The use of traditional medicine to treat infection has been practiced since the origin of mankind, and honey produced by Apis mellifera is one of the oldest traditional medicines considered to be important in the treatment of several human ailments. However, differently processed honeys exhibit different antibacterial activities, thus this study was aimed at investigating the antibacterial activity of fortified honey and comparing the antibacterial activities of fortified with the unfortified and natural honeys on three clinical bacterial isolates (Escherichia coli, Staphylococcus aureus and Salmonella typhi). Confirmatory biochemical tests were carried out to identify these test organisms. The antibacterial activities of four samples of honey (honey fortified with ginger, honey fortified with lemon, unfortified pure honey and natural honey) on the test isolates were determined using the agar ditch diffusion method of sensitivity test as described by Cheesbrough (2006). This study revealed that fortified honey has increased antibacterial activity due to the presence of fortifying agents than the unfortified honey. The antibacterial activity of fortified honey depend on the amount or concentration of the fortifying agents present in the honey samples as honey fortified with ginger produced a contrary result; this implies that the fortifying agents may not necessarily be applied in honey to add to its antibacterial activity rather used as a flavoring agent.
TABLE OF CONTENTS
Title page………………………………………………………………………….….…..…i
Declaration ……………………………………………………….……………….….…….ii
Certification……………………………………………….……….………….…….….…..iii
Dedication………………………………………………………………....….…….…..…..iv
Acknowledgement………………………………………………………….….…….......….v
Abstract ………………………………………………………………………..……..…….vi
Table of contents…………..…………………………………………….………..….…….vii
List of tables…….…………….……….……………………………….……………….…viii
List of figures ……..……….………….………..……….……………..…………….........viv
CHAPTER ONE
1.0 Introduction……………….………..….……………….…..…………..…….……..1
1.1 Background Information……….……………..……………………………….……1
1.2 Justification…………………………...……………………………….…….....……2
1.3 Aim and objectives ……………………..…..………………….…….………….…..2
1.1 Research questions………………….……..……………………….……………..…3
1.4 Scope of the study………………………….…………..…..…….………………….3
1.5 Significance of the study……………………………………….…………….……...3
CHAPTER TWO
2.0 Literature review…………………………………….……….……………….…......4
2.1 Honey………………………………………….………………….….…….………..4
2.1.1 Use of honey in traditional medicine………….….……………..……….…….…....4
2.1.2 Use of honey in modern medicine……..….…..….…….………………………......5
2.1.3 Antibacterial properties of honey………….….….………..…..……………….…...6
2.1.3.1 Hydrogen peroxide……………….……….….……………………………………..7
2.1.3.2 High osmotic effect……………………………………………………..….……….8
2.1.3.3 Acidity………………..…………………………………………………………… 8
2.1.4 Composition of honey……………………….………….……………………..........9
2.2 Honey fortification ………..….………………..………………………….…..….....10
2.3 The fortifying agents …….……...…...……….……………………….…..….…….11
2.3.1 Ginger …………..…...………………………….………………..……………….….11
2.3.2 Lemon ………………………………………….……………….............................12
2.4 Essence of honey fortification………………….……………....……….…….........13
2.5 Profile of test organisms………………………….………………………….…..…13
2.5.1 Staphylococcus aureus…………………….…………………………….………....13
2.5.2 Escherichia coli ………………………….………………………………...............15
2.5.3 Salmonella typhi……………………………….…………….……………………..16
CHAPTER THREE
3.0 Materials and Method……………………………….…………………..…......... 18
3.1 Materials ………………………………………………………......….….…......... 18
3.1.1 Materials/Equipment/Reagents/Media…..…………………………………............18
3.1.2 Glass wares/Sterilization …….……………………..…….…..………....................18
3.1.3 Media preparation/ sterilization ….…………………….….…..….........................19
3.1.4 Source of honey samples……….………………………...……...……..................19
3.2 Test organisms …………………………………….……..……..…..…..................19
3.2.1 Sub culturing and purification of test organisms ………….......……...…......…… 20
3.3 Identification and confirmation of test organisms…………...……………..……. 20
3.3.1 Gram staining and microscopy……………………….……....………….……..... ..20
3.3.2 Biochemical/confirmatory tests…..……….………….………..………..………...... 21
3.3.2.1 Catalase test……..…….……….…….….……………..…….……….….…...…… 21
3.3.2.2 Coagulase test…………………….………………...………...……………......21
3.3.2.3 Indole test……………….……….……………..…...……….….…........……...22
3.3.2.4 Citrate utilization test….……………….………….……….….…….…..……...23
3.3.2.5 MIO test……………………………………….………………………………..23
3.3.2.6 Hydrogen sulphide production test…………….…………..............................24
3.3.2.7 Oxidase test……………………………………….………...............................24
3.3.2.8 Urease test…………………..……………………..……..................................25
3.3.2.9 Methyl red …….……….……………….……….… …….……..…….……….25
3.4 Preparation of stock and standard solution…….……..……………...................26
3.4.1 Preparation of 0.5 McFarland turbidity standard….………….….…..………...26
3.5 Standardization of Inocula……………………..………….………….………...26
3.6 Antibacterial sensitivity test…………………….………….…...........................27
3.6.1 Agar well diffusion method…………..………….……...…..…........................27
CHAPTER FOUR
4.0 Results …………………………………………….……..………….…….……...28
CHAPTER FIVE
5.0 Discussion, conclusion and recommendation.…..…………..….…….……….32
5.1 Discussion…………….………………………….…….…………...………..........32
5.2 Conclusion………………………………….……..………….……………………35
5.3 Recommendation…………………….………………..………….…………….....36
References….……………………………………………………………………...
