PHYTOCHEMICAL ANALYSIS AND ANTIBACTERIAL ACTIVITY OF OCIMUM GRATISSIMUM (SCENT LEAF) ON SOME SELECTED GRAM POSITIVE & GRAM NEGATIVE BACTERIA

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ABSTRACT


Antibacterial properties of Ocimum gratissimum was determined and tested against pure cultures of clinical isolates of Escherichia coli and Staphylococcus aureus.  Water and Ethanol were used for the extraction of the active constituent of the plants. The phytochemical analysis carried out on the plant revealed the presence of Tannin, Saponin, glycoside and alkaloid. Aqueous extract of Ociumum gratissimum showed no inhibitory effect on Escherichia coli. The extract inhibited Staphylococcus aureus only at 100% and 80% concentration with zone of inhibition diameter of 100mm and 9mm the Ethanol extract of the plant showed the mean zone diameter of inhibition. The highest zone of inhibition 30mm and 20mm was recorded against Escherichia coli at 100% and 80% concentration. The least inhibitory effect was observed with Staphylococcus aureus (7mm) at 20% concentration the minimum inhibitory concentration of the different plant extract showed that there was no activity observed with aqueous extract against Escherichia coli, but the minimum inhibitory concentration of the ethanolic extract against Staphylococcus aureus and Escherichia coli was 80%. The result of this study suggests the possibility of using the ethanolic extract in treating the diseases caused by the test organisms.






TABLE OF CONTENTS

Title page

Certification                                                                                                         i

Dedication                                                                                                  ii

Acknowledgement                                                                                                iii

Table of contents                                                                                        iv

List of tables                                                                                               vi

Abstract                                                                                                      vii

                                                                                                          

CHAPTER ONE

1.0 Introduction                                                                                         1

1.1 Aims and objectives                                                                             2

1.2 Objectives                                                                                             3

CHAPTER TWO

2.0 Literature Review                                                                                  4

2.1Antibacterial activity of Ocimum gratissimum                                       12

CHAPTER THREE

3.0 Materials and Methods                                                                         14 

3. 1  Collection of plant materials                                                              14

3.2  Media used and its preparation                                                           14

3.3  Sterilization                                                                                         14

3.4   Sources of test organisms                                                                            14

3.5     Test for Authentication of test organisms                                         15

3.5.0 Gram staining                                                                                   15

3.5.1  Catalase test                                                                                      15   

3.5.2 Coagulase test                                                                                   15

3.5.3 Citrate test                                                                                         16

3.5.4Motility, Indole, Urea                                                                         16

3.5.5  Triple sugar iron test                                                                        17

3.5.6 Oxidase test                                                                                       17

3.6Preparation of plant extracts                                                                          17

3.7 Phytochemical analysis                                                                        18

3.7.0 Test for Alkaloid                                                                               18

3.7.1 Test for glycoside                                                                              18

3.7.2Test for flavonoid           

                                                                   18

3.7.3 Test for tannin                                                                                   18

3.8 Antibiotic sensitivity testing                                                                          19

3.8.0 Determination of minimum inhibitory concentration (MIC)              20

 

CHAPTER FOUR

4.0 Results                                                                                                  21

CHAPTER FIVE

5.0   Discussion, Recommendation and Conclusion                                            27

5.2Recommendations                                                                                 28

5.3Conclusions                                                                                           28

Reference                                                                                                    29                                                                                                   

 

 

 

  

 

 

 

LIST OF TABLES

 

Tables                                              Title                                                           Page

1.     Phytochemical constituents of leaf extract of Ocimum gratissimum            22

2.     Diameter of mean zone of inhibition of aqueous of Ocimum                     

gratissimum tested organisms.                                                                               23         

3.     Diameter of mean zone of inhibition of ethanol extract of Ocimum

gratissimum against tested organisms.                                                          24

4.     Minimum inhibitory concentration of two different extracts of Ocimum

gratissiumum against the test organisms.                                                      25

5.     Biochemical and cultural characteristics of test isolates.                                        26

                                                                                     

                                                  

 


 

 

 


                                                     CHAPTER ONE


1.0  INTRODUCTION

Medicinal plants are of great importance to the health of individual and thecommunities. The medicinal values of some plants lies in some chemical substancesthat produce definite physiological actions in the human body. The most important ofthese bioactive constituents are alkaloids, tannis, flavonoids and phenolic compounds.Many of these indigenous medicinal plants are used as spices and food plants (Okwu, 1999). An Ethno botanical and ubiquitous plant serves as rich resources of naturaldrugs for research and development (Kong et al, 2008). Medicinal plants based drugsowe the advantage of being simple, effective and exhibit broad spectrum activity. Therevival of interest in the use and importance of African medical plants by WHO andmany developing countries has led to intensified efforts on the documentation ofethnomedical data of medicinal efforts. This is because most traditional healers keep norecords and their information is passed on mainly verbally from generation togeneration. . Researchers are increasingly turning their attention to natural productslooking for new leads to develop better drugs against cancer, as well as viral andmicrobial infections. The phytochemical evaluation of Ocimumgratissimumshows thatit is rich in alkaloid, tannins, phytates, flavonoids and Oligosaccharides (Ijehet al., 2004). In the coastal area of Nigeria, the plant Ocimumgratissimumis used in thetreatment of epilepsy, high fever and diarrhea (Sofowora 1993). Ocimumgratissimum(Scent leaf) is a perennial plant which is widely distributed in the tropics of Africa andAsia. It belongs to the family Labiatae and it as the most abundant of the genusOcimum. In the southern part of Nigeria, it is called “Efirinnla” by the Yoruba speakingtribe. “Nichonwu” in Igbo while in the northern part of Nigeria, it is called “Daidoga”(Effraim, et al; 2003). Leaf extract of Ocimumgratissimumand Xylopiaaethiopieawere analyzed against five pathogenic organisms. Staphylococcus aureaus, Escherichiacoli, Streptococcus fecalis, Pseudomonas aeruginosa and Lactobacilli (Ijehet al,2004).The findings justifies the application of Ocimumgratussimumin dermatologicalcream and indicate the effective doses could be achieved at very low concentration andalso shows that the aqueous fractions of both plants have more potential asantimicrobial agents than their ethanolic fractions (Ijehet al., 2004).

Plants are sources of medicine in pharmacopoeia. Herbal medicine can be used as analternative to some commercial drugs. As a result, some people resort to natural leaves forthe treatment of infection. The antimicrobial properties of many plants have beeninvestigated by a number of researchers word wide (Adamuet al.,2005). Medicinal use of plants ranges fromthe administration of the roots, barks, stems, leaves and seeds to the use of extracts anddecoctions from the plants (Ogbulie et al.,2007).

Medicinal plants are used as excellent antimicrobial agent because they posses variety ofchemical constituents in nature. Plants have the ability to synthesize aromatic substancessuch as phenolic, (e.g), phenolic acid, flavonoids, quinines, coumarins, lignans, stibenes,tannins), Nitrogen compound (alkaloids, amines) vitamins, terpenoids (including carotenoids)and some other endogenous metabolites. These substances serve as plants defensemechanism against predation by microbes, insects, and herbivores (Bharathi and Shanmuga.2011).


 1.1 AIMS AND OBJECTIVES

The aim of this is study to investigate the antibacterial and phytochemical composition of Ocimumgratissimum.

 

1.2 OBJECTIVES

1. To determine the antimicrobial effect ofOcimumgratissimumon some selected gram positive and gram negative organisms.

2. To determine the phytochemical components of the leaf extracts.

3. To determine the minimum inhibitory concentration of the leaf extracts on the selected gram positive and gram negative organisms.

 

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