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Antifungal Effects of Breafruit (Artocarpus Camansi) Leaves Extract

Abraham Almerol
Fahad Cuaro
Eulilah Dayanan
Kevin Marc Dioneda
Amerah Jasmine Hosni
Mohammad Ali Ibrahim

Submitted in partial fulfilment of the requirements


in Research II

Iligan City National High School


Iligan City

October, 2009
Ms. Annallee Q. Aron
Research Adviser

Chapter I
Introduction
Background of the Study
Practices, age, compromising immune systems and environmental factors e.g.
climate are the cause of escalating susceptibility of people to fungal infections. These
fungal infections are frequent among Filipinos and ranked as second leading cause of
consultation (Handog and Dayrit, 2005). The present effective drugs for these infections
are becoming more expensive especially for people living in remote areas due to the
current economic crisis.
The effects, too, of fungi to agriculture signal the action of preventing a
widespread parasitism so as to maintain economical standards. A continuous growth of
fungi to agricultural lands adds more expenses o the owners because of crop loss;
lowering quality of plant and products that could greatly affect their exportability; and the
costs for maintenance, i.e. monitoring and applying of fungicide (Palm, 2001).
Resorting to natural remedies such as plants effectively or potentially antifungal
would be a great help to people and/or industries. Artocarpus Camansi, commonly
known as breadfruit or Kamansi, was used by people long ago as a treatment for thrush
(candidiasis), a fungal infection caused by candida yeasts (Quisumbing). Therefore, there
is a need to test the ability of Kamansi on inhibiting the growth of fungi to find an
alternative solution which people could rely on besides commercial products. The test
will be advantageous in the field of medicine and agriculture, humans and plants alike.

Statement of the Problem


This research was conducted to test the effectiveness of Kamansi leaves extracts
on inhibiting the growth of fungi and determine which concentration (50%, 75%, 100%)
is most effective.
Significance of the Study
Because of the current antifungal problems in the society, the study about the
antifungal effectiveness of Artocrapus Camansi will be a very good contribution to the
development of antifungal medicines or drugs in the near future. It will also compare and
determine the effect of the different concentrations of camansi leaves extract to bread
molds (Rhizopus Stolonifer). This research study will also provide a cheaper and easier
solution in treating fungal infections or diseases especially that drugs for fungal
infections are becoming more and more expensive. Thus, itll be very beneficial for
antifungal research and development and also to the people suffering antifungal
problems.
Scope and Delimitation
This study was conducted on October 2009 under ambient room conditions.
Leaves of Artocarpus Camansi were the only source of the extracts. This research used
Rhizopus Stolonifer for the antifungal test.

Chapter II
Review of Related Literature
Currently, herbal plants are the objects of interest in the field of medicine.
Politically speaking, the Philippine government officials seek to find ways as to how
medical drugs could be easily availed by the Filipinos remarkably those who are poor and
also solutions for fungal growth in agricultural lands which could heavily affect the
production of food and etc. thus, having an impact on the economy. These plants are seen
to be cheaper indigenous materials for drug substitutes and fungicides.
The Philippines has bountiful forest resources especially in Mindanao. About
8,000 species of plants in these forests have been known to possess certain medicinal
properties.
One of these is Artocarpus Camansi, commonly known as breadfruit or Kamansi.
It is a member of the family Moraceae (mulberry). Flowering is monoecious with male
and female flowers on the same tree at the ends of branches, with the male inflorescence
appearing first. Male flowers are club-shaped, up to 3 cm (1.2 in) in diameter and 2535
cm (1014 in) long or longer. Thousands of tiny flowers with two anthers are attached to
a central spongy core. Female inflorescences consist of 15002000 reduced flowers
attached to a spongy core. Unlike breadfruit, the individual flowers do not fuse together
along their length. Leaves are alternately arranged, with length 4060 cm (1624 in),
moderately dissected with 46 pairs of lobes and sinuses cut half way to the midrib. New
leaves on young trees can be 76 or more cm (30 in) long. They are densely pubescent,
with many white or reddish-white hairs on upper and lower veins, lower leaf surface, and
petiole. Blade is dull green with green veins. Two large green stipules enclose the bud,

turning yellow before dehiscing. Kamansi trees are now found in cultivation in the
Philippines, where it is typically grown as backyard tree (Ragone, 2006). Kamansi trees
are trees with single trunk and have spreading canopies. They grow in tropical regions.
Trees can grow 10-15 m with a trunk 1 m in diameter or larger. Latex can be found in
every part of the tree.
Kamansi leaves are believed to have antifungal property. Crushed leaves are used
by people long ago to treat thrush. Decoction of leaves are also used for baths in
rheumatism and (yellow leaves) for high blood pressure and asthma. Diluted latex is used
for diarrhea, stomach aches and dysentery.
Test Microorganism
Rhizopus stolonifer (black bread mold) is a widely distributed Mucoralean mold.
Commonly found on bread surfaces, it takes food and nutrients from the bread and causes
damage to the surface where it lives.
Asexual spores are formed within sporangia, which break to release the spores mature.
Germination of these spores forms the haploid hyphae of a new mycelium. R. stolonifer
grows rapidly at temperatures between 15 and 30C.
Rhizopus stolonifer is a heterothallic species (Schipper 1984), in that sexual
reproduction happens only when opposite mating types (designated + and -) come in
contact. Successful mating results in the formation of durable zygospores at the point of
contact. Subsequently, the zygospore germinates and forms a sporangiophore whose
sporangium contains both + and - haploid spores. There are three varieties: R. stolonifer
var. stolonifer produces straight, erect sporangiophores, whereas those of R. stolonifer

var. lyococcos are curved. A closely related species, Rhizopus sexualis, differs primarily
in being homothallic (self-compatible).
Conceptual Framework
Kamansi Leaves Extracts

Inhibition of Growth of Rhizopus stolonifer

Hypotheses
Extracts from Kamansi leaves inhibit the growth of Rhizopus Stolonifer (bread molds).
Different concentrations of Kamansi Leaves extract affect the growth of Rhizopus
Stolonifer.

Chapter III
Methodology
The preparation of materials and the procedures performed in this research are
presented in this section.
A. Preparation of Materials
Twelve Petri dishes, three test tubes, mortar and pestle, metal scalpel, beaker,
graduated cylinder, slides and cover glasses, and funnel were sterilized at 121C and 15
pounds pressure for 15 minutes, stabilized at 13 minutes. A microscope, 0.5% Cotton
Blue, ruler and marking pen were also turned to account for the completion of the study.
All of these materials were taken from the ICNHS Science Laboratory.
B.
Five slices of fresh bread were put in a sterile, wide-mouthed glass container. The
bread was moistened with 10 ml tap water. The container was then left open and was put
in a place with room temperature of 29C. After 4 days, hyphal or plasmodial growth is
visible on the bread as shown in Figure A.
The researchers took a small piece of hypha with the aid of a sterile metal scalpel
and squashed it beneath a coverslip on a slide to provide some clues about the nature of
the fungi. A drop of cotton blue was administered to stain the fungal walls. It was then
placed on the stage of the microscope to observe enough detail.
C. Culturing
A standard medium of potato dextrose agar (PDA) was used to culture bread
molds (Rhizopus Stolonifer). This kind of agar was established from potato infusion and
dextrose (corn sugar). Potato infusion was made by boiling 900g of sliced (washed but

unpeeled) potatoes in water for one and a half hour and then straining the broth to
cheesecloth. Distilled water was added such that the total volume of the suspension was
one litre. Twenty grams of dextrose was then added and the medium was sterilized at 15
psi for 15 minutes.
A small bit of bread molds from the experimented bread was placed in the center
of a Petri dish of PDA. In securing a pure culture, the researchers cut off the tip of a
growing hypha with a sterile blade and transferred it, along with a chunk of agar it was
growing on, to a new dish of PDA. The researchers repeated this procedure until they had
a pure culture.
D. Preparation and Application of Extracts as Antifungal
The researchers gathered matured Kamansi leaves to a mass the needed 150 ml of
extracts will be obtained from for the experiment, which were then washed with water
and drained. The leaves were macerated using a mortar and pestle. Few millilitres of
distilled water was added and 100%, 75%, 50% of extract mixtures were obtained. The
extracts were collected and placed on separate test tubes. Using Completely Randomized
Design (CRD), three of the twelve experimental subjects received one of the three (3)
treatments. The three subjects receiving a particular treatment were considered three
replicates for the treatment. In other words, the twelve subjects, each containing a pure
culture of bread molds were subjected to treatments A, B, C having 50%, 75%, and 100%
concentration of Kamansi leaves extract respectively. Also, through CRD, the researchers
were able to have three experimental subjects that served as the control group.

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