ANTIBACTERIAL ACTIVITIES OF VARIOUS TYPES OF HONEY

ABSTRACT

The study evaluated the antibacterial activity of various concentrations of two honey varieties (Umudike honey and Nsuka honey) against three (3) wound infection isolates; Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Test organisms were obtained from the Microbiology laboratory of the Federal Medical Centre, Umuahia and confirmed by standard techniques based on their morphological appearance, Gram staining reaction, motility and biochemical characteristics. Antibacterial activity of the honey samples against the test isolates were assessed using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and inhibitory zone diameter (IZD) at dilutions of 100%, 50%, 25%, 12.5%, 6.25% and 3.125% respectively. Both honey varieties produced observable in-vitro antibacterial effects on all the organisms at 100% and 50%. Nsuka honey produced better antibacterial effects against the test isolates than the Umudike honey.  The MIC of the Umudike and Nsuka honey ranged from 25-50% and 12.5 - 25% respectively. While the MBC of the Umudike and Nsuka honey ranged from 50-100% and 25-50% respectively. The study concluded that honey even at diluted strength can be used in the treatment of skin infections (burns, scalds and wounds) especially those associated with Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli.

TABLE OF CONTENTS

Cover page                                                                                                                              i

Title page                                                                                                                                ii

Dedication                                                                                                                              iii

Certification                                                                                                                            iv

Acknowledgement                                                                                                                  v

Table of contents                                                                                                                    vi

List of tables                                                                                                                           viii

Abstract                                                                                                                                  x

CHAPTER ONE

1.0  Introduction                                                                                                                1

1.1 Aim of study                                                                                                                     2

CHAPTER TWO

2.0 Review of literature                                                                                                          3

2.1 Historical uses of honey                                                                                                   3

2.2 Economics                                                                                                                        5

2.3 Composition of honey                                                                                                      5

2.3.1 Carbohydrates                                                                                                                5

2.3.2 Proteins, enzymes and amino acids                                                                               6

2.3.3 Vitamins, minerals and trace compounds                                                                      7

2.3.4 Aroma compounds, taste-building compounds and polyphenols                                    8

2.3.5 Free Radicals                                                                                                                 8

2.4 Antioxidants                                                                                                                     10

2.5 Mode of action of antioxidants                                                                                         12

2.6 Health effects of honey                                                                                                     12

2.6.1 Anti-inflammatory effects                                                                                             12

2.6.2 Immunoactivating and immunosuppressive properties                                                 13

2.6.3 Gastroenterological effects                                                                                            13

2.6.4 Cardiovascular effects                                                                                                   14

CHAPTER THREE

3.0       Materials and methods                                                                                                15

3.1       Study area                                                                                                                   15

3.2       Materials                                                                                                                     15

3.3       Sample collection                                                                                                       15

3.4       Test organisms                                                                                                            15

3.5       Confirmation of test isolate                                                                                        15

3.5.1   Morphological appearance                                                                                          16

3.5.2   Gram staining                                                                                                              16

3.5.3   Motility test                                                                                                                 16

3.5.4   Biochemical tests                                                                                                         17

3.6       Preparation of honey samples                                                                                    19

3.7       Antimicrobial susceptibility testing                                                                           19

3.7.1    Agar-well diffusion method                                                                                       19

3.7.2    Minimum inhibitory concentration (MIC) and minimum bactericidal

concentration (MBC) test                                                                                           20

CHAPTER FOUR

4.0       Result                                                                                                                          21

CHAPTER FIVE

5.0 Discussion, conclusion and recommendation                                                                  30

5.1 Discussion                                                                                                                         30

5.2 Conclusion                                                                                                                        32

5.3 Recommendations                                                                                                            32

REFERENCE

CHAPTER ONE

1.0  INTRODUCTION

The continuous use of antibiotics in clinical practice has been the direct cause of the development of multiple antibiotic resistances among bacteria causing human infection (Mandal et al., 2002). To combat such bacterial resistance to antibiotic, scientists discovered natural sources like medicinal plants of non-antibiotic drugs having antibacterial potentiality (Mandal et al., 2010, Amin et al., 2010, Kumar et al., 2009).

Beside the medicinal plants, the antibacterial activity of honey against many different life threatening bacteria has been reported (Tan et al., 2009, Kingsley, 2001 and Chambers, 2006). Wilkinson (2005) determined the activity of 13 different honey samples, including three commercial antibacterial honeys, against Escherichia coli and Pseudomonas aeruginosa. It has been reported that honey showed both bacteriostatic and bactericidal effect against gram positive as well as gram-negative bacteria, and also exhibited anti-fungal activity (Basson and Grobler, 2008 and Nasir et al 2010). Chauhan et al., (2010) reported that the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of honey ranged 0.625-5.000 mg/mL for the clinical isolates of E. coli, P. aeruginosa and Salmonella enterica serovar Typhi (S. enterica serovar Typhi). Another study revealed honey MIC 11% for Pseudomonas isolates (Mullai and Menon, 2005).

Moreover, honey represents the oldest traditional medicines in the treatment of respiratory ailment, gastrointestinal infection and various other diseases. It is being used effectively as a dressing for wounds (including surgical wounds), burns, and skin ulcers to reduce pain and odour quickly. Honey has been reported to maintain moist wound environment that promotes healing, and its high viscosity helps to provide a protective barrier to prevent infection; in addition, the mild acidity and low-level hydrogen peroxide release help in tissue repairing and contribute to the antibacterial activity (Lusby et al., 2005). The study is aimed at evaluating the in-vitro antibacterial activity of two types of against clinical isolates of Staphylococcus aureus, E. coli, and P. aeruginosa.

1.1 AIM OF STUDY

The aim of the study is to evaluate the antibacterial activity of two different honeys on clinical isolates of Staphylococcus aureus, E. coli, and P. aeruginosa.

The specific objectives of the study are;

i.               To determine the Minimum inhibitory concentration of honey

ii.              To determine the Minimum bacteriocidal concentration of honey

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