Antifungal Effects of Breafruit (Artocarpus Camansi) Leaves Extract

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 IIIligan 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 StudyBecause 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 IIReview of Related LiteratureCurrently, 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 FrameworkKamansi Leaves Extracts Inhibition of Growth of Rhizopus stoloniferHypotheses 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 IIIMethodologyThe 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.