THE EFFECTIVENESS OF GREEN LEAVES (PIPER BETLE L.

) IN INHIBITING
THE GROWTH OF STREPTOCCUS MUTANS BACTERIA
*Andi Fairuz Zuraida Eva *, *Mila Febriany **, *Nur Radia Fitri**
*Oral Disease Sciences, Faculty of Dentistry Universitas Muslim Indonesia
** Oral Disease Sciences, Faculty of Dentistry Universitas Muslim Indonesia
***Mahasiswa, Faculty of Dentistry Universitas Muslim Indonesia
Email: nurradiafitri@gmail.com

ABSTRACT
Introduction: Caries is a disease of the hard tissues on the teeth. Dental caries is formed because the

existence of food residue that attached to the teeth. The main microorganism associated with the

development of dental caries is bacteria Streptococcus mutans. It has been confirmed the benefits of

natural ingredients in supporting dental health service programs, especially preventing dental caries.

Several studies have also shown the effect of betel leaf extract (Piper betle L.) on the inhibition of

Streptococcus mutans bacteria. Objective: To determine the effectiveness of green betel leaf (Piper

betle L.) in inhibiting the growth of Streptococcus mutans bacteria. Results: it is found that the betel leaf

extract group with a concentration of 25% contains the highest average inhibition (20.40 (0.214))

compared to other groups. Conclusion: Based on the results of this study, it can be concluded that :

Green betel leaf extract (piper betle L.) with a concentration of 15% is effective in inhibiting

Streptococcus mutans bacteria.

Green betel leaf extract (piper betle L.) with a concentration of 20% is effective in inhibiting

Streptococcus mutans bacteria.

Green betel leaf extract (piper betle L.) with a concentration of 25% was effective in inhibiting

Streptococcus mutans bacteria.

Green betel leaf extract (piper betle L.) with a concentration of 25% is the most effective in

inhibiting the growth of Streptococcus mutans bacteria with p value = 0.000.

Keywords: the concentrations of 15%, 20%, and 25% are effective in inhibiting the growth of

Streptococcus mutans.
 INTRODUCTION
Oral health is a healthy state of the hard and soft tissues of the teeth and related elements in

the oral cavity that help a person to eat, talk, and interact socially without dysfunction,

aesthetic disturbance, and discomfort due to disease, occlusal deviation and loss so that they

can live productively socially and economically. Caries is a disease of the hard tissues of the

teeth that are formed because there is food residue attached to the teeth. As a result, the teeth

become porous, perforated, and even broken. Dental caries is a multifactorial, chronic,

localized infectious bacterial disease that leads to the destruction of the hard tissues of the

teeth. Its development essentially requires three factors that are sustained over time: a

susceptible host, the cariogenic microflora in the biofilm tooth, and an adequate substrate.

The main microorganisms associated with the development of dental caries were most

commonly found, namely Streptococcus mutans, and to a lesser extent Streptococcus

sobrinus and Streptococcus gordonii, as well as Lactobacillus and Actinomyces species.

Currently, many studies have been carried out using natural materials aimed at producing

medicines in an effort to support dental caries prevention programs. Based on research by

Nadya M. Owu. et al (2019) confirmed that betel leaf extract (Piper betle L.) has antibacterial

effectiveness against Streptococcus mutans as evidenced by antibacterial effectiveness testing

at concentrations of 25%, 20% and 15% there was a decrease in absorbance values before and

after incubation which indicated a decrease in the number of bacterial cells. the living.

Meanwhile, at concentrations of 10% and 5% there was an increase in the absorbance value

which indicated that the betel leaf extract could not inhibit the growth of bacteria at these

concentrations.

Based on the above background, the researchers are interested in re-examining the

effectiveness of green betel leaf (Piper betle L.) at concentrations of 25%, 20% and 15% in

inhibiting the growth of Streptococcus mutans bacteria by using a different positive control

using Chlorhexidine 0, 2%.

 METHOD
The study used an experimental design at Fisheries Laboratory of Universitas Hasanuddin

and Laboratory of the Faculty of Pharmacy, Universitas Muslim Indonesia, namely research

conducted by giving treatment to the object of research and the existence of research controls.

This research was conducted at Fisheries Laboratory of Universitas Hasanuddin and

Laboratory of the Faculty of Pharmacy, Universitas Muslim Indonesia. This research was

conducted from September 2021 to May 2022.

RESULTS
The Kruskal-Wallis non-parametric test shows two analytical results. First, the mean ranking

which sorts the groups based on the ranking scores. The results shows that the betel leaf

extract group with concentrations of 20% and 25% have twice score compared to the 0.2%

chlorhexidine positive group. This cannot be concluded directly, because this difference

could be due to sampling error. Therefore we have to look over at the results of the statistical

analysis significance test. In the results it is found that the significance test value of p = 0.000

(p < 0.05). These results indicates that there are significant differences between the treatment

groups. This also explains that betel leaf extract has a better antibacterial effect against

Streptococcus mutans than 0.2% chlorhexidine table 5.1.3.

TaLE 5.1.3 Kruskal-Wallis . Non-Parametric Test

Group N Mean Rank
Aquadest 15 8,00
Concentrate 15% 15 35,70
Concentrate 20% 15 52,50
Concentrate 25% 15 67,47
Chlorhexidine 0,2% 15 26,33
Inhibition: 68,416
Asymp. Sig.: 0,000
 DISCUSSION

These results are in accordance with Rahmawaty Lubis R in her research on the antibacterial

activity of betel leaf extract in inhibiting Staphylococcus aureus, which shows significant

results. In this study, betel leaf extract at concentrations of 0.5% and 1% could have great

potential as a source of therapeutic agents for bacterial infections. in a study explains that the

extracts, essential oils, preparations, and compounds in betel leaf are effective against many

gram-negative and gram-positive bacteria. The bacteria tested includes foodborne pathogens

and other bacteria, including multi-drug resistant (MDR) bacteria that cause severe infectious

disease in humans. Another study using the well-diffusion method shows that the ethanol

extract of betel leaf has a larger zone of inhibition in gram-positive than gram-negative

bacteria. In this study, the antibacterial effect of five types of betel leaf extract extracted from

different solvent polarities. Among these extracts, acetate and ethyl acetate extracts showes

the most remarkable activity against the six bacteria tested with Staphylococcus aureus being

the most susceptible as for research on the antibacterial activity of green betel leaf extract

against two oral pathogenic bacteria and oral opportunistic fungi. Streptococcus mutans is the

most sensitive organism compared to Streptococcus sanguinis and Candida albicans to 0.5%

betel leaf extract with inhibitory diameters of 18.2 mm, 9.9 mm, 16.7 mm, respectively. The

difference in sensitivity of these organisms may be due to the different cell types. Candida

species (blastospores) are different from Streptococcus (Gram bacteria). This causes

differences in sensitivity to betel leaf extract. Streptococcus mutans is more susceptible than

Streptococcus sanguinis. The possible causes are many factors, including virulence.

In 2015, Azizi et al assessed the sensitivity of Streptococcus mutans and Streptococcus

sanguinis with minimum inhibitory concentration (MIC) values of 0.02 and 0.3, respectively

mg/mL which indicates that Streptococcus mutans is more susceptible than Streptococcus

sanguinis. In addition, its sensitivity to Chlorhexidine is also reported by Istighfari et al in

2018, where Streptococcus mutans is more susceptible than Streptococcus sanguinis with
growth inhibition of 23.97 mm and 19.80 mm, respectively.

There are several commercial products available that contain betel leaf such as dietary

supplements, mouthwashes, medicinal products, toothpaste, and others. Current antimicrobial

studies focus on oral pathogens, gram-negative and gram-positive bacteria that are MDR. A

study has also revealed that the potency of betel leaf extract in toothpaste has a significant

increase in the bacterial inhibition zone between 5% - 24% which is observed in the case of

oral pathogens, bacteria and fungi. Research shows that betel leaf extract not only maintains

overall antimicrobial activity, therefore its potential in clinical or commercial applications as

an effective antimicrobial agent requires further research.

Streptococcus mutans is also a normal flora of the oral cavity that acts as a cause of dental

caries and inflammation of the oral mucosa. The acid produced continuously from the

breakdown of the substrate will change the oral environment to become more acidic (pH 5.2

– 5.5) which then causes demineralization of the enamel which then forms caries. Various

actions are taken to prevent oral health, such as brushing teeth and using mouthwash.

Mouthwash contains antimicrobial substances that can reduce the number and adhesion of

bacteria in the oral cavity. The average inhibition zones of Streptococcus mutans bacteria in

various types of mouthwash in Indonesia are 11.6 mm, 9.3 mm, and 8 mm. this can be a

reference for determining the amount of concentration that can be used so that the use of

antimicrobials in mouthwash does not damage other normal flora in the oral cavity. average

inhibition zone of Streptococcus mutans

in various types of mouthwash in Indonesia are 11.6 mm, 9.3 mm, and 8 mm. this can be a

reference for determining the amount of concentration that can be used so that the use of

antimicrobials in mouthwash does not damage other normal flora in the oral cavity. This

study shows results that support the alternative hypothesis (Ha), namely, there is the

effectiveness of betel leaf (Piper betle L.) at concentrations of 15%, 20%, and 25% in
 inhibiting the growth of Streptococcus mutans bacteria. based on the results of research that

has been done, it is found some shortcomings and limitations in the preparation of this thesis.

The drawback is that there are several obstacles in the bacterial inhibition test, so the test is

repeated and causes the results of the research to be quite time-consuming. As for the

limitations in this study, the variables used were less varied which only focused on inhibiting

the growth of one type of bacteria. further research is needed on more applicable betel leaf

extract such as its effectiveness in toothpaste or mouthwash. Besides that, it also considers

the right concentration so as not to damage other normal flora in the oral cavity.

CONCLUSIONS AND SUGGESTION

Based on the results of this study it can be concluded that:

Green betel leaf extract (Piper betle L.) with a concentration of 15% is effective in inhibiting

Streptococcus mutans bacteria.

Green betel leaf extract (Piper betle L.) with a concentration of 20% is effective in inhibiting

Streptococcus mutans bacteria.

Green betel leaf extract (Piper betle L.) with a concentration of 25% is effective in inhibiting

Streptococcus mutans bacteria.

Green betel leaf extract (Piper betle L.) with a concentration of 25% is the most effective in

inhibiting the growth of Streptococcus mutans bacteria with p value = 0.000.

based on the research conducted, it is necessary to do further research on green betel leaf extract

(Piper betle L.) as the basic ingredient of herbal mouthwash with different concentrations or by

using different positive controls.

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