ANTIBACTERIAL ACTIVITY OF SPINACH (SPINACIA OLERACEA)

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Product Code: 00007151

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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. coliS. 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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