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SACE-Larvicide research file
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The Use of Sampaguita (Jasminum sambac) Leaf, Aloe vera (Aloe barbadensis
miller) Leaf and Calamansi (Citrus microcarpa) Fruit Extract as an
Alternative Larvicide against Mosquito Larvae (Culicidae)
A Research Paper Presented to the faculty of
The Senior high, Western Colleges Inc.
In Partial Fulfillment of the
Requirements in Research in Daily Life 1
Presented by:
Chrysler M. Vergara
France William V. Nazareno
Vince Erielle L. Sarmiento
Noel D. Parungao
Marc Francis C. Tambal
2019

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ABSTRACT
Incidents cause by Mosquito-borne diseases are one of the major concerns in
100 countries across the World. The Researchers conducted the study about the use
of “Sampaguita leaves, Aloe vera and Calamansi fruit Extract Larvicide or SACE
Larvicide” against mosquito larvae. They use Completely Randomized Design (CRD)
in their research process, the subjects are randomly assigned to treatments. The
results of the Mortality of the SACE Larvicide shows that 30ml volume of all
concentration (10%, 50% and 100%) of the researcher’s product acquired a 100%
mortality in 31-40 minutes of exposure. Moreover, the Statistical treatment used by the
researchers shows that 30ml volume of 10%, 50% and 100% Concentration of SACE
Larvicide and X-Press Methrin in terms of 31-40 minutes of exposure are equal or
simply because the two different larvicide are both obtained a 100% mortality.
Therefore, the mortality results, statistical measurements and observation to the
effectiveness of SACE Larvicide is clearly proven having a larvicidal activity or
poisoning effect against mosquito larvae in 31-40 minutes of exposure.

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APPROVAL SHEET
In partial fulfillment of the requirements for the Senior High School Program of Chrysler
M. Vergara, France William Nazareno, Noel Parungao, Marc Francis Tambal and
Vince Erielle Sarmiento who is hereby recommended for oral examination has
submitted this Research entitled "The Use of Sampaguita (Jasminum sambac) leaf,
Aloe vera (Aloe barbadensis miller) and Calamansi (Citrus microcarpa) fruit
Extract as an Alternative Larvicide against Mosquito Larvae (Culicidae)".
ENGR. BIENVENIDO B. GARCIA
Adviser
Approved in partial fulfillment of the requirements for the Senior High School Program
by the Oral Examination Committee with a rating of_______________.
RONNIE E. PASIGUI, Ph. D.
Chairman
DANILO H. CABALU, Ph. D. ENGR. BIENVENIDO B. GARCIA
Member Member
Approved in partial fulfillment of the requirements for the Senior High School Program.
DARNIELL C. BALBUENA, MAEd
School Principal

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ACKNOWLEDGEMENT
First and foremost, praises and thanks to the God, the Almighty, for His showers
of blessings throughout our research work to complete the research successfully.
We would like to express our deep and sincere gratitude to our research
supervisor, Engr. Bienvenido Garcia, for giving us the opportunity to do research and
providing invaluable guidance throughout this research. His dynamism, vision,
sincerity and motivation have deeply inspired us. He has taught us the methodology to
carry out the research and to present the research works as clearly as possible. It was
a great privilege and honor to work and study under his guidance. We are extremely
grateful for what he has offered us. We would also like to thank him for his friendship,
empathy, and great sense of humor.
We are extremely grateful to our parents for their love, prayers, caring and
sacrifices for educating and preparing us for our future. Also, we express our thanks
to our siblings for their support and valuable prayers. Our Special thanks goes to the
parents of our groupmates Mrs. Emily Manguiat, Mrs. Neri Nazareno and Mrs.
Glennies Villaluz providing us a place to conduct our study and financial support. We
would like to say thanks to our fellow classmates their constant encouragement.

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We are extending our thanks to our teacher from Western Colleges
Incorporated Ms. Mariztel Escalante and Ms. Marianne Galope for their support during
our research work.
Finally, our thanks go to all the people who have supported us to complete the
research work directly or indirectly.
Sincerely,
Chrysler M. Vergara
France William V. Nazareno
Vince Erielle L. Sarmiento
Noel D. Parungao
Marc Francis C. Tambal

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DEDICATION
This study is wholeheartedly dedicated to our beloved parents, who have been
the source of our inspiration and gave us strength when we thought of giving up to the
challenges that we encounter in conducting our research, they are the one who
continually provide the moral, spiritual, emotional, and financial support for us.
To our brothers, sisters, relatives, mentor, friends and classmates who gave
their words of advice and encouragement to finish and improve our study.
And lastly, we dedicated this Research study to the Almighty God, thank you for
the guidance, strength, power of mind, protection and skills and for giving us a healthy
life. We cannot do all these things without the guidance You gave to us.
Thank you for all the support that we received to all the important persons to
our life. God bless you all.

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TABLE OF CONTENTS
Page
Title Page i
Abstract ii
Approval Sheet iii
Acknowledgement iv
Dedication vi
Table of Contents vii
Chapter 1: The Problem and Its Background
1.1 Introduction
1.2 Background of the Study 2
1.3 Theoretical/Conceptual Framework 5
1.4 Statement of the Problem 6
1.5 Hypothesis 7
1.6 Significance of the Study 8
1.7 Scope and limitation of the Study 9

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1.8 Definition of Terms 11
Chapter 2: Review of Related Literature and Study
2.1 Review of Related Literature 12
2.2 Review of Related Study 19
2.3 Synthesis 24
Chapter 3: Methodology
3.1 Research Design 25
3.2 Research Locale 26
3.3 Research Process 26
3.4 Data Gathering 31
3.5 Statistical Treatment of Data Procedure 35
Chapter 4: Results and Discussion
4.1 Introduction 33
4.2 Discussion
4.2.1 Phytochemical Constituents 33
4.2.2. Physical Properties of SACE Larvicide 35

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4.2.3. Concentration of Solution 36
4.2.4. Tests, Observations and Images
4.2.4.1. Actual Image of the Experiment 38
4.2.4.2. Mortality rate 46
4.2.5. Analysis of Variance (ANOVA) 48
4.3. Results and Discussion 58
Chapter 5: Findings, Conclusion and Recommendations
5.1 Summary of Findings 63
5.2 Conclusion 67
5.3 Recommendations 68
Bibliography
Curriculum Vitae

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CHAPTER 1
THE PROBLEM AND ITS BACKGROUND
1.1 INTRODUCTION
Plants have been known as a major tool in preventing mosquitoes, it is the most
efficient approach to the wide spread of mosquito diseases. To know more about the
floral diversity and to set foot in the field of applying safer insecticides composed of
botanical mixture of chemical compounds are highly recommended because it could
be an alternative solution to synthetic products. This strategy is environmental friendly,
affordable and safe to use, because it is naturally processed. Otherwise, the utilization
of synthetic products in mosquito control program has been restricted, it is because of
concern for the environmental sustainability, harmful effect on human health and other
non-target populations.
Larvicides against mosquito larvae was not typically used by consumers because
of mosquito repellent and mosquito insecticide. In this study, the researcher was
curious on the usage of larvicide so, they assume what if they use larvicide and
improve the product to increase the usage of larvicide like repellents and insecticide.
They also thought that it would be easy if the main target is the larvae of mosquito
because it was not completely formed into a mosquito, and it has soft and sensitive
body so, it is easy to be killed.

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1.2 BACKGROUND OF THE STUDY
Mosquito-borne diseases have been around for many years. These diseases are
dominant in more than 100 countries across the world, infecting over 390 million people
every year globally (WHO, April 2019). It clearly seen that mosquitoes are very
destructive to human health and up until now there is no accurate solution regarding to
this problem. Infected mosquitoes transmit diseases to an individual through its bite, it
carries pathogen which targets the immune system of a person, and it serve as a vector
for most dangerous diseases such as malaria, yellow fever, chikungunya fever,
encephalitis, etc., and most of them came from all tropical and subtropical countries and
other parts of the world. In the year 2018 DOH showed a 21 percent increase in the
number of potentially deadly infection recorded in various government hospitals
nationwide compared to the same period last year. From Jan. 1 to Oct. 6, DOH recorded
a total of 138,444 dengue cases, up from the 114,878 cases during the same period in
2017. On the other hand, the findings from 2016-2018 have shown that mosquito
diseases are consistently increasing and it’s about to cause more suffering to the people
in the community.

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The researcher conducted a study about the use of plant extract and powdered
plant parts that is usually used as insecticide because it contains natural constituents
which has a pathogenic effect to insects and their larvae. The raw material to be used
in this study are: Sampaguita leaves, Aloe vera, and Calamansi which is commonly
seen in the backyard or garden of houses. The researcher uses these ingredients
because it was natural and has a useful effect. These raw materials have the
phytochemicals which has constituents that has a larvicidal activity that will help to kill
mosquito larvae.
The first raw material will be used in this study is the Sampaguita (Jasminum
Sambac) Leaf. It is a spreading or sprawling of shrub, usually less than 2m in height.
Leaves are glossy, ovate or rounded and 6-12 cm long, with short stalks, pointed or
blunt tip and pointed rounded bas. Flowers are white, very scented and borne
individually or in 3s in axillary and terminal inflorence. Calyx teeth are 8-10, very slim,
5-8 mm long, Corolla tube is slim and 1-1.5 cm long, the limb is usually double and
1.5-2 cm in diameter. Stamens, 2, included, ovary, 2-celled, it belongs to oleaceae
family. It blooms every May or every summer and almost continuously in warm
ambiance. Sampaguita is abundant in San Pedro Laguna and in Bulacan.

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The second raw material to be used in the study is the Aloe vera (Aloe
barbadensis miller) Leaf. It is part of the liliaceae family (Asphodelaceae), and is a
pea-green color plant, succulent, xerowphytic, perennial, arborescent or shrubby.
According to Amar Surjushe, et. al, (2008) of Indian Journal of Dermatology, Aloe vera
has triangular, fleshy leaves with edges that are serrated, yellow colored tubular
flowers and fruits that has numerous seeds. Every leaf is composed of three layers: 1)
A clear gel inside that contains 99% water and the remaining is made of
glucomannans, amino acids, lipids, sterols and vitamins. 2) The bitter yellow sap in the
middle layer that contains anthraquinones and glycosides. 3) The rind which is the
outer layer of 15-20 cells has a protective function and synthesizes carbohydrates and
proteins. There are vascular bundles inside the rind that is responsible for
transportation of substances such as xylem (water) and phloem (starch).
And the third raw material to be used is the Calamansi (Citrus microcarpa) Fruit.
This is a smooth and slightly plant, it grows to a height of 3 to 5 meters. The leaflets
are elliptic to oblong elliptic, 4 to 8 centimeters long. Flowers are axillary, solitary, rarely
in pairs, white, and short-stalked. Fruit is yellow when ripe, nearly spherical, 2 to 3.5
centimeters diameter, 6- to 7-celled, and thin-skinned. The skin or peel is green to
yellowish green or yellow, loosely adhering to the flesh. The flesh contains a few light
orange seeds.

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1.3 THEORETICAL/CONCEPTUAL FRAMEWORK
Collection of Dry the

HOT
fresh sampaguita
Wash the
sampaguita leaves
INFUSION collected through
leaves, Aloe
METHOD ingredients sunlight
vera leaves and
Calamansi Fruit (1-2 days)
Extract the juice Dry the Cut the aloe

Strain the
of calamansi strained vera leaves
grinded aloe
fruit and place it leaf skin of into small
vera to get
in a bottle and the aloe pieces and
the extract
put it in a vera in the grind it with
then placed
refrigerator extract the use of
it in a bottle
(2-3hrs) (1 day) blender
Dry the Separately grind Separately place

Boil a
calamansi the dried the pulverized
water in
peel sampaguita sampaguita leaves,
a pot
through leaves, peel of peel of aloe vera
within
sunlight aloe vera leaves leaves and peel of
5-6
(1-2 days) and peel of calamansi fruit in a
mins
calamansi fruit cloth
Cover the
Pour the Ethyl Air-dry the pot and Put the three
Alcohol and the steamed simmer it cloths
extracted aloe vera sampaguita, aloe gently in containing the
to the bottle with vera and 10-20 ingredients in a
calamansi extract calamansi powder mins steam plate on
then cover (1 day) (1 day) the pot
After 1 day put the Let the

powder After 1-2 days. The
powder of sampaguita Larvicide composed of
leaves, peel of aloe dissolve in the
solvent and Sampaguita leaf, Aloe
vera leaves and peel
of calamansi fruit in after 1day vera leaf and Calamansi
the mixture of ethyl strain it to fruit Extract is made or
alcohol, aloe vera and remove SACE Larvicide
Calamansi extract. undissolved
powder

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1.4 STATEMENT OF THE PROBLEM
Generally, this study aims to determine if the larvicide made up of Sampaguita
leaves, Aloe vera leaves and Calamansi fruit Extract (SACE Larvicide) can be used to
kill mosquito larvae.
Specifically, it aims:
1. to determine the Level of effectiveness of SACE Larvicide in terms of:
1.1 Mortality rate in 10%, 50% and 100% concentration with different
amount of the product
1.2 Volume of the product poured (5 ml, 15 ml and 30 ml)
1.3 Fastest time that obtained a higher mortality (10-30 mins, 21-30 mins
and 31-40 mins)

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3. to determine the significant difference between
3.1 5ml volume of 10%, 50% and 100% concentration of SACE Larvicide
and the commercial X-Press Methrin Larvicide.
1.5 HYPOTHESIS
1.5.1 NULL HYPOTHESIS (Ho)
- There is no significant difference between 30 ml volume of 10%, 50% and 100%
concentration of SACE Larvicide and the commercial X-Press Methrin Larvicide.

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1.6 SIGNIFICANCE OF THE STUDY
The result of this study may benefit the following:
Community. This study will give an alternative product which is affordable,
accessible and effective protection for people. And it is much better to use SACE
Larvicide because if the mosquito larva is the main target to be killed, it is appropriate
to do because the mosquito larva is the weakest point of the mosquito life cycle.
Future Researchers. This study will provide reliable and valuable information, facts
and evidences that might be used as a reference to their own research study in the
future.
Nature. This study can also be helpful to the nature, because it is free and fresh.
Since the sampaguita leaves, Aloe vera leaves and Calamansi fruit extract contains
non-toxify constituents and prevents water from contamination unlike other
insecticides or larvicides.
Farmers. The result of the study may help the farmers to give additional profit selling
their crops.
Government. The result of the study may also benefit the government through
exporting the product international when it was approved.

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1.7 SCOPE AND LIMITATION
The scope of this study involves the time and money spent, the tools/material and
techniques used, and the process needed in the experiment to be completed. The
entire study spent 3 months and 2 weeks to achieve the final output of the research
study. The estimated range of the experiment from the extraction of Sampaguita and
Aloe vera leaves, and Calamansi Fruit to the final output of the product ranges from
a minimum time of 30 minutes to 2 hours depending on the amount needed to be
extracted. The general budget of this experiment reaches from the minimum amount
of 300 pesos for the tools and raw materials and with the maximum amount of 3,000
pesos depends on the expenses of materials.
The raw material will be used are sampaguita leaves (Jasminum sambac), Aloe
vera (Aloe barbadensis miller) and Calamansi fruit (Citrus microcarpa). And the tools
to be used are: 48 pcs. Tupperware with (L x W x H= 155 cm x 105 cm x 40 cm), 3
pcs. 500 ml bottles, Ethyl Alcohol, measuring glass, electric blender, knife, strainer,
container and digital weighing scale. Techniques involves following the necessary
process to make sure that the study will be performed well and to achieve the
expected outcome of the alternative larvicide.

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This experiment needs an intense observation to the two sets of trials, with three
treatments in in every concentration of SACE Larvicide and three treatments in SET
B also. At least 10 mosquito larvae will be placed in each treatment so, all in all the
total of larvae will be used is 480 pcs. This study is limited only to the extraction of
sampaguita leaves, aloe vera and calamansi as an alternative larvicide for mosquito
larvae and its physical properties such as Color, Odor, Density, and Texture. And its
limitations are those events, things, etc. that are outside of the scope of this study.

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1.8 DEFINITION OF TERMS
1. Sampaguita - Is the common for 'Jasminum sambac', a flower in the same
family as the common jasmine (Jasminum grandiflores). Sampaguita is one of the
raw material or the main ingredient of this study.
2. Aloe vera- is a succulent plant species of the genus Aloe. An evergreen
perennial, it originates from the Arabian Peninsula but grows wild in tropical
climates around the world and is cultivated for agricultural and medicinal uses. The
species is also used for decorative purposes and grows successfully indoors as a
potted plant. This is one of the raw material or the main ingredient of the study.
3. Calamansi- (Citrus microcarpa, × Citrofortunella microcarpa or × Citrofortunella
mitis), also known as calamansi, is an economically-
important citrus hybrid predominantly cultivated in the Philippines. It is native to
the Philippines and surrounding areas in southern China, Taiwan, Borneo,
and Sulawesi. Calamondin is ubiquitous in traditional Filipino cuisine. It is used in
various condiments, beverages, dishes, marinades, and preserves. This is one of
the raw material or the main ingredient of this study.
4. Ethanol- also known as Ethyl alcohol, a colorless volatile flammable liquid
C2H5OH that is the intoxicating agent in liquors and is also used as a solvent. In
this study, the Ethanol is the solvent of the study.

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5. Larvae - The immature, wingless, and usually wormlike feeding form of those
insects that undergo three stages of metamorphosis. In this study the mosquito
larva will be the population or the specimen of the experiment.
6. Larvicide – is an agent used in killing larvae. In this study, the researcher used
the term larvicide as the product, in which the ingredient of a normal larvicide will
be replaced by a natural raw material such as sampaguita, aloe vera and
calamansi.
7. Phytochemicals- these are the constituents present in Sampaguita leaves,
Aloe vera leaves and Calamansi fruit which has a poisoning effect or larvicidal
activity.
8. Extraction- Extraction in chemistry is a separation process consisting in the
separation of a substance from a matrix. It includes Liquid-liquid extraction, and
Solid phase extraction. The distribution of a solute between two phases is an
equilibrium condition described by partition theory. This is the process will be used
in this study.
9. Dengue- is an epidemic disease that occurs on warmer climates cause by a
virus transmitted by the mosquito. In the Philippines, it is the most common disease
that can get from mosquito.

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CHAPTER 2
REVIEW OF RELATED LITERATURE AND STUDY
This chapter includes the review of related literature and studies that help the
researcher to have a glance on the problem that need intense study.
2.1 RELATED LITERATURE
2.1.1 FOREIGN LITERATURE
According to Anupam Ghosh et al., (2012) of Indian Journal of Medical
Research, that Several groups of phytochemicals such as alkaloids, steroids,
terpenoids, essential oils and phenolics from different plants have been
reported previously for their insecticidal activities.
According to R.F.Rauffaf, Alkaloids, a specific class chemical
compounds found in plants and, occasionally, animals, are notable chiefly for
their physiological activity; many have long histories as poisons, narcotics,
hallucinogens, and medicinal agents. Alkaloids are derived from amino acids
and include an enormous number of bitter, nitrogenous compounds. Generally,
alkaloids are basic, or alkaline, substances.

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According to Kishore Mishra BB et al., (2011), reviewed the effectiveness
of phytochemicals against mosquito larvicidal potentiality of several plant
derived secondary materials, such as, alkanes, alkenes, alkynes, and simple
aromatics, lactones, essential oils and fatty acids, terpenes, alkaloids, steroids,
isoflavanoids, pterocarpans and lignins.
According to Siti Suhaila Harith et al., (2017) of Malaysian Journal of
Analytical Sciences, Phytochemicals or the secondary metabolites in plant
serve as a mean of defense mechanism of the plants to avoid from herbivore
predators and other environmental factors.
According to Anupam Ghosh et al., (2012) of Indian Journal of Medical
Research, mosquitoes are known to transmit more diseases than any other
group of arthropods in the world. World Health Organization (WHO) has
declared that the mosquito is the “number one public enemy”. Diseases coming
from mosquitoes are dominant in more than 100 countries across the world,
infecting over 700,000,000 people every year globally. They serve as a vector
for most dangerous diseases like zika virus, malaria, yellow fever, chikungunya
fever, encephalitis, West Nile virus infection, etc., and most of them came from
all tropical and subtropical countries and other parts of the world.

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According to the members of three WHO regions regularly report the
annual number of cases. The number of cases reported increased from 2.2
million in 2010 to 3.2 million in 2015. Although the full global burden of the
disease is uncertain, the initiation of activities to record all dengue cases partly
explains the sharp increase in the number of cases reported in recent years.
According to Conklin (1989), “How a mosquito develops. Oil in the water
prevents them from breathing through air tubes—one in the larva, two in the
pupa.”
According to World Book M.13 (2013) the mosquito life consists of four
stages: (1) Egg, (2) Larva, (3) Pupa, and (4) adult. At each stage, the mosquito’s
appearance changes completely and the insect lives a different kind of life. In
warm climates, some species develop from newly hatched eggs and to adults
in only a week. In the cold North mosquito eggs may contain dormant form
autumn until late spring. They hatched in May or June and may take a month or
more, to mature that’s why we conducted this study to kill mosquito larvae so
that it cannot spread and cause diseases such as Malaria, Dengue,
Chikungunya and other viruses.

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According to the Illustrated Encyclopedia of Science and of Insect life II
(1994), eggs of the house mosquito (right) are laid on the water’s surface in a
raft. The eggs hatch the following day, the larvae begin the first of four larval
stages, in which they will molt three times. The larval period lasts about 10 days
in water temperature of 250 Celsius or warmer. At the end of the fourth larval
stage, the shell cracks open and the pupa emerges. The pupa continues to live
in water. It eats nothing while the mouth changes from the filter-feeding structure
of the larva to the sucking structure of the adult. After three or four days, the
adult emerges (far right) and flies away.
2.1.2 LOCAL LITERATURE
According to Sahagun (2013), jasminum sambac is a member of the olive
family (Oleaceae). Jasmine oil extracted from sampaguita has a wide range of
medicinal applications and can be used in perfumery, soap, flavoring and the
cosmetic industry. Phytochemical studies show that the compounds
responsible for its unique scentare benzyl acetate, indole, methyl anthranilate,
jasmone, and methyl jasmonate.

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According to Dietmar J. Rummel (2005), Jasminum sambac is a
spreading or sprawling shrub, usually less than 2 m in height. Leaves are
glossy, ovate or rounded and 6-12 cm long, with short stalks, pointed or blunt
tip and pointed or rounded base. Flowers are white, very fragrant and borne
singly or in 3s in axillary or terminal inflorescence.
According to Department of Health Dengue Surveillance (2016) A total
of 101,401 suspect dengue cases were reported nationwide from January 1 to
August 20, 2016. This is 16% higher compared to the same period last year
(87,411). Most of the cases were from the following regions: Region VI (11.1%),
Region IV-A (10.7 %), Region VII (9.8%), Region X (8.8%) and Region XII
(8.6%). Ages of cases ranged from less than 1 month to 100 years old (median
= 13 years). Majority of cases were male (52.4%). Most (38.8%) of the cases
belonged to the 5 to 14 years age group.
According to Department of Health Dengue Surveillance (2017), a total
of 131,827 dengue cases were reported nationwide from January 1 to
December 2, 2017. This is 36.9% lower compared to the same period last year
(208,805). Most of the cases were from the following regions: NCR (17.3%),
Region Ill (15.3%), Region IVA (13.1%), Region Vll (9.6%) and Region VI
(6.8%). Most of the cases were from following provinces: Metro Manila
(17.29%), Cebu (7.16%), Negros Occidental (4.19%), Pampanga (4.10%),

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Bulacan (4.00%) and Cavite (3.67%). Ages of cases ranged from less than 1
month to 98 years old (median = 12 years). Majority of cases were male
(53.5%). Most (22.2%) of the cases belonged to the 5 - 9 years age group.
According to Department of Health Dengue Surveillance (2018) showed
a 21 percent increase in the number of potentially deadly infection recorded in
various government hospitals nationwide compared to the same period last
year. From Jan. 1 to Oct. 6, DOH recorded a total of 138,444 dengue cases,
up from the 114,878 cases during the same period in 2017. The DOH said the
number of deaths due to dengue also rose from 581 in 2017 to 708 this year.?
Central Luzon posted the biggest number of cases with 22,077, followed by the
National Capital Region (NCR) with 18,831 and Calabarzon with 16,177. Other
regions with high cases of dengue include Ilocos region with 11,109 and
Western Visayas with 11,036. While Central Luzon had the most number of
cases, NCR recorded the highest number of deaths with 106 followed by
Calabarzon with 98 and Western Visayas with 69.? Central Luzon recorded 61
deaths and Northern Mindanao, 53. A high 22 percent or 29,874 of the total
cases belong to the 10 to 14 years age group.? More than half or 73,017 of the
cases were males while females accounted for 65,427 or 47 percent of the
dengue cases nationwide.

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2.2 RELATED STUDIES
2.2.1 FOREIGN STUDIES
According to Joseph K Musau et al, (2016) of International Journal of
Mosquito Research, the larvicidal activity was dose dependent and at the
highest concentration of 1mg/ml, all extracts exhibited 100% mortality. This
could be due to presence of secondary metabolites especially alkaloids and
flavonoids. Alkaloids have long history of use as insecticides. They cause
symptoms of poisoning to the insects similar to organophosphate and
carbamate insecticides. The flavonoid rotenone has insecticidal properties
acting as a mitochondrial poison, which blocks the electron transport chain and
prevents energy production. Thus, the presence of alkaloids and flavonoids in
all the plants studied could have contributed to their larvicidal activity. Other
secondary metabolites which have been previously studied and found to have
larvicidal activity include saponins and tannins whose presence in the study
plants could have contributed to larvicidal activity.
According to Phanukit Kunhachan et al, (2012), sampaguita is generally
considered as Safe (GRAS) in the US. The parts that can be used in
Sampaguita are the flower, leaves and roots. Phytochemical analysis of the
ethanolic extract displayed that antioxidants, coumarins, cardiac glycosides,

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essential oils, ﬂavonoids (Rutin), phenolics (Catechol), saponins (Saponin), and
steroids (Steroid) were investigated.
According to Sanjit Kumar Kar et al., (2018), the Aloe vera leaf gel
contains about 98% water 6. The total solid content of Aloe vera gel is 0.66%
and soluble solids are 0.56% with some seasonal fluctuation. On dry matter
basis aloe gel consists of polysaccharides (53%), sugars (17%), minerals
(16%), proteins (7%), lipids (5%) and phenolic compounds (2%) (Fig. 2). Aloe
vera contains 200 potentially active constituents: vitamins, enzymes, minerals,
sugars, lignin, saponins, salicylic acids and amino acids, which are responsible
for the multifunctional activity of Aloe.
According to Amar Surjushe, et. al, (2008) of Indian Journal of
Dermatology, Aloe vera plant has 75 potentially functioning constituents: amino
acids, salicylic. saponins, lignin, sugars, minerals, enzymes, and vitamins.
According to Jayapal Subramaniam et. al, 2012 of Saudi Journal of
Biological Sciences, their study about the “Mosquito larvidal activity of Aloe vera
(Famili: Liliaceae) Leaf Extract and Bacillus sphaericus, gainst Chikungunya
vector, Aedes aegypti”, the Aloe vera is a perennial plant to the family of
Liliaceae, which has about 360 species (Kein and Penneys, 1988). The Aloe
vera is one of the few medicinal plants that can maintain its popularity for a long
period of time. This plant has stiff, gray-green lance-shaped leaves containing

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clear gel in a central mucilaginous pulp and has hypoglycemic (Rajasekaran et
al., 2004)
According to United States FDA and the Center for Food Safety and
Applied Nutrition, (2018), the pH level of Aloe vera (Aloe barbadensis miller) is
about 6.1 acidity.
2.2.2 LOCAL STUDIES
According to the study by Pedro M. Gutierrez, Jr et al., (2014) “Larvicidal
Activity of Selected Plant Extracts against the Dengue vector Aedes aegypti
Mosquito” from International Research Journal of Biological Sciences, the
phytochemical screening revealed the presence of alkaloids, flavonoids and
steroids in the leaf and bark extracts of Jatropha curcas while the leaf and
bark/stem extracts of Citrus grandis and Tinospora rumphii are rich in alkaloids,
saponins, tannins, flavonoids and steroids. These compounds are known to
possess insecticidal and larvicidal properties causing the mortality of insects
and other pests

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According to the study “Larvicidal activity of Four plants against Dengue
virus vector Aedes aegypti (Linn.)” by Oliver Shane Dumaoal, RMT et al, (2012),
the Calamansi (Citrus microcarpa) has able to produce high percent yield of
ethanolic extract with 12.77% while Sampaguita (Jasminum sambac) gave only
percent yield of 5% ethanolic extract.
According to the Philippine Medical Plants, (2012), Preliminary study of
chemical constituents of head space of J. sambac leaves using hydrophobic
resin to absorb the fragrance released from flowers yielded 37 constituents, viz.,
ethyl acetate, ethyl acetate, 3-methyl cyclopentene, 2-methyl hexane, 2, 2, 3,
4-tetramethyl petane, n-heptane, phenyl- 2-propanone, 2-methyl butate, 3-
methyl heptane, butyl acetate, 2-methyl propen-2-1y acetate, n-hexen-l-ol , 6-
methyl-2-hepta- none, 6-methyl 5-hepten-2-one, carbamyl benzoate, β-pinene,
3- hexenyl acetate, limonene, benzaldehyde, ocimene, methyl benzoate,
linalool, trans-linalool oxide, benzyl acetate, 3-hexenyl butate, methyl salicylate,
cyclohexyl formate, indole, 2, 6-dimethyl 5-heptenal, methyl anthranilate, 2, 6-
dimethyl heptenal, β-caryophyllene, β-farnsene, humulene, γ -cadinene, cis-
caryophyllene, trans, trans-farnsol, cyclohexyl benzoate.
According to Philippine Medical Plants, (2005), Calamansi fruit and
leaves yield volatile oil, 0.9 to 1.06%. Rind yields aldehydes; sesquiterpenes;
beta-pinene; linalool; linelyl acetate; tannin; glucoside; cyanogenetic

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substances. Study on the volatile constituents of calamondin peel or whole fruit
essential oil yielded 54 compounds, including 13 monoterpenes, 7 monoterpene
alcohols, 1 monoterpene oxide, 4 monoterpene aldehydes, 2 monoterpene
ketones, 4 monoterpene esters, 12 sesquiterpenes, 3 aliphatic alcohols, 6
aliphatic aldehydes, and 2 aliphatic esters, with limonene and ß-myrcene as
major compounds. In a study of essential oil by hydro distillation, the major
component identified in the peels was limonene (94.0%); minor components
were ß-myrcene (1.8%), linalool (0.4%) and α-terpineol (0.3%). Leaf essential
oil yielded an abundance of sesquiterpene hydrocarbons including hedycaryol
(19%), α-sesquiphellandrene (18.3%). α-eudesmol (14.4%), and ß-eudesmol
(8.6%). Calamondin yields a large quantity of 3',5'-di-C-ß-
glucopyranosylphloretin (DGPP) in the peel, juice sac, and leaves. Study for
carbohydrate and phenolic content yielded 2.342 ± 0.0022 and 0.053 ± 0.0015,
respectively.
According to Engineering ToolBox, (2003), the pH level of Calamansi
(Citrus microcarpa) is about 2.0 acidity.

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2.3 SYNTHESIS
Since there are so many cases of Dengue and other diseases from mosquito,
the researcher wants to create a product that will help to prevent and provide an
alternative protection against mosquito through the way of killing the larvae of
mosquito, because this stage is believed to be the weakest point of mosquito cycle.
This product is composed of different fresh plants, so it will serve as alternative to the
commercial product. With the above mention studies, constituents from the plant such
as Phenolic (Catechol), and Saponins (Saponin) has a pathogenic effect to
mosquito larvae which causes them to die (Musao, 2016). The Sampaguita leaf
(Jasminum Sambac) contain Rutin flavonoids, Catechol phenolic compound, and
Saponin which is known as primary constituents in insecticides (Kunhachan, 2012)
while Aloe vera leaves (Aloe barbadensis miller) has 75 potentially functioning
constituents one of this is the phytochemical saponins (Triterpenoid), (Surjushe, 2012)
and has a 6.1 Ph level (US FDA and the Center for Food Safety and Applied Nutrition,
2018). And Calamansi fruit (Citrus microcarpa) yield volatile oil, 0.9 to 1.06%. Rind
yields aldehydes; sesquiterpenes; beta-pinene; linalool; linelyl acetate; tannin;
glucoside; cyanogenetic substances (Philippine Medicinal Plants, 2005) and has a 2.0
Ph level of acidity (Engineering ToolBox, 2003). All these plants inhabit a larvicidal
properties. Therefore, combination of these components increases the mortality rate
and harm to mosquito larvae.

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CHAPTER 3
METHODOLOGY
In this chapter the researcher describes how the product made and how does it
work.
3.1 RESEARCH DESIGN
In order to provide an alternative larvicide and determine the level of
Effectiveness of Sampaguita and Aloe vera leaves, and Calamansi fruit extract
Larvicide, the researcher will use Completely Randomized Design (CRD) in the
research process. CRD is probably the simplest design for comparative experiments,
as it uses only two basic principles of experimental designs: randomization and
replication. With this design, subjects are randomly assigned to treatments. A
completely randomized design relies on randomization to control for the effects of
extraneous variables. The experimenter assumes that, on average, extraneous factors
will affect treatment conditions equally; so, any significant differences between
conditions can fairly be attributed to the independent variable.

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3.2 RESEARCH LOCALE
The raw materials that will be used in this study are Sampaguita leaves, Aloe
vera and Calamansi. The samapaguita, aloe vera and calamansi will be gather in Naic,
Cavite.
According to the sampaguita vendor, the main supplier of the product is from
Bulacan but according to other studies sampaguita is abundant in San Pedro Laguna
because it has a large amount of sampaguita farms and plantations that’s why the town
claim the sampaguita capital of the Philippines.
3.3 RESEARCH PROCESS
3.3.1 Materials and Methods
Materials and Equipment
-Calamansi fruit (100 pcs.) -Bottle (3pcs.) (500 ml)
-Sampaguita leaves (200 pcs.) -Tupperware (155cm x 105cm x 40cm) (16 pcs)
-Aloe vera leaves (12 pcs.) -Glass bowl
- Ethyl Alcohol (70%) -Strainer
-Knife -Measuring Cup/Glass in ml
- Electric Blender -Container
-Digital Weighing scale -X-Press Methrin Larvicide

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Method
The researcher will use Sampaguita leaves (Jasminum Sambac), Aloe
vera (Aloe barbadensis miller) and Calamansi (Citrus microcarpa) as the
independent variables. Sampaguita leaves is popular and abundant in San
Pedro Laguna, but the researcher chooses to gather the selected (200 pcs)
leaves of Sampaguita, (12 pcs) leaves of Aloe vera and (100 pcs) fruit of
Calamansi in Naic, Cavite. The mosquito larvae would be the subject or the
specimen of the experiment, the researchers will make an incubator with a
dimension of (39cm x 40cm x 44cm) and collect mosquitoes. And wait until the
mosquito lay and hatch egg and form a larva. The 1st, 2nd, 3rd, and 4th instar will
be separated in different Tupperware. The total number of mosquito larvae
needed in the experiment is approximately 480 pieces.
The procedure would take 2-3 days. First is the collection of fresh
Sampaguita leaves, Aloe vera leaves and Calamansi Fruit. Next is wash the
collected ingredients. Dry the sampaguita leaves through sunlight in 1-2 days.
After of it, cut the aloe vera leaves into small pieces and grind it with the use of
blender then strain the grinded aloe vera to get the extract and placed it in an
empty bottle. Next, dry the strained leaf skin of the aloe vera leaves from the
extract in only a day. Then, extract the juice of calamansi fruit and place it in a
bottle and put it in a refrigerator for at least 2-3hrs. Dry the calamansi peel

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through sunlight for 1-2 days. After of it, separately grind and place the dried
sampaguita leaves, peel of aloe vera leaves and peel of calamansi fruit in a
cloth. When the three cloths are done to be contained by the three ingredients,
boil a water in a pot within 5-6 mins. Once the water is already boiling put the
three cloths containing the ingredients in a steam plate on the pot. After of it,
cover the pot and simmer it gently in 10-20 mins. After of 10-20 mins get and
air-dry the ingredients. Next, pour the Ethyl Alcohol and the extracted aloe vera
to the bottle with calamansi extract then cover and wait for at least a day and
when it is done put the powder of sampaguita leaves, peel of aloe vera leaves
and peel of calamansi fruit in the mixture of ethyl alcohol, aloe vera and
Calamansi extract. Wait until the powder dissolved and strain it after to remove
undissolved powder. Then a Larvicide composed of Sampaguita leaf, Aloe vera
leaf and Calamansi fruit Extract is made.

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3.3.2 Applying SACE Larvicide and X-Press Methrin Larvicide
The experiment consists of two sets with different larvicide. The 2 sets
will be named as SET A (SACE Larvicide) and SET B (X-Press Methrin
Larvicide). The solution will be placed in the Tupperware will be 5 ml, 15 ml
and 30 ml. The amount of water is 70 ml including 10 mosquito larvae each, it
is done in all sets of treatments.
In Set A it has 3 treatments, where different amount (5 ml, 15 ml and 30
ml) of every concentration of the SACE Larvicide (10 %, 50% and 100%) will be
poured. In 10% concentration the treatment will be called treatment A-1.1 (5
ml), treatment A-1.2 (15 ml) and treatment A-1.3 (30 ml). In 50% concentration
the treatment will be called treatment A-2.1 (5 ml), treatment A-2.2 (15 ml) and
treatment A-2.3 (30 ml). And in 100% concentration the treatment will be called
treatment A-3.1 (5 ml), treatment A-3.2 (15 ml) and treatment A-3.3 (30 ml).
Each of the 3 Treatments must have four Tupperware’s. The 1st Tupperware
will have 10 pcs of 1st instar larvae, 2nd Tupperware will have 10 pcs of the 2nd
instar larvae, the 3rd Tupperware will have 10 pcs of 3rd instar larvae and the 4th
Tupperware will have 10 pcs of 4th instar larvae.

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While in SET B, it is similar to SET A. This Set has 3 treatments also
where different amount of X-Press Methrin (5 ml, 15 ml, and 30 ml) will be
poured, and it is named Treatment B-1.1 (5 ml), Treatment B-1.2 (15 ml) and
Treatment B-1.3 (30 ml. Like Set A, Set B will also have four Tupperware in its
treatment. The 1st Tupperware will have 10 pcs of 1st instar larvae, 2nd
Tupperware will have 10 pcs of the 2nd instar larvae, the 3rd Tupperware will
have 10 pcs of 3rd instar larvae and the 4th Tupperware will have 10 pcs of 4th
instar larvae.
3.3.3 Monitoring Chemical Effects
Each set of treatments will observe in 10-20 mins., 21-30 mins., and 31-
40 mins. The mortality rate of the solution will be measure based on how many
mosquito larvae died and live after exposure to the larvicide made of
sampaguita and aloe vera leaves and calamansi fruit extract (SACE Larvicide).

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3.4 DATA GATHERING
The researchers will make an experiment containing 2 sets with different
larvicide, in Set A (SACE Larvicide) it has three treatments in every concentration of
the product (10%, 50% and 100%). Where the three treatments of 10% concentration
is named A-1.1 (5ml), A-1.2 (15ml) and A-1.3 (30ml). The three treatments of 50%
concentration is named A-2.1 (5ml), A-2.2 (15ml) and A-2.3 (30ml). And the three
treatments of 100% concentration is named A-3.1 (5ml), A-3.2 (15ml) and A-3.3
(30ml). While in Set B (X-Press Methrin Larvicide), the treatment of this set is similar
to set A. Its three treatments is named B-1.1 (5ml), B-1.2(15ml) and B-1.3(30ml). The
researchers will collect and observe a mosquito until it lay and hatch egg till it become
a larva and the larvae aged (1st, 2nd, 3rd, and 4th instars) will be separated and placed
in a container. Then, 10 mosquito larvae will be placed in each treatment so, all in all
there are 480 pcs of mosquito larvae will be observed. And the changes in treatments
will be observe in 10-20 mins., 21-30 mins., and 31-40 mins. The mosquito larvae will
be considered dead when the criteria of being dead is obtained such as trembling of
its body, not moving, and the position of the larvae is not diagonally than the usually.
The result of the experiment will be the basis of the mortality and effectiveness of the
study.

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3.5 STATISTICAL TREATMENT OF DATA PROCEDURE
The researchers utilized the following statistical techniques to ensure valid and
reliable analysis and interpretation of data.
1. For measuring the concentration of the solution.
mass of Solute (g)

% by mass= ------------------------------------------------------------ X100
mass of solute(g) + mass of Solvent(g)
2. For measuring effectiveness of the concentration of the solution
applied. The control mortalities were corrected by using Abott’s formula (Abott,
1925).
Number of dead larva

Mortality Rate = -------------------------------- X 100
Number of larva
3. Analysis of Variance (ANOVA) - Anova is a statistical test which analyzes
variance. It is helpful in making comparison of two or more means which enable
a researcher to draw various results and predictions about two or more sets of
data (Anova Formula, 2013).

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CHAPTER 4
RESULTS AND DISCUSSION
4.1 INTRODUCTION
In this chapter, the results of the study are presented and discussed with
reference to the aim of the study, which was to determine if the larvicide made up of
Sampaguita leaf, Aloe vera and Calamansi fruit extract can be used to kill mosquito
larvae. It also aims to provide an alternative solution to the existing commercial product
and to assess its effectiveness through conducting a series of experiment.
4.2 DISCUSSION
4.2.1 PHYTOCHEMICAL CONSTITUENTS
Phytochemical Constituents of Sampaguita leaves. The Phytochemical
constituents exist in Sampaguita are: Rutin or Rutoside flavonoids and
Catechol or Pyrocatechol saponins. Rutin flavonoid is a phenolic compound
that can prolong the life cycle of mosquito larvae and cause higher larval
mortality when it is added to the insect diet. Many flavonoids can act as feeding
deterrents for phytophagous insects, even at relatively low concentrations

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(Talita Roberta Ferreira Borges Silva et al, 2016). In one leaf of Sampaguita 0.5
mg/g of Rutin is present. While Catechol or Pyrocatechol is also a phenolic
compound where according to the study of Urbbi Devi (August 2017), the
phenolic fraction revealed that it affected the growth of larvae by decreasing its
life span, fertility and fecundity of the mosquitoes. And Saponin of sampaguita
leaves has insecticidal activities which the effect of this phytochemical is
observed having an increased in mortality, lower the food intake, weight
reduction, retardation in development and decrease in reproduction in insects
(Ellen de Geyter, January 2007).
Phytochemical Constituents of Aloe vera leaves. The Phytochemical
constituents exist in Aloe vera leaves is the Triterpenoid saponins where
according to Ellen de Geyter et al., (January 2007), it is possessed clear
insecticidal activities: they exert a strong and rapid-working action against a
broad range of insects that is different from neurotoxicity. The most observed
effects are increased mortality, lowered food intake, weight reduction,
retardation in development and decreased reproduction. According to the main
hypotheses in literature, saponins exert a repellent/deterrent activity, bear
digestive problems, provoke insect moulting defects or cause cellular toxicity
effects. In one leaf of aloe vera, 3% of Triterpenoid is present. And the Aloe vera
gel has a Ph level of 6.1 acidity which has a larvicidal activity.

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Phytochemical Constituents of Calamansi fruit. The Phytochemical
constituent exist in Calamansi peel and extract is Tannin. The of Tannin can
reduce growth and fecundity of some insect species and it has a Ph level of 2.0
citric acid which is harmful to insects. In one fruit of calamansi 0.28% of Tannin
and 3% citric acid is present.
4.2.2. Physical Properties of SACE Larvicide
Color. The color of the solution of SACE Larvicide in 10% concentration is light
brown, chestnut brown in 50% of concentration and yellow-brown in 100%
concentration.
Odor. The smell of the solution in every concentration is more in ethyl alcohol
and calamansi’s sour flavor.
Texture. The texture of SACE Larvicide is just like an ordinary water with slightly
cold when being touched.
Density. The Density of SACE Larvicide is 1 g/ml in 10%, 50% and 100% of

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4.2.3. CONCENTRATION OF SOLUTION
mass of Solute (g)

Formula: % by mass= ---------------------------------------------------------- X 100
mass of solute(g) + mass of Solvent(g)
1. To get 10% of concentration
20 g
% by mass=------------------- X 100 = 10 %
20 g + 180 g
20 g
% by mass=-------------------X 100 = 50 %
20 g + 20 g
- Pure solvent is used to get 100% concentration.
Table 4.2.3.1. The Mass and Total Mass (Solute and Solvent), Volume and Total
Volume (Solute), and Density of SACE Larvicide in Different Concentration
Concentration Mass Mass Total Volume Total Density

of the Solution (Solute) (Solvent) Mass (Solvent) Volume (g/ml)
10 % 20 g 180 g 200g 191 ml 191 ml 1g/ml
50 % 20 g 20 g 40 g 40 ml 40 ml 1g/ml
100% - 200 g 200 g 211 ml 211 ml 1g/ml

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The table 4.2.3.1 show the how the 10%, 50% and 100% concentration
obtained. In 10%concentration, 20g of solute and 180 g of solvent is needed to obtain
the concentration. While in 50% concentration, 20g of solute and solvent is needed to
obtain the said concentration and in 100% concentration, only 200g of the extract of
the calamansi and aloe vera together with ethyl alcohol as the solvent is needed to
obtain the pure concentration. The table also show that the density of 10%, 50% and
100% concentration of SACE Larvicide are both obtained 1g/ml.

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4.2.4. TESTS, OBSERVATIONS

AND IMAGES

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4.2.4.1. ACTUAL IMAGE OF THE EXPERIMENT

10% CONCENTRATION OF SACE LARVICIDE
Image 1. Actual image of 10% Concentration of Image 3. Observation to the 5ml of 10%
SACE Larvicide together with 4 pcs of Concentration of SACE Larvicide together with 4
Tupperware containing 1st, 2nd, 3rd and 4th pcs of Tupperware containing 1st, 2nd, 3rd and
instar larvae separately. 4th instar larvae separately.
Image 2. Actual image of 5ml of 10% Image 4. Dead mosquito larvae which is exposed
Concentration of SACE Larvicide poured to 4 to 5ml of 10% Concentration of SACE Larvicide.
Tupperwares.

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Image 5. Actual image of 15ml of 10% Image 8. Actual image of 30ml of 10%
Concentration of SACE Larvicide poured to 4 Concentration of SACE Larvicide poured to 4
Tupperwares Tupperwares.
Image 6. Observation to the 15ml of 10% Image 9. Observation to the 30ml of 10%
Concentration of SACE Larvicide together with 4 Concentration of SACE Larvicide together with 4
pcs of Tupperware containing 1st, 2nd, 3rd and pcs of Tupperware containing 1st, 2nd, 3rd and
4th instar larvae separately 4th instar larvae separately.
Image 7. Dead mosquito larvae which is Image 10. Dead mosquito larvae which is
exposed to 15ml of 10% Concentration of SACE exposed to 30ml of 10% Concentration of SACE
Larvicide. Larvicide.

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Image 1. Actual image of 50% Concentration of

SACE Larvicide together with 4 pcs of Image 4. Dead mosquito larvae which is exposed
Tupperware containing 1st, 2nd, 3rd and 4th instar to 5ml of 50% Concentration of SACE Larvicide.
larvae separately.
Tupperwares. Tupperwares.
Image 3. Observation to the 5ml of 50%

Concentration of SACE Larvicide together with 4 Image 6. Observation to the 15ml of 50%
pcs of Tupperware containing 1st, 2nd, 3rd and 4th Concentration of SACE Larvicide together with 4
instar larvae separately. pcs of Tupperware containing 1st, 2nd, 3rd and 4th
instar larvae separately.

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Image 10. Dead mosquito larvae which is

Image 7. Dead mosquito larvae which is exposed exposed to 30ml of 50% Concentration of SACE
to 15ml of 50% Concentration of SACE Larvicide. Larvicide.
Image 8. Actual image of 30ml of 50%

Concentration of SACE Larvicide poured to 4
Tupperwares.

Concentration of SACE Larvicide together with 4
pcs of Tupperware containing 1st, 2nd, 3rd and 4th
instar larvae separately

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Image 1. Actual image of 100% Concentration of Image 4. Dead mosquito larvae which is exposed
SACE Larvicide together with 4 pcs of to 5ml of 100% Concentration of SACE Larvicide.
Tupperware containing 1st, 2nd, 3rd and 4th instar
larvae separately.
Tupperwares. Tupperwares.
Image 3. Observation to the 5ml of 100% Image 6. Observation to the 15ml of 100%
Concentration of SACE Larvicide together with 4 Concentration of SACE Larvicide together with 4
pcs of Tupperware containing 1st, 2nd, 3rd and 4th pcs of Tupperware containing 1st, 2nd, 3rd and 4th
instar larvae separately. instar larvae separately.

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Image 7. Dead mosquito larvae which is Image 10. Dead mosquito larvae which is
exposed to 15ml of 100% Concentration of exposed to 30ml of 100% Concentration of SACE
SACE Larvicide Larvicide.
Image 8. Actual image of 30ml of 100%

Concentration of SACE Larvicide poured to 4
Tupperwares

Concentration of SACE Larvicide together with 4
pcs of Tupperware containing 1st, 2nd, 3rd and 4th
instar larvae separately

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X-PRESS METHRIN LARVICIDE
Image 1. Actual image of X-PRESS METHRIN Image 4. Dead mosquito larvae which is exposed
Larvicide together with 4 pcs of Tupperware to 5ml of X-PRESS METHRIN Larvicide.
containing 1st, 2nd, 3rd and 4th instar larvae
separately.
Image 2. Actual image of 5ml of X-PRESS Image 5. Actual image of 15ml of X-PRESS
METHRIN Larvicide poured to 4 Tupperwares. METHRIN Larvicide Larvicide poured to 4
Tupperwares.
Image 3. Observation to the 5ml of X-PRESS Image 6. Observation to the 15ml of X-PRESS
METHRIN Larvicide together with 4 pcs of METHRIN Larvicide together with 4 pcs of
Tupperware containing 1st, 2nd, 3rd and 4th instar Tupperware containing 1st, 2nd, 3rd and 4th instar
larvae separately. larvae separately.

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Image 7. Dead mosquito larvae which is exposed

Image 10. Dead mosquito larvae which is
to 15ml of X-PRESS METHRIN Larvicide
exposed to 30ml of X-PRESS METHRIN
Larvicide
Image 8. Actual image of 30ml of X-PRESS

METHRIN Larvicide poured to 4 Tupperwares.
Image 9. Observation to the 30ml of X-PRESS

METHRIN Larvicide together with 4 pcs of
Tupperware containing 1st, 2nd, 3rd and 4th instar
larvae separately.

4.2.4.2. MORTALITY RATE
Table 4.2.4.2.1 Mortality of SACE Larvicide in 10%, 50% and 100% Concentration and X-Press Methrin in terms of amount of solution and time of exposure.
(ND- no data because of 100% mortality).
Duration Percentage of
Set (minutes) Total of Dead Mortality of Mosquito
Treatment 10-20 mins 21-30 mins 31-40 mins Mosquito Larvae larvae died
Mosquito Larvae Instar level
1st 2nd 3rd 4th 1st 2nd 3rd 4th 1st 2nd 3rd 4th 1st 2nd 3rd 4th 1st 2nd 3rd 4th
5 10 5 3 2 ND 3 4 0 ND 1 0 5 10 9 7 7 100 90 70 70
ml % % % %
(A-1) 15 10 8 6 3 ND 1 2 1 ND 1 0 2 10 10 8 6 100 100 80 60
or ml % % % %
10% 30 10 9 6 4 ND 1 4 3 ND ND ND 3 10 10 10 10 100 100 100 100
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ml % % % %
5 9 7 1 1 1 1 2 4 ND 1 5 2 10 9 8 7 100 90 80 70
(A-2) ml % % % %
(A) or 15 10 7 5 5 ND 2 3 1 ND 1 2 2 10 10 10 8 100 100 100 80
SACE 50 % ml % % % %
Larvicide
30 10 9 7 6 ND 1 3 2 ND ND ND 2 10 10 10 10 100 100 100 100
ml % % % %
5 2 4 3 2 5 3 3 1 2 2 1 2 9 9 7 5 90 90 70 50
(A-3) ml % % % %

or 15 10 6 3 3 ND 1 3 3 ND 1 0 0 10 8 6 6 100 80 60 60
100% ml % % % %
30 7 6 5 4 3 4 5 2 ND ND ND 4 10 10 10 10 100 100 100 100
ml % % % %
(B) 5 10 10 10 10 ND ND ND ND ND ND ND ND 10 10 10 10 100 100 100 100
X-Press ml % % % %
Methrin
15 10 10 10 10 ND ND ND ND ND ND ND ND 10 10 10 10 100 100 100 100
(B-1)
ml % % % %
30 10 10 10 10 ND ND ND ND ND ND ND ND 10 10 10 10 100 100 100 100
ml % % % %
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Interpretation of Table 4.2.4.2.1.
The results based on the test conducted by the researchers, which is stated on
the table, it shows the percentage of mortality of dead mosquito larvae in terms of time
of exposure (10-20 mins, 21-30 mins and 31-40 mins) and amount of solution (5ml,
15ml and 30ml) for the 10%, 50% and 100% concentration of SACE Larvicide and the
commercial larvicide (X-Press Methrin).
In 10% concentration of SACE Larvicide, records a 100% mortality in 1 st instar
larvae, 90% mortality in 2nd instar larvae and 70% mortality in 3rd and 4th instar larvae
within 40 mins of exposure in 5ml of the solution. In 15ml of the solution it records a
100% mortality in 1st and 2nd instar larvae, 80% mortality in 3rd and 60% mortality in 4th
instar larvae within 40 mins of exposure. And in 30ml of the solution it records a 100%
mortality in all stages of mosquito larvae within 40 mins of exposure.
While in 50% concentration it records a 100% mortality in 1st instar larvae, 90%
mortality in 2nd instar larvae and 80% mortality in 3rd and 70% mortality in 4th instar
larvae within 40 mins of exposure in 5ml of the solution. In 15ml of the solution it
records a 100% mortality in 1st, 2nd and 3rd instar larvae and 80% mortality in 4th instar
larvae within 40 mins of exposure. And in 30ml of the solution it records a 100%
mortality in all stages of mosquito larvae within 40 mins of exposure.

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And 100% concentration records a 90% mortality in 1st and 2nd instar larvae,
70% mortality in 3rd and 50% mortality in 4th instar larvae within 40 mins of exposure
in 5ml of the solution. In 15ml of the solution it records a 100% mortality in 1st instar
larvae, 80% mortality in 2nd instar larvae and 60% mortality in 3rd and 4th instar larvae
within 40 mins of exposure. And in 30ml of the solution it records a 100% mortality in
all stages of mosquito larvae within 40 mins of exposure. While the commercial
larvicide has a 100% mortality in all stages of mosquito larvae within 40 mins of
exposure in 5ml, 15ml and 30ml of the solution.
Table 4.2.5. Analysis of Variance (ANOVA)
The ANOVA is the statistical treatment used by the researcher to
determine the difference between the SACE Larvicide and the commercial
larvicide (X-Press Methrin). The result of the statistical treatment used to
differentiate each concentration of SACE Larvicide and X-Press Methrin in
terms of 10-20 mins, 21-30 and 31-40 mins are shown in table below. (0.05
significance level)

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Table 4.2.5.1. 10% Concentration of SACE Larvicide and X-Press Methrin

Table 4.2.5.1.1 ANOVA Result of dead larvae in every amount of solution between
10% Concentration of SACE Larvicide and X-Press Methrin in 10-20 minutes
Dead Mosquito Larvae
Amount of Solution 10% Concentration of X-Press Merthin
SACE Larvicide (Commercial) S12 S22
5 ml 20 40 400 1600
15 ml 27 40 729 1600
30 ml 29 40 841 1600
TOTAL ∑ S1= 76 ∑ S2= 120 ∑ S12= 1,970 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ + ]-[ ]
𝑛1 𝑛2 𝑛1 +𝑛2
762 1202 (76+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ 3
+ 3 ]-[ 6
] = 322.67
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ 𝑛1
+ 𝑛 ]
2
762 1202
SSW= [1,970+4,800] - [ + ] = 44.67
3 3
Table 4.2.5.1.1.1. ANOVA Table of every amount of solution between 10%

Concentration of SACE Larvicide and X-Press Methrin in 10-20 minutes
Sources of Variance Degree of Freedom Sum of the Square Mean Square Fcomputed
Between Group 2-1= 1 322.67 𝟑𝟐𝟐.𝟔𝟕

= 322.67 𝟑𝟐𝟐. 𝟔𝟕
𝟏
𝟏𝟏. 𝟏𝟕
Within Group 6-2= 4 44.67 𝟒𝟒.𝟔𝟕
= 11. 17
𝟒
Total 6-1= 5 28.89
Fcomputed = 28.89
Ftab = 7.71
Fcomputed = 28.89 > Ftab = 7.71 Reject Ho
Therefore, the variance of 10% concentration and X-Press Methrin in terms of
time exposure (10-20 mins) and different amount of solution (5ml, 15ml and 30ml) are
not equal.

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5 ml 27 40 729 1600
15 ml 31 40 961 1600
30 ml 37 40 1369 1600
TOTAL ∑ S1= 95 ∑ S2= 120 ∑ S12= 3,059 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ + ]-[ ]
𝑛1 𝑛2 𝑛1 +𝑛2
952 1202 (95+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ + ]-[ ] = 104.17
3 3 6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ + ]
𝑛1 𝑛2
952 1202
SSW= [3,059+4,800] - [ + ] = 50.67
3 3
Table 4.2.5.1.2.1 ANOVA Table of every amount of solution between 10%

Concentration of SACE Larvicide and X-Press Methrin in 21-30 minutes.
Between Group 2-1= 1 104.17 𝟏𝟎𝟒.𝟏𝟕

= 104.17
𝟏 104.17
𝟏𝟐. 𝟔𝟕
Within Group 6-2= 4 50.67 𝟓𝟎.𝟔𝟕
= 12.67
𝟒
Total 6-1= 5 8.22
Fcomputed = 8.22
Ftab = 7.71
not equal.

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5 ml 33 40 1089 1600
15 ml 34 40 1156 1600
30 ml 40 40 1600 1600
TOTAL ∑ S1=107 ∑ S2= 120 ∑ S1= 3,845 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ + ]-[ ]
𝑛1 𝑛2 𝑛1 +𝑛2
1072 1202 (107+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ + ]-[ ] = 28.17
3 3 6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ + ]
𝑛1 𝑛2
1072 1202
SSW= [3,845+4,800] - [ + ] = 28.17
3 3

Between Group 2-1= 1 28.17 𝟐𝟖.𝟏𝟕

= 28.17 28.17
𝟏
𝟕. 𝟏𝟕
Within Group 6-2= 4 28.17 28.17
= 7.17
𝟒
Total 6-1= 5 3.93
Fcomputed = 3.93
Ftab = 7.71
Fcomputed = 3.93 < Ftab = 7.71 Accept Ho
Therefore, the variance of 30ml volume of 10% concentration and X-Press
Methrin in terms of time exposure (31-40 mins) are equal.

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Table 4.2.5.2.1 ANOVA Result of every amount of solution between 50%
5 ml 40 18 324 1600
15 ml 40 27 729 1600
30 ml 40 32 1024 1600
TOTAL ∑ S1= 77 ∑ S2= 120 ∑ S12= 2,077 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ 𝑛1
+ 𝑛 ]-[ 𝑛1 +𝑛2
]
2
772 1202 (77+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ 3
+ 3 ]-[ ] = 308.17
6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ 𝑛1
+ 𝑛 ]
2
772 1202
SSW= [2,077+4,800] - [ 3
+ 3 ] = 100.67
Table 4.2.5.2.1.1 ANOVA Result of every amount of solution between 50%

Between Group 2-1= 1 308.17 308.17

= 308.17
𝟏 𝟑𝟎𝟖. 𝟏𝟕
𝟐𝟓, 𝟏𝟕
Within Group 6-2= 4 100.67 100.67
= 25.17
𝟒
Total 6-1= 5 12.24
Fcomputed = 12.24
Ftab = 7.71
not equal.

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5 ml 25 40 625 1600
15 ml 33 40 1,089 1600
30 ml 38 40 1,444 1600
TOTAL ∑ S1= 96 ∑ S2= 120 ∑ S12= 3,158 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ + ]-[ ]
𝑛1 𝑛2 𝑛1 +𝑛2
962 1202 (96+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ + ]-[ ] = 96
3 3 6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ + ]
𝑛1 𝑛2
962 1202
SSW= [3,158+4,800] - [ + ] = 86
3 3

Between Group 2-1= 1 96 𝟗𝟔

= 96
𝟏 𝟗𝟔
𝟐𝟏. 𝟓
Within Group 6-2= 4 86 𝟖𝟔
= 21.5
𝟒
Total 6-1= 5 4.47
Fcomputed = 4.47
Ftab = 7.71
not equal.

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5 ml 34 40 1156 1600
15 ml 38 40 1444 1600
30 ml 40 40 1600 1600
TOTAL ∑ S1= 112 ∑ S2= 120 ∑ S12= 4,200 ∑ S22= 4800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ + ]-[ ]
𝑛1 𝑛2 𝑛1 +𝑛2
1122 1202 (112+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ + ]-[ ] = 10.67
3 3 6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ + ]
𝑛1 𝑛2
1122 1202
SSW= [4,200+4,800] - [ + ] = 18.67
3 3

Between Group 2-1= 1 10.67 𝟏𝟎.𝟔𝟕

= 10.67
𝟏 𝟏𝟎. 𝟔𝟕
𝟒. 𝟔𝟕
Within Group 6-2= 4 18.67 𝟏𝟖.𝟔𝟕
= 4.67
𝟒
Total 6-1= 5 2.28
Fcomputed = 2.28
Ftab = 7.71

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5 ml 11 40 121 1600
15 ml 22 40 484 1600
30 ml 22 40 484 1600
TOTAL ∑ S1= 55 ∑ S2= 120 ∑ S12= 1,089 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ 𝑛1
+ 𝑛 ]-[ 𝑛1 +𝑛2
]
2
552 1202 (55+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ + ]-[ ] = 704.17
3 3 6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ 𝑛1
+ 𝑛 ]
2
552 1202
SSW= [1,089+4,800] - [ 3
+ 3 ] = 80.67

Between Group 2-1= 1 704.17 𝟕𝟎𝟒.17

= 704.17
𝟏 𝟕𝟎𝟒. 𝟏𝟕
𝟐𝟎. 𝟏𝟕
Within Group 6-2= 4 80.67 𝟖𝟎.𝟔𝟕
= 20.17
𝟒
Total 6-1= 5 34.91
Fcomputed = 34.91
Ftab = 7.71
not equal.

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5 ml 18 40 324 1600
15 ml 29 40 841 1600
30 ml 33 40 1,089 1600
TOTAL ∑ S1= 80 ∑ S2= 120 ∑ S12= 2,254 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ + ]-[ ]
𝑛1 𝑛2 𝑛1 +𝑛2
802 1202 (80+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ 3
+ ]-[ ] = 266.67
3 6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ + ]
𝑛1 𝑛2
802 1202
SSW= [2,254+4,800] - [ + ] = 120.67
3 3

Between Group 2-1= 1 266.67 266.67

= 266.67
𝟏 266.67
𝟑𝟎. 𝟏𝟕
Within Group 6-2= 4 120.67 120.67
= 30.17
𝟒
Total 6-1= 5 8.84
Fcomputed = 8.84
Ftab = 7.71
not equal.

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Amount of Solution 100% Concentration X-Press Merthin
of SACE Larvicide (Commercial) S12 S22
5 ml 30 40 900 1600
15 ml 30 40 900 1600
30 ml 40 40 1600 1600
TOTAL ∑ S1= 100 ∑ S2= 120 ∑ S12= 3,400 ∑ S22= 4,800
(∑𝑆1 )2 (∑𝑆2 )2 (∑𝑆1 +∑𝑆2 )2

Ho= 𝝈𝟐𝟏 = 𝝈𝟐𝟐 SSB= [ + ]-[ ]
𝑛1 𝑛2 𝑛1 +𝑛2
1002 1202 (100+120)2

Ha= 𝝈𝟐𝟏 ≠𝝈𝟐𝟐 SSB= [ + ]-[ ] = 66.67
3 3 6
(∑𝑆1 )2 (∑𝑆2 )2
SSW= [∑𝑆12 +∑𝑆22 ] - [ + ]
𝑛1 𝑛2
1002 1202
SSW= [3,400+4,800] - [ + ] = 66.67
3 3

Between Group 2-1= 1 66.67 𝟔𝟔.𝟔𝟕

= 66.67
𝟏 𝟔𝟔. 𝟔𝟕
𝟏𝟔. 𝟔𝟕
Within Group 6-2= 4 66.67 66.67
= 16.67
𝟒
Total 6-1= 5 4
Fcomputed = 4
Ftab = 7.71
Fcomputed = 4 < Ftab = 7.71 Accept Ho

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4.3. RESULTS AND DISCUSSION
The result of mortality rate shows that 30 ml of all concentration (10%, 50% and
100%) of SACE Larvicide obtained a 100% mortality in all instar of mosquito larvae in
31-40 mins of exposure. The use of the SACE Larvicide to the treatments cause
poisoning effect to the mosquito larvae instars. Based on the experiment performed,
mosquito larvae show all the criteria to be considered as dead. The researcher also
considered the used of different volume that they will apply in every treatment to
determine the capability of the product to kill mosquito larvae in terms of the amount
of volume of the solution. The researcher uses 5ml, 15ml and 30 ml of SACE larvicide
in all concentration to determine changes and if it can still achieve a 100% mortality
rate in 10-20mins., 21-30mins., and 31-40mins of exposure.
In the observation of the researcher to the test conducted (see in the table
4.2.4.2.1). In 5ml of 10% concentration of SACE Larvicide 100% mortality is observed
in 1st instar larvae, 50% in 2nd instar larvae, 30% in 3rd instar larvae and 20% in 4th
instar larvae within 10-20 minutes of exposure to the solution. After 21-30 minutes of
exposure, 100% mortality is observed in 1st instar larvae, 80% in 2nd instar larvae, 70%
in 3rd instar larvae and 20% in 4th instar larvae. While in 31-40 minutes of exposure,
100% mortality is observed in 1st instar larvae, 90% in 2nd instar larvae and 70% in 3rd
and 4th instar larvae. In 15ml of the same concentration of SACE Larvicide 100%
mortality is observed in 1st instar larvae, 80% in 2nd instar larvae, 60% in 3rd instar

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larvae and 30% in 4th instar larvae within 10-20 minutes of exposure to the solution.
After 21-30 minutes of exposure, 100% mortality is observed in 1st and 2nd instar larvae,
80% in 3rd instar larvae and 40% in 4th instar larvae. While in 31-40 minutes of
exposure, 100% mortality is observed in 1st and 2nd instar larvae and 80% in 3rd instar
larvae and 60% in 4th instar larvae. In 30ml of the same concentration of SACE
Larvicide 100% mortality is observed in 1st instar larvae, 90% in 2nd instar larvae, 60%
in 3rd instar larvae and 40% in 4th instar larvae within 10-20 minutes of exposure to the
solution. After 21-30 minutes of exposure, 100% mortality is observed in 1st, 2nd and
3rd instar larvae and 70% in 4th instar larvae. While in 31-40 minutes of exposure, 100%
mortality is observed in all mosquito instar larvae.
In 5ml of 50% concentration of SACE Larvicide 90% mortality is observed in 1st
instar larvae, 70% in 2nd instar larvae, 10% in 3rd and 4th instar larvae within 10-20
minutes of exposure to the solution. After 21-30 minutes of exposure, 100% mortality
is observed in 1st instar larvae, 80% in 2nd instar larvae, 30% in 3rd instar larvae and
50% in 4th instar larvae. While in 31-40 minutes of exposure, 100% mortality is
observed in 1st instar larvae, 90% in 2nd instar larvae and 80% in 3rd and 70% in 4th
instar larvae. In 15ml of the same concentration of SACE Larvicide 100% mortality is
observed in 1st instar larvae, 70% in 2nd instar larvae, 50% in 3rd and 4th instar larvae
within 10-20 minutes of exposure to the solution. After 21-30 minutes of exposure,
100% mortality is observed in 1st instar larvae, 90% in 2nd instar larvae, 80% in 3rd

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instar larvae and 60% in 4th instar larvae. While in 31-40 minutes of exposure, 100%
mortality is observed in 1st, 2nd and 3rd instar larvae and 80% in 4th instar larvae. In
30ml of the same concentration of SACE Larvicide 100% mortality is observed in 1 st
instar larvae, 90% in 2nd instar larvae, 70% in 3rd instar larvae and 60% in 4th instar
larvae within 10-20 minutes of exposure to the solution. After 21-30 minutes of
exposure, 100% mortality is observed in 1st, 2nd and 3rd instar larvae and 80% in 4th
instar larvae. While in 31-40 minutes of exposure, 100% mortality is observed in all
mosquito instar larvae.
In 5ml of 100% concentration of SACE Larvicide 20% mortality is observed in
1st instar larvae, 40% in 2nd instar larvae, 30% in 3rd instar larvae and 20% in 4th instar
larvae within 10-20 minutes of exposure to the solution. After 21-30 minutes of
exposure, 70% mortality is observed in 1st and 2nd instar larvae, 60% in 3rd instar larvae
and 30% in 4th instar larvae. While in 31-40 minutes of exposure, 90% mortality is
observed in 1st and 2nd instar larvae and 70% in 3rd instar larvae and 50% in 4th instar
larvae. In 15ml of the same concentration of SACE Larvicide 100% mortality is
observed in 1st instar larvae, 60% in 2nd instar larvae, 30% in 3rd and 4th instar larvae
100% mortality is observed in 1st instar larvae, 70% in 2nd instar larvae, 60% in 3rd and
4th instar larvae. While in 31-40 minutes of exposure, 100% mortality is observed in 1st

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instar larvae, 80% in 2nd instar larvae and 60% in 3rd and 4th instar larvae. In 30ml of
the same concentration of SACE Larvicide 70% mortality is observed in 1 st instar
larvae, 60% in 2nd instar larvae, 50% in 3rd instar larvae and 40% in 4th instar larvae
100% mortality is observed in 1st, 2nd and 3rd instar larvae and 60% in 4th instar larvae.
While in 31-40 minutes of exposure, 100% mortality is observed in all mosquito instar
larvae. This result is one of the basis to determine the level of effectiveness of SACE
Larvicide, because of the 100% mortality that has been observed in 30ml of 10%, 50%
and 100% of SACE Larvicide within 31-40 minutes of exposure, the researcher may
say that the study is effective and can be used as an alternative larvicide against
mosquito larvae instars.
While the other experiment conducted by the researcher, which is the
commercial larvicide (X-Press Methrin) also obtained a 100% of mortality to all instar
of mosquito larvae in exposure to 5ml, 15ml and 30ml of the product in a few minutes.
The Commercial larvicide has a strong odor and chemical which is toxic to our health,
so the observer must use facemask and gloves for safety. According to the observation
the exposure to the commercial larvicide cause extreme trembling of the mosquito
larvae’s body, it took 11mins to kill 1st and 2nd instar, 13 mins in 3rd and 4th instar larvae
in 5 ml of the solution. In 15 ml of solution it took 8mins to kill 1st and 2nd instar, 10 mins
in 3rd and 4th instar larvae in 5 ml of the solution. While in 30 ml of exposure to the

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commercial larvicide have the fastest mortality, it took 6-7 mins to kill all the stages of
instar larvae.
The result of both larvicide shows that the exposure to 5ml, 15ml and 30ml of
X-Press Methrin Larvicide and 30ml of 100%, 10% and 50% of SACE Larvicide cause
100% mortality within a few minutes. And the result of the statistical treatment used
which is the Analysis of Variance, accepted the null hypothesis and shows that the
variance of 10%, 50% and 100% concentration of SACE Larvicide and the commercial
Larvicide X-Press Methrin in terms of time exposure (10-20mins, 21-30mins and 31-
40 mins) and different amount of solution (5ml, 15ml and 30ml) are equal.

lOMoARcPSD|20691372
CHAPTER 5
FINDINGS, CONCLUSIONS AND RECOMMENDATIONS
5.1 SUMMARY OF FINDINGS
1. The Sampaguita leaf, Aloe vera and Calamansi Fruit Extract Larvicide
(SACE Larvicide) is proven that it has a larvicidal activity or simply it can
kill the mosquito larvae which is the dependent variable of the study.
2. In 5ml of 10% concentration of SACE Larvicide, 20 out of 40 mosquito
larvae died or 50% mortality within 10-20 minutes of exposure, 27 out of
40 mosquito larvae died or 67.5% mortality within 21-30 minutes of
exposure while 33 out of 40 mosquito larvae died or 82.5% mortality
within 31-40 minutes of exposure. In 15ml of the same concentration, 27
out of 40 mosquito larvae died or 67.5% mortality within 10-20 minutes
of exposure, 32 out of 40 mosquito larvae died or 80% mortality within
21-30 minutes of exposure while 34 out of 40 mosquito larvae died or
85% mortality within 31-40 minutes of exposure. And In 30ml of the same
concentration also, 29 out of 40 mosquito larvae died or 72.5% mortality
within 10-20 minutes of exposure, 37 out of 40 mosquito larvae died or
92.5% mortality within 21-30 minutes of exposure while 40 out of 40

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mosquito larvae died or 100% mortality within 31-40 minutes of
exposure.
larvae died or 45% mortality within 10-20 minutes of exposure, 26 out of
40 mosquito larvae died or 65% mortality within 21-30 minutes of
exposure while 34 out of 40 mosquito larvae died or 85% mortality within
31-40 minutes of exposure. In 15ml of the same concentration, 27 out of
40 mosquito larvae died or 67.5% mortality within 10-20 minutes of
exposure, 33 out of 40 mosquito larvae died or 82.5% mortality within 21-
30 minutes of exposure while 38 out of 40 mosquito larvae died or 95%
mortality within 31-40 minutes of exposure. And In 30ml of the same
concentration also, 32 out of 40 mosquito larvae died or 80% mortality
95% mortality within 21-30 minutes of exposure while 40 out of 40
exposure.
larvae died or 27.5% mortality within 10-20 minutes of exposure, 23 out
of 40 mosquito larvae died or 57.5% mortality within 21-30 minutes of
exposure while 30 out of 40 mosquito larvae died or 75% mortality within

lOMoARcPSD|20691372
31-40 minutes of exposure. In 15ml of the same concentration, 22 out of
40 mosquito larvae died or 55% mortality within 10-20 minutes of
exposure, 29 out of 40 mosquito larvae died or 72.5% mortality within 21-
30 minutes of exposure while 30 out of 40 mosquito larvae died or 75%
mortality within 31-40 minutes of exposure. And In 30ml of the same
concentration also, 22 out of 40 mosquito larvae died or 55% mortality
90% mortality within 21-30 minutes of exposure while 40 out of 40
exposure.
5. The researcher observed that the age of mosquito also affects their
survival to the exposure in the solution, the higher the instar the stronger
they become while lower instar can be killed easily because of their
vulnerability to pathogens and because they are not fully developed.
6. On the other hand, 50% concentration of solution has a higher potential
than any other concentration.
7. The higher the volume of every concentration of SACE Larvicide the
higher mortality is obtained.

lOMoARcPSD|20691372
8. The time needed to kill mosquito larvae in terms of 5ml and 15ml of 10%
concentration is 40-60 mins to achieve 100% mortality, while in 30 ml of
the same concentration it needed 31-40 mins to achieve 100% mortality.
9. The time needed to kill mosquito larvae in terms of 5ml and 15ml of 50%
concentration is 40-50 mins to achieve 100% mortality, while in 30 ml of
the same concentration it needed 30-35 mins to achieve 100% mortality.
10. And the time needed to kill mosquito larvae in terms of 5ml and 15ml of
10% concentration is 50-90 mins to achieve 100% mortality, while in 30
ml of the same concentration it needed 31-40 mins to achieve 100%
mortality.
11. The researcher accepts the null hypothesis because according to their
statistical treatment, the variance of 30ml volume of 10%, 50% and 100%
concentration of SACE Larvicide and the commercial Larvicide X-Press
Methrin in terms of 31-40 minutes of exposure are equal.

lOMoARcPSD|20691372
5.2 CONCLUSION
Therefore, the result states the effectiveness of the Larvicide made up of
Sampaguita and Aloe vera leaves together with Calamansi Fruit Extract can kill
mosquito larvae in a few minutes just like other commercial larvicide can do to the
mosquito larvae whether the larvae are 1st, 2nd, 3rd or 4th instar.
The only difference of SACE Larvicide to the Commercial Larvicide like the
product used in comparing to the said larvicide is that it is not toxic to human’s health
because it only contains natural constituents which also human used. Experimental
results showed a 100% mortality rate of SACE larvicide within 31-40 minutes of
exposure in all concentration of the product. By all counts and with proven results,
SACE larvicide can be used as an alternative to the commercial product.
The Statistical treatment states that the variance of 30ml volume of 10%, 50%
and 100% Concentration of SACE Larvicide and X-Press Methrin in terms of 31-40
minutes of exposure are equal, because of the 100% mortality obtained in that test
conducted by the researchers.

lOMoARcPSD|20691372
5.3 RECOMMENDATIONS
The Researcher wants to recommend this study to the future researchers to
have some improvements and testing to the existing product to become more effective.
The data of this study will help them to easily improve or add something new to the
product for them to have high level of effectiveness. The researcher examined the
properties of Sampaguita Leaf, Aloe vera Leaf and Calamansi fruit extract to test its
capacity to kill mosquito larvae. However, further research is recommended to assess
the validity of the data and improving the quality of the product, including the following.
1. Developing test methods for evaluating the mortality rate considering the
temperature, volume extracted and concentration of the product.
2. Investigating the influence of combining the plant constituents to
increase the pathogenic effect of the product.
3. Increasing the validity of data using different related mathematical
equation.
4. Developing the process of extracting using different method.
5. Investigating the use of SACE larvicide in different types of water (tap
water, rainwater, well water, etc.)
6. Investigating the use of SACE larvicide in different water surface (barrels,
creeks, ditches, marshy areas, etc.)

lOMoARcPSD|20691372
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SACE-Larvicide research file

Civil Engineering (Emilio Aguinaldo College)

Studocu is not sponsored or endorsed by any college or university

miller) Leaf and Calamansi (Citrus microcarpa) Fruit Extract as an

Alternative Larvicide against Mosquito Larvae (Culicidae)

A Research Paper Presented to the faculty of

The Senior high, Western Colleges Inc.

In Partial Fulfillment of the

Requirements in Research in Daily Life 1

France William V. Nazareno

Vince Erielle L. Sarmiento

Marc Francis C. Tambal

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Incidents cause by Mosquito-borne diseases are one of the major concerns in

Therefore, the mortality results, statistical measurements and observation to the

effectiveness of SACE Larvicide is clearly proven having a larvicidal activity or

poisoning effect against mosquito larvae in 31-40 minutes of exposure.

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ENGR. BIENVENIDO B. GARCIA

RONNIE E. PASIGUI, Ph. D.

DANILO H. CABALU, Ph. D. ENGR. BIENVENIDO B. GARCIA

DARNIELL C. BALBUENA, MAEd

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of blessings throughout our research work to complete the research successfully.

providing invaluable guidance throughout this research. His dynamism, vision,

empathy, and great sense of humor.

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We are extending our thanks to our teacher from Western Colleges

our research work.

research work directly or indirectly.

France William V. Nazareno

Vince Erielle L. Sarmiento

Marc Francis C. Tambal

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our life. God bless you all.

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Approval Sheet iii

Table of Contents vii

Chapter 1: The Problem and Its Background

1.2 Background of the Study 2

1.3 Theoretical/Conceptual Framework 5

1.4 Statement of the Problem 6

1.6 Significance of the Study 8

1.7 Scope and limitation of the Study 9

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1.8 Definition of Terms 11

Chapter 2: Review of Related Literature and Study

2.1 Review of Related Literature 12

2.2 Review of Related Study 19

3.1 Research Design 25

3.2 Research Locale 26

3.3 Research Process 26

3.4 Data Gathering 31

3.5 Statistical Treatment of Data Procedure 35

Chapter 4: Results and Discussion

4.2.1 Phytochemical Constituents 33

4.2.2. Physical Properties of SACE Larvicide 35

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4.2.3. Concentration of Solution 36

4.2.4. Tests, Observations and Images

4.2.4.1. Actual Image of the Experiment 38

4.2.4.2. Mortality rate 46