ANTIBACTERIAL ACTIVITY OF SCENT LEAF OCIMUM GRATISSIMUM (SCENT LEAF) LEAF EXTRACTS ON SPECIES OF MICROORGANISMS ISOLATED FROM A WOUND

ABSTRACT

Medicinal plants are of great importance to the health of individuals and the communities. Leaf extract of Ocimum gratissimum was analysed against 3 pathogenic organisms; Staphylococcus aureus, Escherischia coli, and Pseudomonas aeroginosa. Fresh leaves of Ocimum gratissimum (scent leave) was procured from a local market in Ndoru, Ikwuano, L.G.A. of Abia state and transferred to the laboratory for microbiological analysis. The plant was identified authenticated at the Herbarium of the department of plant science and biotechnology, Michael Okpara University of Agriculture Umudike. Clinical isolates of Staphylococcus aureus, Escherischia coli, and Pseudomonas aeroginosa used in the study were obtained from the Microbiology laboratory, Federal Medical Center Umuahia and confirmed by Gram-staining method, biochemical test, morphological appearance and motility. The crude extract of 20mg/ml was double diluted in ethanol and water to the following concentrations 100%, 50%, 25%, 12.5%, 6.25% and 3.625%%. Antimicrobial susceptibility testing was done by agar well dilution method.    MIC and MBC of the crude extract was done under standard technique. The Minimum Inhibitory Concentration (MIC) of the aqueous extract of Occimum gratissimum against Staphylococcus aureus, 100%, Escherichia coli, 100mg/ml and Pseudomonas aeruginosa was 50% while their Minimum Bactericidal Concentration (MBC) was 50%, 100% and 50% respectively. Conversely the ethanolic extract of Occimum gratissimum  showed a concentration dependent gradient increase in the level of inhibition against the isolates with MIC values of 50mg/ml for each isolate and MBC values of 25%, 50% and 25% against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa respectively This study justifies the use of the leaves in traditional medicine practice as therapeutic agents and can explain the long history use of these plants. This research work has shown that Ocimum gratissimum has potential antibacterial components that are responsible for it’s antibacterial activities.

TABLE OF CONTENTS

Title Page                                                                                                                                 i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgement                                                                                                                  iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          vii

Abstract                                                                                                                                  viii

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1       Aims and Objectives of Study                                                                                    3

CHAPTER TWO

2.0       Literature Review                                                                                                       4

2.1       The Genus Ocimum                                                                                                    5

2.2       Test Organisms                                                                                                           7

2.2.1    Escherichia coli                                                                                                          7

2.2.2    Staphylococcus aureus                                                                                               9

2.2.3    Pseudomonas aeruginosa                                                                               10                                                                   

CHAPTER THREE

3.0       Materials and Method                                                                                                 12

3.1       Study Area                                                                                                                  12

3.2       Materials                                                                                                                     12

3.3       Sample Collection                                                                                                      12

3.4       Test Organisms                                                                                                           13

3.5       Confirmation of Test Isolate                                                                                      13

3.5.1    Morphological appearance                                                                                         13

3.5.2    Gram Staining                                                                                                            13

3.5.3    Motility test                                                                                                                14

3.5.4    Biochemical tests                                                                                                        14

3.6       Preparation of Crude Extracts                                                                                    17

3.7       Preparation of Concentration of Plant Extract                                                           17

3.8       Antimicrobial Susceptibility Testing                                                                         17

3.8.1    Agar-Well Diffusion Method                                                                                     17

3.8.2    Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal

            Concentration (MBC) Test                                                                                         18

CHAPTER FOUR

4.0       Result                                                                                                                          19

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                                          28

5.1       Discussion                                                                                                                   28

5.2       Conclussion and Recommendation                                                                            29

5.3       Recommendation                                                                                                       29

            References                                                                                                                  33

LIST OF TABLES

CHAPTER ONE

1.0       INTRODUCTION

Medicinal plants are of great importance to the health of individual and the communities. The medicinal value of some plants lies in some chemical substances that produce definite physiological actions in the human body. The most important of these bioactive constituents are alkaloids, tannis, flavonoids and phenolic compounds. Many of these indigenous medicinal plants are used as spices and food plants (Okwu, 2001). An Ethno botanical and ubiquitous plant serves as rich resources of natural drugs for research and development (Kong et al, 2008). Medicinal plants based drugs owe the advantage of being simple, effective and exhibit broad spectrum activity. The revival of interest in the use and importance of African medical plants by WHO and many developing countries has led to intensified efforts on the documentation of ethnomedical data of medicinal efforts. This is because most traditional healers keep no records and their information is passed on mainly verbally from generation to generation. . Researchers are increasingly turning their attention to natural products looking for new leads to develop better drugs against cancer, as well as viral and microbial infections. In the coastal area of Nigeria, the plant Ocimum gratissimum is used in the treatment of epilepsy, high fever and diarrhea. Ocimum gratissimum (Scent leaf) is a perennial plant which is widely distributed in the tropics of Africa and Asia. It belongs to the family Labiatae and it as the most abundant of the genus Ocimum. In the southern part of Nigeria, it is called “Efirin nla” by the Yoruba speaking tribe. “Nichonwu” in Igbo while in the northern part of Nigeria, it is called “Daidoga” (Effraim, et al., 2003). Leaf extract of Ocimum gratissimum and Xylopia aethiopiea were analyzed against five pathogenic organisms. Staphylococcus aureaus, Escherichia coli, Streptococcus fecalis, Pseudomonas aeruginosa and Lactobacilli (Ijeh et al, 2004).The findings justifies the application of Ocimum gratussimum in dermatological cream and indicate the effective doses could be achieved at very low concentration and also shows that the aqueous fractions of both plants have more potential as antimicrobial agents than their ethanolic fractions (Ijeh et al., 2004). There is growing interest in exploiting plants for medicinal purposes especially in Africa. This stems from the fact that microorganisms are developing resistance to many drugs and as such created situation where some of the common and less expensive antimicrobial agents are losing effectiveness. Herbal medicine which uses medicinal plants primarily presents as an alternative to such situation. These medicinal plants have immensely contributed to the development of human health and welfare. Concomitantly, there is an increase in data and huge patronage to herbal products round the world (Arekemase et al., 2011). Medicinal plants such as Ocimum gratissimum and Piper guineense have been asserted to provide various culinary and medicinal properties. These medicinal properties exert bacteriostatic and bacteriocidal effects on some bacteria. These effects have been attributed to the peptides, alkalonoids, essential oils, phenols and flavonols which are major components in these plants (Okigbo and Igwe, 2007). O. gratissimum belongs to the family leguminocaeae, commonly known as “alfavaca”. It is naturally used in the treatment of different diseases which include: upper respiratory tract infections, diarrhea, headache, conjunctivitis, skin disease, pneumonia tooth and gum disorder, fever and as mosquito repellants. O. gratissimum is found in the tropical and warm temperature regions such as India and Nigeria (Okigbo and Ogbonnaya, 2006). Some of the vernacular names in Nigeria include: (Ncho-anwu, Ahuji) Igbo, (Efinrin,) Yoruba, (Aramogbo) Edo and (Daidoya) Hausa (Effraim et al., 2000). O. gratissimum has been described to have other species in the flora of tropical West Africa. These include: Ocimum viride Linn, Ocimum suave Linn, Ocimum basilicum Linn and Ocimum canum Sims. Matasyoh et al., (2007) reported their numerous medical uses. The Ocimum oil has been described to be active against several species of bacteria and fungi. These include Listeria monocytogenes, Shigella, Salmonella and Proteus, for fungi Trichophyton rubrum, Trichophyton mentagrophytes, Cryptococcus neoformans, Penicillum islandi cum, and Candida albicans (Akinyemi et al., 2004). From recent findings, O. gratissimum has proved to be useful in the medication for people living with Human Immuno deficiency Virus (HIV), and Acquired Immuno Deficiency Syndrome virus AIDs (Elujoba, 2000). In Congo, O. gratissimum decoction is used for gonorrheal infection, vaginal douches for metritis and vaginitis and used in treatment of mental illness (Effraim et al., 2000).

1.1       AIMS AND OBJECTIVES OF STUDY

·       The aim of this present work  is to evaluate the antibacterial effects of the medicinal plant leaf extract of Ocimum gratissimum on some selected pathogens isolated from wound infection.

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