final preliminaries
TRANSCRIPT
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 1/24
LIST OF FIGURES
Figure Title Page
1 Paradigm of the Study
7
2 Gram-stained Pseudomonas aeruginosa 17
3 Staphylococcus aureus 22
4 Methodological Framework of the Study 36
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 2/24
LIST OF TABLES
Table Title Page
1 The Difference between Manuka Honey and Regular Honey 14
2 Reference value for the zone of inhibtion of Gentamycin 27(+ control for Pseudomonas aeruginosa), Penicillin(+ control for Staphylococcus aureus) and Honey
2.1 Zones of Inhibition of the Different Concentration of Honey 37against (Reference Strain) Staphylococcus aureus
in Mueller-Hinton Broth
2.2 Zones of Inhibition of the Different Concentrations of Honey 38against Staphylococcus aureus (Reference Strain)in Normal Saline Solution
2.3 Zones of Inhibition of the Different Concentration of Honey 38against Pseudominas aeruginosa (Reference Strain)in Mueller- Hinton Broth
2.4 Zones of Inhibition of the Different Concentration of Honey 39
against Pseudominas aeruginosa (Reference Strain)in Normal Saline Solution
2.5 Zones of Inhibition of the Different Concentration of Honey 39against Staphylococcus aureus (Clinical Isolate)in Mueller- Hinton Broth
2.6 Zones of Inhibition of the Different Concentration of Honey 40against Staphylococcus aureus (Clinical Isolate)in Normal Saline Solution
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 3/24
ACRONYMS USED IN THE STUDY
CFU Colony Formation Unit
DNA Deoxyribonucleic acid
LPS Lipopolysaccharides
LOS Lipooligosaccharide
MHB Mueller Hinton Broth
MIC Minimum Inhibitory Concentration
MHA Mueller Hinton Agar
MRSA Methicillin Resistant Staphylococcus Aureus
mm millimeter
NaCl Sodium Chloride
W/v Weight over Volume
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 4/24
TABLE OF CONTENTS
Page Number
TITLE PAGE i
APPROVAL SHEET ii
ACKNOWLEDGEMENT iii
ABSTRACT iv
TABLE OF CONTENTS v
LIST OF TABLE vi
LIST OF FIGURES vii
LIST OF PLATES viii
Chapter
I. THE PROBLEM AND ITS SETTING
1.1 Introduction 1
1.2 Statement of the Problem 3
1.3 Null Hypotheses 4
1.4 Significance of the study 4
1.5 Theoretical Framework 5
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 5/24
1.6 Conceptual Framework 6
1.7Scope and Delimitation 7
1.8Definition of Terms 8
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 6/24
II. Review of Related Literature and Studies
2.1 Honey 9
2.2 Formation 11
2.3 Manuka Honey
2.4 Antibacterial Properties of Honey 14
2.5 The History of Medicinal Use of Honey 16
2.6 Apis negrocincta 16
2.7 Pseudomonas aeruginosa 17
2.7.1 Disease Association 192.7.2 Signs and Symptoms 20
2.8 Staphylococcus aureus 21
2.8.1 Disease Association 22
2.8.2 Duration of illness 24
2.5.4Transmission 24
2.9 Penicillin 25
2.9.1 Mode of Action 25
2.10 Gentamycin
2.10.1 Mode of Action 27
2.11 Related Studies 27
III. METHODOLOGY
3.1 Research Design 31
3.2 Research Locale 31
3.3 Research Procedure 32
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 7/24
IV. PRESENTATIONS, ANALYSIS and INTERPRETATION OF DATA 37
V. SUMMARY, FINDINGS and RECOMMENDATIONS 43
5.1 General Summary of the Study 43
5.1 Summary of Findings 44
5.2 Conclusion 45
5.3 Recommendations 46
REFERENCES 47
APPENDICES 51
CURRICUM VITAE
LIST OF PLATES
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 8/24
PLATES TITLE PAGE
A Preparation of Broth and Media 49
B Sample Honey 49
C Susceptibility Testing 50
D Negative Result of Antimicrobial Susceptibility 51
Testing on Staphylococcus aureus
E Negative Result of Antimicrobial Susceptibility 51
Testing on Pseudomonas aeruginosa
F Negative Result of Antimicrobial Susceptibility 52
Testing on Pseudomonas aeruginosa
(Reference Strain)
G Negative Result of Antimicrobial Susceptibility 52
Testing on Staphylococcus aureus
(Reference Strain)
ACKNOWLEDGEMENT
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 9/24
We would like to express our heartfelt gratitude to the following persons who have
shared the time, knowledge, and expertise throughout the conduct of this research:
To our family, for their constant moral and financial support; raised us, supported us,
taught us, and loved us. To them we dedicate this thesis
To our research professor and panel members Dr. Adorico M. Aya-ay, Mr. Ace Ronald
Sarabia, RMT, Mrs. Judee N. Nogodola, MS Bio, for their encouragement, insightful
comments, and for equipping us with knowledge and guided us in the course of our
study;
To our adviser Ms. Avee Joy B. Dayaganon RMT, MS for, sharing her time, knowledge,
and for motivating us to pursue this study;
To the UIC laboratory headed by Sister Ma. Matilde Omandac RVM for, allowing the
researchers to use the laboratory.
To Mrs. Florence Alzate, RMT, MS for, imparting her knowledge and skill during the
experiment and for her noteworthy advises in the making of the manuscript;
To Mr. Rigil Kent Batistil, Richard Catulong and Shiela Mae Dauz who assisted us in
performing the experiment especially in the preparation of materials needed.
To Mr. Ponciano Almonte, who helped us and supported in this study,
To the Almighty God who is our source of wisdom, without Him none of these would
have been possible.
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 10/24
To those who have contributed their effort, time, knowledge, skill and showed simple
acts of kindness that determined the success of this study, thank you very much.
M.B.II
L.L.A.C
M.A.G
D.C.P
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 11/24
ABTRACT
TITLE Antimicrobial Activity of Honey Against Staphylococcus
aureus and Pseudomonas aeruginosa
RESEARCHERS Breva, Mario II
Crispo, Loigie Loraine
Gayagas, Marwil
Paderog, Danneth
TYPE Experimental Type
ADVISER Avee Joy Dayaganon, RMT, MS
SUBJECT AREA Medical Bacteriology
CONFERRED DEGREE Bachelor in Medical Laboratory Science
Ayurveda a group of Indian surgeon describes honey as the nectar of life and
recommends its use in various ailments. The medicinal properties of honey have been
known since ancient times but there was no recognition of its antibacterial properties, it
was just known to be an effective remedy.
This study was undertaken to extend existing knowledge about the effectiveness of the
antibacterial properties of honey against Staphylococcus aureus and Pseudomonas
aeruginosa. In this study two organisms were used, a Staphylococcus aureus and
Pseudomonas aeruginosa, each bacterium was subjected for testing to different
concentration (40%, 60%, 80%, and 100%) of same type of honey. Each sample has
three replicates to test samples and examined under the supervision of our adviser.
Honey’s efficacy does not only depend on its high osmolality but it also depends on
what type of organism has been healed. The antibacterial action of honey against
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 12/24
Staphylococcus aureus relies on the capacity of bacteria to ferment d-gluconic acid
which is present in high concentration of honey.
The result showed that honey is not effective antibacterial agent against
Staphylococcus aureus. On the other hand antibacterial action of honey against
Pseudomonas aeruginosa does not depend on the fermentation process but it varies on
the ability of Pseudomonas aeruginosa to produce a large number of extracellular
protective and toxic substances. Some of these substances are slime glycolipoprotein,
DNAse, and phospholipase that contribute to the pathogenicity of this opportunistic
microorganism. The researchers highly recommend a more comprehensive study
involving the determination of efficacy of honey as an antimicrobial agent.
CURRICULUM VITAE
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 13/24
Name: Mario B. Breva II
Age: 19
Contact number: 09297142545
Personal Profile:
City Address: #5 Vermillion St. Spring Village Ma-a, Davao City
Birth date: September 17, 1991 Sex: Male
Nationality: Filipino
Religion: Roman Catholic Weight: 60 kg
Civil status: Single
Email Address: [email protected] Height: 5’5
Academic Background
College: University of the Immaculate Conception (2008-Present)
Secondary: University of the Immaculate Conception (2004-2008)
Elementary: University of the Immaculate Conception (1998-2004)
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 14/24
CURRICULUM VITAE
Name: Loigie Loraine A. Crispo
Age: 20
Contact number: 09072595624
Personal Profile:
Provincial Address: Townsite Parang, Maguindanao
City Address; General Luna St. Davao City
Birth date: July 4, 1991 Sex: Female
Nationality: Filipino
Religion: Roman Catholic Weight: 50 kg
Civil status: Single
Email Address: [email protected] Height: 5’1
Academic Background
College: University of the Immaculate Conception (2008–present)
Secondary: Notre Dame of Parang (2003-2007)
Elementary: Notre Dame of Parang (1997–2003)
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 15/24
CURRICULUM VITAE
Name: Danneth Paderog
Age: 24
Contact number: 09077960006
Personal profile:
Provincial Address: Andap New Bataan Comval Province
City Address: Buhangin Davao City
Birth date: October 13,1986 Sex: Female
Nationality: Filipino
Religion: Roman Catholic Weight: 47 kg
Civil status: Single
Email Address: [email protected] Height: 5’4
Academic Background
College: University of the Immaculate Conception
Secondary: New Bataan National High School
Elementary: Cabinuangan Central Elemantary School
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 16/24
CURRICULUM VITAE
Name: Marwil A. Gayagas
Age: 20 years old
Contact number: 09261271787
Personal Profile:
Provincial Address: Matalam North, Cotabato
City Address: Fr. Selga St. Davao City
Birth date: May 31, 1991 Sex: Male
Nationality: Filipino
Religion: Roman Catholic Weight: 62 kg
Civil status: Single
Email Address: [email protected] Height: 5’7
Academic Background
College: University of Immaculate Conception (2008-Present)
Secondary: Matalam National High School (2004-2008)
Elementary: Linao Central Elementary School (1998-2004)
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 17/24
APPROVAL SHEET
This thesis entitled “The Antimicrobial Activity of Honey against Staphylococcus
aureus and Pseudomonas aeruginosa” submitted by Mr. Mario B. Breva II, Ms.
Loigie Loraine A. Crispo, Mr. Marwil A. Gayagas, and Ms. Danneth C. Paderog in partial
fulfillment of the requirements for the degree of Bachelor of Science in Medical
Laboratory has been examined and is recommended for oral examination, acceptance
and approval.
Avee Joy B. Dayaganon, RMT, MS
Adviser
PANEL OF EXAMINERS
Adorico M. Aya-ay, Ph.D.
Chairman
Judee N. Nogodula, MS Bio Ace Ronald C. Sarabia, RMT
Member Member
Accepted and approved in partial fulfillment of the requirements for the degree of Master
of Science in Medical Laboratory.
Avee Joy B. Dayaganon, RMT, MS
Dean of Medical Laboratory Science Program
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 18/24
APPENDICES
Appendix A
Reagents and Media
A. 1 Preparation of Mueller Hinton Agar
Thirty four (34) grams of medium was dissolved in one liter demineralized water by
heating in a boiling water bath and autoclave for 15 minutes at 121°C (Forbes et al,
1998).
A. 2 Preparation of Mueller Hinton Broth
Twenty one (21) grams of Mueller Hinton Broth in 1 liter distilled water was mixed
and heated with frequent agitation and boiled for 1 minute, then sterilized to 121 °C for
15 minutes (Forbes et al, 1998).
A. 3 Preparation of 0.5 McFarland Standard
0.5 mL of 0.048 M BaCl2 (1.175% w/v BaCl2 H2O) was mixed with 99.5 mL 0.3%
N H2SO4 (1%v/v), 5 mL and distributed to sterile screw cap tubes, thightly sealed, and
stored in dark place at room temperature (Forbes et al, 1998)..
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 19/24
Results using Reference Strain
Concentration of Honey
Staphylococcus aureus in
NSS
Interpretation
Penicillin ( +control)
Interpretation
20% 6 mm Resistant 31 mm Susceptible
40% 6 mm Resistant 32.6 mm Susceptible60% 6 mm Resistant 29.6 mm Susceptible80% 6 mm Resistant 29 mm Susceptible100% 6 mm Resistant 31 mm Susceptible
Concentration of Honey
Pseudomonas aeruginosa
in MHB
Interpretation
Gentamycin (+ control)
Interpretation
20% 6 mm Resistant 15 mm Susceptible
40% 6 mm Resistant 15.5 mm Susceptible60% 6 mm Resistant 17 mm Susceptible
80% 6 mm Resistant 16 mm Susceptible100% 6 mm Resistant 15.7 mm Susceptible
Concentration of Honey
Pseudomonas aeruginosa
in NSS
Interpretation
Gentamycin (+ control)
Interpretation
20% 6 mm Resistant 16 mm Susceptible
40% 6 mm Resistant 17.5 mm Susceptible60% 6 mm Resistant 17 mm Susceptible80% 6 mm Resistant 15 mm Susceptible
100% 6 mm Resistant 16.5 mm Susceptible
Concentration of Honey
Staphylococcus aureus in
MHB
Interpretation
Penicillin ( +control)
Interpretation
20% 6 mm Resistant 29 mm Susceptible
40% 6 mm Resistant 31 mm Susceptible60% 6 mm Resistant 29.5 Susceptible80% 6 mm Resistant 32 mm Susceptible100% 6 mm Resistant 32.2 mm Susceptible
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 20/24
Results Using Clinical Isolates
Concentration of Honey
Staphylococcus aureus in
MHB
Interpretation
Penicillin ( +control)
Interpretation
20% 6 mm Resistant 31 mm Susceptible
40% 6 mm Resistant 32.6 mm Susceptible60% 6 mm Resistant 29.6 mm Susceptible80% 6 mm Resistant 29 mm Susceptible100% 6 mm Resistant 31 mm Susceptible
Concentratio
n of Honey
Staphylococc
us aureus inNSS
Interpretatio
n
Penicillin ( +
control)
Interpretatio
n
20% 6 mm Resistant 31 mm Susceptible
40% 6 mm Resistant 32.6 mm Susceptible60% 6 mm Resistant 29.6 mm Susceptible80% 6 mm Resistant 29 mm Susceptible100% 6 mm Resistant 31 mm Susceptible
Approved by:
Ms. Avee Joy B. Dayaganon, RMT, MS
Program Dean/Thesis Adviser
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 21/24
APPENDIX B
A. Preparation of Reagents and Media
B. Sample Honey
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 22/24
C. Susceptibility Testing
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 23/24
D. Negative Result of Antimicrobial Susceptibility Testing onStaphylococcus aureus
DI.
E. Negative Result of Antimicrobial Susceptibility Testing on Pseudomonas
aeruginosa
8/3/2019 Final Preliminaries
http://slidepdf.com/reader/full/final-preliminaries 24/24
F. Negative Result of Antimicrobial Susceptibility Testing on Pseudomonasaeruginosa (Reference Strain)
G. Negative Result of Antimicrobial Susceptibility Testing on Staphylococccusaureus (Reference Strain)