ABSTRACT
The antibacterial activities of the crude extracts from the leaves of Psidium guajava and Ocimum gratissimum were investigated on both gram positive and gram negative bacteria which were isolated from the soil, stream water, cow dung and the skin of humans. These organisms were cultured on different media. Gram staining and biochemical tests were carried out on the organisms for identification. Organisms like E. coli, Pseudomonas, Streptococcus, Staphylococcus aureus, Salmonella, Shigella and Bacillus were identified. The plants were extracted after two weeks of air drying and grinding using the grinder. Two solvents, ethanol and water were used for extraction and about 30g each of the leaves were macerated 200ml of the solvents for 48hrs, which were then evaporated to dryness using the water bath to obtain the crude extract which weighed about 4g each of the extract in ethanol and 2.5g each of the extract in water. The sensitivity, minimum inhibitory concentration and the minimum bactericidal concentration of the extracts were tested on the isolates and the organisms showed susceptibility to the extracts except Pseudomonas. The phytochemical screening of the crude extracts from the plants were also investigated and constituents like tannins, saponins alkaloids and others were seen at different concentrations on different leave extract. The result all showed that these plants can be used in the formulation of oral antibacterial drugs and support the reason why some countries use them as traditional herbal medicine. In conclusion the leave extracts were effective and had a wide range of activity against both gram positive and gram negative bacteria.
TABLE OF CONTENTS
Title
page i
Certification
ii
Dedication iii
Acknowledgement iv
Table of contents v
List
of tables ix
Abstract x
CHAPTER
ONE
INTRODUCTION 1
1.0
BACKGROUND
OF THE STUDY 1
1.1Brief description of Psidium guajava (Guava) 2
1.2 Brief description of Ocimum gatissimum (Scent leave) 2
1.3 Justification for the work 3
1.4 Aims and objectives 3
CHAPTER TWO
LITERATURE REVIEW
4
2.0
Overview 4
2.1 Psidium
guajava 4
2.1.1
Chemical composition 5
2.1.2 The leaves 6
2.1.3 The roots 7
2.1.4 The seeds 7
2.1.5
Antibacterial activity:
7
2.1.6 Anti-diarrheal 8
2.1.7 Anti-inflammatory
effect: 8
2.1.8
Antispasmodic: 9
2.1.9 CNS
activity 9
2.1.10
Conjunctivitis: 10
2.1.11
Coughs 10
2.1.12
Diabetes: 10
2.1.13 Skin
use
10
2.1.14 Vaginal
disorders 11
2.2 Ocimum gratisimum 12
2.2.1 Chemical
composition 12
2.2.2
Antibacterial activity 12
2.2.3
The oil of Ocimum gratissimum 13
2.3 Description of test organism 14
2.3.1 Salmonella sp; 14
2.3.2. Shigella sp 14
2.3.3 Escherichia
coli; 15
2.3.4
Bacillus cereus;
15
CHAPTER THREE
3.0 MATERIALS AND METHOD 16
3.1 Sources of leaves 16
3.2 Sources
of isolation of
organisms 16
3.3 Isolation of the
organisms 16
3.4 Preparation of media and culturing of 16
3.5 Identification of
organisms 17
3.6 Biochemical
identification of the organisms 17
3.6.1 Catalase
test 17
3.6.2 Indole
test 17
3.6.3 Oxidase
test 17
3.6.4 Coagulase
test 17
3.3.5 Motility
test 17
3.6.6 Citrate
test 18
3.7 Preparation of the extracts
18
3.8 Phytochemical screening
18
3.8.1
Flavonoids:
19
3.8.2 Tannins: 19
3.8.3 Test for alkaloids: 19
3.8.4 Test for sterols and steroids: 20
3.8.5
Test for the carborhydrate: 20
3.8.6 Starch 20
3.9 Preparation of the inoculum 21
3.10 Bacterial
sensitivity testing using agar well diffusion method 21
3.11 Determination of the
minimum inhibitory concentration of the plant extracts 21
3.12 Determination of the
minimum bactericidal concentration (MBC) 22
CHAPTER FOUR
4.0 RESULTS 23
4.1 Result from the gram staining reaction 25
4.2 Result from biochemical tests 25
4.3 Result of the phytochemical screening P. guajava and O. gratissimum 26
4.4 Antibacterial sensitivity test 27
4.4.1 P. guajava
27
4.4.2 O.
gratissimum 28
4.5 Determination of the minimum inhibitory
concentrations using tube dilution method 28
4.5.1 MIC of P.
guajava in ethanol
28
4.5.2 MIC of P.
guajava in water 29
4.5.3 MIC of O.
gratissimum in ethanol 30
4.5.4 MIC of O.
gratissimum in water 30
4.6 Determination of MBC 31
CHAPTER FIVE
5.1 Discussions
32
5.2 Conclusion
33
REFERENCES 34
LIST OF TABLES
Table 4.1 Gram staining
reaction 25
Table 4.2 Biochemical
test of different isolates 25
Table
4.3 Phytochemical screening of P. guajava and O. gratissimum 26
Table 4.4
Antibacterial sensitivity test with P. guajava 27
Table 4.5
Antibacterial sensitivity test with O.
gratissimum 28
Table 4.6
MIC of P. guajava in ethanol 28
Table 4.7
MIC of P. guajava in water 29
Table 4.8
MIC of O. gratissimum in ethanol 30
Table 4.9
MIC of O. gratissimum in water 30
Table 4.10
MBC of P. guajava and O. gratissimum in ethanol and water
CHAPTER ONE
INTRODUCTION
1.0
BACKGROUND
OF THE STUDY
The use of plants and
herbs extract in the treatment of human ailments is a very ancient art, a practice
that has been passed on for generations and scientists in Africa and other
developing countries are conducting research into local plants abundant in the
continent for their possible use in traditional medicine (Nneamaka, 1991).
Research into traditional
plants and herbs received further boost due to increasing resistance to many
orthodox medicine and thus a research for new organic molecules of plants with
antimicrobial properties (Sofowara, 1993). Bacteria are responsible for high
mortality rates in numerous developing countries with as many as 50,000 people
dying as a consequence of infection. Plants remedies are increasingly being recognized
by scientists as a very important cost alternative to industrially produced
antibiotics which are not available to all who need them because of their high
price. So present work expresses that plants have great potential as
antimicrobial compounds against microorganisms (Rajendra et al., 2012).
Amongst the various forms
of treatment for diarrhea, the use of traditional plant remedies is common and
widespread. The world health organization (WHO) has categorized more than 20000
plant species with medicinal properties providing treatment for such complaints
as pneumonia, ulcers, diarrhea, bronchitis, colds and diseases of the
respiratory tracts. One method among the many ways in which plants are used in
popular remedies is to extract and consume essential plant oils. Essential oils
are complex chemical mixtures, typically composed of more than a hundred
compounds by and large are responsible for plant aromas. They are obtained from
different parts of the plants. Flowers, leaves, seeds, barks, and tubers of
many plants have medicinal properties (Goncalves et al., 2008).
Increased
developments of resistance to current antibiotics have strengthened scientific
research for discovery of new drugs. However, new leads/hits in drugs discovery
are been developed from natural sources due to growing scientific link between
the folkloric medicinal use of some of these natural products especially of
plant origin, to biological activity. Hence plants continue to provide a good
source for new drugs (Kunle and Egharevba, 2009; Kunle et al., 2013, Begum et al.,
2002).
1.1 Brief description of Psidium guajava (Guava)
Guava
is a small tropical tree that grows up to 35 feet tall; it is grown for its
fruits in tropics. It is a member of the Myrtaceae family, with about 133
genera and more than 3800 species. The leaves and bark of Psidium guajava free have a long history of medicinal uses that are
still employed today (Nwinyi et al.,
2008). The leaves and bark of the guava plant have been used to treat diarrhea,
other gastro intestinal disorders, tooth aches, colds and swelling in areas
such as Tahiti (Weiner, 1971), India (Dutta et
al.,2000), Africa (Rabe and
Vanstaden, 1997;Tona et al., 2000; Len et
al., 2002). Guava is used for skin disorders such as astringent for acne,
rashes and ringworm (Coe and Anderson, 1999). (Duta et al., 2000) tested Psidium
guajava against dermatophytes, Trichophyton
tonsurans, Trichophyton rubrum, Microsporium fulvrum and Candida albicans.
1.2 Brief description of Ocimum gatissimum (Scent leave)
Ocimum gatissimum
is called ‘efinrin’ by the Yorubas of the south western part of Nigeria,
‘nchanwu’ by the Igbos and ‘Daidoya’ by the Hausas. It has been reported to
contain the terpenoids, eugenol, thymol saponins and alkaloids (Gill, 1988). Ocimum gratissimum is germicidal and has
been found widely used in tooth pastes and mouth washes as well as some topical
ointments (Nakamu et al., 1999;
Holets et al., 2003; Pessoa et al., 2003.
Ocimum gratissimum leaf
or the whole herb are popular treatments for diarrhea (Dalziel, 1956). In fact,
the water saturated oil has been shown to be proportional to the thymol content
in preparations where Ocimum gratissimum
is used as cold infusion (El-said et al.,
1969).
1.3 Justification for the
work.
This
work is been embarked upon due to increased resistance by bacteria to many
already synthesized drugs due to changes or modification in the active site of
those drugs and the abuse of orthodox drugs.
Due
to the high price of the orthodox medicines in the market, the antimicrobial
effect of these plant extracts on bacteria will help to reduce the
overdependence on orthodox or synthetic drugs which greater population of the
society cannot afford due to economic/financial restriction.
1.4 Aims and Objectives.
1.
To determine the active components in Ocimium
gratisimum and Psidium guajava
2.
To isolate and identify some bacteria like E.
coli, Staphylococcus aureus, Salmonella and Shigella.
3.
To determine the Minimum inhibitory concentration and the minimum bactericidal
concentration of Ocimum gratissimum
on the isolates.
4.
To determine the Minimum inhibitory concentration and Minimum bactericidal
concentration of Psidium guajava on
the isolates.
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