ANTI-PLASMODIAL PROPERTY OF MORINGA OLEIFERA SEED EXTRACT ON SWISS MICE
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
Title page i
Table of Contentsv
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
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
126.96.36.199Conventional therapeutic agents 18
188.8.131.52 Drug in pipeline 19
2.2 Traditional medicine 21
2.2.1 Control measures 22
2.3 Malaria vaccine 24
2.4 The experimental plant25
2.4.2 Social Economic importance of morning oleifera 29
2.4.3 Ecology and Cultivation 29
3.0 Collection of plant42
3 .1 Control drugs42
3.2 Experimental animal43
3.3 Materials and reagent43
3.4 Extraction from the plant seed43
3.5 Gas chromatography mass spectrometry44
3.6. Experimental Design50
3.7 Collection and inoculation of the parasite 50
3.8 Statistical Analysis 52
3.9 Presentation and statistical analysis of Data 52
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
5.0 Discussion 60
5.2 Recommendation 62
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)
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
2.0 LITERATURE REVIEW
2.1 Definition and History of Malaria
The term malaria was derived from the Italian ‘mala aria’’ meaning foul air (Service and Townson, 2002). It is a protozoal blood infection caused by mosquito-borne apicomplexan parasites of the genus Plasmodium, which are transmitted from one human to another via the bite of infected female Anopheline mosquito species (Carter and Mendis, 2002; Greenwood et al., 2005). United States National Institute of Allergy and Infectious Diseases (U.S. NIAID) defined malaria as: a disease caused by a parasite that lives part of its life in humans and part in mosquitoes (NIAID, 2007).
Malaria is an ancient disease that could be traced back to the very earliest human history. It was accepted as a disease by Hippocrates in the 4th century BC (Krettli and Miller, 2001). In the early 17th Century, the Peruvian bark of Cinchona tree was known to treat fever (CDC, 2016). In 1847, Heinrich Meckel identified black-brown pigment granules in the blood and spleen of insane person (David, 2006). Othmer Zeidler synthesized Dichloro-Diphenyl-Trichloroethane (DDT) in 1874 for his thesis. Alphonse Laveran noticed parasites, he called Oscillaria malariae, in the blood of malaria patient in 1880 (Bruce-Chwatt, 1981). The genus plasmodium was portrayed by Ettore Marchiafava and Angelo Celli in 1885 [Chavatte et al., 2007]. William MacCallum discovered the sexual stages of malaria parasite in 1897. In 1898, Camillo Golgi and others demonstrated that human malaria was transmitted by anopheline mosquitoes (Cox, 2010).
Chloroquine was discovered in 1934 by Hans Andersag (CDC, 2016). The liver stage of malaria parasite was elaborated by Henry Shortt and Cyril Garnham in 1948. In early 1950's, malaria was thought to be eliminated from the U.S. Then after, human infection with P. knowlesi was recognized in 1965.
Artemisinin was isolated from Qinghaosu plant (Artemisia annua) in 1971. Next, dormant stages in the liver were demonstrated in 1982 by Wojciech Krotoski (Cox, 2010; David, 2006; CDC, 2016). Polymerase chain reaction (PCR) based malaria detection was depicted in the early 1990's (Snounou et al., 1993) and mean while malaria rapid diagnostic tests (RDTs) were developed (Dietze et al., 1995)..