ANTIMICROBIAL ACTIVITIES OF OCIMUM GRATISSIMUM (NCHONWU). GONGRONEMA LATIFOLIUM (UTAZI) AND VERNONIA AMYDALINA (ULUBIRI) ON SOME SPECIES OF PATHOGENIC BACTERIA.

ABSTRACT

The emergence of antibiotic resistance as well as the recent undesirable side effect of some of the commercially available antibiotics has led to the screening of plant extract in order to discover new drug that could serve as alternative therapy for the treatment of various infections and diseases. Fresh leaf of Ocimum gratissimum (scent leaf), Gonronema latifoliun(utazi)and Vernonia amydalina (bitter leaf) were collected  air-dried at room temperature and blended to powder using electric blender. The extraction was done using reflux extraction method using ethanol and water  as solvent. The antibacterial activities of the test plants were determined to ascertain the different phytochemicals present in the plant extract. The extract was also tested against some selected Gram negative and Gram positive intestinal pathogenic bacteria; Escherichia coli, Salmonella species,Staphylococcus aureus and Enterococci faecalis  by reconstituting the extract  to obtain different concentration (100,50 and 25 g/ml) and agar well diffusion techniques were used to evaluate the antibacterial susceptibility of the leaf extract. The result of antibacterial analysis showed that the extract of O. gratissimum and V. amydalina has antibacterial activity against E. coli and Salmonella while the extract of G.latifolium has anti microbial activity against S.aureus and E.faecalis. This could be as a result of the presence of various phytochemicals or the interaction of one or more of the identified metabolites against the test organisms. The results obtained from this research, suggest that Escherichia coli and salmonalla was susceptible to the leaf extract of  O.gratissimum and V.amydalina while S.aureus and E.faecalis is susceptible to G.latifolium. Generally the ethanolic extracts of the plant materials shows a higher antibacterial activity than the aqueous extracts.

TABLE OF CONTENTS

Title page                                                                                                                    i

Certification                                                                                                               ii

Dedication                                                                                                                  iii

Acknowledgement                                                                                                      iv

Abstract                                                                                                                      v

Table of content                                                                                                          vi

List of tables                                                                                                               vi      

CHAPTER ONE      

1.0         Introduction                                                                                                                         1

1.1         Aim and Objectives                                                                                                             3

CHAPTER TWO

2.0         Literature Review                                                                                                                4

2.1         Description of Test Materials                                                                                            4

2.1.1     Ocimum gratissimum                                                                                                             4

2.1.2     Vernonia amydalina                                                                                                            7

2.1.2     Gongronema latifolium                                                                                                       7

2.2         Description of Test Organisms                                                                                          8

2.2.1     Escherichia coli                                                                                                                     8

2.2.2     Staphylococcus aureus                                                                                                        9

2.2.3     Salmonella spp                                                                                                                     10

2.2.4     Enterococci faecalis                                                                                                             11

CHAPTER THREE

3.0         Methods and Materials                                                                                                                 13

3.1         Collection and Identification of plant materials                                                                         13

3.2         Materials and reagents                                                                                                                   13

3.3         Preparation of plant extract                                                                                                          13

3.3.1     Preparation of aqueous                                                                                                                   13

3.3.2     Preparation of ethanolic extract                                                                                                     14

3.4         Test for purity and sterilization                                                                                                       15

3.5         bacterial species confirmation                                                                                                        15

3.6       Gram stain                          15

3.7        Biochemical test                                                                                  16

3.7.1     Catalase test                                                                 16

3.7.2      Indole test                                                                       16

3.7.3     Oxidase test                  16         

3.7.4     Coagulase test                                           16

3.7.5      Motility test                                                                    16

3.7.6     Citrate test                                                                 17

3.8        Preparation of different concntraction of extract                                           17

3.9        Test for antimicrobial effects of the test material                          17

3.10      Testing for MIC                                          18

CHAPTER FOUR

4.0        Result                                                              19

CHAPTER FIVE

5.0         Discussion                                        31

5.1       Conclusion                                    32

5.2       Recommendation                         33                         

REFERENCES               34                                                                                                                 

LIST OF TABLES

Table                                          Title                Page

1: Bochemical test result                          17

2:Antimicrobial activity of ocimum gratissimum(scent leaf) ethanol extract against test organism                           18

3: Antimicrobial activity of ocimum gratissimum(scent leaf) aqueous extract against test organism       19                  

4: Antimicrobial activity of vernonia amydalina(bitter leaf) ethanol extract against test organism                     20

5: Antimicrobial activity of vernonia amydalina(bitter leaf) aqueous extract against test organism                                                                   21

6: Antimicrobial activity gongronema latifolium(utazi) ethanol extract against test organism                        22

7: Antimicrobial activity of gongronema latifolium(utazi) ethanol extract against test organism                                                                     23

8: Minimum inhibitory concentration of vernonia amydalina

(bitter leaf) ethanol and aqueous extract                                                               24

9: Minimum inhibitory concentration of gongronema latifolium

(utazi) ethanol and aqueous extract                                                                       25

 10: Minimum inhibitory concentration of ocimum gratissimum

(scent leaf) ethanol and aqueous extract                                                               26

CHAPTER 1

1.0   INTRODUCTION

The flora of tropical West Africa has for centuries provided a wealth of material for healing purposes, and its further investigation presents a challenge to scientists who seek to contribute to the search of new means of alleviating and curing diseases. Tropical forest is a repository of many plants and about 30% of tropical plants have been so far screened for active compounds Wegner( 2005), and the plant kingdom of Nigeria still holds many species of plants of medicinal values  which are yet to be discovered.

More so, the efficacy of many of such products, some which are commercially sold has not been evaluated in the laboratory. It  has been known that the practice of traditional medicine was born out of selection procedure, which has often been trial and error, and at times has been dangerous to the extent of  costing lives. Shellard, (1979). Unfortunately, this practice still persists in most African and third world countries, where majority of the population dwell in rural areas and traditional medicine is the mainstay of health care delivery. Shellard, (1979).

The more subtle and chronic forms of toxicity, such as carcinogenicity, mutagenicity, and hepatotoxicity, may be overlooked by previous generations and it is these types of toxicities that are of most concern  when assessing the safety of herbal.Mattocks, (1986).

The isolation of active compounds from the bulk of unwanted plants parts has in no small measure reduce the toxicity of the plants and much also detailed pharmacological, toxicological and clinical studies have always been carried out on these isolated compounds/medicines in orthox practice all over the world before being marketed. Though temperate plants have provided most of the commercially valuable plants compounds today such as artemisinin, an antimalaria agent from manual Mugwort and taxol from yew trees. Taxol is used for the treatment of ovarian cancer and it is among the 20 best selling drugs. However there are many examples of herbs that also widely studied by many laboratories whose active principles remain unknown even though therapeutics effect has been proven. Some good examples are Valerian (Valeriana officinalis) and Echinacea (Echinacea pallida, Echinacea augustifolia and Echinacea purpurea) Hougton, (1998).

The global trend of revival of interest in the traditional system of medicine especially in the developing countries of the world is also due to the realization of the world need to utilize locally available medicines and therefore to reduce the reliance on expensive and imported drugs and its attendant economic benefit.

Traditional medicine may also provide a solution to the issue of drug resistance. WHO,(1984). Today a worldwide problem is the emergence of multidrug resistant pathogens which cause serious problems in hospitals, long-stayed residential centre and in the community. There are also reports of changing bacterial sensitivities to antibiotics WHO,( 2002). In addition, it is thought that some antibiotics cause the release of endotoxins and other microbial products which can precipitate septic shock. There has also been growing awareness by governments and the scientific and medical communities of the importance of medicinal plants in health care systems in many developing countries. Medicinal plants are potentials sources of new drugs and as starting products for the synthesis of drugs. Moody et al., (1998).   So far, laboratory activities (which involved specimen screening, chemical analysis, pharmacological and toxicological processes, etc.) of some selected plant species, confirm that about 90% of the plants investigated, exhibit active curative properties as assumed by traditional medicinal users. Moody et al., (1998). Increasingly, the world is returning to nature’s cure in the treatment and management of common prevalent diseases afflicting man. It is estimated that up to 80% of the world’s population depend directly or indirectly on traditional medicine for their healthcare needs. Most of the herbal/traditional remedies on sale and in use particularly in non-industrial countries are not standardized and the efficacy as well as safety cannot be readily ascertained. Moody et al., (1998).

1.1 AIM AND OBJECTIVES

The major objective of this work was to verify the traditional uses of these three medicinal plants Ocimum gratissimum , Vernonia amygdalina, and Gongronema  latifolium  as anti-microbial agents.

The objectives are;

1      T o determine the antimicrobial activity of the plant species against a variety of test organisms

2      To determine the minimal concentration of the plant extract  needed to inhibit the growth organisms

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