Research Title

Research Title:
Antifungal Activity of Papaya (Carica papaya) Leaves Using Ethanolic Extract Against Aspergillus niger.

Name of the Researchers: Stella Mae Saluague (Leader)
Lady Danielle Lucas
INTRODUCTION
Use of vegetal extracts with antifungal properties is a common practice for thousands of years, for example, powders or extracts of acacia, garlic, eucalyptus and mint all function as fungicides capable of controlling different diseases. This focus on natural management of phytopathogenic fungi is reflected in extensive research on biological fungicides. However, Medicinal plants being cheap and renewable sources of pharmacologically active substances are known to produce certain chemicals that are toxic to bacteria and fungi (Ahn, 2017).

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Carica papaya is widely grown now and used in different parts of the world not only for food but also for ornamental purpose. Carica papaya L. belongs to the family of Caricaceae, and several species of Caricaceae have been used as a remedy against a variety of diseases. According to the book Healing Foods by DK Publishing House, papaya is “known to have antibacterial properties and promotes good digestion and almost every part of the plant can be used. For many years in pharmaceutical industry plants are used as a source of medicines especially due to the presence of phytochemicals ( Agarwal SG & Goyal S 2013 ). The leaves have high levels of phytochemicals like saponins, tannins, alkaloids, and flavonoids, especially B-Carotene, fungus fighting nutrient falcarinol and plenty of antioxidants, such as beta carotene which will be used as antifungal. There are plenty of antifungal studies but with a comparison on different fungal strains and or, some different part of a certain plant, so with that we propose a new study using only one part of the plant (Carica papaya) which is leaves and to evaluate its effect.

Aspergillus niger is an asexual saprophytic fungus that can grow on dead leaves, stored grain, compost piles and other decaying vegetation.   The spores can be inhaled when simply working with anything A. niger has colonized. It can also be found on marijuana leaves. Although most people are often exposed to aspergillus, infections caused by the A. niger can also affect plants, such as onions and tomatoes. Small animals, such as mice and chicks were fed moldy soybeans with A. niger on them, and the subjects subsequently died after digestion (EPA.gov). Aspergillus niger is not one of those that are so deadly, but can definitely cause sickness and allergic reactions and may be produce compounds that is phytotoxic and a battery of enzymes that destroy the plant’s structures, physical appearance and beauty (Endah Yulia, July 2005). Aspergillus niger contains toxins that can make people with weak immune systems become very sick and can sometimes result in death. These toxins can be inhaled by humans, most commonly people who work around plants or peat, and can cause a lung disease called Aspergillosis, which has infected over 300,000 people worldwide (Keir 2013).One of the most common plant disease is the fungi.

The objective of the study is to determine the effectiveness of the antifungal activity of Ethanolic extract using the papaya (Carica papaya) leaves, to identify what concentration will be more capable to natural sources could be a potential candidate as antifungal agent especially in the current scenario in which plant fungal pathogens have adapted resistance against antifungal antibiotics and to evaluate the zone of inhibition the fungus.

MATERIALS AND METHODS
Preparation of the leaves and the ethanolic extracts
The papaya leaves were collected from the Lucas’s Residence in Purok-3 Sta. Maria, Nabunturan, ComVal Province and Saluague’s Residence in Purok-3 Magading, Nabunturan, Compostela Valley Province. The gathered papaya leaves were air-dried in a shadow room area in two days then powdered the leaves by the use of a blender. The powdered leaves were soaked with the same amount of ethanol about 3 days and were filtered by the use of filter paper. Then the filtered extract was ready to oil-bath. After, the extract was stored in an airtight glass container sealed with parafilm protected from sunlight until the day of experimentation had come.

Preparation of the Aspergillus nigerThe fungi were collected and bought from the DOST 11 Laboratory, Davao City, Davao del Sur Philippines. Only one tube of fungi was used in the whole experimentation. The fungi were stored in the Biotech Laboratory, it was labeled carefully and put it in the refrigerator. The fungi were handled carefully and the researchers make sure that the condition of fungi was all set before the experimentation.

Experimentation
The whole duration of the experimentation was 2-4 weeks. The experimentation was performed in Tagum Doctors Hospital Laboratory. All materials and equipment needed in the experiment was gathered and prepared such as Petri dishes, stirring rod, beaker, inoculation loop, inoculation needle, wire gauze, tripod, graduated cylinder, hot plate, alcohol lamp, hot hands, Erlenmeyer flask, filter paper, and ethanol. All the materials were wrapped with papers putting them in the autoclavable bag to sterilize it in the autoclave for how many minutes. Making and preparing of agar, first is putting 11.5grams of agar and 500mL water in the Erlenmeyer flask and then put it in the hot plate making sure that it will boil. After making the agar solution put it equally in a Petri dishes and wait for it becomes cool.
Each Petri dishes were filled with the same amount of agar and added with the fungi in each Petri dishes. Different concentrations of the ethanolic extract of papaya (Carica papaya) 100% extract, 75% extract solution, 50% extract solution, 25% extract solution applied and the commercial product, serve as the control to every Petri dishes where filter papers were placed. Samples with distilled water, on the other hand, were regarded as the negative control and samples with the mortality data were subjected to profit analysis which determines the minimal inhibition concentration percent of the fungi where there is 80-90 percent of the fungi were inhibited.

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REFERENCES:
Sikandar Khan Sherwani, Tasveer Zahra Bokhari, Kanwal Nazim, Syed Aneelc Gilani, Shahana Urooj Kazmi. Qualitative phytochemical screening and antifungal activity of Carica papaya leaf extract against human and plant Retrieved from: http://dx.doi.org/10.7897/2230-8407.04718
Mello VJ, Gomes MT, Lemos FO, Delfino JL, Andrade SP, et al. (2008) The gastric ulcer protective and healing role of cysteine proteinases from Carica candamarcensis. Phytomedicine 15(4): 237-244.
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NDTV Convergence 2018 Retrieved From: https://food.ndtv.com/food-drinks/9-incredible-benefits-of-papaya-leaf-juice-1672318Megan Ware RDN LD Reviewed by Natalie Butler, RD, LD Last updated Thu 21 December 2017. What are the Health Benefits of Papaya? Retrieved From: HYPERLINK “https://www.medicalnewstoday.com/articles/275517.php” https://www.medicalnewstoday.com/articles/275517.php
Jasmine Zamora March 10, 2014 The Fungal Kingdom: A Program at the Evergreen State College. Retrieved From: http://blogs.evergreen.edu/fungalkingdom/aspergillus-niger-not-your-everyday-mold/

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