ABSTRACT
The emergence of antibiotic
resistance as well as the recent undesirable side effect of some of the
commercially available antibiotics has led to the screening of plant extract in
order to discover new drug that could serve as alternative therapy for the
treatment of various infections and diseases. Fresh leaf of Ocimum gratissimum (scent leaf), Gonronema latifoliun(utazi)and Vernonia
amydalina (bitter leaf) were collected
air-dried at room temperature and blended to powder using electric
blender. The extraction was done using reflux extraction method using ethanol
and water as solvent. The antibacterial
activities of the test plants were determined to ascertain the different
phytochemicals present in the plant extract. The extract was also tested
against some selected Gram negative and Gram positive intestinal pathogenic
bacteria; Escherichia coli, Salmonella
species,Staphylococcus aureus and
Enterococci faecalis by reconstituting the extract to obtain different concentration (100,50 and
25 g/ml) and agar well diffusion techniques were used to evaluate the
antibacterial susceptibility of the leaf extract. The result of antibacterial
analysis showed that the extract of O.
gratissimum and V. amydalina has
antibacterial activity against E. coli
and Salmonella while the extract of G.latifolium has anti microbial activity
against S.aureus and E.faecalis. This could be as a result of
the presence of various phytochemicals or the interaction of one or more of the
identified metabolites against the test organisms. The results obtained from
this research, suggest that Escherichia coli and salmonalla was susceptible to
the leaf extract of O.gratissimum and V.amydalina
while S.aureus and E.faecalis is susceptible to G.latifolium. Generally the ethanolic
extracts of the plant materials shows a higher antibacterial activity than the
aqueous extracts.
TABLE OF
CONTENTS
Title
page i
Certification ii
Dedication iii
Acknowledgement iv
Abstract v
Table
of content vi
List
of tables vi
CHAPTER ONE
1.0 Introduction 1
1.1 Aim and Objectives 3
CHAPTER TWO
2.0 Literature Review 4
2.1 Description of Test Materials 4
2.1.1 Ocimum
gratissimum 4
2.1.2 Vernonia
amydalina 7
2.1.2 Gongronema
latifolium 7
2.2 Description
of Test Organisms 8
2.2.1 Escherichia
coli 8
2.2.2 Staphylococcus
aureus 9
2.2.3 Salmonella
spp 10
2.2.4 Enterococci
faecalis 11
CHAPTER THREE
3.0 Methods and Materials 13
3.1 Collection and Identification of plant
materials 13
3.2 Materials and reagents 13
3.3 Preparation of plant extract 13
3.3.1 Preparation of aqueous 13
3.3.2 Preparation of ethanolic extract 14
3.4 Test for purity and sterilization 15
3.5 bacterial species confirmation 15
3.6 Gram stain 15
3.7 Biochemical test 16
3.7.1 Catalase test 16
3.7.2 Indole test 16
3.7.3 Oxidase test 16
3.7.4 Coagulase test 16
3.7.5 Motility test 16
3.7.6 Citrate test 17
3.8 Preparation of different concntraction
of extract 17
3.9 Test for antimicrobial effects of the
test material 17
3.10 Testing for MIC 18
CHAPTER
FOUR
4.0 Result 19
CHAPTER
FIVE
5.0 Discussion 31
5.1 Conclusion 32
5.2 Recommendation 33
REFERENCES 34
LIST
OF TABLES
Table
Title Page
1:
Bochemical test result 17
2:Antimicrobial
activity of ocimum gratissimum(scent
leaf) ethanol extract against test organism 18
3:
Antimicrobial activity of ocimum
gratissimum(scent leaf) aqueous extract against test organism 19
4:
Antimicrobial activity of vernonia
amydalina(bitter leaf) ethanol extract against test organism 20
5:
Antimicrobial activity of vernonia
amydalina(bitter leaf) aqueous extract against test organism
21
6:
Antimicrobial activity gongronema
latifolium(utazi) ethanol extract against test organism 22
7:
Antimicrobial activity of gongronema
latifolium(utazi) ethanol extract against test
organism
23
8:
Minimum inhibitory concentration of vernonia
amydalina
(bitter
leaf) ethanol and aqueous extract
24
9:
Minimum inhibitory concentration of gongronema
latifolium
(utazi)
ethanol and aqueous extract 25
10: Minimum inhibitory concentration of ocimum gratissimum
(scent
leaf) ethanol and aqueous extract
26
CHAPTER 1
1.0 INTRODUCTION
The
flora of tropical West Africa has for centuries provided a wealth of material
for healing purposes, and its further investigation presents a challenge to
scientists who seek to contribute to the search of new means of alleviating and
curing diseases. Tropical forest is a repository of many plants and about 30%
of tropical plants have been so far screened for active compounds Wegner(
2005), and the plant kingdom of Nigeria still holds many species of plants of
medicinal values which are yet to be
discovered.
More
so, the efficacy of many of such products, some which are commercially sold has
not been evaluated in the laboratory. It
has been known that the practice of traditional medicine was born out of
selection procedure, which has often been trial and error, and at times has
been dangerous to the extent of costing
lives. Shellard, (1979). Unfortunately, this practice still persists in most
African and third world countries, where majority of the population dwell in
rural areas and traditional medicine is the mainstay of health care delivery. Shellard,
(1979).
The
more subtle and chronic forms of toxicity, such as carcinogenicity,
mutagenicity, and hepatotoxicity, may be overlooked by previous generations and
it is these types of toxicities that are of most concern when assessing the safety of herbal.Mattocks,
(1986).
The
isolation of active compounds from the bulk of unwanted plants parts has in no
small measure reduce the toxicity of the plants and much also detailed
pharmacological, toxicological and clinical studies have always been carried
out on these isolated compounds/medicines in orthox practice all over the world
before being marketed. Though temperate plants have provided most of the
commercially valuable plants compounds today such as artemisinin, an antimalaria
agent from manual Mugwort and taxol from yew trees. Taxol is used for the
treatment of ovarian cancer and it is among the 20 best selling drugs. However
there are many examples of herbs that also widely studied by many laboratories
whose active principles remain unknown even though therapeutics effect has been
proven. Some good examples are Valerian (Valeriana officinalis) and Echinacea
(Echinacea pallida, Echinacea augustifolia and Echinacea purpurea) Hougton, (1998).
The
global trend of revival of interest in the traditional system of medicine
especially in the developing countries of the world is also due to the
realization of the world need to utilize locally available medicines and
therefore to reduce the reliance on expensive and imported drugs and its attendant
economic benefit.
Traditional
medicine may also provide a solution to the issue of drug resistance. WHO,(1984).
Today a worldwide problem is the emergence of multidrug resistant pathogens
which cause serious problems in hospitals, long-stayed residential centre and
in the community. There are also reports of changing bacterial sensitivities to
antibiotics WHO,( 2002). In addition, it is thought that some antibiotics cause
the release of endotoxins and other microbial products which can precipitate
septic shock. There has also been growing awareness by governments and the
scientific and medical communities of the importance of medicinal plants in
health care systems in many developing countries. Medicinal plants are
potentials sources of new drugs and as starting products for the synthesis of
drugs. Moody et al., (1998). So far, laboratory activities (which involved
specimen screening, chemical analysis, pharmacological and toxicological
processes, etc.) of some selected plant species, confirm that about 90% of the
plants investigated, exhibit active curative properties as assumed by
traditional medicinal users. Moody et al.,
(1998). Increasingly, the world is returning to nature’s cure in the treatment
and management of common prevalent diseases afflicting man. It is estimated
that up to 80% of the world’s population depend directly or indirectly on
traditional medicine for their healthcare needs. Most of the herbal/traditional
remedies on sale and in use particularly in non-industrial countries are not
standardized and the efficacy as well as safety cannot be readily ascertained. Moody
et al., (1998).
1.1 AIM AND OBJECTIVES
The major objective of this work was to verify the
traditional uses of these three medicinal plants Ocimum gratissimum , Vernonia amygdalina, and Gongronema latifolium as anti-microbial agents.
The objectives are;
1
T o determine the
antimicrobial activity of the plant species against a variety of test organisms
2
To determine the
minimal concentration of the plant extract needed to inhibit the growth organisms
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