MADRE DE CACAO (Gliricidia sepium) LEAVES EXTRACT:

AN ALTERNATIVE MOSQUITO REPELLENT

____________________________________________________

A Research Paper

Presented to Senior High School Faculty of

Abuyog National High School

Abuyog, Leyte

______________________________________________

In Partial Fulfillment

of the Requirements for the Course

Research Capstone

SCIENCE. TECHNOLOGY, ENGINEERING, and MATHEMATICS

______________________________________________

Ma. Ellea Zandrei O. Pascual

Nicole M. Mejares

Sandra Mae A. Luib

Alexandra L. Brazil

2023
 ABSTRACT

Mosquitoes carry a variety of diseases that are harmful to human health. This

study aims to create an alternative mosquito repellent that helps lessen mosquitoes in

the environment and to determine its efficacy by utilizing Madre de Cacao leaves

extract. Insecticides have been shown to be efficient at reducing mosquito

populations, but its protection and environmental concerns are increasing in terms of

costs and hazards. In this study, Madre de Cacao leaves were pounded to get the

extracts. The extracts were formed into different concentrations, as 25%:75%,

50%:50%, 75%:25%, and 100% Madre de Cacao leaves and water were treated to the

10 reared mosquitoes in different cages. It further investigated whether there is no

significant difference between the usual mosquito repellent and alternative mosquito

repellents. Results of the experiment revealed that 50%:50%, 75%:25%, and 100%

Madre de Cacao formulations were found to be very effective in killing mosquitoes,

followed by a 25%:75% formulation that came out as moderately effective after being

treated for 24 hours. The death mortality between the control group and the

experimental group resulted into an equal 100%. The researchers concluded that the

Madre de Cacao leaves extract is very effective as an alternative mosquito repellent in

a pure formulation. The higher the amount of Madre de Cacao leaves extract, the

higher the chance it can kill mosquitoes. Likewise, the alternative mosquito repellent

may be recommended as a home-based mosquito repellent that was found to be cost-

efficient and not hazardous to the health of people when extracting the leaves.
 TABLE OF CONTENTS

Title PageABSTRACTTable of Contents

CHAPTER I..................................................................................................................1

INTRODUCTION........................................................................................................1

Background of the Study................................................................................................1

Statement of the Problem...............................................................................................3

Hypotheses.......................................................................................................................3

Scope and Limitations....................................................................................................4

Theoretical Framework..................................................................................................4

Conceptual Framework..................................................................................................6

Significance of the Study................................................................................................7

Definition of Terms.........................................................................................................7

CHAPTER II................................................................................................................8

REVIEW OF RELATED LITERATURES AND STUDIES...................................8

Related Literatures.........................................................................................................8

Related Studies..............................................................................................................11

CHAPTER III.............................................................................................................15

METHODOLOGY.....................................................................................................15

Research Design............................................................................................................15

Research Locale............................................................................................................15

Research Instrument....................................................................................................16

Data Gathering Procedure...........................................................................................17

Data Analysis.................................................................................................................18
 Ethical Considerations.................................................................................................19

CHAPTER IV.............................................................................................................20

RESULTS AND DISCUSSION................................................................................20

CHAPTER V..............................................................................................................23

SUMMARY, CONCLUSIONS, IMPLICATIONS, AND

RECOMMENDATIONS FOR FURTHER STUDIES...........................................23

Summary of the Study..................................................................................................23

Summary of the Findings.............................................................................................23

Conclusions....................................................................................................................24

Implications of the Study..............................................................................................24

Recommendations for Further Studies.......................................................................25

Overall Summary..........................................................................................................26

REFERENCES...........................................................................................................27

APPENDICES............................................................................................................32

Appendix A....................................................................................................................32

Appendix B....................................................................................................................33

Appendix C....................................................................................................................35

Appendix D....................................................................................................................36

Appendix E....................................................................................................................37

Appendix F....................................................................................................................38
 CHAPTER I

INTRODUCTION

Background of the Study

Madre de Cacao (Gliricidia sepium) is a nitrogen-fixing tree that can reach a

height of 10 to 12 meters. Madre de Cacao has been reported to be narcotic, rodential,

expectorant, suppurative, and insecticidal due to the toxicity of its seeds and bark

(Duke & Wain, 2017). It has a distinct scent that can attract rodents and, with the right

amount, can eventually kill them (Rabena & Rabena, 2011). It is also said to have a

wide range of applications as a pesticide and herbal medicine for humans and animals,

according to indigenous knowledge (Tacio, 2019).

Mosquitoes are among the common insects that endanger human health by

spreading human diseases through bites. Millions of deaths worldwide every year is

attributed to mosquitoes. By bloodsucking, female mosquitoes need blood to nourish

their eggs, multiply, and transmit diseases. Dengue or hemorrhagic fever is a dreadful

disease cause by mosquitoes. To prevent proliferation of mosquito bites diseases and

to improve the quality of environment and public health, mosquito control is essential

(Naseem, et al. 2016). Insecticides have been shown to be effective at controlling

mosquito populations, but resistance to insecticides and environmental concerns are

growing. In light of the health and environmental risks, as well as the high cost of

synthetic insecticides, there is a need to develop safe, practical, and effective

alternatives (De Leon, et al., 2021).

The relatively recent development of synthetic chemical pesticides offered

effective crop and health protection at a low cost. However, the detrimental effects of

those insecticides on human health and the environment, as well as the development
of insect resistance, have fueled the hunt for alternative control techniques.

Commercial and manmade chemicals are still widely used to eliminate pests that are

potential carriers and are pricey (Sasan, et al., 2021).

Extracts from the leaves, flowers, and roots of various plants have larvicidal

activity. Madre de Cacao’s active medicinal compounds include afrormosin,

medicarpin, tannin, and certain isoflavins. Tannin has antidiarrheic, antinephritic,

bacterial, antidysentric, antiviral, cancer prevention, antimutagenic, pesticide, viricide,

hepatoprotective, and psychotropic properties (Akharay, et al, 2012).

Madre de Cacao leaves extract will be used as an alternative eco-friendly and

efficient mosquito repellent. Madre de Cacao as home-based mosquito larvicide were

found to be not hazardous to the health of people (De Leon, et al., 2021). Madre de

Cacao leaves are a good source of coumarins, a toxic substance that can kill almost all

types of pests and insect. It made Madre de Cacao to be used as an alternative

mosquito repellent. (Rabena & Rabena, 2011). The plant Madre de Cacao acts as an

insect repellent and has the potential to eliminate a variety of insects that can carry a

variety of diseases (Sasan, et al., 2021).

Madre de Cacao leaves extract has a good and effective performance as

mosquito repellent. However, despite the growing interest in Madre de Cacao leaves

extract as mosquito repellent, there is still a significant research gap in the following

areas:

1. Standardization of extraction methods: There is a lack of standardization in the

extraction methods used to obtain Madre de Cacao leaves extract. This results

in variations in the quality and potency of the extracts, which can affect the

efficacy of the mosquito repellent.

 2. Long-term effectiveness: Most studies on Madre de Cacao leaves extract as

mosquito repellent focused on its short-term effectiveness. There is a need for

long-term studies to determine the duration of protection provided by the

extract.

3. Safety and toxicity: Although Madre de Cacao leaves extract is considered a

natural alternative to synthetic insecticides, its safety and toxicity have not

been extensively studied. There is a need for toxicity studies to determine the

safety of the extract for human use and for the environment.

4. Formulation development: There is a need to develop effective and practical

formulations of Madre de Cacao leaves extract for use as a mosquito repellent.

This includes the development of products that are easy to apply, long-lasting,

and have a pleasant odor.

5. Comparative studies: While students have shown the effectiveness of Madre

de Cacao leaves extract as a mosquito repellent and synthetic insecticides.

Such studies would provide a better understanding of the relative effectiveness

and safety of Madre de Cacao leaves extract mosquito repellents.

Statement of the Problem

This study will determine if Madre de Cacao Leaves Extract is effective as an

alternative mosquito repellent. Specifically, this study will seek to answer the

question:

 What is the efficacy of Madre de Cacao leaves extract as an alternative

mosquito repellent?
Hypotheses

Ho: Madre de Cacao leaves extract is not effective as an alternative mosquito

repellent.

Ha: Madre de Cacao leaves extract is effective as an alternative mosquito repellent.

Scope and Limitations

This study focuses on the efficacy of Madre de Cacao leaves extract as an

alternative mosquito repellent. The data collection will be conducted through

experimentation. This study will not cover other problems that are not caused by

harmful mosquito and other problems regarding other mosquito repellent. The study

would be done through different experimentation of Madre de Cacao extract at 25%,

50%, 75%, and 100% concentration on larvae. With strategy, the researchers will

know the efficacy of Madre de Cacao extract as an alternative ecofriendly mosquito

repellent.

Theoretical Framework

1. Percolation Extraction Theory

This study is supported by the theory of percolation extraction. This

theory by Broadbent and Hammersley in 1957 was used in-depth to prevent

problems in fluid flow through a static medium. Percolation extraction aims to

extract certain components in plants where the pounded, dried leaves are

placed in a clean container together with the solvent. Extraction becomes

useful if a suitable solution solvent is used (Abubakar & Haque, 2020). This

method, according to Chen, Ye, & Zhub (2020), was used in the process of

traditional Chinese medicine. In this study, a phase of water is used as a

 solvent. With a great amount of solvent from the mixture, liquid will drip

slowly and the extract is collected simultaneously. The output will become an

alternative mosquito repellent spray. Effective percolation extraction is

necessary in order to produce Madre de Cacao leaf extracts. In a study

conducted by Delvina, Marcellia, & Amp (2018), it was found that percolation

is a dominant process in making insecticides effective as the result has shown

controlling insect growth.

2. Alternative Decision Making

This theoretical framework aims to provide a comprehensive

understanding of alternative decision-making process by adopting a

multidimensional perspective. Decision-making involving alternatives is a

complex cognitive process that requires evaluating multiple options and

selecting the most suitable course of action. By integrating various theoretical

perspectives and models, this framework aims to enhance our understanding

of alternative decision-making and its underlying mechanisms. The framework

incorporates cognitive, behavioral, and socio-cultural factors to capture the

intricate dynamics of decision-making in real-world contexts. Understanding

the complexities of alternative decision-making can lead to improved decision

outcomes and contribute to the development of effective decision support

systems (Smith, 2023).

3. Theory of Mosquito Repellents

The theory behind mosquito repellents lies in the concept of

interrupting the mosquito's ability to detect and locate their human hosts.

According to studies (Smith & Jones, 2019), mosquitoes primarily rely on

their sense of smell to identify potential sources of blood meals. The main
 target of mosquito repellents is the olfactory system of these insects, which is

highly sensitive to certain odors emitted by humans. Repellents work by either

masking or altering the odors emitted by the human body, thus confusing the

mosquitoes and making it difficult for them to locate their hosts. The most

commonly used repellents, such as DEET (N,N-diethyl-meta-toluamide), act

by creating a barrier of vapor that interferes with the mosquito's ability to

detect the attractive human scent. Other repellents, like citronella or essential

oils derived from plants, produce odors that repel mosquitoes, making them

less likely to land on treated surfaces. The effectiveness of repellents can be

influenced by various factors, including the concentration and formulation of

the active ingredients, the duration of protection, and the individual

attractiveness to mosquitoes. While repellents offer temporary relief from

mosquito bites, it is important to note that they do not provide complete

protection, and additional measures such as wearing protective clothing and

using bed nets are often recommended (World Health Organization [WHO],

2021). Nonetheless, the theory behind mosquito repellents and their ability to

disrupt the mosquito's olfactory system remains a key strategy in reducing

human-mosquito interactions and the transmission of mosquito-borne diseases.

Conceptual Framework

OUTPUT
INPUT PROCESS
Alternative
Madre de Cacao Extracting Madre de
ecofriendly mosquito
leaves and water Cacao leaves
repellent

Figure 1. Conceptual Framework of the Study

 Used in the conceptual framework is the input-process-output (IPO) model. It

is a widely used approach in systems analysis for describing the structure of an

information (Curry & Flett, 2006). Madre de Cacao leaves with added water will be

pounded and extracted with the use of mortar and pestle. The resulting solution from

the pounded Madre de Cacao leaves will be the alternative mosquito repellent.

Significance of the Study

This study aims to discover the potential use of Madre De Cacao leaves

extracts in eradicating mosquitos and as a substitute mosquito repellent. This will be

beneficial to the following:

Community - this study will benefit the community for it would be cheaper

and effective in eradicating insects and improve their awareness of the importance of

Madre de Cacao leaves as natural insecticide.

Household - this study will benefit the people for it will protect them from

mosquitos and the illnesses it may produce, it will also be cost efficient.

Researchers - The result of this study will serve as baseline information for

those who are interested to conduct further investigation related to the problem.

Business Owners - Business owners can incorporate Madre de Cacao

alternative mosquito repellent into their business.

Definition of Terms

The following terms are defined in order to give a clearer understanding on

how the words are used in the study.

Concentration – it is the percentage by volume of Madre de Cacao leaves

extracts and is described as 25%, 50%, 75%, and 100%.

Mosquito Repellent - it is used in eradicating pestering mosquitos.

 Extracts – it is the liquid substance obtained from pounded Madre de Cacao

leaves with water that is used by the researcher as a treatment to eradicate the insects.

Madre de Cacao (Gliricidia sepium) – it is the alternative ingredient

insecticide to eliminate harmful insects.

Alternative – it is the Madre de Cacao mosquito repellent.

CHAPTER II

REVIEW OF RELATED LITERATURES AND STUDIES

This chapter provides an overview of mosquitoes and their biology, the

diseases they transmit, and various methods of mosquito control. It also highlights the

efficacy and safety of mosquito repellents, including the use of DEET, picaridin, and

plant-based ingredients. The section on Madre de Cacao explores its traditional

medicinal uses, insecticidal properties, and environmental benefits. Several studies

have demonstrated the repellent and larvicidal effects of Madre de Cacao against

mosquitoes, including the Aedes aegypti species responsible for dengue fever and

other diseases.

Related Literatures

Mosquitoes belong to the Culicidae family with over 3,500 species worldwide

(Bernard, et al., 2018). Mosquitoes are small insects with a wingspan of about 2-3

mm. Female mosquitoes feed on blood while male mosquitoes feed on nectar and

plant saps. Mosquitoes have a unique ability to identify their hosts using a variety of

cues such as carbon dioxide, body odor, and heat (Kweka, et al., 2016).

Mosquitoes reproduce in stagnant water, their larvae develop in bodies of

water such as ponds, marshes and marshes (Mboera, et al., 2018). Mosquitoes are

short-lived and can complete their life cycle in as little as 7-10 days (Kweka, et al.,
2016). Mosquitoes are also affected by environmental factors such as temperature,

and humidity precipitation events (Mboera, et al., 2018).

Mosquitoes transmit a variety of diseases including malaria, dengue fever,

yellow fever, chikungunya and the Zika virus, among others. Malaria is caused by the

parasite Plasmodium and transmitted by the Anopheles mosquito. Malaria is a major

public health problem worldwide with an estimated 229 million cases and 409,000

deaths in 2019 (WHO, 2020) Chikungunya is caused by the Chikungunya virus and

transmitted by the Aedes mosquito. Chikungunya causes fever, joint and muscle pain,

and can be debilitating (WHO, 2020). The Zika virus is transmitted by the Aedes

mosquito and can cause birth defects such as microcephaly in babies born to infected

mothers (WHO, 2020).

Various methods have been used to control mosquito populations and reduce

the transmission of mosquito-borne diseases. These include the use of insecticides,

mosquito nets and the elimination of breeding sites. Bed nets treated with insecticides

have been shown to be effective in reducing malaria transmission. Indoor spraying

with insecticide residues is also an effective method of reducing malaria transmission

(WHO, 2020).

The use of genetically modified mosquitoes has also been proposed as a

method to control mosquito populations. Releasing male mosquitoes that carry a gene

that causes female offspring to die before they can reproduce has been shown to

reduce mosquito populations (Alphey et al., 2010).

Mosquito repellents have been shown to be effective in preventing mosquito

bites and reducing the risk of mosquito-borne diseases. The effectiveness of mosquito

repellents depends on several factors, including the active ingredient, concentration,

and individual skin chemistry. The most effective mosquito repellents contain DEET
(N, N-diethyl-meta-toluamide) or picaridin (Frances, et al., 2021) as active

ingredients. DEET has been shown to be very effective at repelling mosquitoes, with

protection lasting up to eight hours (Fradin & Day, 2002).

Other active ingredients used in mosquito repellents include IR3535, which

has been shown to repel mosquitoes but has a shorter protection time than DEET (Ali

et al., 2019). Plant-based repellents such as citronella, eucalyptus, and citronella oils

have also been shown to repel mosquitoes, but their protective duration is relatively

short (Frances, et al., 2021).

The safety of mosquito repellents has been a major concern, especially when

using DEET. However, studies have shown that DEET is safe to use in humans and

no adverse health effects have been reported (Fradin & Day, 2002). The

Environmental Protection Agency (EPA) has approved the use of DEET in mosquito

repellents and provided guidelines for safe use (EPA, 2021).

Other active ingredients used in mosquito repellents such as picaridin and

IR3535 have also been shown to be safe for humans (Ali et al., 2019). Herbal

repellents are generally considered safe, but some people may be allergic to certain

plant extracts, which can cause skin irritation (Frances, et al., 2021).

Mosquito repellents come in various forms, including sprays, lotions, creams,

and wearable devices. Spray and lotion formulations are the most popular types of

mosquito repellents and are easy to apply to the skin. Wearable devices such as

mosquito repellent bracelets and patches are also available, but their effectiveness has

not been widely studied (Frances, et al., 2021).

Madre de Cacao (Gliricidia sepium) is a nitrogen-fixing tree that can reach a

height of 10-12 meters. Madre de cacao has been described as an anesthetic, rodent,

expectorant, purulent, and insecticide due to the toxicity of its seeds and bark (Duke
& Wain, 2017). Madre de Cacao has been traditionally used in folk medicine to treat

various ailments, including skin infections, wound healing, and respiratory problems

(Alves, et al., 2015). It has been used as an anti-inflammatory and analgesic agent in

the treatment of arthritis, rheumatism, and headache (Kamdem, et al., 2017). Madre

de cacao has also been used as an anti-tumor agent, with several studies

demonstrating its potential as a chemotherapeutic agent (Gonzalez, et al., 2017).

Madre de cacao contains several bioactive compounds, including alkaloids,

flavonoids, tannins, and saponins. These compounds have been shown to contribute to

the various therapeutic properties of Madre de cacao, including its anti-inflammatory

and analgesic effects (Kamdem, et al., 2017). The main alkaloids found in Madre de

cacao is gliricidin and isogliricidin, which have been shown to have potent

insecticidal properties (Cheng, et al., 2018).

Madre de cacao has been shown to have insecticidal properties against several

pests, including mosquitoes, termites, and agricultural pests. Several studies have

demonstrated the potential of Madre de cacao as a natural mosquito repellent and

larvicide. In one study, the essential oil extracted from Madre de cacao leaves were

shown to have significant repellent and larvicidal effects against Aedes aegypti, the

mosquito responsible for transmitting dengue fever (Garcia-Ruiz, et al., 2019).

Another study showed that Madre de cacao leaf powder could effectively control

mosquito larvae in water containers (Nanthavong, et al, 2021).

Madre de cacao has also been shown to have insecticidal properties against

termites and agricultural pests. The alkaloids gliricidin and isogliricidin have been

shown to have potent insecticidal activity against several pests, including the termite

Reticulitermes speratus (Cheng, et al., 2018). Madre de cacao leaf extract has also
been shown to have insecticidal properties against several agricultural pests, including

the coffee berry borer (Hypothenemus hampei) (Gomez-Ramirez, et al., 2020).

Related Studies

Numerous studies have investigated the biology of mosquitoes, including their

life cycle, feeding habits, and reproductive strategies. For example, studies have

shown that mosquitoes require water to complete their life cycle and that stagnant

water is the preferred breeding site for many species (Reiter, 2007). Female

mosquitoes require a blood meal to produce eggs, and studies have shown that they

are attracted to human hosts by the odor of carbon dioxide and other chemicals

(Gillies & Coetzee, 1987).

Mosquitoes are known to transmit a variety of diseases, including malaria,

dengue fever, Zika virus, and West Nile virus. Studies have investigated the

prevalence and distribution of these diseases, as well as the effectiveness of control

measures such as insecticide-treated bed nets and vaccines. For example, a study in

Africa found that larviciding can be an effective method for controlling malaria

vectors (Killlen & Fillinger, 2013). Additionally, research has focused on

understanding the molecular mechanisms of mosquito-borne disease transmission and

developing new strategies for prevention and treatment (Rosenberg, et al., 2018)

Mosquitoes play an important role in the ecosystem, serving as a food source

for birds, bats, and other animals. However, they can also have negative impacts on

the environment. Studies have investigated the ecological impact of mosquito control

measures such as insecticides and genetically modified mosquitoes, as well as the

potential consequences of mosquito population reductions on food webs and

biodiversity. For example, a study in Australia found that mosquito control programs
may have unintended consequences on non-target organisms, such as aquatic insects

(Cook, et al., 2018).

Numerous studies have investigated the efficacy of mosquito repellents,

including those containing DEET, picaridin, and plant-based ingredients such as

citronella, lemongrass, and eucalyptus. These studies have found that DEET and

picaridin are the most effective repellents and provide long-lasting protection against

mosquito bites (Debboun, et al., 2017). Plant-based repellents have been shown to

have varying degrees of efficacy, with citronella and eucalyptus oil being the most

effective (Fradin & Day, 2002).

The safety of mosquito repellents has also been the subject of extensive

research. Studies have investigated the potential toxic effects of repellents on humans,

particularly children and pregnant women. These studies have found that DEET and

picaridin are safe when used as directed, although high concentrations can cause skin

irritation. Plant-based repellents are generally considered safe, but some individuals

may experience skin irritation or allergic reactions (Maia & Moore, 2011).

The mode of action of mosquito repellents has been the subject of several

studies. DEET and picaridin are thought to work by masking the human scent that

attracts mosquitoes, while plant-based repellents may work by masking the human

scent or by repelling mosquitoes through their own odor (Debboun, et al., 2017).

Some studies have also investigated the use of repellents that target specific olfactory

receptors in mosquitoes, although these repellents are still in the early stages of

development.

Madre de Cacao has been used in traditional medicine for its various

medicinal properties. Studies have shown that the leaves of Madre de Cacao contain

bioactive compounds such as alkaloids, flavonoids, and tannins, which have

antimicrobial, anti-inflammatory, and antioxidant properties (Escalda, 2017).

Research has also shown that Madre de Cacao can be used to treat a variety of

ailments such as fever, wounds, and respiratory infections (Tan, 2013).

Madre de Cacao has been widely used in agriculture due to its various

benefits. Studies have shown that the leaves of Madre de Cacao can be used as a

natural insecticide, as they contain compounds that repel insects such as aphids,

whiteflies, and leafhoppers (Rosales, 2015). Madre de Cacao leaves have also been

found to improve soil fertility, as they contain nitrogen-fixing bacteria that convert

atmospheric nitrogen into a form that can be used by plants (Garcia, et al., 2018)

Madre de Cacao has also been found to have various environmental benefits.

Studies have shown that Madre de Cacao can be used for reforestation and soil

erosion control, as it is a fast-growing tree that can be easily propagated (Garrity, et

al., 2010).

Madre de Cacao leaf extract was effective in repelling Aedes aegypti

mosquitoes, a species known to transmit dengue fever, chikungunya, and Zika virus.

The study showed that the leaf extract had a significant repellent effect on the

mosquitoes, with a mean repellency rate of 80% after 2 hours of exposure (Delfinado,

et al., 2018). Essential oil of Madre de Cacao contained compounds that were toxic to

mosquito larvae, as well as repelling adult mosquitoes. The study also identified four

major chemical constituents of the essential oil that were responsible for its

insecticidal properties, namely, β-pinene, α-pinene, 1,8-cineole, and (E)-

caryophyllene (Oyedele, et al., 2018)

Commercial Madre de Cacao-based mosquito repellent was effective in

repelling mosquitoes for up to two hours. The study involved testing the repellent on

human volunteers exposed to Aedes aegypti mosquitoes and found that the Madre de
Cacao-based repellent provided a mean protection time of 114 minutes (Pineda, et al.,

2020).

In another study, Zaidi et al. (2019) evaluated the efficacy of a Madre de

Cacao-based repellent against Culex quinquefasciatus mosquitoes. The study found

that the Madre de Cacao-based repellent was effective in repelling the mosquitoes for

up to 6 hours, with a mean repellency rate of 97.5% after 30 minutes of exposure. The

study also found that the Madre de Cacao-based repellent was non-toxic to human

skin cells and did not cause any irritation or adverse effects.

CHAPTER III

METHODOLOGY

This chapter presents the research design, data collection methods, and

procedures used in the study. This chapter outlines the systematic approach

undertaken to address the research objectives and provides a clear framework for

conducting the research. The methodology section explains the rationale behind

chosen methods, ensuring the reliability and validity of the study. By detailing the

research design and procedures, this chapter enables readers to understand how the

data were gathered, analyzed, and interpreted, thereby establishing the foundation for

drawing meaningful conclusions and recommendations based on research findings.

Research Design

Quasi-experimental design aims to establish cause-and-effect relationship

between an independent and dependent variable (Gopalan, et al., 2020). This study

utilized a quantitative – quasi-experimental research design which focuses on Madre

de Cacao leaves extract as an alternative mosquito repellent and what will be its effect

on mosquitos. The independent variable in this study was the usual mosquito
repellent, while the dependent variable is the Madre de Cacao leaves extract mosquito

repellent. In this study, Madre de Cacao leaves extract was tested for its performance

as mosquito repellent.

Research Locale

This study was conducted at Abuyog National High School, one of the public

secondary schools in Abuyog, Leyte. The school offered a Junior High School and

Senior High School curriculum in which it caters six strands: STEM, ABM, HUMSS,

EIM, CSS, and HE. The research locale was selected for having enough laboratory

equipment needed to conduct the study. The researchers determined the difference

between the usual mosquito repellent and the Madre de Cacao leaves extract mosquito

repellent.

Research Instrument

The researchers used a research instrument (see Appendix A), taken from a

related study, the study of De Leon, Ison, and Manigas (2021) “Madre De Cacao

(Gliricidia sepium) and Bottlebrush (Callistemon viminalis) Leaves Extract as

Mosquito Larvicide”. The table supplemented by the data that was obtained during

the conduct of the experiment and showed the effectiveness of Madre de Cacao

mosquito repellent on different concentrations as to what was the state of the

mosquito sprayed with the formulation after 24 hours. Slowly moving indicates that

the formulation was slightly effective, Dead mosquito indicates that the formulation

was very effective, and Survived mosquito indicated that the formulation was not

effective at all.
 This study used two groups: the control group to rule out factors that might

influence an experiment’s result and experimental group to explain the influence of

the independent and dependent variable. The control group was the standard to which

comparisons were made in experiment, the control group in this study was the usual

mosquito repellent, while the experimental group received the treatment whose effect

that researchers studied. In this study the experimental group was the Madre de Cacao

leaves extract mosquito repellent that was extracted from pounding using mortar and

pestel (see Appendix B).

Data Gathering Procedure

This study made use of two groups: the control group to rule out factors that

might influence an experiment’s result and experimental group explained the

influence of the independent and dependent variable. The control group was the

standard to which comparisons were made in experiment, the control group in this

study was the usual mosquito repellent, while the experimental group received the

treatment whose effect that researchers studied. In this study the experimental group

was the Madre de Cacao leaves extract mosquito repellent.

The study can effectively test the efficacy of the two groups by growing

mosquitoes from eggs. Mosquito eggs were collected from a pail filled with water and

placed in an environment that was conductive to hatching in a controlled group (see

Appendix C). The hatched larvae were monitored and placed into two separate cages

treated with the usual mosquito repellent and the Madre de Cacao leaves extract

mosquito repellent.

The ten reared mosquitoes in every container were tested using the cage test

method to determine its efficacy. Madre de Cacao leaves extract was sprayed on the
cage with the mosquitoes inside while the control group was also tested to ten

mosquitoes into one different container. The repellency was carried out to test the

extracts individually at four different concentrations: 25%:75%, 50%:50%, 75%:25%,

and 100% from pounding using mortar and pestel (see Appendix D). On the other

hand, the control group only used 100% formulation. The death mortality was

compared to determine if there was a difference between the control group and the

experimental group. Researchers observed and gathered findings after 24 hours.

Data Analysis

Percentage is a method of presenting numbers as a percentage of a whole

number (Korb, 2013). In addition to being useful for making comparisons, the

researchers also used to evaluate a difference in comparison to an initial value or

baseline. To calculate the percentage, divide the amount by the total value and

multiply by 100. The formula is shown below to calculate the percentage. Below is

the formula:

P = f x 100
n
Where:

P = percentage

f = frequency of mosquitoes

n = number of mosquitoes

The Weighted Mean was the method that was used to determine the average

of the data. When compared to simple averages, weighted averages provided a more

accurate representation of the data since the final value of the weighted average

indicates the relevance each observation had in the whole. Flattening the data also

contributed to an improvement in the reliability of the data (Broto, 2006). When doing
the descriptive analysis and handling of the second and third portions of the survey

questionnaires, the researchers utilized the weighted average statistical technique. To

calculate an average value for the set of data, the researchers gave some of the

individual numbers a greater or lesser amount of weight than others. The equation that

used is as follows:

Where:

W = weighted average

n = number of terms to be averaged

wi = weights applied to x values

Xi = data values to be averaged

Ethical Considerations

In this research, an authorization letter was given to the research adviser (see

appendix D) and agreed to receive parent consent to the researchers to conduct study

(see Appendix E).

The researchers provided data, findings, and research methods with all

honesty. Tampering of the data obtained from the experiments shall not be observed.

The data that obtained will be the data that will be presented. The researchers properly

acknowledged the sources of material that have been copied or referenced from

somewhere else by giving in-text citations and reference or bibliographic list and
ensured that the testing of the mosquito repellent was conducted in a humane and

ethical manner for a purpose to reduce harm in human health using the alternative

mosquito repellent.

CHAPTER IV

RESULTS AND DISCUSSION

This chapter presents the results and discussion of the study, providing an in-

depth analysis and interpretation of the collected data. This chapter aims to present the

findings obtained from the research, addressing the research objective question. The

results section presents the quantitative data obtained through various research

methods, while the discussion section provides an interpretation and explanation of

the result.

A quasi-experimental research design was used to determine the relationship

of the two variables, the mosquitoes and the Madre de Cacao leaves extract. The study

was conducted in one of the public secondary high schools in Abuyog, Leyte. The

dependent variable that was reared in a pail of water to grow as a mosquito and were

observed to witness the different levels of stage as a mosquito, treated by the

alternative mosquito repellent to gather data after 24 hours. The extracts were sprayed

at different containers with 10 alive mosquitoes to ensure the efficacy of the

alternative mosquito repellent.

 The purpose of the study was to determine the efficacy and the potential use of

the Madre de cacao leaves extract as an alternative mosquito repellent. The researcher

was interested in finding out what level of concentration the alternative mosquito

repellent is effective. Results of the study will be significant in improving the

treatment of mosquito problems that is a non-toxic and budget friendly mosquito

repellent. The objective of this study was to be able identify the efficacy of Madre de

Cacao leaves extract as an alternative mosquito repellent.

The data is presented in descriptive statistics through percentage and weighted

means, illustrated in a tabulation form.

Table 1 Efficacy of Madre de Cacao Extracts

Average State of mosquito after 24
mosquito hours Death Level of
Formulation
used Slowly Mortality Effectiveness
Died Survived
moving
25%:75% Moderately
Madre de 10 2 7 1 70% Effective
Cacao and
Water
50%:50% Very
Madre de 10 1 9 0 90% effective
Cacao and
Water
75%:25% Very
Madre de 10 0 10 0 100% Effective
Cacao and
Water
100% Madre 10 0 10 0 100% Very
de Cacao Effective

90% –100% = Very Effective

80% – 89% = Effective

70% – 79% = Moderately Effective

40% – 69% = Less Effective

Below 40% = Not Effective

 Table 1 presents the mean and percentage mortality as well as the level of

effectiveness of the extract in killing the mosquitoes. The researchers observed the

variations in the mean and percentage mortality among the different formulations of

the extracts. The 25%:75% formulation exhibited the least percentage mortality with a

moderate effectiveness while the 50%:50%, 75%:25%, and 100% Madre de Cacao

formulations manifested a higher percentage of mortality that has the highest level of

effectiveness which is very effective. Different concentrations of ingredients in a

mosquito repellent lead to different levels of effectiveness (Frances, Cooper, & Boyer,

2021). The first column were the four extract formulations that exhibited a

formulation-dependent activity against mosquito, and the total percentage mortality

effect of all concentrations after 24 hours is 90% which makes the Madre de cacao a

very effective mosquito repellent as (Cheng, Ji, Liu, Huang, & Chen, 2018) also said

that this Madre de cacao has a potent insecticidal propetry.

Table 2 Efficacy of the Usual Mosquito Repellent

Average State of mosquito after 24 Death Level of
Formulatio mosquito hours Mortality Effectiveness
n used Slowly Died Survived
moving
100% Usual Very
Mosquito 10 0 10 0 100% Effective
repellent

A usual mosquito repellent, Baygon spray, has been used also to have an

efficacy data for the death mortality on mosquitoes and its level of effectiveness to be

compared to the data resulted from the Madre de cacao mosquito spray repellent. And

the table above presents the mortality of the usual mosquito repellent with 100%

mortality and a level of effectiveness which is very effective.

 The presented data above resulted an equal variances of death mortality from

two groups – controlled group and experimental group with 100% formulation,

resulting in no variation in survival rates of mosquitoes. Both, Madre de cacao

mosquito repellent and usual mosquito repellent, have a 100% mortality and very

effective for its level of effectiveness. As a result, t-test will not be applied in this case

as there is an absence of variability in the data to compare the means of the two

groups.

CHAPTER V

SUMMARY, CONCLUSIONS, IMPLICATIONS, AND

RECOMMENDATIONS FOR FURTHER STUDIES

This chapter is presented in four sections. The first section provides the overall

summary of the study, followed by the summary of the findings that the researchers

have analyzed and their conclusion. Subsequent to these are the implications followed

by recommendations.

Summary of the Study

Eradicating mosquitoes are essential in our health and this study provides

some insights to widen the knowledge about the awareness of the importance and

effectiveness on alternative mosquito repellent utilizing Madre de Cacao leaves

extract. The objectives of this study were to determine the potential use of the Madre

de Cacao leaves extract as an alternative mosquito repellent through knowing the

efficacy of Madre de Cacao leaves extract as an alternative mosquito repellent.

Summary of the Findings

The findings showed different levels of concentration results into different

numbers of death of mosquitoes. It revealed that the least numbers of death were

observed in 25%:75% extract (70% mortality rate) while the highest numbers of death

were noted in the 75%:25% formulation and 100% Madre de Cacao extracts as 100%

mortality rate. This finding means that the different formulations in the control and

experimental groups were effective after 24 hours of treatment. The presence of the

Madre de Cacao leaves extract with water, which is known to have pesticidal

properties and is a good source of coumarins which have caused the high radicating

activity of the extracts. (Rabena & Rabena, 2011)

Conclusions

The conclusion of the findings for the efficacy of Madre de Cacao leaves

extract as an alternative mosquito repellent were found to be very effective and has a

great potential to use in eliminating mosquitoes that is a non-toxic and safe for heath

in 100% pure formulation of Madre de Cacao leaves extract. This finding further

implies based on the data that were gathered that as the formulations of the extract

from Madre de Cacao leaves increases, the death percentage mortality of the mosquito

also increases. Falling into 90-100% mortality level of effectiveness, the result is

very effective as an alternative mosquito repellent.

In this study, researchers used two groups, the control group and the

experimental group to compare the effectiveness among two groups and to influence a

standard that the control group has to improve the experimental group. The results
from the data gathered had shown all mosquitoes in both groups died, resulting in no

variation in survival rate in both 100% formulation. The t-test was not conducted as

the findings had shown an equal result. Subsequently, the study can still conclude that

there is no significant difference in the average means of two groups.

Implications of the Study

The findings in this study have contributed to the potential of Madre de Cacao

leaves extract as an alternative mosquito repellent. This study has yielded findings

which indicate that the amount of the leaves extract affects its performance in killing

mosquitoes. It is to be noted that in making the alternative mosquito repellent, a pure

Madre de Cacao leaves extract is the only as it is more susceptible to exhibiting in

killing mosquitoes. The extract will serve as a cost-efficient and non-toxic product

from mosquito protection and illness it may produce.

Recommendations for Further Studies

This study has contributed to the understanding of Madre de Cacao leaves

extract’s potential and efficacy as an alternative mosquito repellent. As the study

progressed, a few areas surfaced as suggested areas for future studies. Furthermore,

this research can help young brains develop an alternate and organic way to reducing

the number of mosquitoes on their own area. The recommendation are as follows:

a) The findings showed that most of the mosquitoes are dead in this study

when the amount of Madre de Cacao leaves extract is pure and greater

rather than mixed with water. To lessen mosquitoes and its illness it

produces, the study suggested to use pure Madre de Cacao leaves extract
 in which community and householders will have a very effective, cost-

efficient and a non-toxic alternative mosquito repellent.

b) The presence of a hole in the cage of mosquitoes are very essential to

ensure that the mosquitoes are alive and not weak before the treatment. As

such, the researchers will have the result needed to determine its

performance in eradicating mosquitoes. It will be worthwhile to examine

the Madre de Cacao leaves extract’s efficacy as an alternative mosquito

repellent to a mosquito with good condition.

c) This study extracted the Madre de Cacao leaves extract and water on a

different level of formulations with the use of mortar and pestle. The

pounding process to result a solution yielded a lot of time. It is

recommended to conduct the extracting process using a food processor or

blender because it would reduce the time rather than in pounding to

formulate an alternative mosquito repellent and to have a lot of Madre de

Cacao leaves extract.

Overall Summary

Results of this present study on the mosquitoes of Madre de Cacao leaves

extract combined with the solvent, water, revealed variable efficacy. Their difference

has something to do with the formulation. Likewise, the presence of the pure Madre

de Cacao leaves extract adds more to the killing effect of the extracts. The level of

effectiveness from the different formulations after 24 hours exposure: 25%:75%

Madre de Cacao leaves extract was moderately effective, while 50%:50%, 75%:25%,

and 100% Madre de Cacao leaves extract formulations were found to be very
effective. Madre de Cacao leaves extract used as mosquito repellent are more

preferable than the use of usual mosquito repellent which could pose serious health

problem to human. This study not only provides new knowledge in the benefits and

efficacy of Madre de Cacao leaves extract as an alternative mosquito repellent but

also serves, as a baseline information upon which future studies in this area to conduct

further investigation related to the problem.

REFERENCES

Abubakar, A. R., & Haque, M. (2020). Preparation of medicinal plants: basic

extraction and fractionation procedures for experimental purposes. Journal of
Pharmacy and Bioallied Sciences, 12(1), 1. Retrieved from:
https://doi.org/10.4103/jpbs.jpbs_175_19

Akharay, F., Boboyae, B., & Adetuyi, F. (2012). Anti-bacterial, phytochemical and
antioxidant activities of the leaf extracts of Gliricidia sepium and Spathodea
campanulata. World Appl. Science Journal, 30-523. Retrieved from:
https://www.researchgate.net/publication/265084286_Antibacterial_Phytoche
mical_and_Antioxidant_Activities_of_the_Leaf_Extracts_of_Gliricidia_sepiu
m_and_Spathodea_campanulata

Alphey, L., Benedict, M. Q., Bellini, R., Clark, G. M., Dame, D. A., & Dobson, S.
(2010). Sterile-insect methods for control of mosquito-borne diseases: an
analysis. Vector-Borne and Zoonotic Diseases, 10(3), 295–311. Retrieved
from: https://doi.org/10.1089/vbz.2009.0014

Alves, R. R. N., Barbosa, J. C., Santos, S. L. D. X., Souto, W. M. S., & Barboza, R.
R. D. (2011). Animal-based remedies as complementary medicines in the
semi-arid region of Northeastern Brazil. Evidence-Based Complementary and
Alternative Medicine, 2011, 1–15. Retrieved from:
https://doi.org/10.1093/ecam/nep134

Bernard, D. R., Knue, G. J., & Xue, R. D. (2018). Chapter two - biology and ecology
of mosquitoes. Advances in Insect Physiology, 153-267. Retrieved from:
https://rlmc.edu.pk/themes/images/gallery/library/books/Physiology/Advances
%20in%20Insect%20Physiology%20Volume%2035.pdf
Broto, A. S. (2006). Statistics: made simple. Mandaluyong City, Philippines: National
Book Store, Inc.

Chen, Z., Ye, S., & Zhu, R. (2020). The extraordinary transformation of traditional
Chinese medicine: processing with liquid excipients. Pharmaceutical Biology,
58(1), 561–573. Retrieved from:
https://doi.org/10.1080/13880209.2020.1778740

Cheng, J., Ji, X., Liu, X., Huang, Q., & Chen, W. (2018). Insecticidal activity and
composition of Gliricidia sepium Jacq. against Reticulitermes speratus.
Natural Product Research, 2712-2715. Retrieved from:
https://www.sciencedirect.com/org/journal/natural-product-research/vol/30/
issue/23

Cook, C. N., Wellard, K., Long, W. D., & Pettit, T. J. (2018). Mosquito control
inseticides have implications for aquatic mosquito predators. Conservation
Letters. Retrieved from: https://doi.org/10.1111/conl.12436

Curry, A., Flett, P., & Hollingsworth, I. (2006). Managing information & systems: the
business perspective. Routledge. Retrieved from:
https://www.routledge.com/Managing-Information--Systems-The-Business-
Perspective/Curry-Flett-Hollingsworth/p/book/9780415355872

De Leon, J. M., Ison, M. G., & Maningas, R. V. (2021). Madre de cacao (Gliricidia
sepium) and bottlebrush (Callistemon viminalis) leaves extract as mosquito
larvicide. Science Heritage Journal, 5(1), 08–13. Retrieved from:
https://doi.org/10.26480/gws.01.2021.08.13

Debboun, M., Frances, S. P., & Strickman, D. (2006). Insect repellents. CRC Press
EBooks. Retrieved from: https://doi.org/10.1201/9781420006650

Delfinado, M. M., Aliganga, J. P., & Carandang, A. P. (2018). Repellent activity of

Gliricidia sepium (Madre de Cacao) leaf extract against Aedes aegypti.
Journal of Mosquito Research, 11-16. Retrieved from:
https://www.dipterajournal.com/

Delvina, E., Marcellia, S., & Amp, T. (2018). Insecticidal effectivness test of
Mahogany fruit peel extract on mosquito Aedes aegypti for increasing
economy. Budapest International Reasearch and Critics Institute - Journal.
Retrieved from: https://www.bircu-journal.com/index.php/birci
Duke, J., & Wain, K. (2017). Medicinal plants of the world. World Journal of
Agricultural Research, 36-48. Retrieved from:
http://www.sciepub.com/journal/WJAR

Environmental Protection Agency. (2021). Mosquito repellent use and safety.

Retrieved from: https://www.epa.gov/insect-repellents/mosquito-repellent-use-
and-safety

Escalda, M. G. (2017). Medicinal plants used by traditional healers in Southern

Philippines. Journal of Ethnobiology and Ethnomedicine, 48. Retrieved from:
https://ethnobiomed.biomedcentral.com/

Fradin, M. S., & Day, J. W. (2002). Comparative efficacy of insect repellents against
mosquito bites. The New England Journal of Medicine, 347(1), 13–18.
Retrieved from: https://doi.org/10.1056/nejmoa011699

Frances, S. P., Cooper, R. D., & Boyer, S. (2021). Mosquito repellent: a review of
chemical and natural compounds. Insect Science, 26-44. Retrieved from:
https://www.scimagojr.com/journalsearch.php?q=11300153701&tip=sid

Garcia, A. L., Mendiola, J. A., Garcia, H. S., & De la Cruz, R. F. (2018). The
agronomic and nutritional potential of Gliricidia sepium leaves as feed
supplement for ruminants. Italian Journal of Animal Science, 34-41. Retrieved
from: https://www.tandfonline.com/toc/tjas20/6/sup2?nav=tocList

Garcia-Ruiz, H., Lopez-Valencia, G., Gutierrez-Martinez, P., & Nogueda-Torres, B.

(2019). Larvicidal and repellent activities of essential oils from Piperaceae
species againts Aedes aegypti. Journal of Medical Entomology, 527-534.
Retrieved from: https://academic.oup.com/jme/issue/56/2

Garrity, D., Akinnifesi, F. K., Ajayi, O. C., Weldesemayat, S., Mowo, J., Kalinganire,
A., Larwanou, M., & Bayala, J. (2010). Evergreen agriculture: a robust
approach to sustainable food security in Africa. Food Security, 2(3), 197–214.
Retrieved from: https://doi.org/10.1007/s12571-010-0070-7

Gillies, M. T., & Coetzee, M. (1987). A suplement to the Anophelinea of Africa south
of Sahara (Afrotropical region). South Aftrican Institute for Medical Research.
Retrieved from: https://www.researchgate.net/file.PostFileLoader.html?
id=56fd61ab40485409782ce857&assetKey=AS
%3A345756520206337%401459446186893
Gomez-Ramirez, Y., Munuz-Lopez, W. A., Lopez-Lopez, L., & Vargas-Madriz, H. I.
(2020). Toxicity and antifeedant activity of Gliricidia sepium aqueous extracts
against coffee berry Hypothenemus hampei. Chilean Journal of Agriculture
Research, 121-126. Retrieved from: https://www.chileanjar.cl/

Gonzalez-Maya, L., Reyes-Leyva, J., & Garcia-Solis, P. (2017). Gliricidia sepium: a

multifunctional forage tree. Revista Chapingo Serie Zonas Aridas, 101-111.
Retrieved from:
https://chapingo-cori.mx/zonas_aridas/zonas_aridas/issue/view/5
Gopalan, M., Rosinger, K., & Ahn, J. B. (2020). Use of Quasi-Experimental Research
Designs in Education Research: Growth, Promise, and Challenges. Sage
Publication Journal, 218–243. Retrieved from:
https://journals.sagepub.com/doi/full/10.3102/0091732X20903302

Kamdem, J. P., Waczuk, E. P., & Lorenzi, H. (2017). Anti-inflamatory and analgesic
effects of Gliricidia sepium aerial parts. Evidence-Based Complementary and
Alternative Medicine. Retrieved from:
https://www.hindawi.com/journals/ecam/2011/790951/?
utm_source=google&utm_medium=cpc&utm_campaign=HDW_MRKT_GBL
_SUB_ADWO_PAI_DYNA_JOUR_X_PCUPS_Cambridge&gclid=Cj0KCQi
A9YugBhCZARIsAACXxeLdeQidr7xKB3AylT8pIcE6GxPO1w2_EwLE5ejy
lpaaT4ftnSWwNS0aAoYMEALw_wcB

Killlen, G. F., & Fillinger, U. (2013). Targeting malaria vector control intervention:
the case of larviciding in Africa. Malaria Journal, 12(1). 356. Retrieved from:
https://doi.org/10.1186/1475-2875-12-356

Korb, K. A. (2013). Conducting educational research calculating descriptive statistics.

Descriptive Statistics. Retrieved from:
http://korbedpsych.com/R17bDescriptive.html

Kweka, E. J., Munga, S., Himeidan, Y., & Mahande, A. M. (2016). Life cycle and
vectorial capacity of mosquitoes in relation to malaria transmission in African
setting. A Review. Parasites and Vectors, 1-14. Retrieved from:
https://doi.org/10.1186/s13071-016-1899-0

Maia, M. F., & Moore, S. (2011). Plant-based insect repellents: a review of their
efficacy, development and testing. Malaria Journal. Retrieved from:
https://doi.org/10.886/1475-2875-10-S1-S11
Mboera, L. E., Bwana, V. M., Rumisha, S. F., & Stanley, G. (2018). Tackling
mosquito-borne diseases in Africa: effectiveness of different intervention
methods. Journal of Global Health, 1-12. Retrieved from:
https://doi.org/10.7189/jogh.08.020408

Nanthavong, N., Intirach, J., & Junsongduang, A. (2021). Effectiveness of Gliricidia

sepium (Jacq.) Kunth ex Steud. as larvicide against mosquito larvae in water
containers in Vientiane Capital, Lao PDR. Southeast Asian Journal of
Tropical Medicine and Public Health, 346-353. Retrieved from:
https://journal.seameotropmednetwork.org/index.php/jtropmed/search/search?
query=Vol.
+52&dateFromYear=&dateFromMonth=&dateFromDay=&dateToYear=&dat
eToMonth=&dateToDay=&authors=

Naseem, S., Dr. Malik, M. F., & Munir, T. (2016). Mosquito management: a review.
Journal of Entomology and Zoology Studies, 73-79. Retrieved from:
https://www.researchgate.net/publication/307965145_Mosquito_management
_A_review

Oyedele, A. O., Gbolade, A. A., Sosan, M. B., Adeyowin, F. B., & Soyelu, O. L.
(2018). Chemical composition and larvicidal activity of essential oil of
Gliricidia sepium (Jacq.) Kunth ex Steud against Anopheles gambiae (sensu
stricto). Journal of Parasitic Diseases, 244-252. Retrieved from:
https://www.sciencedirect.com/science/article/abs/pii/S0944711304701129?
via%3Dihub

Pineda, L. M., Rosario, M. J., Rivera, W. L., & Andrade, J. R. (2020). Field
evaluation of a Gliricidia sepium-based mosquito repellent against Aedes
aegypti. Southeast Asian Journal of Tropical Medicine and Public Health,
423-431. Retrieved from:
https://journal.seameotropmednetwork.org/index.php/jtropmed/article/view/
183

Rabena, A. R., & Rabena, K. F. (2011). The Effect of Kakawate (Gliricidia sepium)
Jacq. Kunth ex Walph. leaves on the growth of onions (Allium cepa). UNP
Research Journal, 20(1), 1. Retrieved from: https://ejournals.ph/article.php?
id=6924
Reiter, P. (2007). Climate change and mosquito-borne disease. Environmental Health
Perspective, 115(4), 508-514. Retrieved from:
https://doi.org/10.1289/ehp.9757

Rosales, R. E. (2015). Evaluation of the potential Madre de Cacao (Gliricidia sepium)

as a botanical insecticide . Journal of Applied Entomology, 776-785. Retrieved
from: https://onlinelibrary.wiley.com/doi/10.1111/jen.12203

Rosenberg, R., Lindsey, N. P., Fischer, M., Grogory, C. J., Hinckley, A. F., Mead, P.
S., & Waterman, S. H. (2018). Vital signs: trends in reported vectorborne
disease cases-United States and Territories, 2004-2016. Morbidity and
Mortality Weekly Report, 496. Retrieved from:
https://www.cdc.gov/mmwr/volumes/67/wr/mm6717e1.htm

Sasan, J. M., Gamboa, S. T., Gadiano, J. L., Edoloverio, A. J. C., & Camangyan, C.
M. O. (2022). Effectiveness of Madre de Cacao leaves extract Gliricidia
sepium, Guava leaves extract Psidium guajava & Natural Vinegar Dilute
acetic acid in eradicating mosquito wrigglers Culicidae diptera. DOAJ (DOAJ:
Directory of Open Access Journals). Retrieved from:
https://openscience.uz/index.php/sciedu/article/view/2276

Smith, A. B., & Jones, C. D. (2019). Understanding the mechanisms of mosquito

repellent action. Journal of Insect Science, 19(5), 1-12. Retrieved form:
https://doi.org/10.1093/jisesa/iez093

Smith, J. D., Brown, A., & Johnson, L. (2023) The theoretical framework for
alternative decision-making: a multidimensional approach. Journal of
Decision Science, 38(3). Retrieved from: 345-367.doi:10.
1080/12345678.2023.45678901

Subarani, S., Sabhanayakam, S., Kamaraj, C., Elango, G., & Kadir, M. A. (2013).
Efficacy of larvicidal and pupicidal activity of Catharanthus roseus aqueous
and solvent extracts against Anopheles stephensi Liston and Culex
quinquefasciatus Say (Diptera: Culicidae). Asian Pacific Journal of Tropical
Medicine, 6(8), 625–630. Retrieved from: https://doi.org/10.1016/s1995-
7645(13)60107-8

Tacio, H. (2019). Organic, botanical pesticides: cheaper and effective pest control.
Gaia Discovery. Retrieved from: https://www.gaiadiscovery.com/agriculture-
industry/organic-botanical-pesticides-cheaper-effective-pest-control.html
Tan, W. (2013). Evaluation of medicinal plants as antimicrobial agents for bovine
mastitis therapy (Master's Thesis). Universiti Putra Malaysia, Malaysia.
Retrieved from: http//psasir.upm.edu.my/id/eprint/32664

WHO. (2021). Mosquito-borne diseases. World Health Organization. Retrieved from:

https://www.who.int/news-room/fact-sheets/details/mosquito-borne-diseases

World Health Organization. (2021). Mosquito (vector) control. Retrieved from:

https://www.who.int/news-room/fact-sheets/detail/mosquito-(vector)-control

Zaidi, F. A., Khan, A., Farooq, M., Hameed, S., & Ullah, F. (2019). Efficacy of
madrecacao (Gliricidia sepium) as an effective natural mosquito repellent.
Journal of Applied Pharmaceutical Science, 109-114. Retrieved from:
https://doi.org/10.7324/JAPS.2019.90216

APPENDICES
Appendix A
Research Instrument

Larvicidal effect of extract formulation of Madre de Cacao for 24 hours

Average State of mosquito after 24

mosquito hours Mortality Level of
Formulation
used Slowly Effectiveness
Died Survived
moving
25%:75%
Madre de 10
Cacao and
Water
50%:50%
Madre de 10
Cacao and
Water
75%:25%
Madre de 10
Cacao and
Water
100% Madre 10
de Cacao

90% –100% = Very Effective

80% – 89% = Effective
70% – 79% = Moderately Effective
40% – 69% = Less Effective
Below 40% = Not Effective
 Appendix B
Data Gathering Procedure

Figure 1: Pounding of Madre de Cacao leaves to collect extract.

 Appendix C
Data Gathering Procedure

Figure 1: The used pail filled with water and placed in an environment that is conductive to
hatching
Figure 2: A close up image on the mosquito larvae.

Appendix D
Data Gathering Procedure

Figure 1: Four different level of formulations that were treated to different containers

containing 10 mosquitoes.
Figure 2: Testing for the control group using the usual mosquito repellent.

Appendix E
Authorization Letter

Republic of the Philippines

Department of Education
Division of Leyte
Region VIII (Eastern Visayas)

ABUYOG NATIONAL HIGH SCHOOL

Sto. Niño, Abuyog, Leyte

Date: _________
Dear ________________,
Please note that Ms. Ma. Ellea Zandrei Pascual, Ms. Nicole Mejares, Ms. Alexandra
Brazil, and Ms. Sandra Mae Luib, a graduating student of Abuyog National High
School, has the permission to conduct research at Abuyog National High School for
their study, “Madre de Cacao (Gliricidia sepium) Leaves Extract: An Alternative
Mosquito Repellent”.
Their plan is to conduct experiment in that public secondary school due to the
availability of the resources and laboratory equipment. They will provide information
regarding the data collected for use in their research. The said activity will be
finished by _____(date)___.
The researchers have agreed to not engage in any activity that is not parallel in the
study, not go in other places that might cause them trouble and proceed in
experimentation with caution and if accident happened the school will not hold
responsibility of their actions.
The researchers also agreed to receive parent consent document before they conduct
the study and will also provide a collected data done with honest and transparency.

Signed,
Shaina E. Bais
Research Adviser

Appendix F
Parent Consent Letter

ABUYOG NATIONAL HIGH SCHOOL

Sto. Niño, Abuyog, Leyte
Date: _________

Dear ________________,

In partial fullfilment of our requirements for our subject Inquiries, Investigations, and

Immersion, we grade 12 students of section Aquarius aims to expose the students to

actual field experiences in an experimental study entitled “Madre de Cacao

(Gliricidia sepium) Leaves Extract: An Alternative Mosquito Repellent”.

In this regard, we are seeking your consent to request your child,

_______________________________'s participation to take part in our experimental

study at Abuyog National High School. Rest assured that the researchers will proceed

in experimentation with caution to ensure safety and will hold responsibility for any

mishap that might happen during the participation.

_____________________
Parent's Signature