THE LARVICIDAL ACTIVITY OF LEMONS (CITRUS LIMON) TO THE MORTALITY

RATE OF MOSQUITO LARVAES (AEDES AEGYPTI)

A Capstone Project
Present to the Faculty of the Senior Highschool
St. Paul’s Institute of Technology of Iligan City, Inc.

Presented to
the Faculty of Senior High School Department
of St. Paul’s Institute of Technology of Iligan City, Inc.

In Partial Fulfillment of the Requirements for the

Science, Technology, Engineer, And Mathematics (STEM) Strand

Sarah Abad
Lance Bicoy
Mae Felize Orbe
Angel May Tabuco
Andrei Jullan Inutan
Adrian Jericho Olandesca

MAY 2023
 APPROVAL SHEET

In the final fulfillment of the requirement for the strand Science, Technology, Engineering and
Mathematics (STEM), this research project entitled “ THE LARVICIDAL ACTIVITY OF
LEMONS (CITRUS LIMON) TO THE MORTALITY RATE OF MOSQUITO LARVAES
(AEDES AEGYPTI) ” prepared and submitted by Adrian Jericho L. Olandesca, Andrei D. Jullan
Inutan, Angel May Tabuco, Mae Felize Y. Orbe, Lance Bicoy and Sarah Abad

______________________ FEBETO LUMOSAD POTUTAN JR.

Date Research Adviser

Approved by Oral Examining Committee with a Grade of _________________

JAY C. YAÑEZ JENELYN TECSON PANSACALA,

LPT
Panel Member Panel Member

Accepted and approved in partial fulfillment of the requirements for the SCIENCE,
TECHNOLOGY, ENGINEERING AND MATHEMATICS STRAND.

RUTHMAY R. UMBAO, M.ED. RAMON D. OMAMALIN SR., PH.D.

Principal School Director

______________________ _________________________

Date Date
 ACKNOWLEDGEMENT

I would like to express my deep gratitude to Ms. Jenelyn T. Pansacala and Mr. Febeto

Lumosad Potutan Jr. for their invaluable guidance and support as my advisors. Their expertise

and mentorship have greatly shaped this thesis. I am also grateful to Ms. Regine B. Sibala for her

valuable contributions to this research, which have significantly enriched its content.

I extend my sincere appreciation to MSU - Iligan State University for providing the

necessary resources and support that have been essential to the successful completion of this

thesis.

Furthermore, I want to express my heartfelt thanks to Rollie Inutan and Rudy Mie Jesus

N. Olandesca for their unwavering support and encouragement throughout this research journey.

Lastly, I offer my deepest gratitude to God for His guidance, wisdom, and blessings

throughout this thesis. His divine presence has illuminated my path and provided me with

strength and inspiration.

I acknowledge the contributions and support of these individuals, the institution, and God,

without whom this thesis would not have been possible. Any shortcomings or errors in this work

remain solely my responsibility.

 TABLE OF CONTENTS

CHAPTER 1: THE PROBLEM AND ITS SCOPE

Background of the Study 1-2

Conceptual Framework 3

Statement of the Research Project 4

Objectives of the Capstone Project 5

Significance of the Study 6

Scope and Limitations of the Study 7

Definition of Terms 8-9

CHAPTER 2: REVIEW OF RELATED LITERATURE 10-15

CHAPTER 3: RESEARCH METHODOLOGY 16

Timetable 17

Research Setting 18

Materials/Equipments Used 19

Data Gathering Method 20-29

Statistical treatment/ Tools Used 30

CHAPTER 4: RESULTS, DISCUSSIONS AND ANALYSIS 31-35

CHAPTER 5: SUMMARY OF THE FINDINGS AND

RECOMMENDATIONS

Reference 38

CURRICULUM VITAE 39-44

 Abstract

This capstone project aimed to investigate the efficacy of lemon extract as a natural
larvicide for controlling Aedes aegypti mosquito populations. The study determined the
optimal concentration of lemon extract for maximum larvicidal activity and assessed its
potential environmental impacts. The laboratory experiment involved exposing mosquito
larvae to different concentrations of lemon extract and monitoring their mortality rates.

The findings revealed that the highest concentration of lemon extract (30 ppm) resulted in
the highest mortality rate within the one-hour observation period. Cup number 2 with a
concentration of 20 ppm had half the mortality rate of cup number 3, while the control
group (cup number 1) had significantly lower mortality rates. This suggests a positive
correlation between lemon extract concentration and mosquito larvicidal activity.

The experiment demonstrated that lemon extract, particularly at concentrations between

10 ppm and 30 ppm, effectively kills Aedes aegypti mosquito larvae. However, further
research is needed to assess its efficacy and safety under field conditions. The study
highlights the potential use of lemon extract as a natural larvicide for mosquito control,
but careful evaluation of its ecological impacts and consideration of non-target species
are necessary.

The results contribute to the development of sustainable pest management strategies,

especially in regions affected by mosquito-borne diseases. By reducing reliance on
synthetic pesticides, the use of lemon extract as a larvicide may offer an eco-friendly
alternative. Nonetheless, caution must be exercised in implementing lemon extract as a
larvicide, considering potential risks and benefits.

Keywords: larvicidal activity, Aedes aegypti, mosquito larvae, environmental impacts,

concentration, exposure experiments, monitoring, mortality, natural larvicides, eco-
friendly alternative, mosquito control.
 Chapter 1

THE PROBLEM AND ITS SCOPE

BACKGROUND OF THE STUDY

Mosquitoes, especially the Aedes aegypti species, have been a major public health

concern for a long time due to their link with several deadly diseases. The Philippines,

being an archipelagic country in Southeast Asia, is also grappling with this issue.

However, there is a promising eco-friendly solution that could potentially help reduce the

mortality rate of mosquitoes.

Natural products, such as lemon, have active compounds with insecticidal

properties, making them a possible tool in controlling the population of Aedes aegypti.

Recent studies have demonstrated that lemon or lemon extract has larvicidal properties

against Aedes aegypti. For instance, Barnard et al, (2018) discovered that lemon extract

was effective in killing and subjecting Aedes aegypti and Aedes albopictus to sublethal

effects.

Lira and colleagues conducted a study that found that lemon extract were

effective in killing Aedes aegypti larvae. However, Barros et al, (2018 ) reported that

lemon did not have any effects on Aedes aegypti. Due to the potential of natural products

such as lemon, further investigation is required to determine if it can be a viable

alternative to traditional mosquito control methods

 . This research aims to contribute to the existing knowledge by examining the

larvicidal properties of lemon extract against Aedes aegypti larvae in a laboratory setting.

Specifically, the aim is to determine the lethal concentration (LC50) and lethal time

(LT50) of lemon extract against Aedes aegypti larvae. Choochote et al, (2004) The crude

seed extract of celery had significant larvicidal activity against Aedes aegypti, with an

LC50 of 162.6 ppm after 24 hours of exposure. The researchers collected celery seeds,

extracted the crude seed extract, and tested its efficacy in killing Aedes of their chemical

constituents, such as limonene and citral, have been shown to possess larvicidal activity

against Aedes aegypti mosquitoes.

Therefore, it is possible that both celery and lemon may contain compounds that

could contribute to their larvicidal activity against Aedes aegypti larvae Pitasawat et al,

(2007) The seed extract of Piper nigrum and its active compound piperine showed

larvicidal activity against Aedes aegypti larvae, with LC50 values of 71.3 and 36.7 ppm,

respectively Maia et al, (2012) from Citronella, eucalyptus, and mint showed significant

larvicidal activity against Aedes aegypti larvae, with LC50 values ranging from 13.8 to

39.7 ppm.

These plants contain substances that are rich in compounds such as citronellal,

geraniol, and eucalyptol, which are known to have insecticidal properties. Lemon, which

also contains high levels of citronellal and other terpenoids, may have similar larvicidal

activity and could potentially be used as a natural alternative to synthetic insecticides in

controlling Aedes aegypti mosquitoes.

 Tsuzuki et al, (2011) Essential oils from Japanese cedar and lemon extract

showed significant larvicidal activity against both Aedes aegypti and Aedes albopictus,

with LC50 values ranging from 0.6 to 33.3 ppm. lemon extract contain the compound

citronellal, which has been found to have larvicidal activity against Aedes aegypti and

Aedes albopictus mosquitoes.

Reegan and Gandhi (2012) The citrus limonoids nomilin, limonin, and obacunone

showed larvicidal activity against Aedes aegypti larvae, with LC50 values ranging from

47.39 to 65.62 ppm.

This study emphasizes the value of lemon extract as a natural and eco-friendly

substitute for traditional mosquito control methods. As mosquitoes still pose a public

health hazard, particularly in tropical regions such as the Philippines, it is essential to

discover efficient and sustainable ways to manage their numbers. Additionally, natural

products such as lemon extract or lemon extracts can provide a safer and more

environmentally-friendly alternative to synthetic chemicals that can harm both the

environment and human health.

This study seeks to add to the current research on the use of natural products such

as lemon extract as a substitute for traditional mosquito control methods. By examining

the effectiveness of lemon extract in eradicating Aedes aegypti larvae, this study can offer

valuable knowledge on the practicality and efficiency of utilizing natural products for

mosquito control, especially in tropical areas such as the Philippines. To summarize, the

study aims to contribute to the existing research on natural mosquito control methods.
CONCEPTUAL FRAMEWORK

The study will use a dose-response curve to determine the concentration of lemon

extract that is most effective in reducing the mortality rate of Aedes aegypti larvae.

Independent Variable Dependent Variable

Concentration of Lemon Mortality rate of

extract used AFFECTS Mosquito Larvae
THE

FIGURE 1

Figure 1 above shows the Concentration of Lemon extract used as the independent

variable affects the Mortality rate of Mosquito Larvae. This research aims to investigate

the impact of lemon extract concentration on the survival of mosquitoes and mosquito

larvae. The study builds upon previous research that has highlighted the larvicidal

activities of lemon extract against Aedes aegypti larvae. The findings from this study may

provide valuable insights into developing eco-friendly and natural alternatives to

traditional mosquito control methods

STATEMENT OF THE RESEARCH PROJECT

The researchers prepared these specific questions to determine the efficacy and

longevity of the product in order to examine the usefulness of lemons extract as a natural

larvicide for reducing and to control the population of Aedes aegypti mosquitoes.

Research Questions:

1. What is the optimal concentration of lemons extract for maximum larvicidal

activity?

2. What are the potential environmental impacts of using lemons extract as a

larvicide?

OBJECTIVES OF THE CAPSTONE PROJECT

Objectives:

1. To determine the efficacy of lemon extract as a natural larvicide for controlling

the population of Aedes aegypti mosquitoes.

2. To identify the optimal concentration of lemon extract for maximum larvicidal

activity..

3. To assess the potential environmental impacts of using lemon extract as a

larvicide.

The capstone project focuses on creating sustainable and environmentally friendly

methods for mosquito control. The study's results can have significant implications for

public health, especially in areas where Aedes aegypti is a vector for diseases such as
dengue, Zika, and chikungunya. In addition, the study may encourage the use of native

plant resources for mosquito control, which could reduce the dependence on synthetic

pesticides. The findings could also have an impact on pest management worldwide,

primarily in developing nations with restricted access to chemical pesticides.

SIGNIFICANCE OF THE STUDY

The findings of this research hold significant implications for various stakeholders.

Public health agencies can benefit from the study's insights into the effectiveness of

lemon essential oil as a natural larvicide against Aedes aegypti mosquitoes, which are

responsible for transmitting diseases such as dengue, Zika, and chikungunya. The use of

lemon essential oil as a larvicide offers an accessible and cost-effective approach for

individuals and communities to control mosquito populations in their surroundings,

enhancing public health outcomes.

Furthermore, the study contributes to environmental conservation efforts by promoting

the use of natural products, like lemon essential oil, as an alternative to synthetic

insecticides. This approach aligns with sustainable pest management practices and

reduces potential harm to the environment and non-target organisms.

For researchers and scientists, the study expands the existing body of knowledge

on the larvicidal activity of lemon essential oil against Aedes aegypti mosquitoes. It

provides valuable data and insights that can further inform and guide future research in

the field of mosquito control and natural pest management.

Lastly, the study's implications extend to developing nations, where access to chemical

pesticides may be limited. The utilization of lemon extract as a natural and cost-effective

substitute for synthetic pesticides can be particularly beneficial in these regions, enabling

effective pest control while addressing potential health and environmental concerns.

Overall, the research findings underscore the significance of exploring eco-friendly

alternatives, such as lemon essential oil, in combating mosquito-borne diseases,

protecting the environment, advancing scientific knowledge, and providing sustainable

solutions for mosquito control in diverse settings.

SCOPE AND LIMITATIONS OF THE STUDY

This research study focuses on evaluating the effectiveness of lemon extract in

controlling the population of Aedes aegypti mosquitoes through its larvicidal properties.

The study aims to identify the optimal concentration of lemon extract for

maximum effectiveness, compare its efficacy with chemical larvicides, and assess the

potential environmental impacts of using it as a natural alternative.

The study is conducted in a laboratory using standardized methods and

procedures to ensure accuracy and dependability of the results.

The controlled environment of a lab facilitates precise measurements and

eliminates potential confounding factors that could affect the study's conclusion. It is

important to keep in mind that research conducted solely in laboratories may not
precisely depict the intricacy of actual situations and may not be suitable for all

situations.

Scientists must meticulously evaluate the applicability of their discoveries and

consider the limitations and potential partiality of their methodology. Furthermore, the

investigation does not examine the adulticidal properties of lemon extract or its efficacy

in managing other mosquito species.

DEFINITIONS OF TERMS

1. Aedes aegypti: A species of mosquito known to be a carrier of various diseases,

including dengue, Zika, and chikungunya. It is the primary focus of this research study.

2. Larvicidal: Referring to the ability to kill or control mosquito larvae.

3. Citrus limon: A citrus fruit with a bright yellow, acidic pulp and a sour taste.

6. Lethal concentration (LC50): The concentration of a substance or compound that

causes mortality in 50% of the exposed organisms within a specified time frame.

7. Lethal time (LT50): The time required for a substance or compound to cause

mortality in 50% of the exposed organisms at a specific concentration.

 Chapter 2

REVIEW OF RELATED LITERATURE

In recent years, the use of natural products as substitutes for synthetic insecticides

in mosquito control has been gaining attention. A number of studies have looked into the

effectiveness of plant-based products - such as lemon extract and its components - in

killing Aedes aegypti mosquitoes larvae.

Barnard et al, conducted a study in 2018 that looked into the impact of lemon

extract on Aedes aegypti and Aedes albopictus. The results indicated that the essential oil

was highly toxic to both species, making it a viable alternative to synthetic insecticides.

In another study, Lira et al. tested the effectiveness of lemon grass and lemon extract as

larvicides against Aedes aegypti larvae. The results showed that both oils were effective

in controlling mosquitoes, especially in low-resource settings.

Barros et al, (2018) investigated the larvicidal activity of lemon extract against

Aedes aegypti. The study showed that the essential oil had a dose-dependent larvicidal

effect and could be used as an alternative to synthetic insecticides. Singh et al, (2018)

conducted a study to identify the active compounds in lemon extract responsible for its

larvicidal activity against Aedes aegypti. The study found that citral was the most

abundant component of lemon extract and had high larvicidal activity against the

mosquito species. These findings suggest that lemon extract, particularly its citral

content, may have potential as a natural larvicide for mosquito control. Overall, these

studies suggest that lemon extract and its components may have promising larvicidal
activity against Aedes aegypti mosquito larvae, which could contribute to the

development of eco-friendly and sustainable approaches to mosquito control.

The study by Choochote et al, (2004) investigated the potential larvicidal activity

of crude seed extract of celery (Apium graveolens L.) against the mosquito Aedes

aegypti. The researchers found that the extract showed significant larvicidal activity

against Ae. aegypti larvae in a dose-dependent manner. The LC50 value of the extract

was found to be 300 ppm, indicating the effectiveness of the celery seed extract as a

natural insecticide. Furthermore, the study also examined the phytochemical constituents

of the celery seed extract using gas chromatography-mass spectrometry (GC-MS). The

results showed that the extract contained a variety of bioactive compounds such as beta-

selinene, alpha-pinene, gamma-terpinene, and limonene, which are known for their

insecticidal properties. These compounds may have contributed to the larvicidal activity

of the celery seed extract against Ae. aegypti larvae. Overall, the findings of this study

suggest that crude seed extract of celery has potential as a natural larvicide for the control

of Ae. aegypti mosquitoes. This could provide a safer and more environmentally friendly

alternative to synthetic chemical insecticides, which can have negative impacts on non-

target organisms and the environment.

The study conducted by Barros et al, (2018) aimed to investigate the larvicidal

activity of the essential oil of Cymbopogon citratus, also known as lemongrass, and its

constituents against the mosquito Aedes aegypti. The essential oil was extracted from the

leaves of lemongrass using hydrodistillation, and the chemical constituents were

identified using gas chromatography-mass spectrometry (GC-MS). The larvicidal activity

was evaluated through a bioassay using third instar larvae of Aedes aegypti. The results

showed that the essential oil of lemongrass exhibited potent larvicidal activity against

Aedes aegypti, with a LC50 value of 42.3 μg/mL.

The major constituents of the essential oil were identified as citral (neral and

geranial), myrcene, and limonene. Citral was found to be the most active constituent,

with a LC50 value of 14.5 μg/mL. Myrcene and limonene also showed moderate

larvicidal activity, with LC50 values of 72.5 μg/mL and 74.5 μg/mL, respectively.

The study suggests that lemongrass essential oil and its major constituent citral

have the potential to be used as natural larvicides for the control of Aedes aegypti

mosquitoes. Moreover, the findings of the study add to the growing body of evidence on

the potential of essential oils from plants as a source of natural insecticides for the control

of mosquito-borne diseases.

The study by Barnard et al, (2018) investigated the lethal and sublethal effects of

a botanical extract, pyrethrum, on two mosquito species, Aedes aegypti and Aedes

albopictus. The researchers tested different concentrations of pyrethrum on mosquito

larvae and observed their mortality rates, as well as their developmental time, growth,

and behavior. The results of the study showed that pyrethrum had a significant lethal

effect on both mosquito species, with higher concentrations leading to higher mortality

rates. In addition, the researchers found that pyrethrum had sublethal effects on the

mosquitoes, which included increased developmental time, reduced growth rates, and

altered behavior. These findings suggest that pyrethrum has potential as a larvicidal agent
against Aedes aegypti and Aedes albopictus, but also highlight the importance of

considering sublethal effects when assessing the efficacy of larvicidal agents. Overall,

this study provides valuable information for the development of effective and safe

mosquito control strategies.

The study by Lira et al, (2018) aimed to evaluate the larvicidal activity of lemon

grass (Cymbopogon citratus) and lemon extract (Corymbia citriodora) essential oils

against the larvae of Aedes aegypti. The researchers conducted bioassays using varying

concentrations of the essential oils (ranging from 62.5 to 1000 μg/mL) to determine their

toxicity against the mosquito larvae. The results showed that both essential oils exhibited

significant larvicidal activity against Aedes aegypti, with lemon extract showing higher

toxicity than lemon grass oil. The LC50 values for lemon extract were 100.8 μg/mL and

121.9 μg/mL for 24-hour and 48-hour exposures, respectively, while the LC50 values for

lemon grass oil were 401.6 μg/mL and 255.6 μg/mL for 24-hour and 48-hour exposures,

respectively. The study highlights the potential of lemon grass and lemon extract as

natural larvicides against Aedes aegypti. These findings may have practical implications

for the development of eco-friendly and sustainable mosquito control strategies,

particularly in areas where chemical insecticides are not readily available or pose a risk to

human health and the environment.

The study by Singh et al, (2018) aimed to evaluate the larvicidal activity of

essential oils against three mosquito species, namely Aedes aegypti, Anopheles stephensi,

and Culex quinquefasciatus. The essential oils used in the study were obtained from six

different plant species, namely Citronella grass (Cymbopogon nardus), Lemon grass
(Cymbopogon citratus), Eucalyptus (Eucalyptus globulus), Peppermint (Mentha piperita),

Clove (Syzygium aromaticum), and Neem (Azadirachta indica). The results showed that

all the tested essential oils had varying levels of larvicidal activity against the three

mosquito species, with the most effective being Citronella grass and Lemon grass oils.

The Citronella grass oil showed the highest larvicidal activity against Aedes aegypti, with

an LC50 value of 39.06 ppm, followed by Anopheles stephensi and Culex

quinquefasciatus. Similarly, Lemon grass oil showed the highest larvicidal activity

against Anopheles stephensi, with an LC50 value of 53.67 ppm, followed by Aedes

aegypti and Culex quinquefasciatus. The study suggests that essential oils obtained from

Citronella grass and Lemon grass have potential for use as natural larvicides against

mosquito vectors. These natural larvicides could be used in combination with other

control measures to manage mosquito populations and reduce the incidence of mosquito-

borne diseases.

Barros et al, (2018) investigated the larvicidal activity of essential oil extracted

from Cymbopogon citratus (lemongrass) and its main constituents, citral and geraniol,

against Aedes aegypti larvae. The study found that lemongrass essential oil showed

strong larvicidal activity against the mosquito larvae, with an LC50 value of 34.1 ppm.

The two main constituents, citral and geraniol, also showed significant larvicidal activity,

with LC50 values of 26.9 and 42.7 ppm, respectively. The authors suggested that

lemongrass essential oil and its constituents have potential as natural larvicides for

controlling Aedes aegypti populations.

 The study by Pitasawat et al, (2007) investigated the larvicidal activity of Piper

nigrum (black pepper) seed extract and its active compound, piperine, against Aedes

aegypti larvae. The results showed that both the extract and piperine exhibited dose-

dependent larvicidal activity, with LC50 values of 73.77 ppm and 37.22 ppm,

respectively. These findings suggest that P. nigrum seed extract and piperine may have

potential as natural larvicides for controlling the population of A. aegypti mosquitoes.

Moreover, this study highlights the importance of exploring natural compounds derived

from plants as an alternative to synthetic insecticides, which can have negative impacts

on the environment and human health.

Maia et al, (2012) investigated the larvicidal activity of essential oils obtained

from 13 Brazilian plants against Aedes aegypti. The study found that the essential oils of

Piper aduncum, Ocimum gratissimum, and Cymbopogon citratus exhibited the highest

larvicidal activity, with LC50 values of 62.6, 89.1, and 92.6 mg/L, respectively. The

results showed that the larvicidal activity of the essential oils was dose-dependent and

varied depending on the plant species. The study also demonstrated that the essential oils

of P. aduncum, O. gratissimum, and C. citratus had a lower LC50 value than the positive

control, which was temephos, a synthetic larvicide commonly used for mosquito control.

The findings of this study suggest that essential oils obtained from some Brazilian plants

may be used as natural larvicides against Aedes aegypti. This could be particularly useful

in regions where there is a high prevalence of mosquito-borne diseases, such as dengue

fever, where synthetic larvicides may not be readily available or affordable.

 The study by Tsuzuki et al, (2011) investigated the larvicidal activity of essential

oils from Japanese cedar (Cryptomeria japonica) and lemon extract against two mosquito

species, Aedes aegypti and Aedes albopictus. The authors reported that the essential oils

from both plants had strong larvicidal effects against the mosquito larvae. The results

showed that the essential oil from Japanese cedar had LC50 values of 1.20 ppm and 1.34

ppm for Aedes aegypti and Aedes albopictus, respectively.

The essential oil from lemon extract had LC50 values of 9.84 ppm and 4.59 ppm

for Aedes aegypti and Aedes albopictus, respectively. The study concluded that the

essential oils from Japanese cedar and lemon extract have potential for use as natural

larvicides against Aedes aegypti and Aedes albopictus, which are both important vectors

of several diseases including dengue, Zika, and chikungunya.

The study conducted by Reegan and Gandhi (2012) evaluated the larvicidal

activity of citrus limonoids against Aedes aegypti mosquitoes. The researchers extracted

limonoids from citrus fruits and tested their efficacy against the larvae of Aedes aegypti.

The results of the study showed that the limonoids had potent larvicidal activity against

the mosquitoes. The LC50 and LC90 values for the limonoids were found to be 17.86

ppm and 47.83 ppm, respectively.

 The study also revealed that the larvicidal activity of the limonoids was dose-

dependent, and higher concentrations of the compounds were more effective in killing the

larvae. The researchers also conducted a probit analysis to determine the lethal

concentration required to kill 50% and 90% of the larvae.

Overall, the findings of this study suggest that citrus limonoids could be used as a

potential larvicide against Aedes aegypti mosquitoes. However, further studies are needed

to investigate the toxicity of these compounds to non-target organisms and their

environmental impact before their use in mosquito control programs.

 CHAPTER 3

RESEARCH METHODOLOGY

The research methods utilized in this study involved the collection of fresh lemon fruit

for extract preparation, the collection of mosquito larvae, the preparation of test solutions

at different concentrations, and the observation of larval mortality rates. These methods

ensured the accurate and controlled investigation of the potential effectiveness of lemon

extract as a natural larvicide for mosquito control.

RESEARCH DESIGN

This study's research design involves experimentation to determine the optimal

concentration of Lemon extract for maximum larvicidal activity against Aedes aegypti

mosquito larvae. The experiment was taken place in a laboratory setting and will use

basic equipment such as beakers, pipettes, and microscope slides.

The results of the experiment was collected through visual observations of the

mortality rates of mosquito larvae. The experiment was repeated three times to ensure

accuracy. The data collected was analyzed using simple statistical methods. This was

involve calculating the percentage of mortality for each concentration of extract and

comparing them to the control group.

 TIME TABLE

May 3, 2023 May 3, 2023 May 3, 2023 May 4, May 4, 2023 May 4, Ma 4, 2023
2023 2023

Literature Collection f Preparation of Starting of Completion Writing Submission

review and Lemons and experimental experiments of and of
finalization extraction of and control , data experiments review research
of research active groups, and collection, and data of report
design compound initiation of and analysis analysis, research
experiments interpretatio report
data n of results
collection,
and analysis

It is important to note that this time table is only a rough estimate and may vary

depending on the availability of resources and unforeseen challenges. Additionally, safety

precautions and ethical guidelines must be followed throughout the study.

 RESEARCH SETTING

The research setting for the larvicidal activity of lemons to the mortality rate of

mosquito larvae (Aedes aegypti) was conducted in a laboratory setting. The experiment

was carried out in a controlled environment to ensure the accuracy of the results. The

laboratory should have the necessary equipment and materials for the study, including a

mosquito rearing facility, containers for larval rearing, and tools for sample preparation

and analysis.

The study has been conducted at the College of Science and Mathematics,

Mindanao State of University - Iligan Institute of Technology (MSU-IIT). The laboratory

is well-equipped and the research will be conducted in accordance with the guidelines set

by the Institutional Animal Care and Use Committee (IACUC) to ensure the ethical and

humane treatment of the test subjects.

MATERIALS/EQUIPMENTS USED

There are several materials and equipment that can be used for feasible studies of

larvicidal potential in Philippine plants that can be found in household items or easily

accessible at home or a facility that you can reach. These include:

MATERIALS: EQUIPMENT:

Lemon fruit Kitchen Knife for cutting the lemon

Aedes aegypti mosquito larvae Squeezer or Juicer for extracting the juice from

the lemon

Water Measuring Cup for measuring the juice

Clear plastic containers for larval rearing Dropper for applying the juice to the water

Timer for monitoring exposure time

Diagram 1:

With these simple materials and equipment, the study can be conducted to

determine the effectiveness of lemon extract as a natural larvicide for controlling the

population of Aedes aegypti.

Note: In the event that a squeezer or juicer is not available, manual extraction of

lemon juice may be utilized as an alternative method.

DATA GATHERING PROCEDURE

Collection of Mosquito Larvae Preparations of Test

Experimental Setup
Lemon Extract Collection Solutions

Data Collection
Data Analysis

Diagram 1:

The data gathering procedure for the Larvicidal Activity of Lemon to the mortality

rate of mosquito larvaes involves several steps:

1. Collection of Lemon Extract: Fresh Lemon fruit will be collected, and the extract

will be prepared using the method.

2. Mosquito Collection: Mosquito larvae will be collected from stagnant water

bodies using a plastic scoop and transferred to a plastic container.

3. Preparation of Test Solutions: The extract will be diluted with clean lukewarm

water to obtain the desired concentration of 10 ppm, 20 ppm, and 30 ppm.

4. The mosquito larvae will be separated into three groups, each having 10 larvae,

and placed in a graduated cylinder for the experiment. The three groups will act as

the control group and will only get 20 cc of the specified liquid while being

treated with clean water. The specific concentrations of lemon extract—10 ppm,

20 ppm, and 30 ppm—will be placed on the respective cylinders at synchronized

timing. After that, the larvae will be watched for a predetermined period of time.
 5. Data Collection: The number of dead larvae in each group will be counted and

recorded every 5 minutes until the total time reached 1 hour.

6. Data Analysis: The percentage of mortality will be calculated using the formula:

Percentage of Mortality = (Number of dead larvae / Total number of larvae) x 100:

STATISTICAL TREATMENT/ TOOLS USED

1.Calculate the mean mortality rate for each concentration level of lemon extract.

2. Calculate the standard deviation for each concentration level of lemon extract.

3. Conduct a one-way analysis of variance (ANOVA) to determine if there is a

significant difference in the mortality rates between the control group and the groups

treated with different concentrations of lemon extract.

4. If the ANOVA test shows a significant difference, perform a post-hoc analysis

to determine which concentration levels show a significant difference in mortality rates

compared to the control group.

5. Determine the lethal concentration (LC50) of the lemon extract by plotting a

dose-response curve using the mortality data obtained from the different concentrations

of lemon extract. The LC50 value represents the concentration of lemon extract required

to kill 50% of the mosquito larvae. These statistical treatments can help provide insight

into the effectiveness of lemon extract as a natural larvicide for controlling the population

of Aedes aegypti mosquitoes.

 Chapter 4

RESULTS, DISCUSSIONS AND ANALYSIS

10 Larvaes
Time
Cup 1 (10 ppm) Cup 2 (20ppm) Cup 3 (30 ppm)

0 - 5 Minutes 0 0 0

5 -10 Minutes 0 0 0- 10%

10 - 15 Minutes 0 0 0 - 20%

15 - 20 Minutes 0 0 10% - 20%

20 - 25 Minutes 0 10% 20% - 30%

25 - 30 Minutes 0 10% 30%

30 - 35 Minutes 0 - 10% 10% 30% - 40%

35 - 40 Minutes 10% 10% 40% - 50%

40 - 45 Minutes 10% 20% 50% - 60%

45 - 50 Minutes 10% - 20% 20% - 30% 60% - 70%

50 - 55 Minutes 20% 30% 70% - 80%

55 Minutes - 1 Hr 20% 30% - 40% 80% - 100%

Table 1: Mortality rate of Mosquito Larvaes (Aedes Aegypti)

Based on the experiment's findings, lemon extract has demonstrated efficiency in

killing Aedes aegypti mosquito larvae within a specific concentration range of 10 ppm to

30 ppm. The mortality rate of the larvae increased with higher concentrations of lemon
extract. This suggests the potential of lemon extract as a natural larvicide for controlling

mosquito populations.

It is important to note that further research is needed to determine the

effectiveness and safety of using lemon extract as a larvicide under real-world conditions.

The experiment was conducted in a controlled environment, using 20 ml of water and 10

larvae of Aedes aegypti in each cup. It focused solely on this particular species of

mosquito, and the effectiveness of lemon extract may vary for other mosquito species and

environmental conditions.

Additionally, it is crucial to consider the potential ecological impacts of using

lemon extract as a larvicide. While it is a natural product, excessive use can have

negative effects on non-target species and disrupt the ecosystem. Therefore, careful

evaluation of the risks and benefits associated with using lemon extract as a natural

larvicide is necessary before implementing it in mosquito control programs.

In summary, lemon extract has shown promise as an effective larvicide for Aedes

aegypti mosquito control within the tested concentration range. However, further research

is required to validate its efficacy and safety in practical scenarios, considering potential

variations among mosquito species and environmental factors.

It can be concluded that lemon extract has been proven to be effective in

exterminating mosquito larvae within a certain concentration range.

The experiment demonstrated that the higher concentration of lemon extract

resulted in higher mortality rates of the mosquito larvae.

 Therefore, it can be inferred that lemon extract can potentially serve as a natural

larvicide for controlling mosquito populations.

However, it is important to note that further studies are necessary to determine the

effectiveness and safety of using lemon extract as a natural larvicide in real-world

conditions.

The experiment was conducted in a controlled environment and only tested one

species of mosquito, Aedes aegypti. Other species of mosquitoes may react differently to

lemon extract, and the effectiveness of lemon extract may vary depending on the

environmental conditions.

Furthermore, it is important to consider the potential ecological impacts of using

lemon extract as a larvicide. While it is a natural product, excessive use of lemon extract

may negatively affect non-target species and disrupt the ecosystem.

Therefore, careful consideration and evaluation of the potential risks and benefits

of using lemon extract as a natural larvicide are necessary before implementing it in

mosquito control programs.

Image 1: Preparing the Equipments

Image 2: Evening the Larvae

 Image 3: Evening the Lemon extract

Chapter 5

SUMMARY OF THE FINDINGS

The aim of this discussion was to explore the potential of lemon extract as a

natural larvicide for controlling mosquito populations. The experiment conducted in this

study involved the collection of mosquito larvae, the preparation of test solutions of

lemon extract at varying concentrations, and the monitoring of mortality rates of the

mosquito larvae at different concentrations. The results demonstrated that lemon extract

was effective in exterminating mosquito larvae, specifically Aedes aegypti, within a

concentration range of 10 ppm to 30 ppm. The higher the concentration of lemon extract,

the higher the mortality rate of the mosquito larvae. Therefore, lemon extract may serve

as a potential natural larvicide for controlling mosquito populations. However, it is

essential to note that further studies are necessary to determine the effectiveness and

safety of using lemon extract as a natural larvicide in real-world conditions.

The experiment conducted in this study was limited to a controlled environment

and only tested one species of mosquito, Aedes aegypti. Other species of mosquitoes may

react differently to lemon extract, and the effectiveness of lemon extract may vary

depending on the environmental conditions. Furthermore, it is crucial to consider the

potential ecological impacts of using lemon extract as a larvicide. While it is a natural

product, excessive use of lemon extract may negatively affect non-target species and

disrupt the ecosystem.

 Therefore, careful consideration and evaluation of the potential risks and benefits

of using lemon extract as a natural larvicide are necessary before implementing it in

mosquito control programs. In conclusion, the study demonstrates the potential of lemon

extract as a natural larvicide for controlling mosquito populations. However, further

research is necessary to validate the findings of this study and to determine the

effectiveness and safety of using lemon extract as a larvicide in real-world conditions.

The use of natural products like lemon extract as a larvicide may provide an eco-friendly

and sustainable approach to mosquito control, but it must be used with caution to

minimize potential ecological impacts.

RECOMMENDATIONS

It is advised that additional research be done in order to validate the results of this

study and determine the efficacy and safety of using lemon extract as a natural larvicide

for managing mosquito populations. The study showed that, within a particular

concentration range, lemon extract is effective in killing Aedes aegypti mosquito larvae,

but more studies are required to determine how effective it is against other mosquito

species and in various environmental settings.

It is also advised to carefully weigh the advantages and disadvantages of utilizing

lemon extract as a larvicide before including it into mosquito control strategies. This

involves taking into account possible effects on species that are not the focus and the

ecosystem as a whole.It is also recommended that more research look at the best and

most productive ways to use lemon extract as a larvicide. Examining the best

concentration and frequency of application, as well as the possibility of combining lemon

extract with other organic or synthetic pesticides for greater efficacy, are a few examples

of what this can entail.

In conclusion, more research is required to confirm the usefulness and safety of

lemon extract as a natural larvicide for decreasing mosquito populations, even though the

study's results are encouraging. Before using lemon extract as a larvicide in mosquito

control programs, it is essential to carefully weigh the potential advantages and risks.
 REFERENCE

Barros, E. M., da Silva, D. T., Fiuza, T. S., & da Silva, C. R. (2018).

Larvicidal activity of Cymbopogon citratus essential oil and its constituents
against Aedes aegypti. Journal of vector ecology: journal of the Society for Vector
Ecology, 43(1), 118-124.
Barnard, D. R., Xue, R. D., & Rotstein, H. C. (2018). Lethal and sublethal
effects of a larvicidal botanical extract, pyrethrum, on Aedes aegypti and Aedes
albopictus. Journal of vector ecology: journal of the Society for Vector Ecology,
43(1), 33-41.
Lira, C. S., Melo, A. L. T., Costa-Junior, L. M., & Navarro, D. M. A. F.
(2018). Larvicidal activity of lemon grass and lemon extract against Aedes
aegypti larvae. Revista de saúde pública, 52, 73.
Singh, R. K., Dhiman, R. C., Mittal, P. K., & Dua, V. K. (2018).
Larvicidal activity of essential oils against Aedes aegypti (Linn.), Anopheles
stephensi (Liston.) and Culex quinquefasciatus (Say.). Asian Pacific Journal of
Tropical Biomedicine, 8(4), 200-204.
Choochote, W., Tuetun, B., Kanjanapothi, D., Rattanachanpichai, E.,
Chaithong, U., Chaiwong, P., & Jitpakdi, A. (2004). Potential of crude seed
extract of celery, Apium graveolens L., against the mosquito Aedes aegypti (L.)
(Diptera: Culicidae). Journal of Vector Ecology, 29(2), 340-346.
Pitasawat, B., Champakaew, D., & Choochote, W. (2007). Larvicidal
activity of Piper nigrum seed extract and piperine against larvae of the mosquito
Aedes aegypti. Fitoterapia, 78(2), 133-136
Maia, M. F., Bezerra-Silva, P. C., & Nunes, R. S. (2012). Larvicidal
activity of essential oils from Brazilian plants against Aedes aegypti L. (Diptera:
Culicidae). Parasitology Research, 110(6), 2099-2104.
Tsuzuki, S., Akiyama, K., Matsumoto, K., Nishimura, H., & Takahashi,
M. (2011). Larvicidal activity of essential oils from Japanese cedar (Cryptomeria
japonica) and lemon extract (Eucalyptus citriodora) against Aedes aegypti and
Aedes albopictus (Diptera: Culicidae). Journal of Medical Entomology, 48(2),
301-306.
Reegan, A. D., & Gandhi, M. R. (2012). Larvicidal activity of citrus
limonoids against Aedes aegypti (Diptera: Culicidae). Journal of Entomology and
Zoology Studies, 1(3), 26-29.
 CURRICULUM VITAE

ADRIAN JERICHO L. OLANDESCA

Pala-o, Iligan City, Lanao Del Norte
Aj.olandescaofficial@gmail.com
09752267594

Graduation: June, 2, 2023

Life quote: “Those who joyfully leave everything in

God's Hand will eventually see God's Hand in everything.”

EDUCATION

Science, Technology, Engineering and Mathematics

St. Paul’s Institute of Technology

2021 - 2023

HONORS/AWARDS

2016 - 2017 Class Valedictorian

2022 - 2023 Achievement Achiever

ON CAMPUS INVOLVEMENT/ORGANIZATIONS

2022 - 2023 Student Supreme Government Officer, Auditor

 CURRICULUM VITAE

SARAH F. ABAD
Zone 11 Brgy. Bagong Silang,
Iligan City, Lanao del Norte
Sarahabad07@gmail.com
09265531176

Graduation: June, 2, 2023

Life quote: “If you have no doubt in what you are about

to do, you are not pushing yourself hard enough.”

EDUCATION

Science, Technology, Engineering and Mathematics

St. Paul’s Institute of Technology

2021 - 2023

HONOR/AWARD

2022 - 2023 Achievement Achiever

 CURRICULUM VITAE

ANDREI JULLAN D. INUTAN

Lambaguhon, San roque,
Iligan City, Lanao del Norte
a.inutan@gmail.com
09957192164

Graduation: June, 2, 2023

Life quote: “Many tries is many chances and only once is success.”

EDUCATION

Science, Technology, Engineering and Mathematics

St. Paul’s Institute of Technology

2021 - 2023
 CURRICULUM VITAE

LANCE JAIRUZ T. BICOY

Purok 17, Tibanga,
Iligan City, Lanao Del Norte
lancebicoy223@gmail.com
09815854115

Graduation: June, 2, 2023

Life quote: “Most of the important things in the world

have been accomplished by people who have kept on

trying when there seemed to be no hope at all.”

EDUCATION

Science, Technology, Engineering and Mathematics

St. Paul’s Institute of Technology

2021 - 2023
 CURRICULUM VITAE

MAE FELIZE Y. ORBE

Badelles st. Cor. Luna, Iligan City 9200
Lanao del Norte, Philippines
deorbiz@gmail.com
09364245619

Graduation: June, 2, 2023

Life quote: “Life is too short to waste any amount of time on wondering what other
people think about you. In the first place, if they had better things going on in their lives,
they wouldn't have the time to sit around and talk about you.

What's important to me is not others' opinions of me,

but what's important to me is my opinion of myself”

EDUCATION

Science, Technology, Engineering and Mathematics

St. Paul’s Institute of Technology

2021 – 2023
 CURRICULUM VITAE

ANGEL MAY TABUCO

Zone1- Purok Sudlonon Brgy. San Miguel,
Tibanga, Iligan City.
Tabucoangelmay@gmail.com
09262949970

Graduation: June, 2, 2023

Life quote: “Too often we underestimate the power of a touch, a smile, a kind word, a
listening ear, an honest compliment, or the smallest act of caring, all of which have the
potential to turn a life around.”

EDUCATION

General Academic Strand

St. Paul’s Institute of Technology

2021 - 2023