ABSTRACT
The antimicrobial activity of the ethanolic and aqueous crude leaf extracts of Guava (Psidium guajava) leaf was evaluated against two clinical isolates namely Methicillin resistant Staphylococcus aureus (MRSA) and Escherichia coli using Agar disc diffusion technique. The result obtained revealed that the ethanolic extract showed the highest zone of inhibition (20mm) against Escherichia coli at 400mg/ml concentration, and also showed the lowest zone of inhibition (11mm) against Methicillin resistant Staphylococcus aureus at 100mg/ml concentration. The phytochemical constituent of the Guava leaves identified were Alkaloids, Crude protein, Fat, Phenol, Saponins and Tannins. The MIC of Psidium guajava extract of Esherichia coli and Methicillin resistant Staphylococcus aureus are 25mg/ml and 12.5mg/ml respectively. The antimicrobial activity of the ethanolic and aqueous solvent extract against known aetiologic agents of diseases traditionally treated with Psidium guajava provide justification of the use of the herb in traditional medicinal practice in Nigeria.
TABLE OF CONTENTS
Cover page
i
Title page ii
Certification iii
Dedication iv
Acknowledgement v
Table of Contents vi
List of Tables vii
List of Figures viii
Abstract xi
CHAPTER
ONE
INTRODUCTION 1
1.1
Aims and Objectives 3
CHAPTER
TWO
LITERATURE
REVIEW
2.1
Historical Background of Antimicrobial Medicinal Plants 4
2.2 Some Medicinal Plants 5
2.3
Medicinal Properties of Psidium guajava (Guava
Pl ant) 7
2.3.1
Origin and Distribution 7
2.3.2
Traditional and Medicinal Uses 8
2.4
Plant Secondary Metabolites Associated With Antimicrobial Effects 9
2.5
Biological and Pathogenicity of Test Organisms 17
CHAPTER
THREE
MATERIALS
AND METHODS
3.1
Plant Collection and Preparation 20
3.2
Preparation of the Plant Extract. 20
3.3
Phytochemical Analysis 20
3.3.1Determination
of Saponins 20
3. 3.2
Determination of Phenol 21
3.3.3 Determination of Protein 22
3.3.4 Determination
of Tannin content 23
3.3.5
Determination of Alkaloid Content 23
3.3.6 Fat Content
Determination 24
3.4 Extraction of
plant material 25
3.5
Media Used 25
3.5.1
Preparation of Media 25
3.6
Sterilization 26
3.7
Collection of the Test Organisms 26
3.8 Biochemical
Identification of the Test Organisms 26
3.8.1 Gram Staining 26
3.8.2 Catalase
Test 27
3.7
Agar Disc Diffusion Method 28
3.8
Minimum Inhibitory Concentration (MIC) of Plant Extracts 29
3.9
Minimum Bactericidal Concentration (MBC) of Plant Extracts 30
CHAPTER
FOUR
4.0RESULT
31
CHAPTER
FIVE
DISCUSSION,CONCLUSION
AND RECOMMENDATION
5.1 Discussion 36
5.2 Conclusion
5.3 Recommendation 36
REFERENCES
APPENDIX
LIST
OF TABLES
|
Table
|
Title
|
Page
|
|
2.1
|
Antimicrobial active compound/class of compounds
from common medicinal plants.
|
7
|
|
4.1
|
Phytochemical composition of the guava (Psidium guajava) leaves extract.
|
32
|
|
4.2
|
Outlines the cultural, morphology and biochemical
confirmation of the bacterial isolates.
|
33
|
|
4.3
|
Diameter zone of (mm) produced by Guava leaves
against Escherichia coli.
|
34
|
|
4.4
|
Diameter zone of (mm) produced by Guava leaves
against Methicillin resistant Staphylococcus
aureus.
|
35
|
|
4.5
|
MIC and MBC values of plants against Escherichia coli concentrations
(mg/ml).
|
36
|
|
4.6
|
MIC and MBC values of plants against Methicillin
resistant Staphylococcus aureus concentrations
(mg/ml).
|
37
|
|
LIST
OF FIGURE
|
Figure
|
Title
|
Page
|
|
1
|
Flow chart showing the preparation of the Ethanolic and Aqueous
extract of the leaves
|
25
|
CHAPTER ONE
INTRODUCTION
In
the recent years, research on medicinal plants has attracted a lot of attention
globally. A large body of evidence has accumulated to demonstrate the promising
potential of medicinal plants used in the treatment of human diseases (Sher,
2009). More than 80% of the world’s population depend upon traditional
medicines derived from plants which have made large contribution to human
health and wellbeing. This is because they are either used directly as
photomedicines for the management of various ailments or they may become the
base and natural blue print for the development of new drugs (Cseke et al.,
2006). Plants have potent components of phytomedicine since time immemorial;
and man is able to obtain from them unmatched availability of chemical
diversity. Any part of the plants like bark, leaves, flowers, roots, fruits,
seeds, many contain natural active components (Gordon and David, 2007).
The
natural substances of plant origin may play fundamental roles in the host.
Pathogen relationship and products from different genera have been reported to
be biologically active, endowed with antimicrobial properties (Cowan, 1999).
Many plant species have been evaluated for their antimicrobial activities in
the past 20 years.
In
1992, it was reported that out of the three hundred thousand (300,000)
different plant species identified to be medical; only about five thousand
(5,000) have been studied for possible medicinal usefulness (Iwu et al., 1999).
Furthermore, the benefits of phytochemicals
cannot be over emphasized, they have low degrees of side effects which have
been one of the fears of using synthetic drugs. They are sedative and have regulating
and balancing effects on the body and thereby preventing disorders and
unbalanced coordination. They have a cleansing effect on the respiratory
system, the blood, skin and live due to its antibiotic and antioxidant content
(Pamptoma – Roger 1999). They are effective, gentle, safer cultural compatible
affordable and suitable for chronic treatments and for the treatment of
infectious disease (Iwu et al., 1999). There are some characteristics
which make phytomedicines unique and different, its principal active
ingredients is frequently unknown.
According
to Iwu (1993), infectious diseases accounts for one half of all deaths in
tropical countries, irrespective of the efforts made in controlling the
incidence of epidemics. Today many countries still rely on the medicinal values
of herbs and use of medicinal plants for their therapeutic practices (Thea et
al., 2008).
In
this vein, Nigeria which is having a vast heritage of knowledge and expertise
in herbal medicines is not an exception. With the recent advances made on using
plant extracts in inhibiting microbial growth, it was observed that
phytomedicines have antimicrobial effect against some human pathogens such as Staphylocuccus aureus, Escherichia coli, Pseudomonas aeruginosa,
and Salmonella typhi. People prefer using photomedicine to their synthetic
counterpart because its natural therapies. In this study Guava plant species
was selected on the basis of the available traditional medicinal information
and the leaves were screened for the antimicrobial properties. Psidium guajava (guava) is a native
of Mexico central American and Northern and Southern America. It comprises of
100 species of tropical shrubs and small trees. Psidium guajava is now
cultivated and naturized throughout the tropical and subtropical in Africa Southeast
Asia and in the Caribbean but can adapt to different climatic condition though
preferably dry climates. It belongs to the family of Myrtaceae. It is found
around town and villages and grows into a small tree 6 – 8 m high with white
flowers, bark is grayfish brown, hard or very rough and resistant to termites.
The fruits are up to 4 inches long fleshy, globosely, ovoid or pear – shaped;
generally yellowish or white when ripe. Ripe containing a reddish edible pulp
and numerous small woody seed embedded in the endocarp though some are seedless
or nearly so. It was reported by Burkhill of its higher amount of Vitamin C
than citrus – 80mg of vitamin C in 10g of fruit and also contain an appreciable
amount of vitamin A. It is also used in the treatment of malaria and skin
diseases (Okunza, 1973).
1.1 Aims and Objectives
To
evaluate the invitro antimicrobial activities of the guava leaves (Psidium
guajava) on two species of bacteria Methicillin resistant Staphylococcus aureus and Escherichia
coli
Specific
objectives
1. Identification
of test organisms.
2. To
screen the guava leaf for the presence of some phytochemical compound.
3. To
determine the Minimum Inhibitory concentration (MIC) and Minimum bactericidal
concentrations (MBCs) of the crude extracts against methicillin resistant Staphylococcus aureus and Escherichia coli.
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