ABSTRACT
Plants have formed the basis of traditional systems of medicine that have been in existence since time immemorial and continued to provide mankind with new remedies. The antimicrobial activity of different parts (stem, leaf and root) of lemon extracts on some pathogens was investigated with different solvents (Methanol and Ethanol) against test organisms (Esherichia coli, Staphylococcus aureuas) using agar well diffusion method. The different extracts were prepared at different concentration (50mg/ml, 100 mg/ml, 150mg/ml and 200mg/ml). Amongst the different parts (stem, leaf and root) of lemon extracts studied, the methanoic extracts have more significant inhibitory effect than the ethanolic extracts. The methanoic extracts of leaf plant part recorded pronounced antibacterial activity against the test pathogens (Staphylococcus aureus) with zones of inhibition varying between 18 mm against Staphylococcus aureus isolate to 13 mm in Escherichia coli at a concentration of 200mg/ml. At the concentration of 200mg/ml, the leaf methanol extract showed the best result with the zone of inhibition of 18mm whereas the stem methanol extract showed the lowest effect with a zone of inhibition of 8mm. The antimicrobial potential of the different parts (stem, leaf and root) of lemon extracts must have resulted due to the presence of bioactive secondary metabolites such as flavonoids, tannins and phenols which have been shown to possess antimicrobial properties. Methanoic extracts and ethanolic extracts of different parts (stem, leaf and root) of lemon have broad-spectrum antimicrobial activity; hence can serve as natural therapeutic agent against some enteric pathogens
TABLE OF CONTENTS
Title
Page i
Certification iii
Dedication iv
Acknowledgements v
Table
of Contents vi
List
of Tables vii
Abstract ix
CHAPTER
ONE
1.0 Introduction 1
1.2 Aim
and Objectives 3
CHAPTER
TWO
2.0 Literature
Review 4
2.1 Lemon 4
2.2 History 6
2.3 Phytomedicine 7
2.4 Risk
of Phytomedicine 7
2.5 Phytochemicals 8
2.6 Classes
of Phytochemical 8
2.6.1 Alkaloids 9
2.6.2 Glycosides 9
2.6.3 Flavonoids 10
2.6.4 Phenolics 10
2.6.5 Saponins 11
2.6.6 Tannins 11
2.7 Mechanism
of Action of Phytochemical 12
2.8 Test Organisms 12
2.8.1 Escherichia
coli 12
2.8.2 Staphylococcus aureus 14
CHAPTER
THREE
3.0 Materials
and Methods 15
3.1 Plant
Materials 15
3.2 Sterilization
Methods 15
3.3 Processing
of the Plant 15
3.4 Preparation
of Crude Extracts 15
3.5 Phytochemical
Screening of Extract 16
3.5.1 Test
for Glycosides 16
3.5.2 Test
for Steroids 16
3.5.3 Test
for Tannins 16
3.5.4 Test
for Alkaloids 17
3.5.5 Test
for Flavonoids 17
3.5.6 Test
for Saponins 17
3.6 Preparation
of Concentration of Plant Extract 17
3.7 Test Organisms 18
3.8 Preparation
of Bacterial Inoculum 18
3.9 Determination
Of Minimum Inhibitory Concentration (MIC) 18
3.10 Minimum
Bactericidal Concentration (MBC) 18
3.11 Antibacterial
Susceptibility Testing of the Extracts with Test the 19
Organisms
CHAPTER FOUR
4.0 Result 20
CHAPTER FIVE
5.0 Discussion, Conclusion and Recommendation 25
5.1 Discussion 25
5.2 Conclusion 27
5.3 Recommendation 27
LIST
OF TABLES
|
TABLE
|
TITLE
|
PAGE
|
|
4.1
|
Phytochemical Constituents
of the different parts (leaf, stem and root) of lemon extracts.
|
21
|
|
4.2
|
Identification and characterization of
Bacterial Isolates
|
22
|
|
4.3
|
Antimicrobial activity
of lemon leaf, stem and root extract against Gram positive and negative
isolate.
|
23
|
|
4.4
|
The Minimum inhibition
and minimum bactericidal concentration value of the Leaf Ethanol and Methanol
extracts.
|
24
|
CHAPTER ONE
1.0 INTRODUCTION
Plants
have formed the basis of traditional systems of medicine that have been in
existence since time immemorial and continued to provide mankind with new
remedies (Jonathan et al., 2007). A
medicinal plant is one which contains substances that can be used for therapeutic
purposes. For a long of time, plants have been valuable and indispensible
sources of natural products for the health of human beings and they have a
great potential for producing new drugs (Nascimento et al., 2000). Plants act generally to stimulate and supplement the
body’s forces; they are the natural food for human beings (Akinnibosun and
Itedjer 2012). For thousands of years before the advent of modern medicine,
herbs and substances derived from plants have been the mainstay of traditional
medicine around the world (Ortuno et al.,
2006). According to World Health Organization, medicinal plants would be the
best source to obtain varieties of drugs. About 80% of individuals from
developed countries have used traditional medicine, which has compounds derived
from plants. Therefore, such plants should be investigated for better
understanding of their properties, safety and efficiency (Chintamunnee and
Mohomoodally 2012). Mouhssen (2013), because
of an increasing interest in plants as source of agents to fight microbial
diseases and the alarming incidence of antibiotic resistance of pathogenic
microbes in particular, there is still constant need for discovering new and
effective therapeutic agents. The control of infectious diseases is badly
endangered by the rise in the number of Microorganism that are resistant to
antimicrobial agents. This is because infectious caused by resistant
Microorganisms often fail to respond to conventional treatment, resulting in
prolonged illness and greater risk of death. Antibiotics resistance is a type
of drug resistance where Microorganisms are able to survive exposure to an
antibiotic. The primary causes of antibiotic resistance are genetic mutation in
bacteria (Fernando and Jose 2013).
Inappropriate and irrational use of antimicrobial medicines provides
favourable conditions for resistant Microorganisms to emerge, spread and
persist. The greater the duration of exposure to the antibiotic, the greater
the risk of the development of resistance, irrespective of the need for the
antibiotic as resistance toward antibiotics becomes more common, a greater need
for alternative treatment arises. However despite a push for new antibiotic
therapies there has been a continued decline in the number of newly approved
drugs (Mankkam et al., 2015). Antibiotics
resistance therefore posses a significant problem. Nevertheless efforts are in
place to solve the problem. Nowadays,
bacteria are considered as the main cause of morbidity and mortality in many
developing countries, especially amongst children. Among the bacterial pathogens,
Escherichia coli, Salmonella specie, and Staphylococcus aureus
are most prevalent. In the recent decades despite developments of different
types of antibiotics by pharmacological industries, resistance to antibiotics
has been increased significantly in many bacterial pathogens. In general,
bacteria have several genetic mechanisms to transmit and acquire resistance to
antibiotics.
The antibacterial
resistance in the bacterial populations is rapidly spreading; this is a serious
threat to successful treatment of infectious diseases. On the other hand, the
side effects associated with the commercial antibiotics are frequently
reported.
In this regard,
there is an urgent and continuous need for finding and investigating novel antimicrobial
compounds. Most of the antibiotics have been developed from microorganisms.
Moreover, the plant materials remain an important resource for finding the
novel antimicrobial compounds. There are many published reports on the
antibacterial effectiveness of the traditional herbs against the gram-positive
and gram-negative bacteria.
Microbial cells
are negatively affected by plant-derived substances via various mechanisms of
actions as these substances attack the phospholipid bilayer of the cell membrane
and disrupt enzymatic systems (Ahmad et al., 2006).
The medicinal herbs have the bacteriostatic effects on the enzymatic activity
associated with energy production, or they can cause denaturation of proteins,
modifying cell wall permeability, or causing the loss of macromolecules.
Therefore, it is difficult for the microorganisms to develop resistance against
these medicinal herbs.
The genus Citrus
is one of the most effective herbs in traditional medicine that belongs to the
family of Rutaceae. Citrus is the native genus of tropical and subtropical
areas in Asia.
The members of
this genus are characterized by many biologically active secondary metabolites
such as flavonoids, limonoids, coumarins and furanocoumarins, sterols,
volatile oils, organic acids, and alkaloids. Many Citrus species are recognized
for their medicinal, physiological, and pharmacological activities including
antimicrobial, antioxidant, anticancer, anti-inflammatory, and hypoglycaemic
activities.
There has been an
increasing interest in looking for the antimicrobial properties of the
plant-derived extracts particularly the essential oils. Essential oils are
rich sources of the biologically active compounds with antibacterial,
antifungal, antiviral, insecticidal, and antioxidant properties.
1.2 AIM AND OBJECTIVES
·
The purpose of this study
was to determine the antibacterial effects of methanoic and ethanolic extract
of three Lemon parts against Escherichia
coli and Staphylococcus aureus by
agar well diffusion method.
·
To carry out
phytochemical screening of the different lemon parts (leaf, root and stem)
using methanol and ethanol solvents.
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