Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
DIPLOMA IN SCIENCE
FACULTY OF APPLIED SCIENCES
UNIVERSITI TEKNOLOGI MARA
OCTOBER 2016
COMPARISON OF THE EFFECTIVENESS OF COMEDONAL ACNE
MEDICINES AGAINST Stapylococcus aureus AND Escherichia coli
OCTOBER 2016
DECLARATION
We hereby declare that the thesis is based on our original work except for quotations
and citations which have been duly acknowledged. We also declare that it has not
been previously or concurrently submitted for any other report at Universiti Teknologi
Genevieonna Pojon
Zebora Dominic
iii
This Mini Project Report entitled “Comparison of the Effectiveness of Comedonal
Acne Medicine Against Staphylococcus aureus and Escherichia coli ” was submitted
by Genevieonna Pojon (2014616516), Muhammad Ilyas bin Rahmat Effandi
(2014662592), Nazaratul Akmam binti Harun (2014898838), Pinaslim binti Sarupin
(2014829152), Siti Noor Haseenah binti Salem (2014849898), and Zebora Dominic
(2014887012) in partial fulfillment for the Diploma in Science, in the Faculty of
Applied Science, and was approved by
_________________________
Dr. Lo Chor Wai
Supervisor
Faculty of Applied Sciences
Universiti Teknologi MARA Sabah
_________________________
Ajis Lepit
Project Coordinator
Faculty of Applied Sciences
Universiti Teknologi MARA (Sabah)
Date: __________________
iv
ACKNOWLEDGEMENTS
many parties. It is a genius pleasure to express our deep sense of thanks and gratitude
to our supervisor and philosopher, Dr. Lo Chor Wai, lecturer of Universiti Tekonologi
Mara Sabah. Her dedication and keen interest above all her overwhelming attitude to
help her students had been solely and mainly responsible for completing our work.
Her timely advice, meticulous scrutiny, scholarly advice and specific approach have
We also not forget to thank profusely to Pn. Anna, the laboratory assistant of
Universiti Teknologi Mara Sabah, for her kind help and co-operation throughout of
Genevieonna Pojon
Muhammad Ilyas bin Rahmat Effandi
Nazaratul Akmam binti Harun
Pinaslim binti Sarupin
Siti Noor Haseenah binti Salem
Zebora Dominic
v
TABLE OF CONTENTS
Page
DECLARATION iii
APPROVAL SHEET iv
ACKNOWLEDGEMENTS v
TABLE OF CONTENTS vi
LIST OF TABLES vii
LIST OF FIGURES viii
LIST OF ABBREVIATIONS ix
ABSTRACT x
ABSTRAK xi
CHAPTER 1 INTRODUCTION
1.1 Background 1
1.2 Significance of study 2
1.3 Objective of study 2
CHAPTER 5 CONCLUSION 17
REFERENCES 18
APPENDICES 20
CURRICULUM VITAE 22
vi
LIST OF TABLES
vii
LIST OF FIGURES
viii
LIST OF ABBREVIATIONS
ix
ABSTRACT
Staphylococcus aureus and Escherichia coli are the most common bacteria that can be
found at our skin and environment. Our study was conducted to compare the
effectiveness of three comedonal acne medicines against Staphylococcus aureus and
Escherichia coli. The comedonal acne medicines were tested for antimicrobial activity
by using the disc diffusion method. The result of the disc diffusion method showed
that 3 types of comedonal acne medicines which are Medicine A, Medicine B and
Medicine C can inhibit the growth of Staphylococcus aureus but it cannot inhibit the
growth of Escherichia coli. Among those, antimicrobial activity showed that
Medicine A produced strong inhibition zone against Staphylococcus aureus. Lastly,
our data indicated that Medicine A had the strongest inhibitory effect on
Staphylococcus aureus and Escherichia coli bacteria.
x
ABSTRAK
Staphylococcus aureus dan Escherichia coli adalah bakteria yang biasanya didapati di
kulit dan persekitaran kita. Kajian kami telah dijalankan untuk membandingkan
keberkesanan tiga ubat jerawat terhadap Staphylococcus aureus dan Escherichia coli.
Ubat jerawat telah diuji untuk aktiviti antimikrob dengan menggunakan kaedah cakera
resapan. Hasil kaedah cakera penyebaran menunjukkan 3 jenis ubat jerawat A, B dan
Staphylococcus aureus. Akhir sekali, data kami menunjukkan bahawa ubat jerawat A
mempunyai kesan yang paling kuat pada Staphylococcus aureus dan Escherichia coli
bakteria.
xi
CHAPTER 1
INTRODUCTION
1.1 Background
Comedonal acne is a mild acne that involves blackheads or whiteheads and usually
found below the oily skin. Comedonal acne medicines are easily to be found
pimples. There are many types of medicine with different ingredients were sold
everywhere but most people do not sure what ingredients have the greater effect on
treating the comedonal acne. The medicine used in this study was a pimple medicine
that has the same major ingredient which is the tea tree oil but the three of it was
being mixed with other different ingredients. The present study was to investigate the
1
1.2 Significance of the study
microbiological techniques. Furthermore, this project also can increase the knowledge
of students on how the mechanism of gram negative and gram positive work between
the antibacterial sources of comedonal acne medicine. Lastly, the result from this
project will be helpful and useful baseline information to other students as reference
for their future research. Nowadays, there are many people suffering skin disease that
been caused by bacteria. Skin disease such as atopic dermatitis have become huge
concern to people as most of the victim that affected by this disease are babies. Our
project is very beneficial ti find which ingredient of each medicines that can help
inhibit bacteria that have potential to cause skin disease especially Staphylococcus
aureus
2
CHAPTER 2
LITERATURE REVIEW
Acne vulgaris is the most common skin disease that affects areas containing
the largest oil glands, including the face, back, and trunk (Leydon, 1997).
Propionibacterium acnes is the one that cause acne which is a type of chronic
Staphylococcus aureus also can cause severe disease to the skin like atopic
colonization and infection on the skin of their patients (Bunikowski and Rita, et
al., 1999).
Not every type of acne medicines works the same for every person; there are 3
and emollients that can smooth the skin and suppler (Overgaard, Olsen L., and G.
tea tree oil (TTO) has wide spectrum antimicrobial activity that can decrease the
and also have significance role in treating cutaneous infection which is another
skin disease (Carson, Christine F., Brian J. Mee, and Thomas V. Riley., 2002).
Staphylococcus aureus. But in these both medicine B and medicine C also contain
tea tree oil (TTO) In Qius’ and Jiazhangs’ results shows that methol might be
4
2.3 Disc diffusion method
Disc diffusion method for antimicrobial testing was carried out according to
the standard method by Bauer et al. (1971) to judge the presence of antibacterial
activities. A bacteria culture was used to lawn Muller Hinton agar plates evenly
using a sterile swab. The plates were dried for 15 minutes and then used for the
sensitivity test. The disc which had been impregnated with a series antibacterial
was placed on the Mueller-Hinton agar surface. Each test plate comprises of four
discs. One of the discs is positive control, which is standard commercial antibiotic
disc and three treated disc. Besides the control each plate had three treated disc
placed about equidistance to each other. The plate was then incubated at 37°C for
24 hours depending on the species of bacteria used in the test. After the
incubation, the plates were examined for inhibition zone. The inhibition zones
were then measured using calipers and recorded. The test was repeated three times
to ensure reliability.
5
2.4 Spread method
Spread plates a technique that spread microbes on the media plate. Microbes
can be dilute or in solution. Then microbes will be transfer to petri dish with
specific media for specific microbes that can grow. Uniformly spread the solution
onto the petri dish by using bent-glass rod. Before using bent-glass rod it must be
sterilize with alcohol and contact few times with red flames of Bunsen burner to
6
CHAPTER 3
Medicine C) at pharmacy store around Sepanggar, Sabah. Three samples are kept at
dissolve mixture at 121oc for 15 minutes, once the agar has been autoclaved, allow it
to cool but not solidify in fume hood. Pour nutrient agar into petri dish and let the agar
solidify. Place the lid of each petri dish and store the plates in refrigerator until use.
Suspend 0.4g of sterile LB broth in 50mL distilled water using 250mL conical flask.
Autoclave the dissolve mixture at 121oC for 15 minute, once the broth has been
autoclaved, allow it to cool down at room temperature. Transfer (5mL) broth into two
separate test tubes (make sure the test tubes are closed before it being used to prevent
contamination). The rest of broth that not used are transferred into sterile container
and keep it in refrigerator and make sure the container is closed tightly to prevent
7
3.3.2 Inoculation of bacteria
Take a loop full of colonies from a bacteria plate of Escherichia coli and inoculate
into LB broth. Do the same as the above for Staphylococcus aureus. Incubate the LB
broth for one day (24 hour) in the incubator at 37oc (make sure the media bottle closed
to prevent contamination).
using micropipette and spread evenly the LB broth using the glass spreader. Pipette
micropipette and spread evenly the LB broth using the glass spreader. The sterile
paper discs which have impregnated with test material acne medicine are placed on
the agar. Each test plate comprises of 2 disc plate (each disc divided into 4 parts
labelled) which is one disc for test bacteria S. aureus and the other on for E. coli. The
aureus and E. coli. Besides the controls, each plate had 3 treated disc placed about
equidistance to each other as shown in figure 1. The plates are incubated at 37 degree
Celsius for 24 hours. (Place the plates in the incubator upside down to prevent
condensation falling into the culture and the lid of the plates should be taped with
parafilm to avoid the risk of pathogens being spread.) After the incubation, the plates
8
Control Medicine B
Agar containing
test bacteria
Medicine C
Medicine A
Measure the diameter of inhibition zone in (mm) by using ruler. Confirm the
inhibition of zone.
9
3.5 List of equipments/facilities:
3.5.1 Autoclave
3.5.2 Incubator
3.5.4 Forceps
3.5.10 Parafilm
10
CHAPTER 4
The present study was to carry out the investigation of antibacterial activities of
pimple medicines against the gram positive Staphylococcus aureus and gram negative
Escherichia coli. Using the proper techniques of disc diffusion method, the presence
of clear inhibition zone was detected on the different agar plates of S.aureus and
E.coli. Disc diffusion method for antimicrobial susceptibility testing was carried out
according to the standard method by Bauer et al. (1966) to assess the presence of
During the disc diffusion method, we must be sure that the three different types of
pimple medicine were applied evenly on the disc made from filter paper being cut and
shaped using paper puncher by using a swab that had been sterilized. The three
medicines also applied on the same agar plate which had been divided into four
regions where one of the regions was filled with the control disc (empty disc). The
disc must be placed about equidistance to each other and incubated at 37oC for 24
hours. This experiment was tested four times on S.aureus and E.coli to ensure the
reliability.
In this study, the three types of pimple medicine had the antibacterial activities against
the S.aureus and E.coli. This is because the three medicines that had been tested
contain the same ingredient which is the tea tree oil. Tea tree oil was known for its
bacteria, fungi, viruses, and mites (Larson et al,.2012). Besides, tea tree oil also had
11
been reported to induce contact sensitization and allergic contact dermatitis reactions
(Larson et al,.2012). From this previous study, all of the three medicine show effect
on S.aureus and E.coli but the different ingredients that mixed with the tea tree oil in
each of the pimple medicine gave different result where it can be seen on how big and
clear the inhibition zone on the agar plates of S.aureus and E.coli.
Besides, this study showed that the medicine A had the strong effect on S.aureus
because it has the ingredient that the both medicine B and medicine C do not have in
it which was the olive oil. Olive oil contains three major antioxidants which are
vitamin E, polyphenol, and phytosterols that help to prevents free radical damage to
the skin. Apart from that, olive oil also penetrates deeply into the skin while providing
12
4.1 Antimicrobial activity of comedonal acne medicine against Staphylococcus
aureus
1 - 25 mm 16 mm 16 mm
2 - 28 mm 14 mm 14 mm
3 - 37 mm 12 mm 10 mm
4 - 14 mm 9 mm 13 mm
Antimicrobial activities for inhibition zone of S. aureus using the three medicines
showed that medicine A had greatest inhibition zone towards S. aureus with the
average of ±26 mm, this was followed by both medicine B and medicine C with the
13
contained more effective ingredients than those present on all the three medicines and
1 - -
2 - - - -
3 - - - -
4 - - - -
Average - - - -
Antimicrobial activities for inhibition zone of E. coli using the three medicines was
tested to find about whether acne medicines are effective in inhibiting E. coli or it did
14
not give out any effect at all (negative result). The result obtained showed that all
result towards the test being carried on. This was probably because of the nature of E.
coli that was usually found in the intestinal tract and not on the skin surfaces. So, all
the acne medicines being tested do not affect the growth of E. coli. To conclude, the
three acne medicines do not affect the antimicrobial activities of E. coli as the result
Furthermore, in this study, there were many errors taking places including the
result with the fixed standard, and wrong selection of agar media (MacConkey agar)
causing unclear result of inhibition zone. The first error was the different
amount measurement was done to the medicines before applying it on each disc.
Then, the second error done in this study was no comparison of result with the fixed
standard. The result obtained should be compared with the standard before stating the
result to make sure the result did not out from the expected results. Lastly, the error
made in this study was the wrong selection of agar media as the result of the
inhibition zone obtained was not clear. The initial agar media chosen for E. coli was
MacConkey agar but after performing the test, the result was unfavourable as the
result of inhibition zone obtained was unclear and the measurement of inhibition zone
was not able to be done as the inhibition zone was unclear. So, the MacConkey agar
was replaced with Nutrient agar which was more suitable for clear result of inhibition
zone. In short, some error made was overcome by changing the material used and
some error was overcome by doing more readings and comparing the expected result.
15
Medicine B
Control
MacConkey agar
(Maroon colour)
Medicine C
Medicine A
16
CHAPTER 5
CONCLUSION
To conclude, the comedonal acne medicine that showed the greatest effect towards
Staphylococcus aureus was medicine A(spot away pimple gel).This was shown by the
final result obtained after performing the disk diffusion method. Medicine A (spot
away pimple cream) show the highest average for inhibition zone for Staphylococcus
gel) with the same inhibition zone (±13mm). In short, Medicine A has greater effect
for Staphylococcus aureus. This is because Medicine A contains olive oil that has
excellent spreading and skin absorption. It also prevents moisture loss and restores
skin suppleness which might be advantages among the other two acne medicines. This
is also due to medicine A contain malic acid that able to penetrate the skin more
deeply than the other two products, due to its chemical composition, and consequently
can greatly help in improving skin smoothness. As for Escherichia coli, it shows the
negative result. This is because Escherichia coli is a bacteria that found that
coli gram -negative bacteria and Staphylococcus aureus by measured the inhibition
zone. The antibacterial activity of comedonal acne medicine against Escherichia coli
17
REFERENCES
Bunikowski and Rita, et al.(1999) "Prevalence and role of serum IgE antibodies to the
Staphylococcus aureus–derived superantigens SEA and SEB in children with
atopic dermatitis." Journal of Allergy and Clinical Immunology 103(1)
Edward. (2015). Benefits of Olive Oil for Skin. Accessed 5 September 2016 from
www.globahealingcenter.com/natural-health/benefits-of-olive-oil-for-skin/
Kim, Seong-Jin and Young-Ho Won(1998) ‘The effect of glycolic acid on cultured
human skin fibreblasts: cell proliferative effect and increased collagen
synthesis’.The journal of dermatology 25(2) 85-89
Larson David Jacob and Sharon E. (2012). Tea tree oil. Volume 23 :48–49.
18
Mandal, A MD. Staphylococcus aureus. Accessed 15 August 2016 from
www.news-medical.net/health/What-is-Staphylococcus-Aureus.aspx
Marler Clark. (2016). E. Coli food poisoning. Accessed 15 August 2016 from
www.about-ecoli.com/
Williams, Hywel C., Robert P. Dellavalle, and Sarah Garner(2012) "Acne vulgaris.
The Lancet 379(9813)
Zaidan, M.R.S, Noor Rain, Badrul, A.R and Adlin A. (2005). In vitro screening of
five local medicinal plants for antibacterial activity using disc diffusion
method. Vol 22(2): 165–170
19
APPENDICES
20
Appendice (e): The experiment was conduct in the laminar floor
21
CURRICULUM VITAE
A. Personal
Citizenship: Bumiputera
Gender: Female
Address: No 232, lot 3, Taman Jumbo petagas. 88200, Kota Kinabalu Sabah
B. Academic qualifications
(PASS)
Rendah (PASS)
C. Related experiences
2. SUKANUN (2015)
22
A. Personal
Citizenship: Bumiputera
Gender: Female
B. Academic qualifications
Malaysia (PASS)
(PASS)
Rendah (PASS)
Rendah (PASS)
C. Related experiences
23
A. Personal
Citizenship: Bumiputera
Gender: Female
B. Academic qualifications
(PASS)
Rendah (PASS)
Rendah (PASS)
C. Related experiences
2. SUKANUN (2015)
24
A. Personal
Citizenship: Bumiputera
Gender: Female
Telephone no : 0168056208
B. Academic qualifications
Rendah (PASS)
Rendah (PASS)
C. Related experiences
2. SUKANUN (2015)
25
A. Personal
Citizenship: Bumiputera
Gender: Male
Address: Taman Indah Jaya, Batu 4, Lot AC1727 Blok 168, Lorong 7,
B. Academic qualifications
(PASS)
Rendah (PASS)
Rendah (PASS)
C. Related experiences
26
A. Personal
Citizenship: Bumiputera
Gender: Female
Address: Kampung Lahanas Jalan Kem Paradaise 89150 Kota Belud, Sabah
B. Academic qualifications
sabah (PASS)
Rendah (PASS)
C. Related experiences
2. SUKANUN (2015)
27