PHYTOCHEMISTRY AND EVALUATION OF ANTIMICROBIAL ACTIVITY OF GARCINIA KOLA ON METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS AND ESCHERICHIA COLI

ABSTRACT

The antibacterial activity of Garcinia kola seeds varieties extracted in ethanol and water  was tested against some selected clinical bacteria  isolates; Staphylococcus aureus, and  Escherichia coli  at concentrations of 10, 20, 30 ,40 mg/ml. Disc diffusion method was employed to determine the antimicrobial activity of the extracts against test microorganisms. The results showed that the seed ethanol extracts exhibited more antimicrobial activity at a concentration of 40 mg/ml, with zones of inhibition ranging from 12 to 22mm for ethanol. The aqueous (water) extracts showed zones of inhibition ranging from 12 to 20mm.The Minimum Inhibitory Concentration (MIC) showed antimicrobial activity at lowest concentration against Staphylococcus aureus. There were presence of phytochemical compounds such as Tannins, Saponins, Alkaloid and Phenol. The results imply that the ethanol and aqueous extracts of Garcinia kola seed posses strong antibacterial properties in the treatment of bacterial infection, and hence its potential as a useful chemotherapeutic agent in the treatment of bacterial infections in humans.

TABLE OF CONTENTS

Title page                                                                                                                                       i

Certification                                                                                                                                  ii

Dedication                                                                                                                                    iii

Acknowledgements                                                                                                                      iv

Table of Contents                                                                                                                          v

List of Tables                                                                                                                                viii

Abstract                                                                                                                                         ix

CHAPTER ONE

1.0 Introduction                                                                                                                              1 

CHAPTER TWO                                                                                                                            

2.0 Literature review                                                                                                                       5

2.1 Garcinia kola                                                                                                                            5

2.2 Scientific classification                                                                                                             15

2.3Common names                                                                                                                         16                                                                                                                                               

2.4 Traditional uses and medicinal values                                                                                      16                                                                       

2.5 Scientific research on Garcinia kola                                                                                         17

2.5.1 Chemical constituents                                                                                                            18

2.5.2 Anti-microbial properties                                                                                                       17                                                                                                                                     

2.5.3 Anti-diabetic properties                                                                                                         18

2.5.4 Hepatoprotective and Anti-oxidant Activities                                                                      18

2.5.5 Effects on fertility                                                                                                                 19

2.5.6 Other studies on Garcinia kola                                                                                              19

3.0 CHAPTER THREE

3.1 Preparations of plant extract                                                                                                     21

3.2 Media used                                                                                                                               21

3.3 Sterilization                                                                                                                               21

3.4 Isolation of test organism                                                                                                         22

3.5 Phytochemical analysis                                                                                                             23

3.5.1 Test for Alkaloid                                                                                                                    23

3.5.2 Test for Tannin                                                                                                                                                                23                                                                                

3.5.3 Determination for Saponins                                                                                                  23

3.5.4 Determination of Total Phenols                                                                                             23           

3.6 Identification of bacterial isolates                                                                                            24           

3.6.1 Cefo Oxitin test                                                                                                                     24

3.7 Gram staining                                                                                                                            25           

3.8 Biochemical cultural characteristics                                                                                        25

3.8.1 Catalase test                                                                                                                          25

3.8.2 Coagulase test                                                                                                                       25

3.9 Antimicrobial susceptibility testing                                                                                         25

3.9.1 Disc diffusion method                                                                                                          25

3.9.2 Determination of Minimal Inhibitory Concentration                                                            25

CHAPTER FOUR

4.0 Results                                                                                                                                     27

CHAPTER FIVE

5.1 Discussion                                                                                                                                32                                                                          

5.2 Recommendation                                                                                                                     33

5.3 Conclusion                                                                                                                               33

Reference

LIST OF TABLES

 TITLE                                                                      PAGES

 1: Morphology and biochemical identification of isolate                                                      28                                                 2: Phytochemical composition of Garcinia kola from different extracts                               29

3: Antimicrobial activity of bacterial isolates of water and ethanol extract of Garcinia kola measured by zone of inhibition                                                                                         30                                                                                              4: The minimum inhibitory concentration (MIC) exhibited by Garcinia kola extracts against some selected bacterial isolates                                                                                                31

CHAPTER ONE

1.0       INTRODUCTION

Antimicrobial agents are chemotherapeutic agents employed to kill micro-organisms or prevent their growth for the treatment of diseases. These are classified according to their application and spectrum of activity, as germicides that kill micro-organisms, whereas micro-biostatic agents inhibit the growth of pathogens and enable the leucocytes and other defense mechanism of the host to cope up with static invaders. The germicides may exhibit selective toxicity depending on their spectrum of activity. They may act as viricides (killing viruses), bacteriocides (killing bacteria), algicides (killing algae) or fungicides (killing fungi). They are produced by micro-organisms or they might be fully or partly prepared by chemical synthesis. They inhibit the growth of micro-organisms in minimal concentrations. Antibiotics may be of microbial origin or purely synthetic or semi synthetic. The beginning of modern chemotherapy has largely been due to the efforts of Dr. Paul Ehrlich (1910), who used salvarsan, as arsenic derivative effective against syphilis. Paul Ehrlich used the term chemotherapy for curing the infectious disease without injury to the host’s tissue, known as chemotherapeutic agents such as antibacterial, antiprotosoal, antiviral, antineoplastic, antitubercular and antifungal agents. Later on, Domagk (1953) prepared an important chemotherapeutic agent sulfanilamide. Antibiotics are used as inhibitory chemicals and they were believed to lead in the complete eradication of infectious diseases (Rosina et al., 2009). Despite the progress made in introducing new antibiotics, emergence of drug resistant strains cause failure of infectious disease treatment (Matthias et al., 2000; Gibbons, 2005). It is believed that consumption of antibiotics in livestock, agriculture and poultry is one of the factors that have caused an increase in the development of drug resistance. New effective strategy is therefore necessary for the management of resistance bacteria: one of which includes medicinal plants (CourvalinandTreu-Cuot, 2006) which stands partly as the objective of this comparative study, evaluating the potency of Garcinia kola (Bitter kola).

Although there is a wide range of antibiotics for the treatment of bacterial infections, the development of resistance to chemotherapeutic agents such as erythromycin and penicillin is increasingly becoming a pressing problem (Abimbola et al., 2003). Other limitations of modern chemotherapeutic drugs are their high costs and non-availability, especially in rural areas. As a consequence, it is necessary to search new organic molecules with antibacterial activity; which, in addition, could be potential sources for starting materials for the semi-synthesis of new drugs.

It is estimated that more than two thirds of the world’s plant species have medicinal value; and about 80% of people rely on herbs for their medicines. Medicinal plants are believed to be an important source of new chemical substances with potential therapeutic benefits. They contain many biologically active compounds with medicinal properties and largely employed in developing countries (Rabe et al., 2007). Compounds isolated from natural origin such as medicinal plants are believed to have less side effects, tolerated by patients and can be afforded by most people since they are sold at a cheaper, reasonable price (Bakkali et al., 2008).

African plants, in particular, medicinal plants, constitute a rich but still largely untapped pool of natural products (Hostettmann, et al., 2006). WHO estimates indicate that 80% of the population (mostly in developing countries) still relies on plant-based medicines for primary health care (Fox, 2004).Bitter kola have been used in Cameroon for such clinical conditions as bronchitis, partial impotence, cough, laryngitis, etc. by the local population, but without supporting scientific evidence (Surh et al., 1998). Several findings on chemotherapeutic potentials of plants have shown that they can be sources of antimicrobial compounds of value and a typical example of such plant is Garcinia kola (Njume et al., 2011).

G. kola plant has been investigated largely due to its diverse bioactivities. Garcinia kola is a species of flowering plant belonging to the family Guttiferae. It is found mostly in Central and Western Africa; in countries such as Cameroon, Benin, Democratic Republic of Congo, Ivory Coast, Gabon, Ghana, Liberia, Nigeria, Senegal and Sierra Leone. This plant has a popular acronym “wonder’’ plant among the South-Western Nigerian people because every part of it has been found to be of medical importance. It consists of fruits, leaves, roots, barks, stems and twigs and an average of four seeds is contained in a fruit. All parts of the plant are used in traditional medicine for various therapeutic purposes. The fruits are reddish or yellowish in colour and each fruit contain two to four seeds and pulp that have a sour taste (Nzegbule and Mbakwe, 2001). The seeds are smooth, oval shaped with yellow pulp and covered with a brown seed coat. They have been reported to possess antibacterial activity, antidiabetic, antiviral and antihepatotoxicity potentials (Akoachere et al., 2002). They are used to prevent or relieve colic pains, cure head and chest colds. Essential oils produced by plants are not only used in agriculture or in food industries as food preservatives or additives, but also used pharmaceutically for their therapeutic activities in the treatment of various diseases (Vila et al., 2010; Sibandaand Okoh, 2008).

Presently there are global problems of antibiotic resistance to infections coupled with the emergence of new and re-emerging diseases. There is also a belief that the use of plants for medicinal purposes has been associated with less side effects (Rabe et al., 2007; Sibanda et al., 2010). There is therefore a need to search for more efficacious and cost-effective antimicrobial agents of natural origin to complement the existing synthetic antimicrobial drugs that are becoming less potent against pathogenic microorganisms (Ncube et al., 2008). And this quest for a more effective antibacterial agent, even that with little or no side effect is the paramount reason for this research work.

AIM

Phytochemical and antimicrobial activity of Garcinia kola on methicilin resistant Staphylococcus aureus and E.coli.

OBJECTIVES

1. To determine the antimicrobial activity of Garcinia kola on methicilin resistant Staphylococcus aureus and E.coli.

2. To determine the phytochemical composition of Garcinia kola.

3. To determine the minimal inhibitory concentration of Garcinia kola.

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