UNIVERSITY OF DAR ES SALAAM

COLLEGE OF NATURAL AND APPLIED SCIENCE (CONAS).

DEPARTMENT OF MOLECULAR BIOLOGY AND
BIOTECHNOLOGY (MBB).
PROPOSAL ON RESEARCH PROJECT: BL 390.
TITTLE: ANTIFUNGAL ACTIVITY OF CRUDE LEAF EXTRACT
OF MELOCHIA CORCHORIFOLIA (MALVACEAE, MALVALES).

NAME: LYIMO LEODGARD A.

REG NO. 2014-04-03009.
DEGREE PROGRAM: Bsc General.
SUPERVISOR: Prof. F.S. Magingo

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Contents
No table of contents entries found.
1. INTRODUCTION..3

1.1 General introduction3

1.2 Statement of the research problem..4

1.3 Objectives.4

1.3.1 General objectives.4

1.3.2 Specific objective..4

1.4 Significance of the study..5

1.5 Literature review.5

1.6 Hypothesis...6

2. METHODOLOGY....6

2.1.1 Study area......6

2.1.2 Study site...6

2.2 Research design....6

2.2.1 Collection and extraction of plant sample.........6

2.2.2 Microbial stain..7

2.2.3 Preparation of media.7

2.2.4 Testing procedure..7

2.3 Timeframe and plan.8

3. REFFERENCES9

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INTRODUCTION

1.1 General introduction.

Within the earth there are many kind of plant covered the earth surface. These plants have grown
in different geographical areas depending on the nature and condition of a certain geographical
location, example in warm region, cold region and so on. Different plants are made up of
different chemical important composition which can interact

With biological system and cause an important biological response. Some of these plants
chemical composition can interact with other organism and inhibit growth, kill them, and to
prevent their effect. If so, we can use these plants to produce drugs which can be used to cure
different diseases caused by microorganism.

Short description about Melochia corchorifolia.

Melochia corchorifolia is a plant observed to grow in tropical areas of Africa, and other part of
the world such as Asia, Australia, United state and so on .The plant has been mostly frequently
seen to grow in open areas such as highways. Morphological has ovate leaves, and linear
stipules, it has been utilized as a homeopathic remedy, source of fiber for making dillybags and it
is known to contain phytochemical feature , its leaves have been analysed to have triterpenes
(friedelin, friedelinol and amyrin)

Classification of the plant.

Kingdom- Plantae.

Order- Malvales.

Family- Malvaceae.

Genus- Melochia.

Species- M. corchorifolia.

The image of Melochia corchorifolia taken from udsmmain campus.

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 Fungal infection
Are infections which are caused by
microscopic organism, that invade the epithelial tissues. Diseases caused by fungal
include superficial infection of the skin and can be transmitted from organism to
organism by direct contact of each other, also systemic infection are caused by inhalation
of spores and cause fungal pneumonia, this cannot transmitted from human to human.

Antifungal chemotherapy
Fungal cells are complex and differ from eukaryotes cell, therefore the agents interact
with fungal target and not the eukaryotic cell. The fungal cell wall is unique and differs
greatly from bacterial cell wall and hence fungal is not affected by antibacterial cell wall
inhibitor.
There are three mechanism of action for antifungal agent:
Cell membrane disruption.
Inhibition of cell division and,
Inhibition of cell wall formation.

1.2. Statement of the research problem.

Aspergillus and Penicillium both belongs to same kingdom, fungi found in various climates
worldwide. Some species can cause infection in human, animals, and stored crops. Many drugs
have been developed so as to eradicate the infection of the species although they tend to develop
resistant against those drugs. The plant is known to contain tripetenes (friedelin, friedelinol and
amyrin) which would have an important action on these two species.

The aim of the study is to investigate the presence of antifungal activity of crude leaf extract of
Melochia corchorifolia to Aspergillus and Penicillium spicies.

1.3 Objectives

The main objective of this study is to determine the antifungal activity of crude leaf
extract of Melochia corchorifolia against the species of fungus, Aspergillus and
Penicillium.

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 1.3.2 Specific objectives.

To determine the effectiveness of Melochia corchorifolia compared to commercial

antifungal agents.
To determine the resistance of Aspergillus and Penicillium to Melochia corchorifolia
crude leaf extract.
1.5 Significance of the study.

Due to an increase of fungal infection, it make sense for scientists to struggle in order to fight
against this situation. The use of traditional drugs ,indicate higher demand of the society also
worldwide, to fight against microscopic infection including fungal infection.At the end of study
we will be able to decrease at higher level the infection caused by Aspergillus and penicillium
through the extract from Melochia corchorifolia.

Also the study will create a margin and allow judgment to other scientist to think about the
subject and other related topics about antifungal activity.

Further research will find out the way to improve effectiveness of this drug, through combination
with other plant extract in order to counteract with those other species which have been
developed a resistant against Melochia corchorifolia.

1.6 Literature review

Russell E. Lewis, May 1998:clinical antifungal susceptibility testing for Candida spicies, With
the emergence of fungi as important nososcomial pathogen, increasing report of antifungal
resistance, and expanding drug therapy options, the need for clinically relevant antifungal
susceptibility testing is evident. Over the last decade the National Committee for Clinical
Laboratory Standard (NCCLS) worked to standardize procedures for in vitro susceptibility
testing of Candida spicies agaist fluconazole, intraconazolewith the establishment of
arepredicible methodology, correlation of antifungal susceptibility in vitro with clinical outcome
is a priority.

M.cuenca-Estrella, J. L. Rodriguez- Tudela July 2001, status of the detection of antifungal

resistance. The NCCLS reference methodology for antifungal susceptibility testing is a new
milestone of the evolution of medical mycology. The use of this method how-ever is not problem
free. At present, major limitation are a trailing phenomenon with azoles, unreliable detection of
resistance to amphotericin B, poor growth of some organisms and unpractical procedures for the
clinical laboratory.

Michael E. Klepser, August 2001 Antifungal resistance among Candida species. Isolation of
Candida spicies less susceptible to traditional therapies and recovery of increasingly resistant
isolate during antifungal therapy are growing problems. It is important for clinician to be aware
of trends and mechanism responsible for the expression of resistance.

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1.4 Hypothesis.

Does the crude leaf extract of Melochia corchorifolia have any antifungal activity
to the Aspergillus and Penicillium.spicies of fungal.
Does the Melochia corchorifolia more effective than commercial antifungal
agents.

2 METHODOLOGY.

2.1.1 Study area.

The research will be conducted at University of Dar es Salaam main campus, which is located at
eastern part of Tanzania, its climate is warm up to 25 degree of Celsius, and The area is greenish
due to presence of many herbaceous, trees and other kinds of plant.

2.1.2. Study site.

The study site also will be University of Dar es Salaam behind the botany laboratory where the
herb of interest Melochioais corchorifolia is grown. Microbiology laboratory and botany
laboratory will be included because almost all these practicals will be done there.

2.2 Research design

2.2.1 design research

Because all tools and reagents are available at the laboratories, all practices will be done there,
such facilities which are available include reagents like, chloroform, methanol, distilled water,
media, and tools like wire loops, tubes, measuring cylinder, plates, and incubators and the
organism under the study.

2.2.2 Collection and extraction of plant

The plants under the study Melochioais corchorifolia are located at University of Dar es Salaam
main campus ,backside of the botany laboratory. Fresh leaves of the plant will be collected and
dried with sunlight, air drying may take 3-7 days, to month and up to year depending on the type
of sample dried, then will be crushed into powder form using mortar and pestle, this increase the

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surface contact between sample and extraction solvent. This pre-preparation is important as for
efficient extraction to occur; the solvent must make contact with the target analytes and particles
size smaller than 0.5 mm is ideal for efficient extraction.

Extraction method

Extraction is the separation of active medicinal portion of plant using selective solvent through
standard procedure. The solvent which will be used is chloroform and methanol in ratio of 1:1,
the purpose of all extraction is to separate the soluble plant metabolites, leaving behind the
insoluble residue. Some of the initial obtained extract may be ready for use as medicinal agent in
the form of fluid extract but some need further processing.

The sample will be extracted with petroleum ether for eight hours, at a temperature of 60 -70
degree of Celsius, the already plant extracted material will re-extracted with chloroform and
methanol. The concentration of the extract will carried out using a rotator.

2.2.3 Microbial strain

Microorganism under study Aspergillus and Penicillium spicies will be isolated from two
sources, rotten bread and other will be provided from the laboratory.

2.2.4 Media preparation.

A wide range of media are used for growing fungi, most mycologist develop preference for
certain type of media based on experience and peculiarities of the type of fungi that are routinely
grown. Potato Dextrose Agar is mostly used for growing fungi, specifically yeast and moulds. It
is composed of 200g of potato infusion,20gb of dextrose, 15g of agar and Ph range from 5.4 -5.8.

2.2.5 Testing procedure

Agar well diffusion method will be used to test for infectivity of Melochioais corchorifolia to
Aspergillus and Penicillium, where a clean and well sterilized plate with a solidified PDA
medium will prepared with microorganism. With the use of sterile bore, three holes will made
on the agar plate contained microorganism and then by using micropipette, each hole will be
filled with different amount of solution from crude leaf extract and then incubated at 37 degree
of Celsius for about 3 to 7 days and the zone of inhibition will be determined and recorded.

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2.3 Time plan

EVENT TIME

Title design 16 Dec 2016

Proposal submission 29 Dec 2016

Proposal presentation 2 Jan 2017

Research activity After proposal presentation

Report writing After research activity

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3 REFERENCES

1. Russell E. Lewis, Dimitrios P, August 2001 Rationale for combination Antifungal

Therapy.
2. Holly L. Hoffman, Michael A. Pfaller August 2001, in vitro Antifungal susceptibility
testing.
3. Michael E. Klepser June 2006 Candida resistance and its clinical relevance.
4. Russell E. Lewis, Michael E. Klepser, Michael A. Pfaller, 6 May 1998, update on clinical
Antifungal susceptibility testing for candida spicies.

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TITTLE: Antifungal activity of crude leaf extract of Melochia corchlifolia against
Aspergillus and Penicillium fungal spicies.

INTRODUCTION.

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