TAGAYTAY CITY SCIENCE NATIONAL HIGHSCHOOL

Sungay West, Tagaytay City

Acceptability of Excelsa Coffee (Coffea Excelsa) in Combination of Wild Cane

(Saccharum Spontaneum) Against Enterobacteriaceae

In Partial Fulfillment of the Requirements in Research III

Prince Nelson J. Aguinaldo Jr.

Jandin Joshua B. Aseron

Kyla Rose S. Diaz

Lia Margaret A. Villanueva

Proponents

Christian C. Luna

Research Adviser

February 2020
 CHAPTER I

THE PROBLEM: RATIONALE AND BACKGROUND

INTRODUCTION

Bacteria is one of the most common things that is on Planet Earth. Bacteria is used in

many ways. They are used in fermentation processes like brewing, baking, and cheese

manufacturing. Bacteria is also known to be in common diseases including pneumonia

(Streptococcus pneumoniae), meningitis (Haemophilus influenzae), and food poisoning

(Escherichia coli and Salmonella).

But there is one family of bacteria that this research wants to accomplish in killing.

And it is the Enterobacteriaceae family. The group of researchers have found some

ingredients that could kill a bacterium that is in the Enterobacteriaceae family. One ingredient

is the Excelsa Coffee (Coffea Excelsa). They have found that Liberica Coffee (Coffea

Excelsa) could kill the bacteria at a rate of 50%. Another ingredient is a Wild Cane

(Saccharum spontaneum). Although they could not determine the rate which it could kill the

bacteria, it is said to be really effective in killing the bacteria.

Wild Cane (Saccharum Spontaneum) is said to be an effective specie to kill

enterobacteria most studies say that it contain Anti-microbial, Antibacterial, Antidiarrheal

and Anti-inflammatory substance.

Medicine is one of the most common research topic used in Research. With that idea,

the researcher have found out that the combination could wipe out the entire bacteria, called
Enterobacteriaceae. Of course, through the research they will conduct, they will find out if

enterobacteriaceae will die or it will spread and multiply throughout the organism or specie.

If the research succeeds it will be a good source of medicine to the diseases caused by

enterobacteriaceae. This research could make a difference in the world of medicine. It could

cure many diseases that could lower the death rate due to disease. Humans wouldn’t have to

suffer the symptoms of the disease. Depending on the time that it will take to kill the bacteria,

it could lessen the infected people with the bacteria. With this, more people will be healthy

than sick. People will live their life to the fullest and they will be more fit than before.

STATEMENT OF THE PROBLEMS

The main purpose of the study is to determine the evaluation and characterization of

antibacterial activity of Liberica Coffee (Coffea Excelsa) in combination of wild cane

(Saccharum spontaneum) against enterobacteriaceae. Specifically, it attempts to answer the

following research questions:

1. What is the evaluation of the antibacterial activity of liberica coffee (Coffea Excelsa)

in combination of wild cane (Saccharum spontaneum) against enterobacteriaceae?

2. What is the characterization of the antibacterial activity of liberica coffee (Coffea

Excelsa) in combination of wild cane (Saccharum spontaneum) against

enterobacteriaceae?

3. Is there a significant difference in the evaluation of the antibacterial activity of

liberica coffee (Coffea Excelsa) in combination of wild cane (Saccharum

spontaneum) against enterobacteriaceae?

 4. Is there a significant difference in the characterization of the antibacterial activity of

liberica coffee (Coffea Excelsa) in combination of wild cane (Saccharum

spontaneum) against enterobacteriaceae?

HYPOTHESES

NULL HYPOTHESIS

1. The evaluation of the antibacterial activity of liberica coffee (Coffea Excelsa) in

combination of wild cane (Saccharum spontaneum) against enterobacteriaceae is zero.

2. The characterization of the antibacterial activity of liberica coffee (Coffea Excelsa) in

combination of wild cane (Saccharum spontaneum) against enterobacteriaceae is zero.

3. There is no significant difference in the evaluation of antibacterial activity of liberica

coffee (Coffea Excelsa) in combination of wild cane (Saccharum spontaneum) against

enterobacteriaceae.

4. There is no significant difference in the characterization of the antibacterial activity of

liberica coffee (Coffea Excelsa) in combination of wild cane (Saccharum

spontaneum) against enterobacteriaceae.

ALTERNATIVE HYPOTHESIS

1. The evaluation of the antibacterial activity of liberica coffee (Coffea Excelsa) in

combination of wild cane (Saccharum spontaneum) against enterobacteriaceae is not

zero.
 2. The characterization of the antibacterial activity of liberica coffee (Coffea Excelsa) in

combination of wild cane (Saccharum spontaneum) against enterobacteriaceae is not

zero.

3. There is a significant difference in the evaluation of antibacterial activity of liberica

coffee (Coffea Excelsa) in combination of wild cane (Saccharum spontaneum) against

enterobacteriaceae.

4. There is a significant difference in the characterization of the antibacterial activity of

liberica coffee (Coffea Excelsa) in combination of wild cane (Saccharum

spontaneum) against enterobacteriaceae.

OBJECTIVES OF THE STUDY

This study aims to determine whether Liberica Coffee in combination of wild cane can be

used against the bacteria enterobacteriaceae. Specifically, it aims to:

1. Determine the evaluation of antibacterial activity of liberica coffee in combination

of wild cane against enterobacteriaceae in terms of effectiveness,

efficiency and sustainability.

2. Determine the characterization of antibacterial activity of liberica coffee in

combination of wild cane against enterobacteriaceae in terms of purity, surface

chemistry and stability.

3. Determine the significant difference on the evaluation of antibacterial activity of

liberica coffee in combination of wild cane against enterobacteriaceae in terms of

effectiveness, efficiency and sustainability.

 4. Determine the significant difference on the characterization of antibacterial activity

of liberica coffee in combination of wild cane against enterobacteriaceae in terms of

purity, surface chemistry and stability.

CONCEPTUAL FRAMEWORK

Process Output
Input
Production of liberica
Gathering of materials: coffee in combination of
wild cane against The evaluation and
Antibacterial activity of
enterobacteriaceae: characterization of
liberica coffee in
combination of wild cane antibacterial activity of
Evaluation of antibacterial liberica coffee in
against enterobacteriaceae activity of liberica coffee
made out of their extracts. combination of
in combination of wild
cane (effectiveness, wild cane against
efficiency and enterobacteriaceae.
sustainability) Through
tests or examinations.

Figure 1.1 Input, Process, Output Model of the Research Study

Interpretation

Figure 1.1 shows the materials and process in making the antibacterial activity against

enterobacteriaceae out of Liberica coffee in combination of wild cane. We would use

different methods for getting the extract for the antibacterial activity to use it against the

enterobacteriaceae using statistic for the evaluation and characterization of quality attributes.
RESEARCH PARADIGM

Gather the
Information

Plan for Expected

Outcomes

Conduct a Survey
for the Acceptability
of the Idea

Compute the data

Figure 1.2. Process made during the duration of the Research

SIGNIFICANCE OF THE STUDY

There are many uses of Excelsa Coffee (Coffea Excelsa) and Wild Cane (Saccharum

Spotaneum). The reason why the researchers chose the study to help people to kill and avoid

Bacteria easier. Coffea Excelsa could kill bacteria rate of 50% . The Coffee is full of vitamins

that could help to avoid and kill a bacteria.

SCOPE AND LIMITATIONS

This study entitled “Evaluation and Characterization of Antibacterial Activity of

Liberica Coffee (Coffea Excelsa) in Combination of Wild Cane (Saccharum Spotaneum)

against Enterobacteriaceae” was conducted at Pasong Langka, Silang Cavite within school

year 2019-2020. Coffea Excelsa and Saccharum Spotaneum were collected for its

acceptability in bacteria. The scope of this study is to know the Evaluation and

Characterization of Antibacterial Activity of Liberica Coffee (Coffea Excelsa) in

Combination of Wild Cane (Saccharum Spotaneum) against Enterobacteriaceae.

DEFINITION OF TERMS

1. Liberica Coffee - is a species of flowering plant in the family Rubiaceae from which

coffee is produced. It is native to western and central Africa from Liberia to Uganda and

Angola, and has become naturalized in the Philippines. It has a great anti-bacterial property

that can eliminate different types of bacteria.

2. Enterobacteria - are bacteria from the family Enterobacteriaceae, which are primarily

known for their ability to cause intestinal upset. And also are responsible for a variety of

human illnesses, including urinary tract infections, wound infections, gastroenteritis,

meningitis, septicemia, and pneumonia.

 CHAPTER II

REVIEW OF RELATED LITERATURE

Related Legal Basis

Republic Act No.8423

This act is known as the “Traditional and Alternative Medicine Act (TAMA) of 1997”

have set the way for the production of alternative medicines from certain herbs proven safe,

effective, and affordable. This Act also created the Philippine Institute of Traditional and

Alternative Health Care (PITAHC) which the purpose is to “accelerate the development of

traditional and alternative health care in the Philippine, providing for a traditional and

alternative health care development fund and for other purposes”.

The foregoing production of alternative medicines conveys that traditional medicines

or natural products that are proven and tested are helpful to many Filipinos especially to
poverty-stricken people because it will lessen their expenses and they don't need to go to a

hospital and pay a tremendous amount of money.

Republic Act No.8479

This act is known as the “Philippine Clean Air Act of 1999”. The State shall promote and

protect the global environment to attain sustainable development while recognizing the

primary responsibility of local government units to deal with environmental problems. The

State recognizes that the responsibility of cleaning the habitat and environment is primarily

area-based. The State also recognizes the principle that “polluters must pay”. Finally, the

State recognizes that a clean and healthy environment is for the good of all and should,

therefore, be the concern of all.

This act commits to have a clean environment and to eliminate air pollution, it can be

helpful in the health of every people to prevent having health problems or air-caused

pollution diseases.

Related Literature

Wild Cane

Husaqqin, Raza, Ullah and Aziz(2014), Saccharum spontaneum (Family: Poaceae),

was evaluated for its antibacterial potential against human pathogenic bacterial strains. In-
vitro antibacterial tests were performed by disc diffusion method on nutrient agar, in order to

analyze the percentage zone of inhibition. Whole plant’s extract showed the significant zone

of inhibition (mm), against Staphylococcus aureus (17.00), Streptococcus pneumoniae

(16.50), Bacillus cereus (15.90), Bacillus pumilus (15.45), Escherichia coli (18.00),

Klebsiella pneumoniae (17.10), Pseudomonas aeruginosa (15.20) and Citrobacter freundii

(14.00), with relative percentages of inhibition of 76.90, 71.60, 57.40, 56.85, 70.40, 69.90,

61.05 and 54.30 respectively. Modified agar well diffusion method was used to measure the

minimum inhibitory concentration (MIC) and MIC values lies within the range of 75 to 300?

g /ml for the G+ve strains while 75 to 600?g /ml for G-ve. Due to presence of tannins and

flavonoids, it inhibits the growth of bacteria on most regulatory levels such as peptidoglycan,

DNA, RNA and protein synthesis.

RELATED STUDIES

Coffee

The in vitro antimicrobial activity of commercial coffee extracts and chemical compounds

was investigated on nine strains of enterobacteria. The antimicrobial activity investigated by

the disc diffusion method was observed in both the extracts and tested chemical compounds.

Even though pH, color, and the contents of trigonelline, caffeine, and chlorogenic acids

differed significantly among the coffee extracts, no significant differences were observed in

their antimicrobial activity. Caffeic acid and trigonelline showed similar inhibitory effect

against the growth of the microorganisms. Caffeine, chlorogenic acid, and protocatechuic

acid showed particularly strong effect against Serratia marcescens and Enterobacter cloacae.

The IC(50) and IC(90) for the compounds determined by the microtiter plate method

indicated that trigonelline, caffeine, and protocatechuic acids are potential natural
antimicrobial agents against Salmonella enterica. The concentrations of caffeine found in

coffee extracts are enough to warrant 50% of the antimicrobial effect against S. enterica,

which is relevant to human safety. (Almeida, Ana Amélia & Farah, Adriana & Silva, Daniela

& Nunan, Elzíria & Gloria, Maria Beatriz A. (2006).

Wild Cane

In this study, Saccharum spontaneum (Family: Poaceae), was evaluated for its antibacterial

potential against human pathogenic bacterial strains. In-vitro antibacterial tests were

performed by disc diffusion method on nutrient agar, in order to analyze the percentage zone

of inhibition. Whole plant’s extract showed the significant zone of inhibition (mm), against

Staphylococcus aureus (17.00), Streptococcus pneumoniae (16.50), Bacillus cereus (15.90),

Bacillus pumilus (15.45), Escherichia coli (18.00), Klebsiella pneumoniae (17.10),

Pseudomonas aeruginosa (15.20) and Citrobacter freundii (14.00), with relative percentages

of inhibition of 76.90, 71.60, 57.40, 56.85, 70.40, 69.90, 61.05 and 54.30 respectively.

Modified agar well diffusion method was used to measure the minimum inhibitory

concentration (MIC) and MIC values lies within the range of 75 to 300?g /ml for the G+ve

strains while 75 to 600?g /ml for G-ve. Due to presence of tannins and flavonoids, it inhibits

the growth of bacteria on most regulatory levels such as peptidoglycan, DNA, RNA and

protein synthesis. . ( Hussaqin, Musaddique & Ullah, Razi & Raza, Shahid & Aziz, Abdul

& Bakhsh, Hazoor & Majeed, Abdul & Mumtaz, Faiza. (2014).)
 CHAPTER III

METHODOLOGY

A. RESEARCH DESIGN

The research design used in the study entitled Evaluation and Characterizations of

Antibacterial Activity of Liberica Coffee (Coffea Excelsa) in Combination of Wild Cane

(Saccharum Spontaneum) Against Enterobacteriaceae is randomized control trial,

specifically randomized block experimental design. This will involve determining the

enterobacteriaceae level before and after the administration of each treatment per group.

B. MATERIALS

The materials used in this study were 5.25 ml of Liberica Coffee, 5.25 ml of Wild

cane to have 10.5 ml to be used for the group of treatments or experiment

C. EQUIPMENTS AND UTENSILS

 The equipments and utensils used in this study is petri dish, mortar and pestle, knife

for chopping, a cloth and some materials for the filtering method.

D. PRODUCING THE PRODUCT

The groups or treatments will be divided into 4 groups. The four groups have the

same amounts of Enterobacteriaceae in a petri dish but has different amounts of Liberica

Coffee with Wild Cane extract. The group 1 will be receiving 1 ml of Liberica Coffee

with wild cane extract. The group 2 will be receiving 1.5 ml of Liberica Coffee with wild

cane extract. The group 3 will be receiving 3 ml of Liberica Coffee with wild cane

extract. The group 4 will be receiving 5 ml of Liberica Coffee with wild cane extract. The

groups’ antibacterial activity will be evaluated using different kinds of measurements to

identify the effects and using other kinds of tests to prove the researchers’ observation

stated.

E. RESEARCH INSTRUMENT

The researchers used rating scale in testing the evaluation and characterization of the

antibacterial activity and in collecting data. Rating Scale is a format where the answers to

the question are in form of scale. The hierarchy or gradation of the intensity of response,

according to the sets of ordered categories. This instrument is gives more specific,

accurate and reliable data. Compare to the questionnaire, this instrument is more qualified

and an advantage to mathematical analysis. This instrument can give the respondents a
 particular agreement or disagreement. The researchers used 9-point Hedonic Scale. 9

stands for extremely acceptable; 8 for very much acceptable; 7 for moderately acceptable;

6 for slightly acceptable; 5 for either acceptable; 4 for neither unacceptable; 3 for

moderately unacceptable; 2 for very much unacceptable and 1 for extremely

unacceptable.

Sampling Design

The sampling design used in this study is stratified sampling. This sampling technique

is wherein the researcher divides the entire population into different subgroups or strata, then

randomly selects the final subjects proportionally from the different strata.

Respondents

The respondents would be from Tagaytay City Science National High School.

F. DATA GATHERING PROCEDURES

The researcher gathers the data by means of observation. The researcher will use

observation method for Data Gathering Procedure because observation method spent less

time and less cost.The researcher will observe if the extract of Liberica Coffee combined

with Wild Cane could kill enterobacteria. Observation is the means by which you collect

knowledge of the researched phenomenon by observing the phenomena as how and when

they occur.Scientific observation is “a systematic description of events,behaviors,and

artifacts in the social setting chosen for study”(Kawulich,2005)

 G. STATISTICAL TREATMENT

The statistical tool used in analyzing and interpreting the data and testing the

null hypothesis of the study includes the percentage,mean, and T-test. T-test is

applicable beacause it is commonly used when the test statistic would follow a normal

distribution if the value scaling term in the test statistic were known.

GANTT CHART

ACTIVITIES WEEKS
1 2 3 4 5 6 7 8
Choosing
the Study
Researching
Resources
Calculating
for Sampling
Preparing
Questions
Conduct a
Survey
Compute
using T-Test
Combine all
Datum
Finalizing
the Research
 Chapter 5

SUMMARY OF FINDINGS, CONCLUSONS, and RECOMMANDATIONS

Summary of Findings

1.)