the prophylactic property of calabur on selected pathogenic bacteria (preliminaries)
TRANSCRIPT
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 1/25
i DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
The Prophylactic Property of Calabur ( Muntingia calabura) Extract on
Selected Pathogenic Bacteria
A Science Investigatory Project
Presented to
The Faculty of Science and Technology Department
Dagupan City National High School
Fourth Year
Proponent:
April Antonette F. Velasco
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 2/25
ii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
ABSTRACT
Title: The Prophylactic Property of Calabur (Muntingia calabura) Extract on
Selected Pathogenic Bacteria
Proponent: April Antonette F. Velasco
School: Science, Technology and Engineering (STE) Program – Dagupan City National
High School
The study entitled, “The Prophylactic Property of Calabur (Muntingia calabura)
Extract on Selected Pathogenic Bacteria” was conducted to determine if Calabur exhibits
a prophylactic property on Staphylococcus haemolyticus, Staphylococcus aureus and Escherichia coli. Specifically, the study aimed to determine the effect of Calabur on the
selected pathogenic bacteria based on the zone of inhibition (mm).
Specifically, the study sought to determine the prophylactic property of Calabur
leaves extract on the selected pathogenic bacteria based on their zones of inhibition. It
also sought to determine whether there is a significant difference between the effect of
Calabur leaves extract and the standard antibiotics used.
One hundred grams of Calabur leaves were gathered, washed with water and
soaked in 300ml ethyl alcohol for 24 hours. Then, the plant material was separated with
the extract. The obtained extracts were subjected to evaporation until the ethyl alcohol
has completely evaporated. The obtained pure extract undergone Antimicrobial
Sensitivity Testing.
Results revealed that Staphylococcus haemolyticus is moderately susceptible to
Calabur extract; susceptible to Vancomycin; and resistant to Oxacillin based on the
11.7mm, 12.6mm, and 0mm zones of inhibition respectively. Staphylococcus aureus is
intermediate to Calabur extract; intermediate to Vancomycin; and resistant to Oxacillin
based on the 10.9mm, 10.5mm and 0mm zones of inhibition respectively. Escherichia
coli is resistant to Calabur extract; and intermediate to Chloramphenicol based on the0mm and 10.0mm zones of inhibition respectively.
The researcher had concluded that Calabur leaves extract exhibits a prophylactic
property on Staphylococcus haemolyticus and Staphylococcus aureus. It was also found
out that there is no significant difference between the effect of Calabur leaves compared
with the effect of standard antibiotics used.
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 3/25
iii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
APPROVAL SHEET
The investigatory project entitled “The Prophylactic Property of Calabur
(Muntingia calabura) Extract on Selected Pathogenic Bacteria” prepared by April
Antonette F. Velasco in partial fulfillment of the requirements, has been examined and
approved by the Scientific Review Committee (SRC) on Oral Examination.
Adviser: ______________________
Critic Reader: ______________________
SRC: ______________________
______________________
Chairperson: ______________________
Accepted in partial fulfillment of the requirements of Research II.
Cherry A. Cayabyab ______________________
Head Teacher VI, Science
Erly G. Datario ______________________
Principal IV
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 4/25
iv DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Republic of the Philippines
Department of Education
Region I
Division of City Schools
DAGUPAN CITY NATIONAL HIGH SCHOOL
Dagupan City
RESEARCH PLAN (IA)
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 5/25
v DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
The Prophylactic Property of Calabur (Muntingia calabura ) onSelected Pathogenic Bacteria
A Research Plan Presented to the
Scientific Research Committee of the Science, Technology and Engineering Program
Dagupan City National High School
By:
April Antonette F. Velasco
June 24, 2013
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 6/25
vi DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Statement of the Problem
This study aims to determine the prophylactic property of Calabur extract on
selected pathogenic bacteria Staphylococcus haemolyticus, Staphylococcus aureus
and Escherichia coli. Specifically, it will seek to answer the following questions:
a) Does Calabur (Muntingia calabura) exhibit a prophylactic property based on
the zone of inhibition of the selected pathogenic bacteria (Staphylococcus
haemolyticus, Staphylococcus aureus and Escherichia coli)?
b) Is there a significant difference between the effect of Calabur (Muntingia
calabura) leaves extract on the selected pathogenic bacteria (Staphylococcus
haemolyticus, Staphylococcus aureus and Escherichia coli) compared to the
effect of the standard?
c) Among the selected pathogenic bacteria, on which of the following is Calabur
extract most effective based on their zones of inhibition?
Hypothesis
The hypothesis will be tested at 0.1 level of significance.
Ho – Calabur (Muntingi calabura) does not exhibit a prophylactic property based
on the zone of inhibition of the selected pathogenic bacteria (Staphylococcus
haemolyticus, Staphylococcus aureus and Escherichia coli).
Ho – There is no significant difference between the effect of Calabur (Muntingia
calabura) leaves extract on the selected pathogenic bacteria (Staphylococcus
haemolyticus, Staphylococcus aureus and Escherichia coli) compared to the
effect of the standard.
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 7/25
vii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
I. Materials and Equipment
Plant Collection and Extraction
Calabur leaves
beaker
graduated cylinder
funnel
weighing scale
300 ml 95% ethanol
filter paper
Erlenmeyer flask
parafilm
thin white cloth
Preparation of Culture Media and Sensitivity Test
Staphylococcus aureus
Escherichia coli
Staphylococcus haemolyticus
Mueller Hinton Agar plates and Nutrient Broth (NB)
forceps
cotton swabs
inoculating loop
standard (commercial antibiotic)
petri dish
alcohol lamp
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 8/25
viii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
A
• Gathering of Laboratory Equipment and Collection of PlantMaterial
B• Extraction of Plant Material
C• Collection and Preparation of Culture Media
D• Sterilization of Materials
E
• Subculture of Pathogenic Bacteria and Incorporation in NutrientBroth
F• Antimicrobial Sensitivity Test
G
• Computation of Antimicrobial Indices of the Extract on thePathogenic Bacteria
H• Interpretation of Results
I• Summary of Findings and Conclusion
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 9/25
ix DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
II. Extraction of Plant Material
The plant extract will be obtained by maceration method. The Calabur leaves
will be cut into smaller portions and will be placed inside an Erlenmeyer flask.
Ninety-five percent ethanol will be measured to 300 ml using the graduated cylinder
and is to be placed in the Erlenmeyer flask containing the Calabur leaves. The flask
will be covered with parafilm to prevent the evaporation of ethanol. The botanical
material will be soaked in ethanol for 24 hours at room temperature. The plant extract
will be obtained by separating the botanical material using a clean white cloth. Then,
the mixture will be filtered using the filter paper. The obtained extracts will then be
subjected to evaporation of ethanol to obtain the pure extract.
III. Collection and Preparation of Culture Media
The test organisms: Staphylococcus haemolyticus, Staphylococcus aureus and
Escherichia coli will be obtained and cultured at the medical laboratory science of
University of Pangasinan – PHINMA.
First, the inoculating loop and the opening of the screw cap tube containing
the nutrient broth will flame-sterilized using alcohol lamp to avoid contamination and
spread of fungus. The bacteria in nutrient broth will be inoculated across the surface
of the agar with the use of sterile inoculating loop. After this, the bacteria in the plate
will incubated for about 24 hours at 38 degree Celsius. The process will be done on
each of the selected pathogenic bacteria.
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 10/25
x DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
IV. Sterilization of Materials
The treatments, forceps, paper discs, and cotton swabs will be wrapped in
coupon bond and will be sterilized using an autoclave machine; the Mueller Hinton
Agar plates, Nutrient Broth (NB) will be refrigerated to avoid contamination and
growth of other microorganisms.
V. Subculture of Pathogenic Bacteria
Inoculating loop, alcohol lamp and Nutrient Broth (NB) will be prepared in
subculturing the test organisms. The inoculating loop and the opening of the screw
scrap tube containing the Nutrient Broth (NB) will be flame sterilized by the use of
alcohol lamp. The bacteria on the plate after incubation will be incorporated on the
Nutrient Broth (NB) with the aid of the inoculating loop. Right after this, subculture
of bacteria will be incubated prior to the sensitivity test.
VI. Antimicrobial Sensitivity Test
Paper disc method will be followed in conducting the sensitivity test. The
subculture of bacteria will be applied using sterile cotton swabs on the agar plates.
The plates will be incubated overnight and will be observed thereafter.
After incubation, the absence or presence of circular inhibition zones will be
used as bases in determining the activity of the sample being tested. The length of
zones of inhibition will be recorded and interpreted using “The Interpretative
Guidelines for Non-fastidious Aerobic bacteria Zone Diameter Interpretative
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 11/25
xi DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Standards” published in the book Introduction to Diagnostic Microbiology by Maria
Damnessa Delost. The results were given descriptive equivalents of Susceptible,
Moderately Susceptible, Intermediate and Resistant.
VII. ANTIMICROBIAL INDEX OF THE EXTRACT
For the antimicrobial index of the extract, the following formula will be used:
Antimicrobial Index =
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 12/25
xii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Bibliography
Madrid, Domingo A., A Dictionary of Philippine Plant Names; pg. 58
Delost, Maria D., Introduction to Diagnostic Microbiology; pg. 72-73
Rummel, Dietmar J., Useful Plants of the Philippines Vol. 1
Philippine Medicinal Plants – Aratiles/Calabur;
www.stuartxchange.com/Aratiles.html
Staphylococcus aureus, www. wikipedia.com
Zone of Inhibition test: Antimicrobial activity screening;
www.antimicrobialtestlaboratories.com
Staphylococcus aureus, Escherichia coli; Todar’s Online Textbook of
Bacteriology; 2011
Staphylococcus haemolyticus, www.microbewiki.kenyon.edu (July 15, 2013)
Staph infections: Treatments and drugs, www.mayoclinic.com (July 3, 2013)
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 13/25
xiii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
CONTENT
Title Page i
Abstract ii
Approval Sheet iiiResearch Plan ivContent xiii
CHAPTER 1 - INTRODUCTIONBackground of the Study 1
Statement of the Problem 2
Statement of Hypothesis 2
Significance of the Study 3Scope and Delimitations 3
Definition of Terms 4
CHAPTER 2 - REVIEW OF RELATED LITERATURE
Related Literature 5
Related Studies 8
Related Readings 10Relevance of the Study 12
CHAPTER 3 - METHODOLOGY
Materials Used in the Experiment 13
Procedures 14
CHAPTER 4 - RESULTS AND DISCUSSION 17
CHAPTER 5 - SUMMARY, CONCLUSION AND RECOMMENDATIONS
Summary of Findings 19
Conclusion 21Recommendations 21
Bibliography xivPlates xv
Appendices
Antimicrobial Computation xix
Acknowledgment xxv
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 14/25
xiv DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
BIBLIOGRAPHY
Books
Madrid, Domingo A., A Dictionary of Philippine Plant Names; pg. 58
Rummel, Dietmar J., Useful Plants of the Philippines Vol. 1
Internet Sources
Philippine Medicinal Plants – Aratiles/Calabur;
www.stuartxchange.com/Aratiles.html
Republic Act No. 9502 “Universally Accessible Cheaper and Quality Medicines
Act of 2008”; www.lawphil.net
Staphylococcus aureus, www. wikipedia.com
Zone of Inhibition test: Antimicrobial activity screening;
www.antimicrobialtestlaboratories.com
Staphylococcus aureus, Escherichia coli; Todar’s Online Textbook of
Bacteriology; 2011
Staphylococcus haemolyticus, www.microbewiki.kenyon.edu (July 15, 2013)
Staph infections: Treatments and drugs, www.mayoclinic.com (July 3, 2013)
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 15/25
xv DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
PLATES
Preparation of Extract and Subculture of Pathogenic Bacteria
Materials used for the extraction of plantmaterial. Soaking of Calabur leaves in 95%ethanol in an Erlenmeyer flask
covered with parafilm.
Obtained extract is filtered using filter
paper.
Extract is subjected to water bathuntil all of the ethanol has
evaporated.
The obtained pure extract
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 16/25
xvi DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Paper discs are soaked in the pure extractand placed in hot air oven at 00 C for 10
minutes to dry.
Agar is prepared to be placed in
petri dishes.
Petri dishes were labeled for the different bacteria.
Subculture of bacteria is inoculated into the
nutrient broth and is incubated overnight.
ot air oven at 00 C is used to drythe paper discs with pure extract for
10 minutes.
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 17/25
xvii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Standard antibiotic is prepared for each
bacteria. Vancomycin 30mcg andOxacillin 1mcg for Staph. aureus and
Staph. haemolyticus; Chloramphenicol
30mcg for E. coli.
The nutrient broth containing
the bacteria after incubation.
The bacteria in the nutrient broth will beaseptically swab on the agar plates usingsterile cotton swabs. The extract paper
discs and positive control will be
incorporated on the agar. The plates will
be incubated for 24 hours prior tointer retation of results.
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 18/25
xviii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Antimicrobial Susceptibility Test Results
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 19/25
xix DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 20/25
xx DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 21/25
xxi DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 22/25
xxii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
APPENDICES
Computation of Data
Staphylococcus haemolyticus
Antimicrobial Index of Calabur leaves extract Zone of Inhibition: 11.7 mm
Against Staphylococcus haemolyticus
Antimicrobial Index =
=
= 0.93
Antimicrobial Index of Vancomycin 30mcg Zone of Inhibition: 12.6 mm
Against Staphylococcus haemolyticus
Antimicrobial Index =
=
= 1.00
Antimicrobial Index of Oxacillin 1mcg Zone of Inhibition: 0 mm
Against Staphylococcus haemolyticus
Antimicrobial Index =
=
= 0
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 23/25
xxiii DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Staphylococcus aureus
Antimicrobial Index of Calabur leaves extract Zone of Inhibition: 10.9 mm
Against Staphylococcus aureus
Antimicrobial Index =
=
= 1.04
Antimicrobial Index of Vancomycin 30mcg Zone of Inhibition: 10.5 mm
Against Staphylococcus aureus
Antimicrobial Index =
=
= 1.00
Antimicrobial Index of Oxacillin 1mcg Zone of Inhibition: 0 mm
Against Staphylococcus aureus
Antimicrobial Index =
=
= 0
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 24/25
xxiv DAGUPAN CITY NATIONAL HIGH SCHOOL
Science, Technology and Engineering (STE) Program
School Year 2013 - 2014
Escheri chia coli
Antimicrobial Index of Calabur leaves extract Zone of Inhibition: 0 mm
Against Escherichia coli
Antimicrobial Index =
=
= 0
Antimicrobial Index of Chloramphenicol 30mcg Zone of Inhibition: 10 mm
Against Escherichia coli
Antimicrobial Index =
=
= 1.00
7/23/2019 The Prophylactic Property of Calabur on Selected Pathogenic Bacteria (Preliminaries)
http://slidepdf.com/reader/full/the-prophylactic-property-of-calabur-on-selected-pathogenic-bacteria-preliminaries 25/25
xxv DAGUPAN CITY NATIONAL HIGH SCHOOL
ACKNOWLEDGMENT
I would like to extend my profound gratitude and appreciation to the following
who extended their generous support and contribution allowing the completion of this
study:
Mr. and Mrs. Pilar F. Velasco, my parents, for all their unwavering concern,
financial and moral support, and understanding;
Mrs. Monica E. Cayabyab, my mentor, who guided me throughout the research
process, unselfishly imparted her ideas, time and knowledge and for all her valuable
suggestions, comments and contributions in every aspect of the study which inspired me
to make this study as successful as it can be;
Mrs. Cherry A. Cayabyab, science department head of DCNHS, for letting me
borrow laboratory materials for the extraction process of my project;
Dr. Hannah A. Balanon, head of the medical technology department of UPang-
PHINMA, for allowing me to use their laboratory and facilities in the conduct of my
experiment.
Mr. David Contad, for his assistance in sub-culturing the bacteria and the
experiment in general;
The Almighty God, for the wisdom, strength, and vision he has given me to be
able to create this masterpiece;
And for those whose names were not mentioned who had helped me in a way or
another in making this science investigatory project possible.