on media.

(Pratiwi, 2008)

The result of this study is accordance

with the Morsi’s research who reported Black Cumin

(Nigella satia) extract can inhibit gram positive

bacteria growth such as streptococcus and

staphylococcus.

(Asniyah, 2009)

Beside that, Wasim

Raja who reported black cumin extract can inhibit

gram positive bacteria like Bacilus subtilis with the

result of inhibition zone measurement is 9 cm.

(Raja,

2016)

There is an increase inhibition zone value as the

concentration value increase. This result is as same as

the results of the Arici study, that showed there is an

unidirectional relationship between both.

(Arici, 2007)

An experiment of Asniyah about antibacterial effect

of black cumin extract to gram negative bacteria like

Escherichia coli showed the average measurement of

inhibition zone at 50% concentration is 0.9 cm, then

at 100% concentration is 1.3 cm.

(Asniyah, 2009)

The

other experiment of Wasim Raja about antibacterial

effect of black cumin extract to gram positive bacteria

like Enterobacter showed the average measurement

of inhibition zone at 50% concentration is 5.6 cm,

then at 100% concentration is 11.15 cm.

(Raja, 2016)

There is a difference in the average inhibition zone

value between gram positive and negative bacteria.

The average value in gram positive bacteria is greater

than gram negarive bacteria. Etiology of this situation

is the cell wall structure of gram positive bacteria is

more simple than gram negative. The cell wall

structure of gram positive consists of peptidoglycan

and teichoic acid. In conclusion, gram positive

bacteria is easier inhibited by antibacterial substances

than gram negative bacteria.

control 50% 75% 100%

Inhibitionzonemeasurement

Inhibitio

nzone

measure

ment

ICOSTEERR 2018 - International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches

906

5 CONCLUSION

Based on the results of research and data analysis, it

can be concluded the lowest inhibition zone value at

50% concentration and the highest value at 100%

concentration. The p-value from Kruskall wallis

statistical test is p<0.05, it means black cumin is

effective in inhibiting the growth of Staphylococcus

aureus bacteria. The content of thymoquinone, tanin,

thymol, and dithymoquinone in black cumin having a

function as an antibacterial. This inhibition zone is

formed caused by the active ingredient of black

cumin. They are thymoquinone, tanin, thymol, and

dithymoquinone. Thymoquinone and

dithymoquinone can form irreversible complexes

with bacterial proteins and resulting in protein

inactivation. While tanin and thymol worked by

inactivating enzymes and cell wall proteins that

disrupt bacterial growth.

(Aprilia, 2016)

REFERENCES

Aprilia E, Setiawati EM, Kurnia S. 2016 The effectiveness

of nigella sativa extract to reduce subgingival plaque

bacteria. Indonesian Dent J 23(3) 60-2.Fragiskos D

2007 Oral surgery ( Leipzig: Springer) p. 205-20

Arici M, Sagdic O, Gecgel U 2007 Antibacterial of turkish

black cumin (Nigella sativa) oils. J Of Plant Develop

Sci 3(4) 259-62.Kohli M, Mathur A, Siddiqui SR 2009

In vitro evaluation of microbiological flora of orofacial

infection. J Maxillofac Oral Surg 8(4) 329-33.

Asniyah 2009 Efek antimikroba minyak jintan hitam

terhadap pertumbuhan Escherchia coli in vitro. J of

Biomedic 1(1) 25-9.Syed R, Prasad G, Deeba F, Rani

D, Jamil K, Alshatwi AA 2011 Antibiotic drug

resistance of hospital acquired Staphylococcus aureus

in Andra Pradesh: A Monitoring Study. Afr J Microbiol

Res 5(6) 671-4.

Kohli M, Mathur A, Siddiqui SR 2009 In vitro evaluation

of microbiological flora of orofacial infection. J

Maxillofac Oral Surg 8(4) 329-33.

Pratiwi ST 2008 Mikrobiologi farmasi (Jakarta: Erlangga)

p.188-91

Raja W, Dewangan T 2016 Determination of antibacterial

activity of nigella sativa seed extract against some

human pathogenic bacteria. American J of Bio and

Pharm Res 3(2) 65-69.

Syed R, Prasad G, Deeba F, Rani D, Jamil K, Alshatwi AA

2011 Antibiotic drug resistance of hospital acquired

Staphylococcus aureus in Andra Pradesh: A

Monitoring Study. Afr J Microbiol Res 5(6) 671-4.

Effectivity Test of Black Cumin Extract (Nigella Sativa) on Growth of Staphylococcus Aureus Isolates Odontogenic Infection Pus

907