Antimicrobial Activity Of Essential Oil From Citrus Paradisi (Grape) Against Isolates From The Respiratory Tract

ABSTRACT

The antimicrobial activity of essential oil of citrus paradise feel has been demonstrated against a range of organism (Streptococcus pneumoniae and Klebsiella pneumoniae). The oil was obtained from the peels of grape fruits by hydro-distillation procedure and analyzed using gas chromatography coupled by mass spectrometer. 18 compounds were identified namely, cyclohexene, 1-methyl-4-(1-methylethenyl), Bicyclo (3,1,1) hept-3-en-2-01,4,66,-trimethyl, limonene oxide, bicycle (4,1,0) heptanes,-3-cycloprophyl, -7- hydroxymethyl, trans,  2-cy- clohecene-1-01,2-methyl-5-(1-methylethonyl)-cis, 2-cycloheven-1-one,2 7-oxabicyclo(4.1.0)hept-ane, 1-methyl -4- (2-methyloxivanyl), 1,2-cyclohexanediol, 1-methyl -4- (1-methyl-ethenyl),2,7-octadiene-1,6-diol,2,6-dimethyl, Limonene oxide,cis, Z,Z,Z-4-6.9-Nonadecatriene, Bicyclo(3.1.1) heptanes-2,3-diol, 2,6,6,-trimethyl.Hexadeianoic acid,  Z,Z- 2,5-pertadecadien-l-ol-  2- Butyl-3-methyl -5- (2-methylprop-2-enyl) cyclohexanone  Alpha-Farnesene, Oleic acid and octadecanoic acid.The oil extracts exerted different degree of inhibitions on the test organism or isolates was dependent on the dissolution concentrations used. Klebsiellla pneumoniae had the highest zone diameter of inhibition 2.0mm while the Streptococcus pneumonia had 10.5mm. The undiluted had 13.5mm for Klebsiella pneumonia and 11.5m for Streptococcus pneumiae. The MIC value of the essential oil against Klebsiella pneumoniae was at 12.5% concentration. However, the essential oil exhibited a bactericidal effect against the organism Streptococcus pneumoniae and Klebsiella pneumoniae at 50% concentration. This study finally has shown that grape  peels from Nigeria contains some antibiotic principles which may be explored for use in the treatment of certain diseases (respiratory tract infections).

TABLE OF CONTENTS

Title Page ﾿ i

Certification ﾿ ii

Dedication ﾿ iv

Acknowledgement ﾿ v

Table of Contents ﾿ vi

List of Tables ﾿ viii

Abstract ﾿ ix

CHAPTER ONE

1.0   INTRODUCTION ﾿ 1

1. 1 Aim ﾿ 2 ﾿

1.2 Objectives ﾿ 3

CHAPTER TWO

2.0 Literature Review ﾿ 4

2.1 General Concept ﾿ 4

2.2 ESSENTIAL OIL COMPOSITION ﾿ 5

2.3 Factors That Influence the Determination of the Antimicrobial Activity ﾿ 6

2.3.1The Assay Technique ﾿ 7

2.3.2 The Test Microorganism ﾿ 11

2.3.3 Culture Conditions ﾿ 12

2.4 Mechanism of Action of Plant Essential Oil (Eos) ﾿ 12

2.4.1 Cell Wall and Membrane Disturbance        ﾿ 13

2.4.2 ATP Production ﾿ 14

2.4.3 Protein Synthesis  ﾿ 15

2.4.4 PH   Disturbance  ﾿ 16

2.4.5 Intracytoplasmic Changes  ﾿ 17

2.4.6 DNA ﾿ 17

2.4.7 Quorum  Sensing ﾿ 18

2.5 Citrus Paradisi and importance of EO ﾿ 20

CHAPTER THREE

MATERIALS AND METHODS 

3.1 Plant Collection and Identification ﾿ 23

3.2 Essential Oil Extraction from Grape Fruit ﾿ 23

3.3  Isolation of test organism ﾿ 23

3.4 Characterization/ Identification      ﾿ 24            

3.4.1 Gram Staining ﾿ 24

3.4.2 Catalase Test ﾿ 25

3.4.3 Bile Solubility Test ﾿ 25

3.4.4 Citrate Test ﾿ 25

3.4.5 Voges Proskauer Test ﾿ 26

3.5 Antimicrobial Susceptibility Using Disc Diffusion Method ﾿ 26

3.5.1 Preparation of Disc ﾿ 26

3.5.2 Sensitivity Assay ﾿ 26

3.5.3 Minimum inhibitory Concentration (MIC) and Minimum 

        Bactericidal Concentration (MBC) ﾿ 26

3.5.4 Minimum Inhibitory Concentration ﾿

3.5.5 Minimum Bactericidal Concentration ﾿ 27

3.6   Analysis of Plant Extract (Citrus Paradisi) Using Gas 

       Chromatography-Mass Spectrometry (GC-MS) ﾿ 27

CHAPTER FOUR

4.1 RESULTS ﾿ 29 ﾿

CHAPTER FIVE

DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1Discussion ﾿ 36

5.1Conclusion ﾿ 37

5.2Recommendation ﾿ 37

References

Appendix