Final Preliminaries

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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



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



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



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



1.6 Conceptual Framework 6

1.7Scope and Delimitation 7

1.8Definition of Terms 8



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



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



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



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.



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



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



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



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)



CURRICULUM VITAE

Name: Loigie Loraine A. Crispo

Age: 20


Personal Profile:

Provincial Address: Townsite Parang, Maguindanao

City Address; General Luna St. Davao City

Birth date: July 4, 1991 Sex: Female





Academic Background

College: University of the Immaculate Conception (2008–present)

Secondary: Notre Dame of Parang (2003-2007)

Elementary: Notre Dame of Parang (1997–2003)



CURRICULUM VITAE

Name: Danneth Paderog

Age: 24


Personal profile:

Provincial Address: Andap New Bataan Comval Province

City Address: Buhangin Davao City

Birth date: October 13,1986 Sex: Female





Academic Background

College: University of the Immaculate Conception

Secondary: New Bataan National High School

Elementary: Cabinuangan Central Elemantary School



CURRICULUM VITAE

Name: Marwil A. Gayagas

Age: 20 years old


Personal Profile:

Provincial Address: Matalam North, Cotabato

City Address: Fr. Selga St. Davao City

Birth date: May 31, 1991 Sex: Male





Academic Background

College: University of Immaculate Conception (2008-Present)

Secondary: Matalam National High School (2004-2008)

Elementary: Linao Central Elementary School (1998-2004)



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



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)..



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


Pseudomonas aeruginosa

in MHB

Interpretation

Gentamycin (+ control)

Interpretation


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


Pseudomonas aeruginosa

in NSS

Interpretation

Gentamycin (+ control)

Interpretation


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



MHB

Interpretation


Interpretation


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



Results Using Clinical Isolates



MHB

Interpretation


Interpretation



Concentratio

n of Honey

Staphylococc

us aureus inNSS

Interpretatio

n

Penicillin ( +

control)

Interpretatio

n



Approved by:

Ms. Avee Joy B. Dayaganon, RMT, MS

Program Dean/Thesis Adviser



APPENDIX B

A. Preparation of Reagents and Media

B. Sample Honey



C. Susceptibility Testing



D. Negative Result of Antimicrobial Susceptibility Testing onStaphylococcus aureus

DI.

E. Negative Result of Antimicrobial Susceptibility Testing on Pseudomonas

aeruginosa



F. Negative Result of Antimicrobial Susceptibility Testing on Pseudomonasaeruginosa (Reference Strain)

G. Negative Result of Antimicrobial Susceptibility Testing on Staphylococccusaureus (Reference Strain)

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