ETHNOBOTANICAL STUDY AND ANTIMICROBIAL SCREENING OF MEDICINAL PLANTS FOUND IN CHEREBES AND ENDO VILLAGE KEIYO COUNTY
PRESENTED BY: KOSGEY JANET CHERUIYOT
SC/PGB/049/09
A RESEARCH PROPOSAL WRITTEN AND SUBMITTED TO THE SCHOOL OF SCIENCE, DEPARTMENT OF BIOLOGICAL SCIENCE AS A PARTIAL COMPLETION OF A DEGREE COURSE IN MASTER OF SCIENCE IN MICROBIOLOGY
2011
DECLARATION
I, Kosgey Janet Cheruiyot, duly declare that this is my original work and has not been presented in any other university for any other awards. Works from other sources have been dully acknowledged.
STUDENT
NAME: KOSGEY JANET CHERUIYOT
SIGN:……………………………………,
DATE: 22nd NOV, 2010
UNIVERSIITY SUPERVISOR
NAME: Dr. NJENGA
HEAD DEPARTMENT OF BIOLOGICAL SCIENCE
SCHOOL OF SCIENCE, MOI UNIVERSITY
SIGN:………………………………
DATE: 22nd NOV, 2010
FIELD SUPERVISOR
NAME: Dr. MUTAI
CENTRE FOR TRADITIONAL MEDICINE,
KENYA MEDICAL RESEARCH INSTITUTE (KEMRI),
SIGN:…………………………
DATE: 22nd JUNE, 2011
TABLE OF CONTENTS
DECLARATION 2
TABLE OF CONTENTS 3
ABSTRACT 4
CHAPTER ONE 5
1.0 INTRODUCTION 5
1.1 PROBLEM STATEMENT 5
1.2 JUSTIFICATION 6
1.3 OBJECTIVES 7
1.4 HYPOTHESIS 8
CHAPTER TWO 9
2.0 History of Drug Development 9
2.1 Medicinal Plants 9
2.2 Plants as an Alternative Source of Therapeutic Agents 10
2.3 Plants and Plant Parts Used 11
2.4 Documentation 11
2.5 Plant Parts Used 12
2.6 Cultivation of Medicinal Plants 12
2.7 Synergsm 13
2.8 Activity of Some Medicinal Plant 13
3.0 METHODOLOGY 14
3.1 Preliminary experimental design 14
3.2 Collection of the plants from the field 14
3.3 Reagents 14
3.4 Extraction of plant materials 15
3.5 Microbial Test Organisms 15
3.6 Test strains 15
3.7 Preparation of test organisms 17
3.8 Preparation of McFarland Standard 17
3.9 Antimicrobial Screening Test 18
3.10 Determination of Minimum inhibitory concentration 18
3.11 Cell toxicity 19
3.14 Statistical data analysis 20
3.15 Conservation 20
EXPECTED OUTPUT 20
BUDGET 21
WORKPLAN 24
REFERENCES 26
ABSTRACT
Traditional herbal remedies are an important component in the provision of primary health care. They serve as an alternative to conventional medicines which are normally too expensive for most Kenyans, but the rate at which these herbal remedies are disappearing from their natural habitat is alarming. There is little or no documentation on the uses of these plants. Moreover, the indigenous knowledge on medicinal properties of these plants is secretly guarded by the herbalists and is not available to most Kenyans. The methods that were previously used to pass the information are presently not applicable. The research seeks to validate the claims attributed to the medicinal plants found in Keiyo County based on the indigenous knowledge and also to document the medicinal plants found in the same area. The roots, stems and leaves of these plants will be collected from Kerio valley Cherebes and Endo village in Keiyo County. The samples will be extracted using water, ethyl acetate, hexane and methanol solvent. The crude extract’s phytochemical composition will be determined. The extracts will be tested for antimicrobial activity using Staphylococcus aureus, Klebsiella pneuformans, Escherichia coli, Salmonella typhi, Cryptococcus neoformans, Candida albicans, Microsporum gypsium and Trichopyton metangropyte. The minimum inhibition concentration of the herbs will be determined by incubating microorganisms in different concentrations of the extract, for 24 hours at 37oC then cultured in solid media for bacteria and yeast and at 27oC for 48 hours for moulds. The data obtained will be analysed using two way ANOVA. Means will be separated using Tukey’s HSD test at 5 % level of significance. The results will show the rich biodiversity we have and the need to conserve it.
CHAPTER ONE
1.0 INTRODUCTION
Traditional medicine has and still remains the main source for a large majority(80%) of people in Ethiopia for treating health problems and medicinal consultancy including consumption of the medicinal plants has a much lower cost than modern medicine attention(Tildhun and Giday, 2007). Traditional medicine is used throughout the world as it is dependent on locally available plants, which are easily accessible, and capitalizes on traditional wisdom-repository of knowledge, simple to use and affordable. (Tesfaye and Demissew, 2009). The traditional methods, especially the use of medicinal plants, still play a vital role to cover the basic health needs in the developing countries too and more over, the use of herbal remedies has increased in the developed countries in the last decades. In this connection, plants continue to be a rich source of therapeutic agents. The remarkable contribution of plants to the drug industry was possible because of the large number of phytochemical and biological studies all over the world. The Indian subcontinent is endowed with rich and diverse local health tradition, which is equally matched with rich and diverse plant genetic source. A detailed investigation and documentation of plants used in local health traditions and ethno-pharmacological evaluation to verify their efficacy and safety can lead to the development of invaluable herbal drugs or isolation of compounds of therapeutic value. (Kesaran et al, 2007)
1.1 PROBLEM STATEMENT
The documentation of medicinal plants in Kenya is very poor .A few of documentation have been done on Plants from Central Kenya as Veterinary Medicine by Ngugi. Also few plants have been studied from Kakamega rain forest; hence there is need to document medicinal plants found in Keiyo County. Resistance to drugs by microorganisms has increased. This resistance have been attributed to overdose and under dose of drugs due to over counter prescription of drugs, ability of microorganisms to undergo genetic variability (mutation), use of antibiotics in food preservation and general misuse of drugs (Gislene, et a1 2000) hence there is need to come up with sensitive and effective drugs. Keiyo County provides a myriad of the medicinal plants. However, there is need to carry out proper identification of the medicinal plants, their antimicrobial activity and know their phytochemical composition. This information will be necessary later for conservation purposes and cheap drug manufacturing in Kenya.
1.2 JUSTIFICATION
Natural products have provided biologically active compounds for many years and many of today’s medicines are either obtained directly from natural source or were developed from a lead compound originally obtained from a natural source (Graham 2001).
In Kenya 75 plants species from 34 families are used to cure 59 ailments in traditional medicine of central Kenya, 80% of South Africans use herbal remedies for their physical and psychological health care at different stages of their life. Also in United State, 36 of the 101 plants species implicated in drugs discovery are weedy species found mainly in disturbed habitant (Lewis, 2009). There is good reason to be optimistic about the potential future usefulness of plants based on natural products as a continued source of potential lead compounds. Within many thousands of years of trial-and –error by evolution on her side, Mother Nature is a vastly superior experimentalist to any mere human organic chemist (Thomas, 2005). Many of this lead compounds are useful drugs in themselves e.g. morphine and quinine, while other have been the basis for synthetic drugs e.g. local anaesthetics developed from cocaine. Plants still remain a promising source of new drugs and still continue to do so. Occasionally useful drugs which have recently been isolated from plants include the anticancer agents, toxol, from the jaw tree and the anti malarial agents artemisinin from Chinese plant (Graham, 2001.) In South Africa different plant species are used to treat several diseases especially among the rural population where western medicine is either not accessible or affordable. Today, about 200,000 traditional healers practice herbal medicine in South Africa and a high percentage of the population use traditional medicine as their primary source of health care (Lewis, 2009). Most biological active natural products are secondary metabolites with quite complex structures. This has the advantage that they are extremely novel lead compounds. In general natural products are particularly good at providing many new chemical structure which no chemist would dream of synthesizing. For example, the anti-malarial drug artemisinin is one such example, containing an extremely unstable-looking trioxane, ring-one of the most unlikely structure to have appeared in recent years (Grahams, 2001). Plant evolution has culminated in a wide variety of bio-molecules that affect any animal that may choose to eat them; it is biologically advantageous for other plants to produce noxious chemicals to decrease the likelihood of their being eaten. Because of these diverse biological activities, any of these non-human biosynthetic molecules could in principle, be a lead compound for human drugs discovery (Thomas, 2005). Considering the debt medicinal chemistry owes to natural world, it is soberly to think that very few plants have been fully studied, the vast majority have not been studied at all. The rainforests of the world are particularly rich in plant species which have still to be discovered, let alone studied. Who knows how existing new lead compounds await discovery for the fight against cancer, AIDs or other many of human afflictions? This is why the destruction of rainforest and other ecosystems is so tragic. Once these ecosystems are destroyed, unique plants species are destroyed with them. Medicine has lost potentially useful plants for ever. For example, siphion a plant cultured near Cyrene Is now extinct. It is almost certain that many more useful plants have become extinct without medicine ever being aware of them (Graham, 2001)
1.3 Specific objectives
1 To identify and document traditional medicinal plants found Endo and Cherebes villages in Keiyo County.
2 To determine the antimicrobial activity of these medicinal plants.
3 To determine the phytochemical constituents of medicinally active plants.
4 To determine toxicity of these medicinal plants
5 To provide a scientific rationale to validate the claimed therapeutic properties of the medicinal plants.
1.4 HYPOTHESIS
NULL HYPOTHESIS
1. Medicinal plants from Keiyo county has no significant antifungal activity and antibacterial activity
2. Medicinal plants from Keiyo county high toxicity level
3. Medicinal plants from Keiyo county has very high minimum inhibition concentrations
4. Medicinal plants from Keiyo county has no active ingredient
CHAPTER TWO
2.0 History of Drug Development
In primitive times, surgical ‘therapies’ for epilepsy included ‘trephining’ holes through the patients’ skull in order to release ‘evil humors and devil spirits’. The ancient also employed ad hoc ‘medicinal’ therapies, ranging from rubbing the body of an epileptic with genitals of a seal, to inducing episodes of sneezing at sunset. Also human blood was widely regarded as curatives.
Charlatans would massage the heads of epileptic, thereby re-aligning the bone plates of the skull, taking pressure off the brain and alleviating the curse of epilepsy.
By the Middle Ages, scientific foundations of epilepsy therapy were formed. They used ‘magical prescriptions’, which ranged from taking dogs bile to human urine. During Renaissance magical treatment was subjected by medicinal profession in favor of ‘rational and scientific’ Gaelic therapies, which relied upon forced vomiting and bowel purging with concomitant oral administration of peony extracts. During this time epileptic were treated by castration, circumcision and clitoridectory since epilepsy was perceived as secondary hyper-sexuality. This mode of treatment failed or killed unfortunate patients. During late Renaissance inorganic salts were used as putative therapies.
Antiepileptic copper salts were introduced in the first century with reported success; this led to attempts of lead, bismuth, tin, silver, iron and mercury giving rise to metallotherapy. This led to widespread failure due to lack of efficiency and excessive toxicity. In mid 19th century quackery prospered as a treatment of epilepsy. Phlebotomy was adopted and king CharlesII was among the unfortunate patient treated by this method, he bled to death. (Nogrady and Weaver, 2005)
2.1 Medicinal Plants
The economic crisis, high cost of industrialized medicines, inefficient public access to medicinal and pharmaceutical care, in addition to the side effects caused by synthetic drugs are more of the factors contributing to the control role of medicinal plants in health care. There is currently an increased in number of immune-compromised individuals due to advances in medical technology and a pan-epidemic of HIV infections with the rise in at-risk patients, the number of invasive fungal infections has dramatically increased in both in developed and developing countries. (Susana et al, 2007).
Also many infectious microorganisms are resistant to synthetic drugs, hence an alternative therapy is much needed (Swapna and Kannabiran, 2006). In the present scenario, an emergence of multiple drug resistance in human pathogens and small numbers of antimicrobials classes available stimulates research directed towards the discovery of novel antimicrobial agents from other sources. (Susana et al, 2007)
Among the 887 medicinal plant species in Ethiopia, about 26 species are endemic and are becoming increasingly rare and are at verge of extinction. (Tesfaye and Dimissew, 2009) the solution is to practice cultivation and conservation of endangered species. Another factor that contributes to the problem is the explosive growth in the use of plant-base products in the last ten years. Hundreds of plant species are being converted into dozens of different product types, for use across a wide variety of markets. These plants and derived plant products are traded locally, regionally and internationally, as neuraticals, dietary supplements, phytomedicines, homeopathic drugs, aromatherapy, oils, flavors, fragrance and food additive. Because of the vast growing multi-million trade, there is need to protect the medicinal plants by forming laws that governs harvesting of medicinal plants.
2.2 Plants as an Alternative Source of Therapeutic Agents
Ethno-botanical studies are often significant in revealing locally important plant species especially for the discovery of crude drugs. Documentation of traditional knowledge, especially on the medicinal use of plants has provided many important drugs of modern days (Tildhun and Giday, 2007). Over 50% of all modern clinical drugs are of natural product origin and natural products play an important role in drug development programs of pharmaceutical industry. In developing countries, especially in rural contexts people usually turn into traditional healers when in diseased conditions and plants of ethno-botanical origin are often presented for use. (Olatunde, 2003)
Also in developing countries, the world health organization (WHO) estimates that about 80% of the population relies on plant based preparations used in their traditional medicine system and as basic needs for human primary health care. In recent years there is need to study the plants having different values in their medicinal plants have been evaluated for possible antimicrobial activity and potential cures from a variety of ailments especially of microbial origin (Kesaran et al, 2007)