The anti-bacterial effect of

natural plant extracts on e-coli

and common bacteria found on
currencies

Presented by Group V-Formation Ngs-Seet

Group Members: Justin Ng (Leader)
Edmund Ng (Time Keeper)
Ian Ng (Secretary)
Nicholas Seet (Research Leader)
Introduction

Nowadays, soap can easily be found on the

market. This anti-bacterial soaps normally contain
triclosan or EDTA (ehtylenediaminetetraacetic
acid).

Overuse of such chemicals could cause some

bacteria to evolve and create resistance against
anti-bacterial action, and the two chemicals are
also toxic to the body.

In addition, this soap may cause the skin to dry

after frequent use. Hence, this means not
everyone can use the soap, especially those with
sensitive skin.
Goals of the research

Our goal of the project is very simple: To

create a soap that is able to have anti-
bacterial effects, and a soap that is made
from a natural addictive/extract with the least
amount of preservatives added.

This soap will be used when one touches a

note/currency; This is because research has
shown that money/currency is the dirtiest
item in the world.

Hence, another goal is to be able to identify

the bacteria on currencies and destroy it.
 Reasons for a natural extract
soap instead of a normal soap

Disadvantages of Advantages of making

commercial soaps
They normally contain
triclosan or EDTA (Ethylene
Diamine Tetraacetic Acid)
which are toxic to our body.
These substances can cause
redness and irritation to the
skin.
soaps with natural additive
They can improve the
antibacterial effect of the
soaps.
They are less likely to cause
redness and irritation to
human skin.

Bacteria can probably evolve

Bacteria are less likely to be
resistance to these
resistant to them.
substances.
Plant extracts chosen…
Hence, having decided on natural extracts, we have
chosen 4 different plants to experiment on and find out
their anti-bacterial properties.

These plants are :

Clitoria (Flowers and Plants)

Pandan

Aloe Vera

Mint

Hence, there must be a reason why we chose this

plants. The following slides will explain why.
Background:
Reasons why these plants were
chosen

Aloe Vera Mint

P-menthane
Acemannan components

Clitoria flower Clitoria leaves

Protein (Finotin) Protein (Finotin)

Pandan
Roots and Leaves
Background
Clitoria Flower, Clitoria Leaves, Pandan Leaves,
Aloe Vera and Mint Leaves; all have been used for
their medical properties.

For example, previous studies have shown that the

p-methane components of mint have variable
degree of anti-bacterial activities.

Clitoria plant (Clitoria ternatea) was also previously

tested for anti-bacterial properties and such, and it
had shown that it had significant antibacterial
activity against both Gram-positive and Gram-
negative bacteria. Clitoria plant also has a protein
called finotin, which has broad and potent
Background

Acemannan in Aloe Vera was also

tested and it showed anti-bacterial
activities.
Pandan plants’ roots has also shown
anti-bacterial properties, and its leaves
are commonly used to treat skin
diseases.
Thus these samples with their anti-
bacterial properties were chosen for
our experiment and research.
Background

Hence, as we’re making soap, we needed to

have a soap base.

Therefore, glycerin was chosen.

Glycerin is a humectant which attracts

moisture to your skin. It is also which is good
for tender skin.

Glycerin soap is colourless and can be made

into various colours. This will allow people a
greater choice when picking our soap as
many colours can be available to the public.
 Purpose of our experiment

Lastly, with our results, we will form a soap with the strongest
anti-bacterial extract; The soap will mainly be to destroy the
bacteria that come from currencies.
Procedure for experiments
Filter paper
discs are
dipped into
sample
solutions.

Each of the
experiment was
carried out for 2 sets
and incubated

Effectivenes
s of sample
Zone of extracts is
inhibition is determined .
measured.
Preparations of extracts
Samples of Aloe Vera, Pandan and Mint were bought from
the market.

Samples of Clitoria flowers and leaves were obtained

from the Eco-garden in the Primary School.

Aloe Vera, Pandan, Clitoria leaves, Clitoria flowers and

Mint were cut into small pieces and grounded. Aloe Vera
was peeled and macerated in a blender.

Samples of Aloe Vera, Pandan, Clitoria leaves and flowers

and Mint were numbered from 1 to 5 respectively.

The substances were stored in the refrigerator at 2

degrees Celcius until use.
Identification of bacteria on
currency
Hence, taking 3 $2 notes, we swiped the bacteria off
from the note.

Using gram staining, we were able to identify the

bacteria.

The bacteria was E-Coli, although other unknown

bacteria that were not of a significant amount were
present as well.

Escherichia Coli (E-Coli) is a gram negative bacterium

that is commonly found in the lower intestine of warm-
blooded animals. Most E.Coli strains are harmless, but
some can cause serious food poisoning in humans, and
are occasionally responsible for costly product recalls.

A reason why E-Coli, a skin bacteria, may be present

may be because of improper washing of hands in the
 Steps taken for Gram Staining
1. Heat-fixed smear of bacteria was prepared on a slide.

2. The slide was stained with Hucker’s crystal violet for one minute and
then washed with water.

3. The slide was stained with gram’s iodine solution for one minute and
gently washed with water.

4. The slide was decolourised by adding gram’s decolorizer drop by drop

and washed immediately with water.

5. The slide was stained by safranin for two minutes and washed with
water.

6. The slide was blottled dry and observed using a microscope

7. Steps 1-6 were repeated with different samples of currencies.

8. This was how we arrived at the identity of the master plate bacteria
obtained from currencies
Experiment 1 & 2; Plating of petri
dish

Round pieces of filter paper that

will be dipped into the extract solution.

Container with
prepared extract
solution

Petri Dish with 100

micro litres of e-coli
bacteria.
Experiment 1

Plant extract were contaminated.

E-coli bacteria was not spread evenly.

 Experiment 1

This experiment was most likely a failure due to our

lack of experience and lack of accuracy.

The results that we obtained from our first

experiment was that the agar plate that we grew the
colonies on were flooded with bacteria; so much so
that we were even unable to locate the piece of
paper which was soaked in the leaves extracts.

The first experiment, (1) did not show the anti-

bacterial effect of the extracts. This was most
probably due to contamination. The extracts may be
contaminated before we had obtained them.
 Experiment 1

The spreading method used for E.coli in the first

experiment was carried out by gently shaking the
plate in an attempt to even the E.coli out.

Also, we had left our experiment in the incubator for

over a period of one week. This was a huge mistake
as the bacteria were able to grow so much that even
the anti-bacterial properties of the natural extracts
were unable to prevent the bacteria from growing
around it and on it.
Discussion on
Improvements…
Hence, to prevent anymore failure
experiment, we took extra precaution in the
second experiment.

we used fresh extracts every time we

conducted the experiment and we washed
each extract cleanly.

Also, we took special measures like replacing

the inoculation loop every time we did
plating. The plating method of bacteria was
also modified with the use of a bent glass rod
for more even spreading.
 Experiment 2 Results
Herbs/Natural Set 1; Zone of Set 2 ; Zone of
Extracts inhibition inhibition
(Measured in (Measured in
mm^2) mm^2)
Clitoria Leaves 2.5 2.3

Clitoria Flower 2.4 2.5

Pandang Leaves 0.5 0.5

Mint Leaves 0 0.3

Aloe Vera 1 1.3

 Experiment 2 Discussion
This time, the second experiment was a success. We were able
to obtain the results that were expected.

Also, the problem of contamination and spread of bacteria was

solved as there was a zone of inhibition for all the extracts.

Clitoria leaves and flower, from the same plant, apparently was
the most effective and had the most anti-bacterial effect
against e-coli as it had the largest average zone of inhibition.
(2mm)

Hence with the anti-bacterial effects and results, we went on to

combine two extracts together, so as to see if they would be
able to produce a greater anti-bacterial effect. Also, we decided
to do away with the extracts that were unable to produce a
Zone of inhibition of 1mm and more.
 Experiment 3 Results
Combined Set 1; Zone of Set 2; Zone of
Extracts/Herbs Inhibition (Measured inhibition (Measured
in mm^2) in mm^2)

Clitoria Leaves and 3.4 3.7

Flower

Clitoria Leaves and Aloe 3.2 3.0

Vera

Clitoria Flower and Aloe 2.9 3.1

Vera
Discussion

Through this experiment, it is obvious that

Clitoria leaves and Clitoria Flowers has
properties that compliment each other,
therefore being the most effective anti-
bacterial substance.

Hence, with this result, we are able to create

our soap now with Clitoria flowers and leaves.
Soap Making

Clitoria Flowers
& Clitoria Leaves
 Testing and Surveys
5 people were randomly picked and asked to smell the
soap

All 5 were asked to use the soap on their hand and

asked to smell their hand.

Their feedback is recorded.

The general feedback and comments on the soap

would be that the soap could be improved if a
fragrance was added. Also, there was feedback that
the appearance of the soap was appealing due to the
clitoria flower giving the soap a blue colour. Hence,
this combination is the best choice for a soap due to
the fact that there was anti-bacterial properties, a
 Future Research
In the future, hypoallergenic soap with no fragrance can be
made for sensitive skin or people with skin condition. A soap
containing fragrance can be made for normal population use.
The fragrance can be also from another natural extract that
has anti-bacterial properties that agree with the current
combination and has a fragrance.

In addition, clitoria leaves showed relatively consistent anti-

bacterial effects and thus soap with clitoria leaves can be
made. This soap will appear green due to the chlorophyll
content of the Clitoria leaves. This soap will also be available to
those with sensitive skin, skin condition or the normal
population.

As a result, both blue and green soaps as well as the soap both
with fragrance and no smell can be sold. This gives customers
Acknowledgements
Thank
You
For
Listening!!!