Anti-Plasmodial Property Of Moringa Oleifera Seed Extract On Swiss Mice

Medical and Health Science Project and Seminar Material

Anti-Plasmodial Property Of Moringa Oleifera Seed Extract On Swiss Mice


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


Table of Contents


  • Contents pages
  • Title page
  • Certification
  • Dedication
  • Acknowledgements
  • Table of Contents
  • Abstract

Chapter One

1.0 Introduction

  • 1.1 Background Study
  • 1.2 Statement of the problem
  • 1.3 Justification
  • 1.4 Aim and Objectives of Study

Chapter Two

2.0 Literature Review

  • 2.1 Definition and history of Malaria
  • 2.1.2 Etiologic and vectors of malaria
  • 2.1.3 Epidermiology of malaria
  • 2.1.4 Life cycle of malaria parasite
  • 2.1.5 Molecular cell biology and pathogenesis
  • 2.1.6 Diagnosis of malaria
  • 2.1.7 Management of malaria
  • 2.1.7.1 Conventional therapeutic agents
  • 2.1.7.2 Drug in pipeline
  • 2.2 Traditional medicine
  • 2.2.1 Control measures
  • 2.3 Malaria vaccine
  • 2.4 The experimental plant
  • 2.4.1Moringa Oleifera
  • 2.4.2 Social Economic importance of morning oleifera
  • 2.4.3 Ecology and Cultivation

Chapter Three

3.0 Collection of plant

  • 3 .1 Control drugs
  • 3.2 Experimental animal
  • 3.3 Materials and reagent
  • 3.4 Extraction from the plant seed
  • 3.5 Gas chromatography mass spectrometry
  • 3.6. Experimental Design
  • 3.7 Collection and inoculation of the parasite
  • 3.8 Statistical Analysis
  • 3.9 Presentation and statistical analysis of Data

Chapter Four

4.0 Result

  • 4.1 Parasite density at different concentration of the extract of Maringa oliefera seed
  • 4.2 Percentage difference in parasitaemia inhibition at different concentration among seed

Chapter Five

5.0 Discussion, Conclusion and Recommendations

  • 5.1 Discussion
  • 5.2 Conclusion
  • 5.3 Recommendation
  • References

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:

  1. To assess the phytochemical components of the extracts from the seed of Moringa Oleifera
  2. To investigate the in-vivo anti-plasmodium activities of extracts from Moringa Oleifera seedon on laboratory animals at different concentration
  3. To evaluate the percentage parasitaemia inhibition at different concentration among Moringa olifera seed extract administration

1.5 Research Hypothesis (Null)

  • Ho: Extracts from the seed of Moringa Oleifera does not contain ligands
  • Ho: Extracts fromthe seed of Moringa Oleifera does not contain phytochemical components
  • Ho: Extracts from the seed of Moringa Oleifera shows no significant difference in percentage parasitaemia inhibiton

Chapter Five


5.0 Discussion, Conclusion and Recommendations

This study investigated in-vivo antiplasmodium of Moringa Oleifera seed extract. Related literature review was made considering scholars explanation of the subject matter. Relevant data for the study was generated through laboratory experiments conducted by the researchers. Three hypotheses were postulated and tested for the purpose of the study. The hypotheses were tested in this study using Analysis of Variance (ANOVA) and Duncan Multiple Test. All the hypotheses were tested at 0.05 level of significance.

This chapter contains the discussion of findings, conclusion of the study, recommendations for relevant stakeholders and suggestions for further studies.


5.1 Discussion of Findings

This study investigated in-vivo antiplasmodium of Moringa Oleifera seed extract. The findings revealed that extracts from the seed of Moringa Oleifera contain phytochemical components.Benzene-l-ethyl-2, 3-dimethil-P-Cymene O-Cymene has retention rate of 13.869 and percentage area of 2.57. Dodeane has retention rate of 18.504 and percentage area of 3.39, n-Hexadecanoic acid has retention rate of 39.250 and percentage area of 77.95, while 9-Octa decanoic acid-2-hydroxylethyl ester Oleeic anhydride Oleoyl Chloride has retention rate of 44.507 and percentage area of 16.09 respectively. The retention rate of 9-Octa decanoic acid-2-hydroxylethyl ester Oleeic anhydride Oleoyl Chloride was found to be higher than that of other peaks. With respect to percentage area, n-Hexadecanoic acid was higher than others.

The result of this study also revealed that anti-plasmodium activities of moringa olifera seed extract on laboratory animals shows significant effect at different concentration. Significant reduction in parasite density was also observed across the days from day 1 to day 3 for all the concentrations except for positive control that showed increase with days. The same level of reduction was noticed at 60% concentration, 80% concentration, 100% concentration and negative control (the group that was given Chloroquine). However, increase in parasite density was noticed in positive control (group that received distilled water). By implication, extracts from the seed of Moringa Oleifera contain anti-plasmodium components that can be used in the treatment of malaria like the way Chloroquine is being used. This result supported the earlier work of Mitchel (2002) who reported miring to effective in the treatment of malaria.

On the final note, the result of this research work revealed a significant difference in parasitaemia inhibition at different concentration among olifera seed extrat administration. Negative control recorded 100% inhibition, 100% concentration recorded 70% inhibition, both 60% and 80% concentration recorded 67% inhibition, 40% concentration recorded 56% inhibition, while positive control has no record of parasitaemia inhibition. This result is in line with Kang et al. (2004) who found that high concentration of oleifera extract was effective for the treatment of malaria.


5.2 Conclusion

This study investigated in-vivo antiplasmodium of Moringa Oleifera seed extract. Therefore based on the findings from this study, it was concluded that;

  1. Extracts from the seed of Moringa Oleifera contain phytochemical components.
  2. The anti-plasmodium activities of moringa oleifera seed extract on laboratory animals shows significant effect at different concentration.
  3. There is significant difference in parasitaemia inhibition at different concentration among oleifera seed extract administration.

5.3 Recommendation

Based on the results of data study and tested hypotheses, it is hereby recommended that:

  1. There should be awareness campaign on the effectiveness of Oleifera extract in the treatment of malaria.
  2. Pharmaceutical companies should device means of making malaria drugs with extracts from moringa.
  3. There should be research grants for medical relate students so as to enhance the discovery of ideas that could assist in medical practices.

5.4 Suggestions for Further Studies

This study investigated in-vivo antiplasmodium of Moringa Oleifera seed extract. Based on the findings of the study, the researcher recommends;

  1. That the topic of this study should be replicated to cover more sample/ specimen in the state, Geopolitical zones and the nation (Nigeria) at large.
  2. Antioxidant and anticancer activities of Moringa Oleifera leaves
  3. Some physio-chemical properties of Moringa oleifera seed oil extracted using solvent and aqueous methods

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