Bagundol, Jamie Gellamucho September 19, 2015

BSED-BIO III Remarks:

Activity 4

Anti-fungal Properties of Selected Medicinal Plants

I. INTRODUCTION

The use of medicinal plants as a source for relief from illness can be traced back over

five millennia to written documents of the early civilization in China, India and the Near east,

but it is doubtless an art as old as mankind. Neanderthals living 60, 000 years ago in present

day Iraq used plants such as holly back, these plants are still widely used in ethno medicine

around the world [1, 2].

The potential of higher plants as source for new drugs is still largely unexplored. Among

the estimated 250,000-500,000 plant species, only a small percentage has been investigated

phytochemicals and the fraction submitted to biological or pharmacological screening is even

smaller. Thus, any phytochemical investigation of a given plant will reveal only a very narrow

spectrum of its constituents. Historically pharmacological screening of compounds of natural or

synthetic origin has been the source of innumerable therapeutic agents. Random screening as

tool in discovering new biologically active molecules has been most productive in the area of

antibiotics [3, 4]. Even now, contrary to common belief, drugs from higher plants continue to

occupy an important niche in modern medicine. On a global basis, at least 130 drugs, all single

chemical entities extracted from currently in use, though some of them are now being made

synthetically for economic reasons [5].

Medicinal plants represent a rich source of antimicrobial agents. Plants are used

medicinally in different countries and a new a source of many potent and powerful drugs. A

wide range of medicinal plant parts is used for extract as raw drugs and they possess varied
medicinal properties. Although, hundreds of plant species have been tested for antimicrobial

properties, the vast majority of have not been adequately evaluated [6, 7]. Considering the vast

potentiality of plants as sources for antimicrobial drugs with reference to antibacterial and

antifungal agents, a systematic investigation was undertaken to screen the local flora for

antibacterial and antifungal activity from guava, asunting, madre de cacao, sambong and

mayana leaves. Sambong (Blumea balsamifera) is widely used in the Philippines as herbal

medicine. Sambong leaves are known for its ngai or Blumea camphor that is used as herbal

medicine to treat kidney stones, wounds and cuts, rheumatism, anti-diarrhea, anti-spasms, colds

and coughs and hypertension. Mayana (Coeus blumei Benth) is one of the traditionally used

folklore medicine and is primarily used for pain, sore, swelling and cuts and in other instances

as adjunct medication for delayed menstruation and diarrhea. Bayabas (Psidium guajava) or

guava is used in herbal medicine as antiseptic, anti-inflammatory, anti-spasmodic, antioxidant

hepatoprotective, anti-allergy, antimicrobial, anti-plasmodial, anti-cough, antidiabetic, and

antigenotoxic. Asunting/ Ringworm Shrub (Cassia Alata) leaves are also known to be sudorific,

diuretic and purgative, usedto treat intestinal problems including intestinal parasites. Asunting/

Ringworm Shrub (Cassia Alata) is also used as herbal medicine to treat bronchitis and asthma.

Because of Asunting’s anti-fungal properties, it is a common ingredient in soaps, shampoos, and

lotions in the Philippines. Madre de Cacao e. Madre de Cacao (Gliricidia sepium), the Kakawate

is scientifically considered as the second most important multi-purpose legume tree with wide

healing properties. As a medicinal plant, the Kakawate is known in the Philippines as a topical

solution to cure scabies in human. Now, a group of dog lovers, the Philippine Pug Lovers Club,

recommends it for dog's skin problems a well. Traditionally, the juice extract of the kakawate

leaves, roots or bark are used to wash and treat scabies and fungal infections. For dogs with

skin irritations, the Kakawate's anti-mange characteristics can help heal scabies and eliminate

bad odor and itching. Eventually, the Kakawate solution also promotes smoother and softer

coat on dogs, the PPLC says (http://www.metro-pets.com/health--wellness/treat-dogs-scabies-

with-kakawate-madre-cacao).
 II. OBJECTIVE

At the end of the exercise, the students shall have:

1. Observed the anti-fungal activity of different medicinal plants; and

2. Compared the Anti-fungal activities of different medicinal plants.

III. MATERIALS

1. Gelatin with Beef cubes 13. Distilled Water

2. Nutrient agar 14. Medicinal Plants:

3. Sterile cotton swabs a. Mayana (Coeus blumei

4. Alcohol lamp Benth)

5. Erlenmeyer flask b. Guava (Psidium guajava)

6. Mortar and Pestle c. Gabon/Sambong (Blumea

7. Petri dishes balsamifera)

8. Ethanol d. Asunting/ Ringworm Shrub

9. Forceps (Cassia Alata)

10. Paper discs (Sterilized) e. Madre de Cacao (Gliricidia

11. Micro Pipette sepium)

12. Molds (Bread and Tabon-tabon)

IV. PROCEDURES

1. Based on literature and related studies, identify three plants with potential anti-fungal

activities/properties.

2. Prepare the plant extracts. Different extraction methods may be used depending on the

availability of materials/equipment crude extracts, ethanolic, or methanolic extracts.

3. Prepare the culture media using Sabouraud glucose agar or potato dextrose agar (refer

to procedures on agar preparations). The number of plates to be prepared will depend

on the number of trials that will be used per plant extract. Common practice requires

two replicates for each extract and for each extract and for each test organism.
4. Perform the standard procedure in the analysis of the plant extracts against molds using

the paper disc diffusion method. Refer to the procedures in your previous experiments.

5. Read the assay plates and record the results (measuring zone of inhibition using a

Vernier Caliper).

V. ILLUSTRATIONS

Schematic Diagram of the Procedures

 Pictures

Mayana (Coeus blumei Benth)

Gabon/Sambong (Blumea balsamifera)

Asunting/ Ringworm Shrub (Cassia Alata)

Guava (Psidium guajava)

 Madre de Cacao (Gliricidia sepium)

VI. OBSERVATIONS

1. Explain briefly the paper disc diffusion method.

Paper Disc Diffusion Method

When a filter paper disc impregnated with a chemical is placed on agar the chemical will

diffuse from the disc into the agar. This diffusion will place the chemical in the agar only around

the disc. The solubility of the chemical and its molecular size will determine the size of the area

of chemical infiltration around the disc. If an organism is placed on the agar it will not grow in

the area around the disc if it is susceptible to the chemical. This area of no growth around the

disc is known as a “zone of inhibition”.

2. What is the difference between a crude extract and an ethanolic extract?

 Because of heavy hydrogen bonding, water has a higher freezing point (o) than ethanol

(-114), same is also true for boiling point. It is the angular (rather than linear) structure

of water along with the network of hydrogen bonding that gives water a higher freezing

point than ethanol.

  In solids, there should be some minimum distance for molecular binding. As the

temperature is lowered, the bonds become shorter. In case of water it happens earlier,

while ethanol molecules come closer at a much lower temperature.

3. Which test plant/s yielded the highest zone of inhibition? Explain your answer.

Zone of Inhibition of Mayana, Gabon, Asunting, Bayabas, and Madre de Cacao

Zone of Inhibition (mm)

Dish 1 Dish 2
Plant Disc Disc Disc Disc Disc Disc Disc Disc
1 2 3 4 Ave. 1 2 3 4 Ave.
Mayana 4 5 6 3 4.5 5 4 2 5 4
5.62
Gabon 7 5 5 7 6 4 5 6.5 7 5
Asunting 4 5 2 3 3.5 4 5 3 4 4
2.62
Bayabas 2 3 1.5 4 5 2 3 2 5 3
Madre de 6.37
Cacao 7 7 6.5 5 5 5 4 3 5 4.25

The table above shows the zone of inhibition of the five different plant species namely

the Mayana, Gabon, Asunting, Bayabas, and Madre de Cacao.

The results of the investigation clearly indicate that the antibacterial and antifungal

activity vary with the species of the plants.

4. After knowing the potential antifungal activities of the test plants, what steps will you do

in order to promote the use of these plants for treating fungal infections?

To promote the use of these plants for treating fungal infections, first is to implement it

at home. Why? Well, it because we could never implement anything without proving it

ourselves. Though investigation through laboratory activities is more scientific than experience-

based activities, it is more likely believable if we try it ourselves and show to the community the

effectiveness of these plants as anti-fungal and antibacterial plants. GOD made our world

perfect for everything could be seen in our environment. Even those things that we thought

worthless are unconsciously full of worth to animals including humans.

VII. CONCLUSIONS

The ultimate conclusion of this study supports the traditional medicine use of different

plant extracts in treating different infections caused by pathogenic fungi in either by using a
single or combined extracts. It also suggests that a great attention should be paid to medicinal

plants which are found to have plenty of pharmacological properties that could be sufficiently

better when considering a natural food and feed additives to improve human and animal health.

Several test such as paper disc diffusion was used to know the effectiveness of the anti-

fungal and anti-bacterial properties of some medicinal plants. The measurement of the zone of

inhibition or the area of no growth will determine how susceptible it is to other external factors.

Everything was made perfect through GOD ALMIGHTY that HE created nature as a

natural medicine for us humans and even other organisms affected by it.

VIII. REFERENCES

http://www.metro-pets.com/health--wellness/treat-dogs-scabies-with-

kakawate-madre-cacao