ABSTRACT
Human beings have used plants to treat common infectious disease and some of these traditional practices are still included as part of the habitual treatment of various maladies. Today, researches are focusing on the discovery of new therapeutic substances of natural origin with possible low or no toxicity to humans, animals and the environment based on the ethnomedicinal practices. Antimicrobial activity of the V. amygdalina (Bitter leaf) aqueous and ethanolic extracts against the test isolates, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa were assessed using minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and inhibitory zone diameter(IZD) at the dilutions of 100mg/ml, 50mg/ml, 25mg/ml, 12.5mg/ml 6.25ml/ml and 3.125mg/ml respectively . Both the water and ethanolic extract concentrations of V. amygdalina leaf produced observable in-vitro antibacteria effects on all the organisms at 100mg/ml and 50mg/ml. Ethanolic extract produced better antibacterial effects against the test isolate than that of aqueous extract. This research has shown that V. amygdalina leaves have great antibacterial activities. This can be applied in the treatment of the disease caused by the test organisms.
TABLE OF CONTENTS
Title Page i
Certification ii
Dedication iii
Acknowledgements iv
Table of Contents v
List of Tables viii
Abstract ix
CHAPTER ONE: INTRODUCTION
1.1 Background 1
1.2 Statement
of Result 4
1.3 Aims and Objectives of Study 4
CHAPTER TWO: LITERATURE REVIEW
2.0 Bitter
Leaf (Vernonia amygdalina) 6
2.1 Medicinal
Properties 6
2.2 Botanical
and Ecology Description of Vernonia
amygdalina 6
2.3 Pharmatology 7
2.4 Ethnobotanical
Uses 8
2.5 Ethnoveterinary
and Zoopharmacognostic uses of V.
amygdalina 9
2.6 Bioactivities
of V. amygdalina 10
2.6.1 Anticancer
and cytotoxic activity of V amygdalina: 10
2.6.2 Anti
bacterial activity 11
2.6.3 Antidiabetic
activity 11
2.6.4 Antimalarial/Antiplasmodial
Activity 11
2.6.5 Antioxidant
Activity 12
2.6.6 Phytochemistry 12
2.6.7 Toxicity 12
CHAPTER THREE: MATERIALS AND METHODS
3.1 Reagents
And Apparatus/Equipment 14
3.1.1 Apparatus used for the experiment 14
3.1.2 Reagents used for the Experiment 14
3.2 Sample
Collection 14
3.3 Test Organisms 15
3.4 Confirmation
of the Test Isolates 15
3.4.1 Morphology 15
3.4.2 Gram staining 15
3.4.3 Mortility
test 16
3.4.4 Biochemical
tests 16
3.5 Procedure of Preparation of Leaf Extract 18
3.5.1 Preparation of Stock Solution of Plant
Extract 19
3.5.2 Antibacterial Activity Test 20
3.6 Agar
Well Diffusion Method 20
3.7 Minimum Inhibitory Concentration (MIC) 21
3.8 Minimum Bactericidal Concentration (MBC) 22
CHAPTER FOUR: RESULTS AND DISCUSSION
4.1 Results 23
4.2 Discussion 30
CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATIONS
5.2 Conclusion 32
5.3 Recommendations 32
References 34
LIST
OF TABLES
Tables Title Page
4.1 Morphological Characteristics
of the isolates 24
4.2 Biochemical Identification of
Isolates 25
4.3 Diameter of zones of inhibition
(mm) of V. amygdalina against
P. aeruginosa, S. aureus and E. coli using water extracts 26
4.4 Antimicrobial activity of V. amygdalina against P. aeruginosa,
S. aureus and E.
coli using ethanolic extracts 27
4.5
MIC, MBC of Vernonia amygdalina bitter leaf
water extract on Staphylococcus aureus, Pseudononas 28
aeruginosa and Escherichia coli
4.6:
MIC, MBC of Vernonia amygdalina bitter leaf ethanolic
extract on Staphylococcus aureus, Pseudononas aeruginosa
and Escherichia coli 29
CHAPTER
ONE
INTRODUCTION
1.1 BACKGROUND
During the last century, the practice of herbal medicine
became mainstreams throughout the world. In spite of great advances in modern
medicine, plants still make important contributions to healthcare.
According to WHO a medicinal plant is a plant that contains
chemical compounds that possess established therapeutic activity .studies have
shown that such activity is due to mainly to the antimicrobial properties of
the plant . Any chemical substance inhibiting the growth or causing death of a
microorganism is known as antimicrobial activity.
Over the years, plant have been used as valuable sources of
natural products for maintaining human health .plants have been reported to
contain large varieties of chemical
substances that possess important
preservatives and curative therapies. About 80% of individuals from developing
countries use traditional medicines which have compounds derived from medicinal
plants (Igbinosa et al., 2009).
Due to its multidirectional promising aspects, the interest
in nature product continue to this day
(Kaul and Joshi, 2001; Kroll, 2001; Marriott, 2001; Bhattaram et al., 2002;Holt and Chandra, 2002).The use of herbal
drugs is once more escalating in the
form of complementary and alternative medicine (CAM) .
Mechanical plants are
of great importance to the health of
individuals and communities because they contain physiologically active
components which over the years have been exploited in the traditional medical
practices for the treatment of various ailments .it was the advent of
antibiotics in the 1950s that led to the use of plant derivatives as
antimicrobial (Agedah et al., 2010).
Despite the presence of various approaches to drug descovery, plants still
remain reservoir of natural medicine, clinical microbiologists have two reasons
to be interested in the topic of antimicrobial plant extracts, first, it is
very likely that these phytochemicals will find their way into the arsenal of
antimicrobial drugs prescribed by physicians; several are already being tested
in humans. It is reported that, on average, two or three antibiotics derived
from microorganisms are launched each year .After a downturn in that pace in
recent decades, the pace is again quickening as scientists realize that the
effective life span of any antibiotic is limited. Worldwide spending on finding
new anti-infectivee agents (including vaccines) is expected to increase 60%
from the spending levels in 1993. New sources, especially plant sources are
also being investigated. Second, the public is becoming increasingly aware of
problems with the over prescription and misuse of traditional antibiotics. In addition,
many people are interested in having more autonomy over their medical care.
Also interest in plants with antimicrobial properties has
been revived as a result of antimicrobial resistance. This resistance could be
attributed to indiscriminate use of commercial drugs or not taking an
antiboitic prescription according to the instruction, for example not taking
all the prescription in the treatment of infectious diseases (Aliero and
Afolayan, 2006). In addition, certain antibiotics present undesirable side
effects such as nausea, depression of bone marrow, thrombocytosis purpura and
agranulocytosis leading to the emergence of previously uncommon diseases. This
has given scientists the impetus to search for newer and alternative microbial
compounds from medicinal plants. Besides ,the high cost of conventional drugs,
particularly in resource limited communities has led to the increased use of
plants as an aternative for treatment of infectious disease (Marchese and
Shito,2001;Poole,2001).
Plant extracts and physiochemicals with antimicrobial
properties are of great important significance in therapeutic treatments. Their
antimicrobial properties are due to compounds synthesized in the secondary
metabolism of the plant. The screening of plant extracts and plant products for
antimicrobial activity has shown that plants represent a potential source of
novel antibiotic prototype (Afolayan, 2003).
The practice of herbal medicine in modernized form is now
gaining momentum in Nigeria, with various health officials and other persons
coming to realize the pontencies and efficacies of some of the indigenous
plants . Among the diseases that have been successfully managed traditionally
include malaria, epilepsy, indigestion, scurvy, diarrhea , dysentery ,
gonorrhea, worm infection and mental illness. Medical uses of these plants
ranges from administration of the plants root, barks, stem, leaves and seeds,
to the use of extracts from a whole plant.
The World Health Organization (WHO) estimates that
approximately 80% of the world’s inhabitants rely on traditional or herbal medicines for their
primary health care and plants
have long formed
the basis of sophisticated traditional
medicine systems and purportedly provide
excellent leads for
new drug developments (Sofowora,
1993; Pravi, 2006; Akinjogunla et
al., 2009). Herbal
medicine is the oldest form of healthcare known to
mankind and over50% of all
modern clinical drugs
are of natural products origin
and natural products
play important roles in
drug development in the pharmaceutical industry (Preethi et al.,
2010).
With the recent
trend of high
percentage resistance of
multiple drug resistant microbial
strains, efforts have
been intensified by researchers to
search for possible
alternatives (Adetunji et al., 2013). Medicinal
plants and traditional
preparation with
antimicrobial activities have
been used extensively
in the West African
regions. These plants of
medicinal important have been proven to be very effective even where treatments
with antibiotics failed (Oshim et al., 2016).
The rediscovery of the connection between
plants and health
is responsible for the
launching of a
new generation of
multicomponent botanical drugs,
dietary supplements and plant
produced recombinant proteins
(Akinjogunla et al., 2011). However, the
increasing problems of multidrug
resistant (MDR) bacteria
is of great
concern to both the
clinicians and pharmaceutical industries
and this has made it significant to search for newer drugs that are
highly effective, affordable,
acceptable and available (Akinjogunla
et
al., 2011).
1.2 STATEMENT OF
PROBLEM
The discovery and
development of antibiotics are among the most powerful and successful
achievements of modern science and technology for the control of infectious
diseases. However, the rate of resistance of pathogenic microorganisms to
conventionally used antimicrobial agents is increasing at an alarming rate.
Survey have revealed
that almost no group of antibiotics has been introduced to which resistance had
not been observed (Eloff, 2000)
1.3 AIMS AND OBJECTIVES OF STUDY
The major aim of this project work is to determine the
antimicrobial activity of the leaf extract of Vernonia amygdalina (bitter leaf) on Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa.
Objectives
The specific
objectives include:
•
To obtain the leaf extract of Vernonia amygdalina
•
To determine the zone diameter of
inhibition of the leaf extract of vernonia
amygdalina on some selected microorganisms
•
To determine the minimum inhibitory
concentration (MIC) of the leaf extract
•
To determine the minimum bactericidal
concentration (MBC) of the leaf extract.
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