ANTI-PLASMODIAL PROPERTY OF MORINGA OLEIFERA SEED EXTRACT ON SWISS MICE

TABLE OF CONTENTS

Contents                                                                                                                     pages

Title page                                                                                                                    i                                                                                  

Certification                                                                                                                ii         

Dedication                                                                                                                  iii        

Acknowledgements                                                                                                    iv        

Table of Contents                                                                                                       v         

Abstract                                                                                                                      vi

CHAPTER ONE

1.0 Introduction                                                                                                          1

1.1 Background Study                                                                                                            1

1.2 Statement of the problem                                                                                     2

1.3 Justification                                                                                                           3

1.4 Aim and Objectives of Study                                                                               4

CHAPTER TWO

2.0 Literature review                                                                                                   6

2.1 Definition and history of Malaria                                                                         6

2.1.2 Etiologic and vectors of malaria                                                                        7

2.1.3 Epidermiology of malaria                                                                                  8

2.1.4 Life cycle of malaria parasite                                                                             10-13

2.1.5Molecular cell biology and pathogenesis                                                            13

2.1.6Diagnosis of malaria                                                                                           14

2.1.7 Management of malaria                                                                         18

2.1.7.1Conventional therapeutic agents                                                                      18

2.1.7.2 Drug in pipeline                                                                                              19

2.2 Traditional medicine                                                                                             21

2.2.1 Control measures                                                                                               22

2.3 Malaria vaccine                                                                                                     24

2.4 The experimental plant                                                                                          25       

2.4.1Moringa Oleifera                                                                                                 26       

2.4.2 Social Economic importance of morning oleifera                                              29

2.4.3 Ecology and Cultivation                                                                                    29       

CHAPTER THREE                                                            

3.0 Collection of plant                                                                                                            42       

3 .1 Control drugs                                                                                                       42       

3.2 Experimental animal                                                                                             43       

3.3 Materials and reagent                                                                                            43       

3.4 Extraction from the plant seed                                                                             43       

3.5 Gas chromatography mass spectrometry                                                              44       

3.6. Experimental Design                                                                                            50       

3.7 Collection and inoculation of the parasite                                                            50

3.8 Statistical Analysis                                                                                                52

3.9 Presentation and statistical analysis of Data                                                         52

CHAPTER FOUR

4.0 Result                                                                                                                    53      

4.1 Parasite density at different concentration of the extract of Maringa oliefera seed 55

4.2 Percentage difference in parasitaemia inhibition at different concentration among   

      seed                                                                                                                          58

CHAPTER FIVE

5.0 Discussion                                                                                                             60

5.1 Conclusion                                                                                                            61     

5.2 Recommendation                                                                                                  62

ABSTRACT

Malaria is an increasing worldwide threat, with more than three hundred million infections and one million deaths every year. Due to the emergence of antimalarial drug resistance, the continuous search for antimalarial agents. This study was conducted to determine the antimalarial efficacy of Moringa oleifera Seed extract in Swiss albino mice infected with Plasmodium berghei .After extraction, phytochemical screening and gas chromatographic mass spectrometry (GC-MS) screening of the extract, the mice were grouped into six groups, five per group. Designated  as 40% treated with 40mg/kg of the Maringa oliefera seed extract, 60% treated with 60mg/kg, 80% treated with 80mg/kg,100% treated with 100mg/kg and positive control treated with distilled water while negative control was given choloroquone.  For the period of 3 days at 12 hours interval.  Parasite density was determine by preparing of thick and thin blood film, stain with Giemsa stain and view under microscope to determine the antiplamodial activity of the extract

CHAPTER ONE

1.0 INTRODUCTION

1.1 Background of the study

 Since the beginning of human civilization, medicinal plants have been used by mankind for its therapeutic value. Nature has been a source of medicinal agents for thousands of years and an impressive number of modern drugs have been isolated from natural sources. Many of these isolations were based on the uses of the agents in traditional medicine. The plant-based, traditional medicine systems continues to play an essential role in health care, with about 80% of the world’s inhabitants relying mainly on traditional medicines for their primary health care (Owolabi et al., 2007). Medicinal plants are plants containing inherent active ingredients used to cure disease or relieve pain (Okigbo et al., 2008). The medicinal properties of plants could be based on the antioxidant, antimicrobial antipyretic effects of the phytochemicals in them (Cowman, 1999; Adesokan et al., 2008). The ancient texts like Rig Veda (4500-1600 BC) and Atharva Veda mention the use of several plants as medicine. The books on ayurvedic medicine such as Charaka Samhita and Susruta Samhita refer to the use of more than 700 herbs (Jain, 1968). According to the World Health Organization (WHO, 1977) “a medicinal plant” is any plant, which in one or more of its organ contains substances that can be used for the therapeutic purposes (Okigbo, 2009). The term “herbal drug” determines the part/parts of a plant (leaves, flowers, seed, roots, barks, stems, etc.) used for preparing medicines.

1.2 Statement of the problem

Malaria is a potentially deadly parasitic disease of global public health relevance. The infection is known to cause death and illness in children and adults, especially in tropical countries. In Nigeria, malaria is termed to be endemic and perennial in all parts, with seasonal variations more pronounced in the Northern part (Caraballo, 2014).   According to the 2010 national census, 24.2 million Ghanaians are at risk of malaria infection. Children under five years and pregnant women however stand a higher risk of severe illness due to declined immunity (WHO, 2014). The control of malaria requires an integrated approach, including prevention, which deals primarily with vector control and prompt treatment with effective anti-malarial (WHO, 2014).

Management of malaria has seen a lot of changes, mainly as a result of resistance development of P. falciparum against anti-malarials in use. For instance, Chloroquine, which used to be one of the most effective drugs, has now been proven to be ineffective in malaria treatment (Greenwood et al., 2010). Currently, WHO recommends a combination therapy involving any of the artemisinins and other classes of antimalarials for the treatment of uncomplicated malaria (WHO, 2014).

Some of the recommended combinations include, Artesunate -Amodiaquine, Artemether - Lumefantrine, Atovaquone-Proguanil, Chloroquine-Proguanil, and Mefloquine– Sulphadoxine-Pyrimethamine (CDC, 2016).

A school of thought holds that, the solution to plasmodial resistance development rests in the use of traditional medicinal plants (Liu et al., 2010). Several authors have documented medicinal plants that are used in the treatment of malaria in Ghana and other African countries (Cox, 2010). The story behind the discovery of the artemisinins, as an example, seeks to provide a head way in the discovery of bioactive constituents from medicinal plants for combating malaria (Cox, 2010). Armed with information from successful traditional treatments of malaria, it is possible to discover novel compounds from plants that could be developed into potent antimalarials. This study was thus carried out to determine the antiplasmodium activities of extract from the seed of Moringa oleifera Lam (Moringaceae).

1.3 Justification of the study

In sub-Saharan Africa, infectious diseases remain the predominant cause of illness and death. Plasmodium falciparum malaria alone causes an estimated 1 million deaths annually (Lopez et al., 2009). Malaria remains the most serious and widespread protozoal infection of humans. Over 40% of the world’s population is at risk of contracting malaria, which is endemic in 91 countries, mostly developing. The disease is widespread in tropical and subtropical regions that are present in a broad band around the equator, (Caraballo, 2014).  This includes much of Sub-Saharan Africa, Asia, and Latin America. The World Health Organization estimates that in 2012, there were 207 million cases of malaria. That year, the disease is estimated to have killed between 473,000 and 789,000 people, many of whom were children in Africa, (WHO, 2014). Malaria is commonly associated with poverty and has a major negative effect on economic development, (Worrall et al., 2009). In Africa it is estimated to result in losses of $12 billion USD a year due to increased healthcare costs, lost ability to work and effects on tourism, (Greenwood et al., 2010). However drug resistance to malaria has been a major challenge to public health. Many authors have documented drug resistance strains of plasmodium falciparun (WHO, 2010). However many countries such as Mali, China, Vietnam, Sri Lanka and India has integrated herbal products into their health care delivery system for effective treatment (Kazambe and Munyarari, 2006). But in Nigeria, natural products is yet to gain wider acceptance by the physicians due to the facts that most natural products does not have a biochemical explanation to their mode of action. Also there is paucity of information on the anti-plasmodium properties of Moringa Oleifera seed extracts, against the background this study was carried out.

1.4 Aim and objectives

1.4.1 Aim of the study

This study aims at investigating the ligands and in-vivo anti-plasmodium study of Moringa Oleifera seed extract.

1.4.2 Specific objectives

The specific objectives of this study where: 

·         To assess the phytochemical components of the extracts from the seed of Moringa Oleifera

·         To investigate the in-vivo anti-plasmodium activities of extracts from Moringa Oleifera seedon on laboratory animals at different concentration

·         To evaluate the percentage parasitaemia inhibition at different concentration among Moringa olifera seed extract administration

Research Hypothesis (Null)

H_o: Extracts from the seed of Moringa Oleifera does not contain ligands

H_o: Extracts from the seed of Moringa Oleifera does not contain phytochemical components

H_o: Extracts from the seed of Moringa Oleifera shows no significant difference in             percentage parasitaemia inhibiton.

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