Physicochemical and Phytochemical Analysis of Honey and Shea Butter Samples and Their Antibacterial Effects on Staphylococcus aureus AND Klebsiella pneumoniae

ABSTRACT

This research work was carried out to examine the physicochemical and phytochemical constituents of honey and Shea butter samples respectively and their antibacterial effect on Staphylococcus aureus and Klebsiella pneumoniae. The physicochemical screening of honey showed that the honey samples used have low water activity, low moisture content, low pH value below 4.0 and Ash while the phytochemical analysis of Shea butter revealed the presence of Saponnins, Tannins, Alkaloids, Steroid and Phenol. The organisms Staphylococcus aureus and Klebsiella pneumoniae were isolated and identified by standard microbiological procedures. The antibacterial screening protocol was carried out by disc diffusion method. The result obtained showed that the honey alone was able to give an inhibition zone that is up to 29mm and the Shea butter gave up to 26mm. The synergistic effect of honey and Shea butter showed higher inhibitory effect of 42mm at 100% concentration. The standard antibiotics; Ciprofloxacin and Streptomycin had the highest mean of 36mm. The synergism is hereby recommended for its high antibacterial properties for therapeutic purposes and can be used as an alternative medicine against Staphylococcus aureus and Klebsiella pneumoniae infections. It may be even more conventional to use this natural means as it has very low antibiotic resistance compared to synthetic antibiotic.

TABLE OF CONTENT

Title page ﾿ i

Certification ﾿ ii

Dedication ﾿ iii

Acknowledgement ﾿ iv

Table of Content ﾿ v

List of tables ﾿ vi

Abstract ﾿ x

CHAPTER ONE ﾿ 1

1.0 INTRODUCTION ﾿ 1

 Aims and Objectives ﾿ 2

CHAPTER TWO ﾿ 3

2.0 LITERATURE REVIEW ﾿ 3

2.1 Honey ﾿ 4

2.1.1 Extraction ﾿ 6

2.1.2 Local test for real honeys ﾿ 6

2.1.3 Classification of honey ﾿ 7

2.1.3.1 Crystallized honey ﾿ 7

2.1.3.2 Pasteurized Honey ﾿ 7

2.1.3.3 Raw honey ﾿ 7

2.1.3.4 Ultrasonicated honey ﾿ 7

2.1.4 Mode of action of some of the antibacterial substances in honey. ﾿ 7

2.1.4.1 High osmotic pressure ﾿ 8

2.1.4.2 Low water activity ﾿ 8

2.1.4.3 Glucose oxidase enzyme ﾿ 8

2.1.4.4 Low pH/ acidic environment ﾿ 8

2.2 Medical Uses of Honey ﾿ 8

2.2.1 Remedy for Diarrhea ﾿ 8

2.2.2 Gastritis/gastric Ulcers ﾿ 9

2.2.3 Wound healing ﾿ 9

2.2.4 Treatment for arthritis: ﾿ 9

2.2.5 For post-operative wound infection: ﾿ 9

2.2.6 Adverse reaction of honey ﾿ 10

2.3 Shea Butter Tree ﾿ 10

2.3.1 Extraction of Essential Oil from Shea Kernel ﾿ 11

2.3.2 Composition of Shea butter ﾿ 12

2.3.3 Shea Butter Phenolics ﾿ 12

2.3.4 Therapeutic uses of Shea Butter ﾿ 12

2.3.5 General uses of shea butter ﾿             13 ﾿

2.4 Antimicrobial Agent ﾿ 13

2.5 Antimicrobial synergy ﾿ 14

2.6 Antimicrobial Susceptibility and Resistance ﾿ 15

 2.7 An overview of the test organisms ﾿ 15

2.7.1 Staphylococcus aureus ﾿ 15

2.7.2 Klebsiellapneumoniae ﾿ 16

CHAPTER THREE

3.0 MATERIALS AND METHODS ﾿ 17

3.1 Collection of test samples ﾿ 17

3.1.1 Collection of clinical samples ﾿ 17

3.2 Physicochemical Analysis of Honey ﾿ 17

3.2.1 Colour determination ﾿ 17

3.2.2 PH determination ﾿ 17

3.2.3 Moisture content ﾿ 18

3.2.4 Ash content ﾿ 18

3.2.5 Water activity ﾿ 18

3.3 Phytochemical Analysis of  Shea Butter ﾿ 18

3.3.1 Detection of alkaloids ﾿ 18

3.3.2 Detection of flavonoids ﾿ 19

3.3.3 Detection of saponins ﾿ 19

3.3.4 Detection of tannins ﾿ 19

3.3.5 Detection of phenol and polyphenol ﾿ 19

3.3.6 Detection of steroids ﾿ 19

3.4 Preparation of media ﾿ 20

3.5 Isolation and identification of bacteria from upper respiratory tract ﾿ 20

3.5.1 Gram staining ﾿ 20

3.5.2 Indole Test  ﾿ 21

3.5.3 Catalase Test ﾿ 21

3.5.4 Citrate test ﾿ 21

3.5.5 Oxidase test ﾿ 21

3.5.6 Coagulase test               

3.6 Antibacterial activity testing ﾿ 22

3.6.1 Preparation of test samples ﾿ 22

3.6.2 Sensitivity test ﾿ 23

3.7 Synergistic antimicrobial assays ﾿ 24

3.8 Determination of Minimum Inhibition Concentration (MIC) ﾿ 25

3.9 Determination of Minimum Bactericidal Concentration ﾿ 25

CHAPTER FOUR

RESULTS ﾿ 26

CHAPTER FIVE

5.0 DISCUSION, CONCLUSION AND RECOMMENDATION ﾿ 35

5.1 Discussion ﾿ 35

5.2 Conclusion ﾿ 37

5.3 Recommendation ﾿ 37

REFERENCES ﾿ 38

LIST OF TABLES

Table 1 Composition of honey ﾿ 5

Table 2 Physico-chemical analysis of honey ﾿ 28

Table 3 Phytochemical analysis of shea butter 29

Table 4 Identification of Staphylococcus   aureusand Klebsiella pneumonia ﾿ 30

Table 5 ﾿ Effects Of Honey And Shea Butter On Staphylococcus Aureus ﾿ 31

Table 6 ﾿ Antibiotics used on Staphylococcus aureus ﾿ 32

Table 7 ﾿ Effects Of Honey And Shea Butter On Klebsiella pneumonia ﾿ 33

Table 8 ﾿ Standard Antibiotics used on Klebsiella pneumoniae ﾿ 34