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ICROEST IOP Publishing
IOP Conf. Series: Earth and Environmental Science 343 (2019) 012145 doi:10.1088/1755-1315/343/1/012145

Antimicrobial activity of moringa leaf (Moringa oleifera L.)

extract against the growth of Staphylococcus epidermidis

Ervianingsih1, M Mursyid1, R N Annisa2, I Zahran1 , J Langkong3 and I

Kamaruddin3
1
Department of Pharmacy, Faculty of Health Agriculture and Marine Science,
Universitas Muhammadiyah Palopo, South Sulawesi, Indonesia
2
Department of Marine Science, Faculty of Health Agriculture and Marine Science,
Universitas Muhammadiyah Palopo, South Sulawesi, Indonesia
3
Departement of Agricultural Technology, Faculty of Agriculture, Universitas
Hasanuddin, South Sulawesi, Indonesia

E-mail: [email protected]

Abstract. The study was aimed to determine the antimicrobial activity of Moringa oleifera L.
leaf extract againts the growth of Staphylococcus epidermidis. This study was conducted at the
Microbiology Laboratory of the Department of Pharmacy, Ministry of Health Polytechnic
Makassar, South Sulawesi. Three different concentrations of Moringa leaf extract were used,
namely 2% b/v, 4% b/v and 8% b/v which were tested on S. epidermidis bacteria using the
paper disk diffusion method. The results showed that Moringa oleifera L. extract could inhibit
the growth of S. epidermidis by showing the inhibition zone around the extract. The biggest
inhibition zone at a concentration of 8% b/v was 14 mm, for a concentration of 4% b/v was
10.8 mm and the smallest zone at a concentration of 2% b/v was 9.3 mm.

1. Introduction
Infection disease is the most suffered disease by the population of developing countries including
Indonesia. One of the causes is bacteria [1]. The term infection describes the growth of replication of
microorganism in the host’s body. The disease grows when the infection produces changes to the
normal body physiology [2].
The research conducted by the WHO in 14 countries around the world shows that 8.7% of the
hospital patients have an infection during hospital treatment [3]. Bacteria that often cause the infection
are Escherichia coli, Staphylococcus aureus, Pseudomonas, and S. epidermidis. The treatment of
patients in bacterial infections is antibiotics. The selection of antibiotics used as an infectious therapy
has an important role in patient recovery. Furthermore, in the National Nosocomial Infection
Surveillance (NINS) was explained that therapy with dose and duration that is not appropriate, or the
dose that is not educate, can cause the change of the bacteria pattern and resistance to various
antibiotics [1].
S. epidermidis is one of the genus staphylococcus that is most commonly found in clinical interest.
These bacteria are gram-positive bacteria in white or yellow colonies that are facultative anaerobes
and include staphylococcus with negative coagulation [1]. Most of these bacteria are normal flora on
the skin and mucous membranes in human. Previously, these organisms rarely caused significant
infection. But, by the increasing use of catheter or implants and prosthetic devices [4]. S. epidermidis

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ICROEST IOP Publishing
IOP Conf. Series: Earth and Environmental Science 343 (2019) 012145 doi:10.1088/1755-1315/343/1/012145

becomes an important agents that cause nosocomial infections. The treatment for this bacterium
becomes very difficult due to the increased resistance to various microbial agents and its ability to
form biofilms. About 70% of isolated S. epidermidis has been resistance to penicillin, oxacillin,
cifazolin, ciprofloxacin, and ciprofloxacin. The high rate of resistance will be difficult in the treatment
of the infections [5].
One of the alternative treatments for infections caused by bacteria is by using natural ingredients,
such as Moringa oleifera L. plants. This plant is referred to as the world’s most valuable multipurpose
and miracle tree because all parts of the plant are useful for food, medicine, cosmetics, or water
purified [6]. Moringa oleifera L. leaf pharmalogically has benefits as antimicrobial, antifungal,
antihypertensive, antihyperglycemic, antitumor, anticancer, anti-inflammatory [7]. One of the most
prominent effect is antioxidants. Based on phytochemical tests, Moringa oleifera L. leaves contain
tannins, steroids, triterpenoids, flavonoids, saponins, interquinones, and alkaloids that all are
antioxidants [8]. Based on a research by Fuglie, fresh moringa leaves has antioxidant strength seven
times higher than vitamin C [9]. One of the flavonoid group that moringa has, is quercetin. It contains
antioxidant 4 to 5 times higher than vitamin C and vitamin E [10].
It is known that tannin and flavonoids are active compounds in Moringa oleifera L. moringa leaf
extract with ethanol solvent are able to inhibit the formation of S. aureus, and have antibacterial
properties against Pseudomonas aerugenosa and E. coli. However, there is no scientific evidence yet
that supports the effectiveness of Moringa oleifera L. ethanolic extract against S. epidermidis. Based
on these description, an antimicrobial activity of Moringa oleifera L. leaf extract was conducted on S.
epidermidis.

2. Methodology
The study was an experimental research. It used static-group comparison design. It was conducted in
May 2018 at microbiology laboratory Pharmacy Department, The Ministry of Health Polytechnic
Makassar, South Sulawesi.

2.1. Extraction procedure

Moringa oleifera L. leaves extract was produced by maceration method. The simplicia weighed as
much as 200 gram, then put into the maceration container and soaked with 70% ethanol, then stirred
until homogeneous. Subsequenlty, it closed immediately then stored in a room that is protected from
the sun light for five days while stirring it occasionally. After soaking for five days, the extract is
filtered by using filter paper to obtained filtrate (liquid extract). Next, the liquid extract is collected in
a container and then evaporated. The Moringa leaves extract were made in three levels, namely 2% b/v
and 8% b/v. Moringa oleifera L. leaf extract was weighed 0.2 gram for 2% b/v, 0.4 gram for 4% b/v,
and o,8 for 8% b/v in the petri dish, then each level dissolved with Na CMC up to 10 ml and stirred
until it is homogeneous, then labelled.

2.2. Bacterial preparation

The bacteria used was S. epidermidis. The pure bacterial stock was taken from NA slant
media and then incubated for 1x24 hours at 37ºC. The tested bacterial were taken from one
culture and then suspended with 10 ml of sterile aquadest.

2.3. Antimicrobal activity of moringa leaf extract

The antimicrobial activity test of Moringa oleifera L. leaf extract was carried out by using paper disk
diffusion. The suspended bacterial test was inoculated on the surface of a sterile NA medium using
sterile swab. After that the paper disk which has been soaked in Moringa oleifera L. leaves extract
with each concentration of 2% b/v, 4% b/v, and 8% b/v is placed aseptically using sterile tweezers on
the surface of the medium with 2-3 cm space among the paper disk from the edge of the petri dish.
Likewise for the positive control (Tetracycline 30bjp) and negative control (Na CMC). Then, it was
inoculated for 1x24 hours at 37ºC.

2
ICROEST IOP Publishing
IOP Conf. Series: Earth and Environmental Science 343 (2019) 012145 doi:10.1088/1755-1315/343/1/012145

2.4. Observation of antimicrobial activity

Antimicrobial activity was determined by measuring Diameter of Inhibition Zones (DIZ) in mm after
incubation for 24 hours. The inhibition zone formed was measured using a caliper The data obtained
were then analyzed using the ANOVA test and followed by the LSD test.

3. Results and discussion

The research result showed that from the three concentrations of Moringa oleifera L. leaf extract can
inhibit the growth of S. Epidermidis which is characterized by the formation of transparent and
circular inhibition zones around the paper disk for incubation period along 24 hours at 37ºC (figure1).
The transparent circle around the reservoir is caused by the extract of Moringa oleifera L. leaf which
inhibits the growth of bacteria that cause infection.

a b

c d

Figure 1. The antimicrobial activty of Moringa oliefera L. leaf extract against S. epidermidis growth
at incubation time 1×24 hours : (a) 8% b/v, (b) 4% b/v, (c) 2% b/v, (d) positive control and
(e) negative control
The measurement of inhibition zone of Moringa oliefera L. Leaf extract against S. Epidermidis
growth can be seen in table 1.
Table 1. The measurement result of inhibition zone diameter of Moringa oliefera L. Leaf extract
against S. Epidermidis growth

Inhibition zone diameter (mm)

Replication Concentration (% b/v) Positive Total
Negative control
2 4 8 control
I 0 9 11 13 32 65
II 0 11 11 17 30 69
III 0 8 10 12 28 58
Total 0 28 32 42 90 192
Average 0 9.3 10.6 14 30 64.3

Table 1 shows the average resistance diameter for the concentration of 2% b/v is 9.3 mm, for the
concentration of 4% b/v is 20.8 mm, for the concentration 8% b/v is 14 mm. Then, the positive control
(tetracycline) is 30.11 mm while in the negative control using Na CMC there is no no inhibition zones
was formed. This result is appropriate and greater than the study of the antimicrobial activity of water

3
ICROEST IOP Publishing
IOP Conf. Series: Earth and Environmental Science 343 (2019) 012145 doi:10.1088/1755-1315/343/1/012145

extract of moringa leaf at 8.75 mm [11] and 7 mm indicated by moringa leaf hexane extract [10]
against S. Aureus bacteria [12]. Based on the observation, the average of moringa leaf extract diameter
of 8% b/v concentration was greater than the concentration of 4% b/v, and 2% b/v. Although the
diameter difference was not too large, but the significance showed the differences between the three
concentrations.
The calculation obtained using statistical calculations show the effectiveness of Moringa oleifera
L. leaf extract in inhibiting the growth of bacteria where the calculated value (Fh) is 127.946 greater
than the table value (Ft) which is 3.48 at the α = 0.05 level.
As a follow-up test to support the result of statistical data using LSD shows a significant difference
in level (α = 0.05) between moringa leaf extract concentration 2% b/v, 4% b/v, 8% b/v, or negative
and positive control. From the results of LSD, there is a significant difference between the
concentration of 2% b/v, 4% b/v, 8% b/v and positive control. It shows that the three concentrations
had different inhibition zones. So, it could be concluded that the greatest concentration in inhibiting S.
epidermidis growth is the highest concentration of 8% b/v ad positive control using tetracycline.

4. Conclusion
Based on the research result, it could be concluded that Moringa oleifera L. leaf extract can inhibit the
growth of S. epidermidis. For the concentration of 2% b/v it can inhibit the growth of S. epidermidis in
the amount of 9.3 mm, for the concentration of 4% b/v in the amount of 10.8mm, for the concentration
of 8% b/v in the amount of 14 mm, while for positive control it is 30.11 mm. the biggest power
inhibition is in the concentration of 8% b/v in the amount of 14mm. the statistical result shows a
significant difference among treatments.

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