ABSTRACT
Plants contain a wide range of natural drugs. Thus the present study investigated the effect of different solvent extracts of Spinacia oleracea on some test organisms. The well diffusion method was used in determining the antibacterial efficacy of plant extract against test organisms. Here two polar solvents namely ethanol and methanol were used. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were evaluated for all solvent extracts of Spinacia oleracea against each pathogen that recorded antibacterial activity and found the significant bactericidal MIC ranged between 0.152 to 1.250 mg/ml and MBC ranged between 1.250 to 5.000mg/ml against test bacteria. These results predict that the polar solvent extract of this plant can be used as natural antibacterial agent to cure E. coli, S. aureus and Klebsiella pneumonia which are test organisms.
TABLE OF CONTENTS
Title
page i
Certification ii
Dedication iii
Acknowledgements iv
Table
of Contents v
List
of Tables vii
List
of Plates viii
Abstract ix
CHAPTER ONE
1.0
INTRODUCTION 1
1.2 Aim and objectives of the study 3
CHAPTER TWO
2.0 LITERATURE
REVIEW 4
2.1 Description 4
2.1.1 Scientific Classification 5
2.2 Brief history of Spinach 5
2.2.1 Horticulture and Plant Specifics 7
2.3 Types
of Spinach 8
2.3 Nutritional
profile of Spinach 8
2.3.1 Micro nutrients 8
2.3.1.1 Magnesium 9
2.3.1.2 Iron 9
2.3.1.3 Zinc 10
2.3.1.4 Selenium 10
2.3.2 Phytochemicals 10
2.3.2.1 Antioxidant activity 11
2.3.2.2 Phenolic content 11
2.3.2.3 Flavonoid content 12
2.3.2.4 Carotenoid content 12
2.3.2.5 Vitamin C 13
2.3.2.6 Other
Phytochemicals in Spinach 13
2.4 The
health benefits of Spinach 14
2.4.1 Protection against ageing and improves brain function 15
2.4.2
Cancer-fighting antioxidants
16
2.4.3 Promotion of heart health 16
2.4.4 Assistance in fetal development 16
2.4.5 Promotion of body health 17
2.5 Chemical constituents of Spinach 17
2.5.1 Flavonoids 17
2.5.2 Phenolic Compounds 17
2.5.3 Carotenoids
17
2.5.4 Vitamins
17
2.5.5 Minerals 17
2.6 Traditional
uses of Spinach 18
2.7 Pharmacological
activities of Spinach 19
2.7.1 Protection against Gamma Radiation 19
2.7.2 Antioxidant
Activity 20
2.7.3 Inhibition of Mammalian DNA Polymerases 21
2.7.4 Sulphite Oxidase Activity
21
2.7.5 Hepatoprotective Activity 22
2.7.6 Inhibition of Clastogenisity 22
2.7.7 Anticancer Activity 23
2.7.8 CNS Depressant Effect 23
2.7.9 Inhibition of Proliferation of Human Gastric
Adenocarcinoma Cells 24
2.7.10 Anthelmintic
Activity 24
CHAPTER
THREE
MATERIALS
AND METHODS 26
3.1 Sample
Collection 26
3.2 Plant extraction 26
3.3 Preparation of the media 26
3.4 Qualitative phytochemical
screening 27
3.4.1 Test for saponin 27
3.4.2 Test for tannins 27
3.4.3 Test for flavonoids 27
3.4.4 Test for alkaloids 27
3.4.5 Test for steroids 27
3.4.6 Test for terpenoid 28
3.4.7 Test of phenol 28
3.5 Gram staining technique 28
3.6 Identification test 28
3.6.1 Catalase test 29
3.6.2 Citrate test 29
3.6.3 Motility test 29
3.6.4 Coagulase test 29
3.6.5 Indole test 29
3.6.6 Oxidase test 30
3.7 Sugar fermentation 30
3.8 Preliminary phytochemical
Screening 30
3.9 Screening of Antibacterial
Activity 31
3.10 Determination of Minimum
Bactericidal Concentration (MBC) 31
3.11 Determination of Minimum
Inhibitory Concentration (MIC) 32
CHAPTER
FOUR
RESULTS 33
CHAPTER
FIVE
DISCUSSIONS,
CONCLUSION AND RECOMMENDATIONS
5.1 Discussion 40
5.2 Conclusion 42
5.3 Recommendations 42
REFERENCES
LIST
OF TABLES
Table Title Page
1: Qualitative phytochemical constituents
of spinach leaf extract 34
2: Biochemical characterization of test
organisms 35
3: Antimicrobial activity of methanol
extract against test organisms 36
4: Antimicrobial activity of ethanol
extract against test organisms 37
5: Minimum Inhibition Concentration Values
of Spinach Leaf Extract 38
6: Minimum Bactericidal Concentration
Values of Different Extract of
Spinach 39
LIST OF FIGURES
Figure Title Page
1: Spinach (S. oleracea) 47
CHAPTER
ONE
1.0
INTRODUCTION
Plants are wealthy
sources of antimicrobial/antibacterial agents as they contain a different
variety of phyto-constituents. The past few decades have seen increasing
scientific interest in the both growth of plant tissue culture and the
commercial development of this technology as means of producing that valuable phytochemicals
(Ergene et al., 2006). These types of
natural drugs are always a better substitute of synthetic drugs (Ergene et al., 2006). For centuries, medicinal
plants have been used all over the world for the treatment and prevention of
various ailments, particularly in developing countries where infectious
diseases are endemic and modern health facilities and services are inadequate.
Elsewhere, many potent drugs have been purified from medicinal plants including
anti-malarial, anti-cancer, anti-diabetic and antibacterial compounds. The
medicinal value of a plant like spinach lies on bioactive phytochemical
constituents that produce a definite physiological action on the human body
(Akinmoladun et al., 2007). These phyto-constituents
work with nutrients and fibers to form an integrated part of defense system
against various diseases and stress conditions. The most important of these
bioactive constituents of plants are alkaloids, tannins and flavonoids (Edeoga et al., 2006).
However,
the increasingly high numbers of bacteria that are developing resistance to
classical antibiotics drive much of the current interest on natural
antimicrobial molecules in hope that they may provide useful leads into
anti-infective drug candidates. Pathogenic bacteria such as Salmonella, Escherichia coli, Listeria monocytogens, Bacillus cereus, and Staphylococcus aureus continue
to be a common and serious threat to public health (Centre for Disease Control
and Prevention, 2009) (and found in diarrhea cases and different foods) (El-Gamel,
2015; El-Gamel et al., 2015; Hosny et al., 2011). The potential of
developing a new antibacterial from plants are rewarding towards different uses
for the benefit of mankind (Merina and Souvik, 2013).
Spinacia oleracea (Spinach) is an edible flowering
plant. It is an annual plant having medicinal property native to central and
southwestern Asia (Kavitha and Ramadas, 2013). Spinacia oleracea belongs
to the family Chenopodiaceae and it is widely distributed, cultivated in India (Rao
et al., 2015). It is a vegetable
cultivated Worldwide, contains such nutritive components as vitamin C and iron.
It has a high nutritional value and is extremely rich in antioxidants,
especially when fresh, steamed, or quickly boiled. It is a rich source of vitamin
A (and especially high in lutein), vitamin C, vitamin E, vitamin K, magnesium,
manganese, folate, betaine, iron, vitamin B2, calcium, potassium,
vitamin B6, folic acid, copper, protein, phosphorus, zinc, niacin,
selenium and omega-3 fatty acids (Merina and Souvik, 2013).
Spinacia oleracea is
also packed with a number of antioxidants components like polyphenols,
flavonoids and carotinoids which are shown to possess anti-inflammatory
effects, anti-mutagenic potential, antineoplastic effects, as well as chemo-preventive
activities (Boivin et al., 2009;
Hait-Darshan et al., 2009). Spinach
as also contains photochemicals as tannins and alkaloids which have been found
to possess antibacterial activity against some organisms (Banso and Adeyemo,
2007).
Lanter-Marquez
et al. (2005) cited that S. oleracea is known to be rich in
flavonoids, phenolic acids and pigments such as lutein and chlorophyll which
are also antioxidants. It is a very good source of dietary fiber, protein and
Omega-3-fatty acids, zinc and vitamin B1, while this mixture of
conventional nutrient gives spinach a unique status in the antioxidant and
anti-inflammatory department (Mldadul, 2011).
S.
oleracea is a widely distributed annual plant, ranging from temperate
to tropical zones, and has a long history of use as both a medicinal and an
edible plant (Nebel and Heinrich, 2009). It has certain antibacterial
properties, as well as a contraceptive effect (Dong et al., 2010). It has been observed in use as diuretic, circulation
and lactation stimulants, and has also been recommended for diabetes, obesity,
acne, kidney stones and eye problems (Kuti and Konoru, 2004).
1.2 AIM AND OBJECTIVE OF THE STUDY
The
aim of the study is to determine the antibacterial activity of Spinach (S. oleracea). The specific objectives
include:
i)
To evaluate the antibacterial activity of
the different extracts of the leaves
ii)
To determine the phytochemical
constituents of the leaf extracts.
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