ANTIBACTERIAL ACTIVITY OF AQUEOUS AND METHANOL EXTRACT OF MORINGA OLEIFERA LEAVES ON SOME BACTERIAL PATHOGENS

ABSTRACT

This study was carried out to investigate the antibacterial potential of the aqueous and methanol extract of Moringa oleifera leaves  on some target organisms. This study was carried out using the aqueous and methanol extracts of Moringa oleifera leaves. The Both extracts were used to test for antibiotics sensitivity using the agar well diffusion method. The organisms (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Salmonella typhi) were inoculated in a prepared Mueller Hinton Agar by spread method and a cork borer of diameter 6mm was used to make wells on the agar.`The results evaluated by zones of inhibition showed that  methanolic extract at 500mg/ml, 250mg/ml and 125mg/ml were  inhibitory to staphylococcus arueus (12.0+1.41mm, 10.5 + 0.71mm, 8.0+0.0mm),  Escherichia  coli (11.5+0.71mm, 9.5 + 0.71mm, 7.5+0.71mm),respectively. Salmonella typhi was inhibited at 500mg/ml and 250mg/ml only with inhibition zone of (11.0+1.41mm and 8.5 + 0.71mm) respectively. Pseudomonas aeruginosa and Enterococcus faecalis were the least susceptible. Inhibition was observe at concentration of 500mg/ml with the zone of inhibition at (9.5+0.71mm and 9.0+0.0mm), respectively. The aqueous extract showed inhibition only at concentration of 500mg/ml on  staphylococcus aureus (9.0+1.41mm), Escherichia coli (10.5+1.41) and Salmonella typhi (8.5+0.71mm) whereas, Pseudomonas aerouginosa and Enterococcus faecalis were not susceptible. Minimum inhibitory concentration was also carried out using the Mueller Hinton broth and the control was Gentamycin. The result obtained for aqueous extract, MIC/MBC was at 250mg/ml  concentration for other isolates except E. coli that had MIC at 125mg/ml, and its MBC at 250mg/ml. For the methanolic extract, MIC and MBC result for all isolates were 125mg/ml  and 250mg/ml  respectively. The results revealed that the methanol extract has more inhibitory effect than the aqueous extract. The phytochemical ansalysis result revealed that Alkaloid, Saponin, Terpenoid, and Flavonoid are in the composition of 0.96%, 1.84%, 1.69%, and 0.46% respectively. The consequences of this study suggests that Moringa oleifera  leaves has a bactericidal potential which can be used to discover antibacterial agent for developing new pharmaceuticals to control studied bacterial pathogens responsible for illnesses.

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                            ii

Dedication                                                                                                                              iii

Acknowledgements                                                                                                                iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          vi

Abstract                                                                                                                                  viii

CHAPTER ONE

Introduction                                                                                                                            1

1.1  Background of Study                                                                                                       1

1.2  Aims and Objective                                                                                                          2

CHAPTER TWO

2.0  Literature Review                                                                                                                        3

2.1 Moringa Tree                                                                                                                     3

2.2 Moringa-the Miracle Tree                                                                                                 4

2.3 Traditional, medicinal and non-medicinal use of Moringa                                               6

2.4  Phytochemical Constituents of Moringa                                                                         8

2.5 Antibacterial Activity of Moringa                                                                                    8

CHAPTER THREE

MATERIALS AND METHODS                                                                                          11

3.1 Collection of Plant Material                                                                                              11

3.2 Preparation of Dried Leaf Powder                                                                                   11

3.3  Aqueous Extraction of Leaf Powder                                                                               11 

3.4  Methanol Extraction of Leaf Powder                                                                              11

3.5  Test Organism                                                                                                                  12

3.6  Preparation of Stock Solution of Extract                                                                        12

3.6.1  Preparation of Culture Media                                                                                       12

3.7  Reactivation of Stock Culture of Test Organisms                                                           12

3.8 Antibacterial Testing using the Agar well Diffusion Method                                          13

3.8.1 Determination of Minimum Inhibitory Concentration                                                  13

3.8.2 Determination of Minimum Bactericidal Concentration                                               14

3.9 Determination of Phytochemical Composition of Moringa olefiera leaves                      15

3.9.1 Alkaloids                                                                                                                        15

3.9.2 Terpenoid                                                                                                                       15

3.9.3 Flavonoid                                                                                                                       15

3.9.4 Saponin                                                                                                                          16

CHAPTER  FOUR

RESULTS                                                                                                                               17

CHAPTER FIVE

5.1     Discussion                                                                                                                     23

5.2     Conclusion                                                                                                                    25

References

LIST OF TABLES

Table                                                                                                                                       pages

1a Mean diameter zone of inhibition (mm) produced by Aqueous  extracts of

 Moringa oleifera  leaves.                                                                                                        18

1b.Mean diameter zone of inhibition (mm) produced by Methanol extract of

 Moringa oleifera leaves .                                                                                                        19

2a MIC and MBC values of Aqeous extract of Moringa oleifera

Lam leaves against test organisms.                                                                                         20

2b MIC and MBC values of methanol extract of Moringa oleifera

Lam leaves against test organisms.                                                                                         21

3 Quantitative Phytochemical Composition of Moringa oleifera leaves.                               22

CHAPTER ONE

INTRODUCTION

1.1 Background of the study

The frequency of life threatening infectious diseases caused by pathogenic microorganisms has increased worldwide and is becoming an important cause of morbidity and mortality among immune compromised  patients in developing countries (Al-Bari et al., 2006).

 The increasing prevalence of multi-resistant strains of bacteria and the recent appearance of strains with reduced susceptibility to antibiotics raised the specter of untreatable bacterial infections and added urgency to the search for new infection fighting strategies (Zye et al., 2005; Rojas et al 2006). Since human civilization, medicinal plants have been used by mankind for its therapeutic value, nature has been a source of medicinal agents for thousand years and an impressive number of modern drugs have been isolated from natural sources. Many of these isolations were based on the uses of the agents in traditional medicine. The plant based traditional medicine systems continues to play an essential role in health care, with about 80% of the world’s inhabitants relying mainly on traditional medicine for their primary health care (Owolabi et al., 2007). Medicinal plants are plants containing inherent active ingredients used to cure diseases or relieve pain (Okigbo et al., 2008).

The medicinal properties of plants could be based on the antioxidant, antimicrobial, antipyretic effects of the phytochemicals in them (Cowman, 1999; Adesokan et al., 2008). The practice of complementary and alternative medicine is now on the increase in developing countries in response to world health organization directive culminating in several preclinical and clinical studies that have provided the scientific basis for the efficacy of many plants used in folk medicine to treat infections (Vijaya and Anathan, 1997; Dihuydy and Patients, 2003).

Despite the existence of potent antibiotics and antifungal agent, resistant or multi-resistant strains are continuously appearing imposing the need for a permanent search and development of new drugs (Silver, 1993). It is therefore, very necessary that the search for newer antibiotic source be a continuous process. plants are the cheapest and safer alternatives and sources of antimicrobials (Pretorius and Watt 2001, Shariff and Banik, 2006; Doughari et al., 2007).

 Hence, this present study is under taken specifically to investigate the role of aqueous methanol extracts of Moringa oleifera leaves as potential antimicrobial agent against bacterial pathogens.

1.2 Aims and objectives of the study

1.     To evaluate the methanolic and aqueous extract of Moringa oleifera for its antibacterial activity on some selected bacterial pathogens .

2.     To determine the minimum inhibitory concentration and minimum bactericidal concentration of the extract against the test organism.

3.     To determine the Phytochemical Composition of Moringa oleifera.

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