Evaluation of Antibacterial Activity of Different Extract of Ipomoea

aquatic Leaves against Staphylococcus aureus and Streptococcus

pyogenes

Mohamed Rasny Mohamed Razik

1*

, S. Angielina

1

, Reyadh Al- Rashidi

1

, Samer Al-Dhalli

1

,

Jiyauddin Khan

1

, Kiran Chanabasappa Nilugal

1

, Santosh Fattepur 1, Kaleemullah, M

1

, Shariq

Baber

1

, Chen Jie

1

, Fadli Asmani

1

, Eddy Yusuf

2

1

School of Pharmacy, Management and Science University, 40100 Shah Alam, Selangor, Malaysia;

2

International Center for Halal Studies, Management and Science University, 40100 Shah Alam, Selangor, Malaysia

Keywords: Ipomoea aquatica, Staphylococcus aureus, Streptococcus pyogenes.

Abstract: The skin is the largest organ in the body and can be vulnerable to various microbial infection. Although

antibiotics are clinically proven to be useful in the treatment of bacterial skin infections, they are largely

subjected to antibiotic resistance and adverse effects. This has led to the screening of several medicinal

plants for their potential antimicrobial activity since they are less expensive, has reduced occurrence of

adverse effects and widespread availability. The aim of this research will focus on evaluating the

antibacterial activity of different extracts of Ipomoea aquatica leaves against Staphylococcus aureus and

Streptococcus pyogenes that causes skin infections. Leaves were extracted separately with 95% methanol

and 95% ethanol using maceration process. Phytochemical screening was done for each extract and the

minimum inhibitory concentration (MIC) was determined for each extract against both bacteria using 10

different concentrations ranging from 10mg/ml up to 100mg/ml via disc diffusion method in triplicates.

Two concentrations above the MIC from each extract were selected and antibacterial assay of the different

extracts against the two bacteria respectively was performed using disc diffusion method in triplicates. MIC

for methanolic extract against both bacteria was 10mg/ml, while MIC for ethanolic extract was 10mg/ml

against Staphylococcus aureus and 30mg/ml against Streptococcus pyogenes. Methanolic extract of the

plant at a concentration of 90mg/ml and 100mg/ml was statistically significant against Streptococcus

pyogenes with a significance value of 0.00 (p<0.05), with 100mg/ml having larger mean inhibition zone of

17.00mm ± 0.00mm than 90mg/ml (14.33mm ± 0.58mm). Statistical analysis was performed using one-way

ANOVA (Tukey’s Test). Both methanolic and ethanolic extract of the leaves has positive antibacterial

activity against both Staphylococcus aureus and Streptococcus pyogenes at different concentrations.

1 INTRODUCTION

The skin is the largest organ in the body and acts as

a static, stationary or inert wrapping for the body.

Large numbers of microorganisms are present in the

various components of the skin. For example, the

number of bacteria on the skin surface can range

from 1000 organisms per square centimetre to more

than 10 million. The principal members of the

normal skin flora are Diphtheroids, Staphylococci

and Fungi (Hall, 2001). Skin infections are clinical

entities comprising of many etiology, manifestations

and severity that involves microbial invasion of the

layers of the skin (Ki and Rotstein, 2008). Many

types of bacteria can cause infection to the skin, in

which the most common ones are Staphylococcus

and Streptococcus.

According to CDC, Staphylococcus aureus (S.

aureus) is a type of bacteria that about 30% of

people carry in their noses and often found on the

surface of the skin. It is usually harmless, but

invasive staphylococcus infections can lead to life

threatening medical complications in as little as 12

hours (Dr. . Tom Frieden, MD, 2013). On the other

hand, Streptococcus pyogenes (S. pyogenes) can

cause a variety of diseases in immunocompetent

individuals, from pharyngo-tonsillitis to life-

threatening invasive diseases, such as streptococcal

Razik, M., Angielina, S., Al-Rashidi, R., Al-Dhalli, S., Khan, J., Nilugal, K., Fattepur, S., Kaleemullah, M., Baber, S., Jie, C., Asmani, F. and Yusuf, E.

Evaluation of Antibacterial Activity of Different Extract of Ipomoea aquatic Leaves against Staphylococcus aureus and Streptococcus pyogenes.

DOI: 10.5220/0009845100002406

In Proceedings of BROMO Conference (BROMO 2018) - Symposium on Natural Product and Biodiversity, page 1

ISBN: 978-989-758-347-6

Copyright

c

1

toxic shock syndrome, and rapidly progressing deep-

tissue infections, such as necrotizing fasciitis.

(Johansson et al., 2010).

Although antibiotics have been clinically proven

to be useful in the treatment of bacterial skin

infections, they are largely subjected to limitations

such as antibiotic resistance and adverse effects. The

progressing failure of chemotherapeutics and

resistance to antibiotics has led to the screening of

several medicinal plants for their potential

antimicrobial activity (Oyewole and Kalejaiye,

2012). Unlike conventional medicines or treatments,

herbal treatments have several advantages in that

they are less expensive, more effective in certain

chronic conditions, has reduced occurrence of

adverse effects as well as widespread availability.

In this regard, one of the plant which is being

evaluated for its therapeutic efficacies is Ipomoea

aquatica (I. aquatica). In the ancient science of

Indian medicine and homeopathy, extracts of I.

aquatica leaves are administered orally to alleviate

antioxidant related disorders.

The plant is also used effectively against

nosebleed and high blood pressure. Furthermore, its

leaf extract can be used to reduce blood sugar levels

and as an antibiotic against Escherichia coli,

Pseudomonas aeruginosa and Bacillus subtilis. The

floral buds are used as an anthelmintic (Prasad et al.,

2005).

Water spinach is a perennial herb found

throughout India, Sri Lanka, Tropical Asian

countries, Africa and Australia. It is grown as weed

in India and USA, while in Malaysia, China,

Singapore and Hong Kong, it is grown commercially

(Mbatchou and Dawda, 2012). It is also known with

its common name which is swamp morning glory or

‘kangkung’ in Malaysia. I. aquatica, a green leafy

vegetable which is a rich source of amino acids and

vitamins, has been explored for the isolation and

identification of its bioactive compounds that

provides many health benefits. The leaves of I.

aquatica contains 90% moisture, 4.3%

carbohydrates, 3% protein, 2% mineral matter, 0.9%

fibre, 0.4% fat, 0.6mg/100g of nicotinic acid,

120mg/100g of riboflavin, 137mg/100g of Vitamin

C and 11mg/100g of Vitamin E (Mbatchou and

Dawda, 2012).

Plants are potential sources of natural bioactive

compounds such as primary and secondary

metabolites. Flavonoids are one of the secondary

metabolites produced by plants and are present in

most plant tissues and often in vacuoles. The basic

structures of flavonoid molecules are composed of

three rings with various substitutions, including

glycosylation, hydrogenation, hydroxylation,

methylation and sulfation. Flavonoids are

hydroxylated phenolic substances known to be

synthetized by plants in response to microbial

infection and they have been found to act as

antimicrobial agents against a wide array of

microorganisms in vitro. Their activity is probably

due to their ability to complex with extracellular and

soluble protein and to complex with bacterial cell

wall (Yadav and Agarwala, 2011). The type of

flavonoids present in I. aquatica leaves are quercetin

3-methyl ether and quercetin 4-methyl ether.

Therefore, the aim of this research will focus on

evaluating the antibacterial activity of different

extracts of I. aquatica leaves against S. aureus and

S. pyogenes that causes skin infections.

2 METHODOLOGY

2.1 Preparation of Plant Extract using

Maceration Process

2kg of fresh leaves of I. aquatica obtained was first

weighed and washed thoroughly using running tap

water to remove all adhering foreign materials and

soil particles. The leaves were then dried under

shade and sun for seven days. The amount of dried

leaves was weighed again in order to calculate the

percentage of moisture content in the plant. The

weight obtained was 101.27g. After that, the dried

leaves were coarsely powdered using a mechanical

blender. The amount obtained was weighed and

equally separated into two portions. One part (50g)

was macerated with 95% ethanol and the other part

(50g) with 95% methanol.

Both was allowed to stand at room temperature

for 7 days with occasional shaking. The final

extracts obtained was clarified by filtration using

filter papers. The filtrates were then concentrated

under vacuum in a rotary evaporator in order to

remove the solvent and obtain a solid mass. The

solid mass of both methanolic and ethanolic extracts

was weighed and the percentage yield of the plant

obtained after extraction was calculated.

2.2 Phytochemical Screening of Leaf

Extracts

Both methanolic and ethanolic extracts of I.

aquatica leaves were evaluated for qualitative

determination of primary and secondary metabolites

by preliminary phytochemical screening respectively

BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity

2

(Yadav and Agarwala, 2011). The tests done for the

presence primary metabolites were Molisch’s Test

for the presence of carbohydrates, Millon’s Test for

the presence of proteins, Ninhydrin Test for the

presence of amino acids and Filter Paper Test for the

presence of fats and oil. The tests done to detect

secondary metabolites were Alkaline Reagent Test

for the presence of flavonoids, Liebermann’s Test

for the presence of glycosides, Mayer’s Test for the

presence of alkaloids, Foam Test for the presence of

saponins, Salkowski’s Test for the presence of

steroids as well as Ferric Chloride Test for the

presence of phenols and tannins.

2.3 Determination of Minimum

Inhibitory Concentration (MIC)

using Disc Diffusion Method

The minimum inhibitory concentration was

determined for both methanolic and ethanolic

extracts of I. aquatica leaves against both S. aureus

and S. pyogenes respectively. This was done by

preparing different concentrations of each extract in

w/v (100mg/ml, 90mg/ml, 80mg/ml, 70mg/ml,

60mg/ml, 50m/ml, 40mg/ml, 30mg/ml, 20mg/ml and

10mg/ml). Empty sterile discs were impregnated in

each concentration of the extracts for a sufficient

time and then they were placed on agar plates that

has been inoculated with the selected bacteria

strains. The control group for this assay was empty

sterile discs that has been impregnated in distilled

water. The plates were then incubated at 37

0

C for 24

hours in an incubator. After 24 hours, each plate was

observed and the zone of inhibition of each sample

was measured and recorded. MIC was determined

by observing the lowest concentration of plant

extract that was able to inhibit the bacteria growth.

From this, a suitable concentration of the plant

extract was used for Antibacterial Assay using Disc

Diffusion method for both methanolic and ethanolic

extracts of I. aquatica leaves.

2.4 Antibacterial Assay using Disc

Diffusion Method

Strains: Strains of S. aureus and S. pyogenes

Medium: Mueller Hinton Agar (MHA) for S.

aureus

: Nutrient Agar (NA) for S. pyogenes

Samples: Concentrations of 30% and 40% of

methanolic extract of I. aquatica leaves against S.

aureus and concentrations of 90% and 100% against

S. pyogenes.

Concentrations of 70% and 90% of ethanolic extract

of I. aquatica leaves against S. aureus and

concentrations of 80% and 90% against S. pyogenes.

Positive control (Antibiotic): Vancomycin disc

(30mcg/disc)

Negative control (Solvent): Distilled water

The antibacterial activity of I. aquatica leaves were

tested using Disc Diffusion method. A suitable

concentration of both methanolic and ethanolic

extracts of I. aquatica leaves as stated above were

prepared respectively, in which sterile paper discs

were impregnated for a sufficient time. The positive

control, Vancomycin discs (30mcg/disc) kept in

refrigerator was taken out and left to cool to room

temperature before use.

The negative control, distilled water was

prepared and empty sterile discs were impregnated

in them respectively. S. aureus and S. pyogenes was

inoculated on the agar plates that were prepared and

stored earlier, respectively.

The discs impregnated in methanolic extract,

ethanolic extract, distilled water as well as

Vancomycin discs was then placed on each agar

plates with appropriate distance between each disc.

All the plates were incubated in an incubator at 37

0

C

for 24 hours. After incubation, the agar plates were

observed and the diameter of zone of inhibition of

each and every agent and disc used was measured.

These procedures were performed in triplicates in

order to obtain the mean and standard deviation

(n=3, mean ± standard deviation) zone of inhibition

for each agent used.

2.5 Statistical Analysis

The results obtained were statistically analysed

using One-Way Analysis of Variance (ANOVA) and

Tukey’s Test via Statistical Package for the Social

Science (SPSS) software. ANOVA was followed by

Tukey’s Test for control, standard and test group

comparisons for statistical evaluation. p value less

than 0.05 was considered statistically significant.

3 RESULTS AND DISCUSSION

The moisture content of I. aquatica leaves used in

this research was 94.95%, while its percentage yield

obtained after extraction with methanol and ethanol

was 89.16% and 96.80% respectively. Maceration of

the leaves with methanol produced more amount of

extract compared to ethanol.

Evaluation of Antibacterial Activity of Different Extract of Ipomoea aquatic Leaves against Staphylococcus aureus and Streptococcus

pyogenes

3

Percentage of Moisture Content (MC) in I. aquatica

leaves

% Moisture Content = Initial Weight (IW) – Dried Weight

(DW) X 100%

Initial Weight

(IW)

= 2000g – 100.91g X 100%

2000g

= 94.95%

Percentage of Yield obtained after Extraction of I.

aquatica leaves

Methanolic Extract of I. aquatica leaves

% Yield after Extraction = Initial Weight (IW) – Final

Weight (FW) X 100%

Initial

Weight (IW)

= 50.0g – 5.42g

X 100%

50.0g

= 89.16%

Ethanolic Extract of I. aquatica leaves

% Yield after Extraction = Initial Weight (IW) – Final

Weight (FW) X 100%

Initial

Weight (IW)

= 50.0g – 1.60g

X 100%

50.0g

= 96.80%

Both methanolic and ethanolic extracts of I.

aquatica leaves were tested for the presence of

primary and secondary metabolites respectively.

According to the results, methanolic extract of the

leaves contained carbohydrates, amino acids,

flavonoids, glycosides, alkaloids, saponins, steroids,

phenols as well as tannins. Meanwhile, the ethanolic

extract of the leaves showed positive results for the

presence of carbohydrates, proteins, amino acids,

flavonoids, glycosides, alkaloids, saponins, steroids,

phenols and tannins. According to a study conducted

in 2011 by Yadav & Agarwala, it was found that

flavonoids were responsible for the antimicrobial

activity of I. aquatica leaves. Flavonoids are

hydroxylated phenolic substances known to be

synthetized by plants in response to microbial

infection and they have been found to act as

antimicrobial agents against a wide array of

microorganisms in vitro. Their activity is probably

due to their ability to complex with extracellular and

soluble protein and to complex with bacterial cell

wall (Yadav and Agarwala, 2011). Therefore, the

presence of flavonoids in both methanolic and

ethanolic extracts of I. aquatica leaves in this study

can be said to be accountable for its antibacterial

activity.

Table 1: Results of phytochemical screening for both methanolic & ethanolic extracts of I. aquatica leaves.

METABOLITES

TEST

OBSERVATIONS

INTERPRETATION

(METHANOLIC

EXTRACT)

INTEPRETATION

(ETHANOLIC

EXTRACT)

Carbohydrates

Molisch’s Test

Appearance of a

violet ring at the

interphase.

Present

Proteins

Millon’s Test

Turning of white

precipitate to red

upon gentle heating.

Absent

Present

Amino Acids

Ninhydrin Test

Appearance of violet

colour.

Present

Fats & Oil

Filter Paper Test

No permanent

staining of the filter

paper.

Absent

Flavonoids

Alkaline

Reagent Test

Intense yellow

changed to

colourless.

Present

Glycosides

Liebermann’s

Test

Colour changed from

violet to blue / green.

Present

Alkaloids

Mayer’s Test

Formation of yellow

coloured precipitate.

Present

Saponins

Foam Test

Formation of stable

foam.

Present

Phenols &

Ferric Chloride

Appearance of blue-

Present

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Tannins

Test

black coloration.

Steroids

Salkowski’s Test

Red colour produced

in the lower

chloroform layer.

Present

According to Shamli et al 2015, phytochemical

analysis of acetone and petroleum ether extract of

I.aquatica showed that proteins, carbohydrates,

tannins, phenols and terpenoids were present in both

exctarct and glycoside and flavonoids were absent in

petroleum ether extract. Whereas, Steroids, alkaloids

and saponin were absent in both extract (Shamli,

Chandra and Nadu, 2015).

Methanolic Extract of I. aquatica Leaves

Ethanolic Extract of I. aquatica Leaves

Molisch’s Test for Carbohydrate

Millon’s Test for Protein

Evaluation of Antibacterial Activity of Different Extract of Ipomoea aquatic Leaves against Staphylococcus aureus and Streptococcus

pyogenes

5

Ninhydrin Test for Amino Acids

Filter Paper Test for Fats & Oil

Alkaline Reagent Test for Flavonoids

BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity

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Liebermann’s Test for Glycosides

Mayer’s Test for Alkaloids

Foam Test for Saponins

Evaluation of Antibacterial Activity of Different Extract of Ipomoea aquatic Leaves against Staphylococcus aureus and Streptococcus

pyogenes

7

Ferric Chloride Test for Phenols &Tannins

Salkowski’s Test for Steroids

Figure 1: Results of Phytochemical Screening

The determination of MIC for both methanolic

and ethanolic extracts of I. aquatica leaves were

determined using ten different concentrations of

both extracts, ranging from 10mg/ml up to

100mg/ml via Disc Diffusion method, which was

done in triplicates (Table 2).

As for methanolic extract of the leaves, the

lowest concentration that showed and were able to

inhibit the growth of S. aureus was 10mg/ml with a

mean zone of inhibition of 6.00mm (n=3, mean),

while for S. pyogenes, the MIC was also 10mg/ml,

but with a mean zone of inhibition of 4.00mm (n=3,

mean).

On the other hand, as for ethanolic extract of the

leaves, the lowest concentration that showed and

were able to inhibit the growth of S. aureus was

10mg/ml with a mean zone of inhibition of 3.33mm

(n=3, mean), while for S. pyogenes, the MIC was

30mg/ml, with a mean zone of inhibition of 4.00mm

(n=3, mean). After the determination of MIC for

both methanolic and ethanolic extracts of the leaves,

two concentrations above the MIC with the highest

mean value were chosen to be tested for antibacterial

activity against S. aureus and S. pyogenes

respectively.

Table 2: Minimum Inhibitory Concentration (MIC) for methanolic and ethanolic extracts of I. aquatica leaves after tested

against S. aureus and S. pyogenes respectively.

Concentration

(mg/ml)

10

20

30

40

50

60

70

80

90

100

(-)

Control

Extract /

Bacteria

Mean Zone of Inhibition (mm)

Methanolic

Extract

S. aureus

6.00

4.67

6.33

5.67

0.00

3.33

0.00

S. pyogenes

4.00

5.33

6.00

6.67

5.33

6.33

3.00

9.33

9.00

0.00

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Ethanolic

Extract

S. aureus

3.33

2.00

6.33

6.67

6.00

7.67

5.67

6.67

5.33

0.00

S. pyogenes

0.00

4.00

7.33

5.00

7.00

5.67

9.67

10.67

9.33

0.00

From the determination of MIC for methanolic

extract of I. aquatica leaves, concentrations of

30mg/ml and 40mg/ml of the extract were observed

to have the highest mean values for the zone of

inhibition of the growth of S. aureus, and therefore

they were chosen to perform antibacterial testing

against S. aureus using disc diffusion method. The

results obtained showed that both concentrations had

antibacterial activity against S. aureus with a mean

and standard deviation of 7.33mm ± 0.58mm (n=3,

mean ± standard deviation). According to one-way

ANOVA test, both the concentrations did not show

significant effects against S. aureus since the p value

was 1.00 (p > 0.05) when compared to the positive

control, Vancomycin (30mcg/disc) which had a

mean and standard deviation of 23.67mm ± 1.15mm

(n=3, mean ± standard deviation), with a p value of

0.00 (p < 0.05) (Table 4).

One-way ANOVA using Post-hoc Tukey’s test

was performed in order to evaluate the significance

of antibacterial activity of the concentrations

obtained from both methanolic and ethanolic

extracts of I. aquatica leaves against the test bacteria

(S. aureus and S. pyogenes). Based on the

interpretation from ANOVA test, p values lower

than 0.005 (p<0.05) were considered significant for

antibacterial activity and vice versa.

On the other hand, concentrations of 90mg/ml

and 100mg/ml of methanolic extract of I. aquatica

leaves were prepared to test its antibacterial activity

against S. pyogenes. The results obtained showed

that the extract of 100mg/ml had larger zone of

inhibition, with a mean and standard deviation of

17.00mm ± 0.00mm (n=3, mean ± SD) than

90mg/ml of extract.

At the same time, comparison between each

group using ANOVA test (Table 5) showed that

each group had significant effects against S.

pyogenes, since the significance value was 0.00 (p <

0.05), in which it can be said that 100mg/ml of the

extract has higher significant antibacterial effect

than 90mg/ml of the extract since 100mg/ml has

higher mean zone of inhibition against S. pyogenes.

For ethanolic extract of I. aquatica leaves, the

concentrations of the extract used were 70mg/ml and

90mg/ml in order to test the susceptibility of S.

aureus towards the extract. The higher concentration

(90mg/ml) of the extract had larger zone of

inhibition compared to 70mg/ml of the extract.

However, there were no significant difference

between the two concentrations against S. aureus

according to ANOVA test (Table 6). This is because

the p value for each concentration when compared to

one another was 0.754 (p > 0.05).

Meanwhile, the concentrations of 80mg/ml and

90mg/ml of ethanolic extract was used to test its

antibacterial activity against S. pyogenes. Clearly,

the higher concentration of the extract had bigger

zone of inhibition with a mean and standard

deviation of 7.67mm ± 0.58mm (n=3, mean ±

standard deviation) compared to 80mg/ml of the

extract. Nonetheless, ANOVA test showed that there

was no significant difference in the antibacterial

activity between the two concentrations against S.

pyogenes since the p value was 0.754 (p > 0.05)

when both concentrations were compared to each

other (Table 7).

When we compare the antibacterial activity

between methanolic and ethanolic extract of I.

aquatica leaves against S. aureus and S. pyogenes, it

can be said that both extract require different

concentrations in order to inhibit the bacteria’s

growth (Figure 1 and 2). However, when we

compare the antibacterial activity of methanolic and

ethanolic extract of I. auatica to the standard

positive control (Vancomycin); both methanolic and

ethanolic extract of I. aquatica got low antibacterial

activity than vancomycin.

Evaluation of Antibacterial Activity of Different Extract of Ipomoea aquatic Leaves against Staphylococcus aureus and Streptococcus

pyogenes

9

Table 3: Mean and SD of zone of inhibition for methanolic and ethanolic extracts of I. aquatica leaves and controls after

tested against S. aureus and S. pyogenes respectively.

Bacteria / Concentration

of Extract (mg/ml)

Mean Zone of Inhibition (mm) ± Standard Deviation (SD)(mm)

Sample Extract

(-) Control

(+) Control (Vancomycin

30mcg/disc)

Methanolic Extract

S. aureus / 30

7.3 ± 0.6

0.0 ± 0.0

23.7 ± 1.2

S. aureus / 40

7.3 ± 0.6

0.0 ± 0.0

23.7 ± 1.2

S. pyogenes / 90

14.3 ± 0.6

0.0 ± 0.0

35.0 ± 0.0

S. pyogenes / 100

17.0 ± 0.0

0.0 ± 0.0

35.0 ± 0.0

Ethanolic Extract

S. aureus / 70

5.0 ± 0.0

0.0 ± 0.0

24.3 ± 1.2

S. aureus / 90

5.7 ± 1.2

0.0 ± 0.0

24.3 ± 1.2

S. pyogenes / 80

7.3 ± 0.6

0.0 ± 0.0

30.0 ± 0.0

S. pyogenes / 90

7.7 ± 0.6

0.0 ± 0.0

30.0 ± 0.0

Table 4: Significance of each methanolic extract of I. aquatica Leaves compared to one another against S. aureus according

to their zone of inhibition.

BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity

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Table 5: Significance of each methanolic extract of I. aquatica Leaves compared to one another against S. pyogenes

according to their zone of inhibition.

Figure 2: Evaluation of the mean zone of inhibition exhibited by different concentrations of methanolic extract of I.

aquatica leaves against both S. aureus and S. pyogenes.

0

2

4

6

8

10

12

14

16

18

30 90 40 100

Mean Zone of Inhibition (mm)

Concentration of Methanolic Extract (mg/ml)

S. aureus S. pyogenes

Evaluation of Antibacterial Activity of Different Extract of Ipomoea aquatic Leaves against Staphylococcus aureus and Streptococcus

pyogenes

11

Table 6: Significance of each Ethanolic extract of I. aquatica Leaves compared to one another against S. aureus according

to their zone of inhibition.

Table 7: Significance of each Ethanolic extract of I. aquatica Leaves compared to one another against S. pyogenes

according to their zone of inhibition.

BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity

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Figure 3: Evaluation of the mean zone of inhibition exhibited by different concentrations of ethanolic extract of I. aquatica

leaves against both S. aureus and S. pyogenes.

4 CONCLUSION

Based on the conducted study, it can be concluded

that both methanolic and ethanolic extracts of I.

aquatica leaves contain flavonoids that is thought to

be responsible for its antimicrobial properties. At the

same time, both type of the extracts showed positive

antibacterial activity and were effective against both

notorious S. aureus and S. pyogenes that are known

to cause skin infections. However, each extract

require different concentrations in order to inhibit

the bacteria’s growth. Further studies such as

determination of the total flavonoid content of I.

aquatica leaves and fractionation of the extract of I.

aquatica leaves can be carried out in order to isolate

the leaves’ constituents as well as improving its

antibacterial properties. Moreover, future research

can be done on developing a formulation using

ethanolic extract of I. aquatica leaves in order to

heal and fight infection on the skin caused by S.

pyogenes, since in this research, ethanolic extract of

the leaves showed a significant effect against the

mentioned bacteria.

ACKNOWLEDGEMENT

The author is thankful to all the research committees

and lecturers of School of Pharmacy, Management

and Science University (MSU), Malaysia for their

endless support, teaching and guidance as well as in

providing all the required materials, equipment and

laboratory facilities throughout the completion of

this research.

CONFLICT OF INTEREST

The author confirms that there is no conflict of

interests.

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