Antifungal Effect Of Pineapple Skin Extract To Saccharomyces cereviciae Essay

This study was conducted by the researchers from Calamba City Science High School. Let us know the following researchers:

Zaira May De Castro is a third year high school student from the section of Hemingway. She was born on July 30, 1998, and presently 15 years of age living with her parents, Dolorosa and German De Castro in Asia 2, Canlubang. She graduated elementary from San Ramon Elementary School.

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Raylene Audrey Diolata is a third year high school student from the section of Hemingway. She was born on July 18, 1999, and presently 15 years of age living with her parents, Marlene and Ramon Diolata in Ma. Leonora Subdivision, Mayapa, Calamba City. She graduated elementary from Mayapa Elementary School.

Aila Marie Ilustre is a third year high school student from the section of Hemingway. She was born on November 7, 1998, and presently 14 years of age living with her parents, Marissa Ilustre and Reynaldo Barraquia in Brgy. Sanpiruhan, Calamba City. She graduated elementary from Jose Rizal Memorial School.

John Mark Makahiya is a third year high school student from the section of Hemingway. He was bor on August 8, 1999, and presently 14 years of age living with his parents, Vivien and Apolonio Makahiya in Makiling, Calamba City. He graduated elementary from Makiling Elementary School. Hazel Anne Quirao is a third year high school student from the section of Hemingway. She was born on September 26, 1998, and presently 15 years of age living with her parents, Mary Jane Quirao and Jose HaryQuirao in Villa de Calamba. She graduated elementary from Calamba Elementary School. Ma. Angelika Silapan is a third year high school student from the section of Hemingway. She was born on March 5, 1999, and presently 14 years of age living with her parents, Alicia Silapan and Rolando Silapan in Ilaya, Parian, Calamba. She graduated elementary from Liceo de Calamba.

ACKNOWLEDGEMENT
The success of this study required the help of various individuals. Without
them, the researchers might not meet their objectives in doing this study. The researchers want to give gratitude to the following people for their invaluable help and support:

Foremost, we would like to express our sincere gratitude to our Research adviser Ms. Christine Solares for the continuous support of our study and research, for her patience, motivation, enthusiasm, and immense knowledge. Her guidance helped us in the time of doing this research. Besides our Research advisor, we would like to express our gratitude to Mrs. Maria Renee Rose A. Elarco for her support, for lending some instruments that are needed during the experimentation. Of course, this project would not have been possible without the participation of the group members. Last but not the least, we would like to thank our families for their unconditional support.

Submitted to Ms. Christine Solares, Research III adviser at Calamba City Science High School, Calamba City, Laguna

Zaira May Samiano De Castro
Raylene Audrey Alegre Diolata
Aila Marie Datu Ilustre
John Mark Fabreag Makahiya
Hazel Anne Carog Quirao
Ma. Angelika Alipio Silapan

Abstract

This research is about the antifungal effect of the pineapple’s skin extract on Saccharomyces cerevisiae. The researchers’ null hypothesis is that pineapple skin extract is not effective to be anti-fungal to Saccharomyces cerevisiae. The researchers wanted to know if the pineapple skin extract is in anyway effective to kill Saccharomyces cerevisiae.The researchers’ alternative hypothesis is that pineapple skin extract is effective to be anti-fungal to Saccharomyces cerevisiae.Yeast, a species of Saccharomyces cerevisiae, was used in testing the effectiveness of the pineapple skin extract. The yeasts were group into 3 set ups: Set A, with 96.1% of
pineapple skin extract; Set B, with 63.3%; Set C, with 52.4%.Each set, was given 20mL of pineapple skin extract depending on the designated concentration to them. The yeast with pineapple skin extract solution was observed for an hour. After observation, each set ups, was transferred in a 150mL beaker with 5g of glucose and 150mL of water in 45°C. The solution accumulated in the middle and formed foamy bubbles.After 15 minutes, the height of the foamy bubbles was measured by aruler.The researchers used F- test (ANOVA). The results will be considered statistically significant at 95% confidence interval of p< 0.05. According to the results 57.35(value of F) > 9.55(value of cr), therefore, using the three different concentrations of pineapple skin extract has significant differences on the death of Saccharomyces cerevisiae. In other words, using pineapple skin extract is not really effective to be antifungal in Saccharomycescerevisiae. Based from the review of related literature found, if the yeast is already dead or not active, if placed in warm water with glucose, it will not form the foamy bubbles. Since from the experiment itself, it can concluded that the pineapple skin extract is not effective to be antifungal to Saccharomnycescerevisiae. The researchers recommend doing the same study but with different kind of fungi, with knowing the exact numbers of colony that may grew or die. The researchers also prefer to use different variable that have a desirable trait to be antifungal and have longer observation to gain better result. INTRODUCTION

Background of the Study
Yeasts are eukaryotic microorganisms classified into kingdom Fungi. They are chemoorganotrophs for they use organic compounds as their source of energy and do not require sunlight to grow. Unlike bacteria, no known yeast grows only by anaerobically. That is because yeasts either require oxygen for aerobic cellular respiration or are anaerobic, but still have some aerobic methods included for their energy production. (http://en.wikipedia.org/wiki/Yeast) Saccharomyces cerevisiaeis a species of yeast that is widely used in biology and cellular researches for improvements or discoveries in the industry. It is useful instrument is winemaking, baking and brewing since ancient times. In nature, yeast cell are found primarily on ripe fruits such as grapes, and with adequate
nutrients, they can double in 100 minutes. (http://en.wikipedia.org/wiki/Saccharomyces_cerevisiae) The pineapple (Ananascomosus) is a tropical plant with edible multiple fruit consisting of coalesced berries, and the most economically significant plant in the Bromeliaceae family. Pineapples may be cultivated from a crown cutting of the fruit, possibly flowering in 20–24 months and fruiting in the following six months. Pineapple does not ripen significantly post-harvest. Pineapples are consumed fresh, cooked, juiced, and preserved, and are found in a wide array of cuisines. In addition to consumption, in the Philippines the pineapple’s leaves are used to produce the textile fiber piña- employed as a component of wall paper and furnishings, amongst other uses. (http://en. wikipedia.org/wiki / Pineapple) Bromelain is an extract derived from the stems of pineapples, although it exists in all parts of the fresh plant and fruit, which has many uses. The extract has a history of folk and modern medicinal use. As a supplement it is thought to have anti-inflammatory effects. Bromelain also contains chemicals that might interfere with the growth of tumor cells and slow blood clotting, however there is no peer-reviewed research showing any efficacy against tumours. As a culinary ingredient it is used primarily as a tenderizer.The term “bromelain” may refer to either of two protease enzymes extracted from the plants of the family, Bromeliaceae, or it may refer to a combination of those enzymes along with other compounds produced in an extract. (http://en.wikipedia.org/wiki/Bromelain)

Statement of the Problem
The study is about the antifungal effect of pineapple skin extract to Saccharomyces cerevisiae. The researchers wanted to know if the pineapple skin extract is in anyway effective to kill Saccharomyces cerevisiae. Hypotheses

The study deals with the possibility of the pineapple skin extract’s effectiveness to be anti-fungal to Saccharomyces cerevisiae. The researchers’ null hypothesis is that pineapple skin extract is not effective to be anti-fungal to Saccharomyces cerevisiae.

The researchers’ alternative hypothesis is that pineapple skin extract is effective to be anti-fungal to Saccharomyces cerevisiae. Significance of the Study
The study tested about the effectiveness of pineapple skin extract as a possible anti-fungal to Saccharomyces cerevisiae.This will give an insight about the possibility of pineapple skin extract to be anti-fungal to Saccharomyces cerevisiae.

Scope and Limitation
The study was conducted to find out the possibility of pineapple skin extract to be antifungal in Saccharomyces cerevisiae, a type of yeast. The researchers did not use other types of yeast as a subject and solely focused on Saccharomyces cerevisiae. The researchers also used the type of pineapple that was bought from the supermarket of the same brand(DOLE).

Time Series Experimental Design

Table 2.1 The Conceptual Framework
REVIEW OF RELATED LITERATURE
According to the formulation and evaluation of anti- bacterial and anti- fungal activity of herbal ointment containing Aloe- Vera, Azadarichiaindica and Curuma- longa. In the last few decades there has been an exponential growth in the field of herbal medicine. It is getting popularized in developing and developed countries owing to its natural origin and lesser side effects. The scientific evidence has brought about the possibility of utilization of herbal plant in the treatment of fungal and bacterial infections and the development of anti-bacterial and anti-fungal products. That’s why the researchers come up to a study about the anti- fungal effect of pineapple’s skin extract in Saccharomyces cerevisiae. Since pineapple fruit has some active principles with important effect on skin – sugars and a-hydroxy-acids (AHA) as well as enzymes. These active principles are responsible for its moisturizing and anti-ageing action, and also have an antifungal and purifying effect on the skin. The extract of pineapple also gives the skin the needed vitamin C and mineral. (http:// www. Skipschool/pineapple/elements.html) How to Proof Yeast

Use 1/2 cup water, 1 tablespoon white sugar, and one package of yeast for this test. This test is effective for either compressed fresh cake yeast or with dry active yeast. Heat the water to approximately 100 degrees F (40 degrees C). We recommend testing the water temperature using a thermometer. In a nutshell, yeast eats various sugars and excretes alcohol and carbon dioxide. Alcohol and carbon dioxide are what the pockets in bread are filled with as bread bakes. Whisk the sugar into the water to help it to dissolve quickly. Once the sugar has been evenly distributed throughout the water, add the yeast, and stir. After 5 or 10 minutes, the yeast should begin to form creamy foam on the surface of the water, the higher it accumulates, the better. You can now proceed to combine the yeast mixture with the flour and other dry ingredients. If there is no foam in the bowl, the yeast is dead and you should start over with a new packet of yeast. (http:// www. Allrecipes.com/ howto/ proofing- yeast.html)

Methodology
Location of the Study
The study was conducted in Calamba City Science High School,Brgy. 3,Calamba City. The study would test the antifungal effect of the pineapple’s skin extract on Saccharomyces cerevisiae. Instruments

The researchers used yeast, a species of Saccharomyces cerevisiae. The fungus, then, was divided into four setups: First, the control set up which contains 5g of yeast in three different petri dishes and the others are, Set A, Set B, and Set C. Each set contains 5g of yeast placed in three different petri dishes. These sets were given different concentration of the pineapple skin extract; Set A was given 96.1%, Set B was given 63.3% and Set C was given 52.4%. The researchers used an analytical weighing scale to measure the weight of the yeast and the pineapple skin extract solution, alaboratory thermometer for measuring the water’s temperature in testing the extract’s effectiveness and the researchers also usedruler for determining the measurement of the height of the bubbles. Data Gathering Process

Set A, B and Cwas given 20mL of pineapple skin extract depending on the
concentration designated to them. The yeast was observed for an hour. After observation, each set ups was transferred in a three different 150mL beaker with 5g of glucose and 50mL of water in 45°C and stirred. After 15 minutes, the yeast with glucose solution accumulated and formed foamy bubbles at the top. The height of the foam per set up was measured.

Statistical Analysis
The researchers measured the height of the foamy bubbles in each set up in two trials. After measuring the height of the foam, the researchers used ANOVA (F- Test). The results will be considered statistically significant at 95% confidence interval of p; 0.05.

Results and Discussion
Table 4.1 shows the measurement of the height of the foam in the three concentrations in two trials. The height of the foams was measured by a ruler. On the first trial, Set A was 2.5cm; Set B was 3 cm; Set C was 3.5 cm. Meanwhile, on the second trial, Set A was 2.5 cm; Set B was 2.5 cm; Set C was 2.6 cm. Yeast with Pineapple Skin Extract Concentration of

1st Trial
2nd Trial
Set A: 96.1%
2.5 cm
2.5 cm
Set B: 63.3%
3 cm
2.5 cm
Set C: 52.4%
3.5 cm
2.6 cm
Table 4.1 The measurement of the height of the foams in two trials
Table 4.2 shows the measurement of the height of the foams in the controlled set up which doesn’t have any pineapple skin extract. Controlled Set Ups
Measurement of the Height of the bubbles formed (cm)
1
5
2
5.5
3
7.5
Table 4.2 The measurement of the height of the foams in the controlled set up
Table 4.3 shows the mean height of the foam in Set A was 2.5 cm, while the mean height of the foam in Set B was 2.75 cm, and the mean height of the foam in Set C was 3.05 cm.
Set A
Set B
Set C
x
2.5 cm
2.75 cm
3.05 cm
N
3
3
3
?(x1-x)2
0
0.12
0.40
Critical Value
9.55
F
57.35
Table 4.3 The summary of the computation
Conclusion and Recommendation
This study was conducted in Calamba City Science High School, Brgy. 3,Calamba City. It deals with antifungal effect of pineapple skin extract on Saccharomyces cerevisiae. This study would give an insight of the effectiveness of the pineapple skin extract to be antifungal inSaccharomyces cerevisiae. F- Test or ANOVA was used to compare the mean height to each
other to determine and analyze the pineapple skin extract antifungal effect show any significant difference. Based from the F-test (ANOVA), was used to get a result of a significant difference which can be determined if, by computation, the value of F is higher than the critical value of the data. Since, according to the results 57.35(value of F) ;9.55(value of cr), therefore, using the three different concentrations of pineapple skin extract has significant differences on the death of Saccharomyces cerevisiae. In other words, using pineapple skin extract is not really effective to be antifungal in Saccharomyces cerevisiae. Based from the review of related literature found, if the yeast is already dead or not active, if placed in warm water with glucose, it will not form the foamy bubbles.Since from the experiment itself, it can concluded that the pineapple skin extract is not effective to be antifungal to Saccharomnyces cerevisiae. The researchers recommend doing the same study but with different kind of fungi, with knowing the exact numbers of colony that may grew or die. The researchers also prefer to use different variable that have a desirable trait to be antifungal and have longer observation to gain better result.

REFERENCES
Freeman, Scott. (2005). Biological Science. 2nd Edition Vol. 1. Upper Saddle River, NJ: Pearson Education.

Madigan, Michael T., and John M. Martinko.(2006) Biology of Microorganisms.11th Edition Vol. 1.Upple Saddle River, NJ: Pearson Education,.

Osley, M A., and D Norris.(2007). The Two Gene Pairs Encoding H2A and H2B Play Different Roles in the Saccharomyces Cerevisiae Life Cycle. Molecular and Cellular Biology 7 (1987): 3473-3481. Biological Sciences.IIlumina.Murphy Library, La Crosse, WI. 25.

Stella, A V., R Paratte, L Valnegri, et. al., (2007).Effect of Administration of Live Saccharomyces Cerevisiae on Milk Production, Milk Composition, Blood Metabolites, and Faecal Flora in Early Lactating Dairy Goats. Small Ruminant
Research 67: 7-13. Web of Science.ISI Web of Knowledge.Murphy Library, La Crosse, WI.

Stewart, Graham G.(1987). Biological Research on Industrial Yeasts.1st ed. Vol. 3. Boca Raton, Fl: CRC P.

Stolc, Viktor, and Sigrid Reinsch.(2007). Saccharomyces Cerevisiae Yeast Strain. National Aeronautics and Space Administration.Retrieved Septemer 2013 from http://www.nasa.gov/centers/ames/news/releases/2004/yeast/yeast_prt.htm.

APPENDICES

The process of tearing up the pineapple skin into smaller pieces.

The researchers placed the pineapple skin in a 150mL beaker. Set A had 70g of pineapple skin; Set B had 120g; Set C had 170g. The researchers also put 100mL of water.

Transferring the pineapple skin extract in the 5g of yeast in Set A.

Transferring the pineapple skin extract in the 5g of yeast in Set B.

Transferring the pineapple skin extract in the 5g of yeast in Set C.

Transferring the yeast with pineapple skin extract in the 150 mL beaker.

After 3 hours, each set ups were transferred in a 500mL beaker with 5g of glucose and 150mL of water in 45°C. After few minutes, the solution is starting to accumulate in the middle and form foamy bubbles.

The three set ups before 15 minutes and the solution already forms foamy bubbles and starting to rise up.

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