Inquiry Chapter

University of Perpetual Help System GMA Campus
San Gabriel, GMA, Cavite
Senior High School Department
EFFECTIVENESS OF BAMBOO LEAVES WITH NATURAL OIL-BASED

INSECTICIDE INFUSION AS A LETHAL OVITRAP TO MOSQUITO
AMONG SELECTED RESIDENTS OF BRGY. SAN GABRIEL
GMA, CAVITE
A Quantitative Research
Presented to
THE FACULTY
SENIOR HIGH SCHOOL DEPARTMENT
UNIVERSITY OF PERPETUAL HELP SYSTEM JONELTA-GMA
San Gabriel, General Mariano Alvarez, Cavite
In Partial Fulfillment of
The Requirements for the Subject of
INQUIRIES, INVESTIGATION AND IMMERSION
By
BALDEMORO, KURT ZEDRICK M.

BRAZA, TERENCE MARINELLE S.
CERBO, REYNOLD A.
DON, MANNILYN A.
GULAY, VERA CLAIRE M.
JANIER, LYNN JANE L.
LIU, LICHEEL C.
LONTOC, MARK ARGIEL D.
NOLLORA, AALIYAH CHRISTINE M.
SARMIENTO, GAIL YVANA D.
2018
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Chapter 1
INTRODUCTION
Background of the Study
Imagine an individual after a long day of working and/or studying, nothing could
be more satisfying for him/her than simply resting on the sofa indoor or a chair by the
porch to enjoy a tranquil ambience that is void of stressors. After a while, that person’s
calf began itching, he/she scratched and upon a quick glance, has seen a swollen large
area of red, raised bump—a mosquito bite, imperfectly interfering with an almost perfect
relaxation. As a tropical country, the Philippines abounds with mosquito and the diseases
related to it. It is because they are vector of diseases such as dengue, Chikungunya,
Japanese encephalitis, malaria and filariasis (Gonzales, 2016).
Incidence of dengue, a mosquito-borne disease and a leading cause of childhood
hospitalization in the Philippines, has increased 30-fold over the last 50 years, is endemic
in more than 100 countries and causes an estimated 50 million infections annually, as
stated by the World Health Organization or the WHO (2012) in its published guideline.
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In the country alone, more than 300 deaths have been recorded for the year 2016 due to
dengue out of about 90, 000 cases (Geronimo, 2016). This is alarming, for those cases
and even death are preventable by the implementation of the appropriate approach
towards the main problem, the population of mosquito.
According to Benelli (2015), the current control strategies mainly rely on
synthetic pesticides, insect growth regulators and microbial control agents. However,
synthetic chemicals lead to a number of negative implications, including high operational
costs, development of resistance and toxic effects on non-target organisms and human
health. Natural pesticides offer such benefits such as being clean, safe, eco-friendly and
cost-effective. Angelis (2016) posited that boric acid, neem oil, canola oil, rotenone,
cedar oil, spinosad, pyrethrum and diatomaceous earth are among the first choice for
most home and garden pest control needs.
It is important to know the different stages of the mosquito’s life cycle in
mitigating the adverse effects associated with it and choosing the appropriate pesticide to
be used against it. All species of the insect vector undergoes four (4) distinct stages: egg
stage, hatches when exposed to water; larva stage, or wriggler lives in water; pupa stage,
or tumbler does not feed; and adult stage (Pruitt, 2016). The adult flies shortly after
emerging and only the female ones bite and feed on blood of humans or animals.
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Recent studies have shown that leaves of particular plants and trees can be used to
attract gravid mosquito as indicated in the studies “Identification of Bacteria and
Bacteria-associated Chemical Cues that Mediate Oviposition Site Preferences by Aedes
aegypti” by Apperson et.al. (2008) and “Characteristics of Grass Infusions as
Oviposition Attractants to Aedes (Stegomyia) (Diptera: Culicidae)” by Eiras et.al.
(2016). The aforementioned studies proved that bamboo and white-oak leaf, along with
grass can be used a mosquito oviposition attractant. Regarding the usage of natural oils,
Bogran et.al. (2011) have suggested that canola oil, clove oil, cotton seed oil, garlic oil,
neem oil, rosemary oil, sesame oil and soybean oil, have the potential for acting as an
insecticidal agent in their publication “Using Oils as Pesticides.”
With the growing concern pertinent to mosquito-borne diseases and urgency of
the need to intensify vector studies and improve vector-control methods, a novel method
must be contrived to satisfy both sides of the problem. Upon such premise, the
researchers was inspired to conduct a study about making a lethal ovitrap that can attract
and kill mosquito in a multi-stage manner. This quantitative and descriptive research will
determine the effectiveness of bamboo leaves with natural oil-based insecticide infusion
as a lethal ovitrap to mosquito among selected residents of Barangay (Brgy.) San Gabriel
in General Mariano Alvarez (GMA), Cavite.
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This study entitled "Effectiveness of Bamboo Leaves with Natural Oil-based
Insecticide Infusion as a Lethal Ovitrap to Mosquito Among Selected Residents of Brgy.
San Gabriel GMA, Cavite" was conceived by the researchers out of the prevalent
predicament in the country in relation to mosquito population. Because of its tropical
climate, the Philippines is home to several species of the foresaid insect vector. One of
those species is Aedes aegypti, which is responsible for the spread of dengue, malaria and
yellow fever among others. With the researchers' concern for the further proliferation of
the foregoing ailments, they decided and desired to create an innovative stratagem to deal
with the aforementioned matter.
The Philippines is an archipelagic and tropical country, with numerous bodies of
water and frequent rainfalls, plagued with mosquito-borne diseases. As a consequence,
several initiatives have been set out by the public and private sectors of the country
against the foresaid disorders, these includes Dengvaxia, a controversial dengue vaccine
still under trials. This reflects the utmost need for the investigation of a novel method to
control mosquito population. In line with that, the researchers deem it imperative and
paramount to conduct this quantitative and descriptive study.
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Statement of the Problem
The general objective of this quantitative research is to determine the
effectiveness of bamboo leaves with natural oil-based insecticide infusion as a lethal
ovitrap to mosquito among selected residents of Brgy. San Gabriel in General Mariano
Alvarez (GMA), Cavite. By that ground, the researchers will aim to answer specifically
the following questions:
1. What are the respondents’ numbers of hour/s upon which the bamboo leaves with
natural oil-based insecticide infusion was utilized?
2. What are the respondents’ Likert scale scoring of the bamboo leaves with natural
oil-based insecticide infusion effectiveness?
3. Is there a significant relationship between the numbers of hour/s upon which the
bamboo leaves with natural oil-based insecticide infusion was utilized and its
Likert scale scoring for effectiveness by the respondents?
4. What is the correlation between the numbers of hour/s upon which the bamboo
leaves with natural oil-based insecticide infusion was utilized and its Likert scale
scoring for effectiveness by the respondents?
Objectives of the Study
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This quantitative study will help in devising a technique for controlling Aedes aegypti
population. To align with this study’s primary objective, the following objectives are
enumerated:
1. The researchers will determine the respondents’ numbers of hour/s upon which
the bamboo leaves with natural oil-based insecticide infusion was utilized.
2. The researchers will determine the respondents’ Likert scale scoring of the
bamboo leaves with natural oil-based insecticide infusion effectiveness.
3. The researchers will determine if there is a significant relationship between the
numbers of hour/s upon which the bamboo leaves with natural oil-based
insecticide infusion was utilized and its Likert scale scoring for effectiveness by
the respondents.
4. The researchers will determine the correlation between the numbers of hour/s
upon which the bamboo leaves with natural oil-based insecticide infusion was
utilized and its Likert scale scoring for effectiveness by the respondents.
Hypothesis of the Study
The hypothesis serves as an intellectual tentative answer regarding the study’s
specific questions. This study has used the null type of hypothesis.
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1. The researchers believe that the respondents’ numbers of hour/s upon which the
bamboo leaves with natural oil-based insecticide infusion was utilized cannot be
determined.
2. The researchers believe that the respondents’ Likert scale scoring of the bamboo
leaves with natural oil-based insecticide infusion effectiveness cannot be
determined.
3. The researchers believe that there is no significant relationship between the
numbers of hour/s upon which the bamboo leaves with natural oil-based
insecticide infusion was utilized and its Likert scale scoring for effectiveness by
the respondents
4. The researchers believe that there is no correlation between the numbers of hour/s
upon which the bamboo leaves with natural oil-based insecticide infusion was
utilized and its Likert scale scoring for effectiveness by the respondents.
Significance of the Study
This study must be conducted for dengue, a mosquito-borne disease, incidence
has increased 30-fold over the last 50 years, is endemic in more than 100 countries and
causes an estimated 50 million infections annually (World Health Organization, 2012).
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The importance of this research can be reflected from the impact it has on its
beneficiaries and donees, which are listed as the following:
Respondents. The 15 respondents from General Mariano Alvarez (GMA), Cavite will be
benefitted by firsthand knowledge and experience of the effect of bamboo leaves with
natural oil infusion as a lethal ovitrap to mosquito.
Filipino Citizens. People from the Philippines will be benefitted as they do live in a
tropical country where mosquito population and diseases related to it are major concerns.
Department of Health or DOH. In adherence to the national agency’s mission of
improving the health system in the whole country, this study will contribut to their
serious exertion regarding diseases transmitted and spread by mosquitoes.
Brgy.San Gabriel. This study will help in devising strategies and techniques to control
mosquito population upon the said barangay, possibly aiding in the Local Government
Units (LGUs) fight against tropical diseases.
Future Researchers. The ultimate goal of research is to add upon the fund of knowledge
to improve human life. Likewise, this study will procure enrichment about the usage of
bamboo leaves with natural oil-based insecticide infusion as a lethal ovitrap. The
researchers do hope that future researchers would find this study relevant to their own
study regarding a related topic.
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Scope and Limitation
This study entitled "The Effectiveness of Bamboo Leaves with Natural Oil-based
San Gabriel GMA, Cavite," will be conducted to determine the effectiveness of bamboo
leaves with natural oil infusion as a lethal ovitrap to mosquito. The said study will be
conducted in the year 2018 at the municipality of General Mariano Alvarez (G.M.A.),
Cavite. There will only be 15 respondents from the aforementioned municipality.
To assess the effectiveness of bamboo leaves with natural oil-based insecticide
infusion as a lethal ovitrap, a survey questionnaire will be used. This study will only
focus on the (1) respondents’ numbers of hour/s upon which the bamboo leaves with
natural oil-based insecticide infusion was utilized, and the (2) respondents’ Likert scale
scoring of the bamboo leaves with natural oil-based insecticide infusion effectiveness.
Also included in this study’s scope is the (3) significance of the relationship between the
numbers of hour/s upon which the bamboo leaves with natural oil-based insecticide
infusion was utilized and its Likert scale scoring for effectiveness by the respondents, and
the (4) correlation between the numbers of hour/s upon which the bamboo leaves with
natural oil-based insecticide infusion was utilized and its Likert scale scoring for
effectiveness by the respondents.
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The demographic profile of the respondents that includes age, sex, marital status and
such will not be determined. Additionally, the isolation and identification of the particular
chemical/s that attract/s and/or kill/s mosquito will not be pursued by the researchers.
Any other variables or factors that can overtly and/or covertly affect the result of the
study, aside from the foregoing ones, will not be determined and studied in this study.
Likewise, acquiring the profile and personal life of the respondents delimits this study.
Definition of Terms
The following terms will be operationally defined for a clearer understanding of
the study:
Bamboo Leaves. From the endemic bamboo in the Philippines, it has potential application
as a mosquito attractant when used as a solute in an aqueous solution.
Natural Oil. A liquid substance acquired from natural and organic sources, usually out of
plants that uses it for their own defense, which makes them environmental-friendly and
safer to use as a insecticide.
Natural Oil-based Insecticide. A mixture consisting of canola oil and egg yolk as an
emulsifier mixed with bamboo leaves infusion as an insecticide.
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Infusion. The process done to the bamboo leaves, which is defined as allowing a
substance, as a solute, to be permeated with water for extraction, it is also the product out
of such procedure.
Lethal Ovitrap. Alternatively, oviposition trap is used to capture mosquito by using an
attractant; it also incorporates pesticides to kill larvae and/or the adult mosquito.
Mosquito. A small flying insect widely distributed in tropical regions and is considered a
vector of various diseases including dengue and malaria.
Theoretical Framework
This portion discusses the theories that provided the foundation of this study. The
principal aim of this research is to provide information about the effectiveness of bamboo
leaves with natural oil-based insecticide infusion as a lethal ovitrap to Aedes aegypti
mosquito among selected residents of Brgy. San Gabriel, General Mariano Alvarez
(GMA), Cavite for the benefactors of this study using the theory below as the
fundamental structure.
The Mosquito Theory of Life. This theory, proposed by Okechukwu Ofili (2015), states
that “the man who kills mosquitoes without killing its eggs will soon die of malaria.”
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Principally, if a person or group of person hacks at a problem without attacking its root, it
will only lead to a futile and endless cycle of hacking until the predicament itself
overcomes the solution. Parenthetically, mosquitoes’ life cycle consists of four stages:
egg larva, pupa and adult (Pruitt, 2016). In connection to the aim of this study, the
bamboo leaves with natural oil infusion as a lethal ovitrap works as an ovicide, larvicide
and adulticide, thus greatly curbing mosquito population and its harmful corollaries.
Theory of the Natural Nidality of Diseases. A theory by Pavlovsky adopted by Codeco
et.al. (2012) in their work “Spatial Studies on Vector-Transmitted Diseases and Vectors,”
which depicts three (3) axioms regarding spread of diseases. The first principal dictates
that diseases tend to be limited geographically. While the second and third one states that
this spatial variation arises from underlying variation in the physical and/or biological
conditions that support the pathogen and its vectors and reservoirs; and that if those
abiotic and biotic conditions can be delimited on maps, then both contemporaneous risk
and future change in risk should be predictable, respectively. As what could be derived
from the three foregoing notions, the potency of diseases is determined by the physical
and/or biological variables in a locality, with those in mind, future risks can be effectively
precluded.
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Ecological Theory to the Environmental Risk Evaluation of Natural Pesticide. Posited by
Chandler et.al. (2008) in the article “Microbial Biopesticides for Integrated Crop
Management: An Assessment of Environmental and Regulatory Sustainability,” this
theory discusses points regarding the development of an environmentally sustainable
systems for controlling pests that are less reliant on chemical pesticides as the primary
management tool.
The best way is through Integrated Pest Management (IPM) and Integrated Crop
Management (ICM), which combine a range of complementary methods to reduce a pest
population below its economic injury level while minimizing impacts on other components of
the agro-ecosystem. This takes into account the needs of producers, wider society and the
environment. In summary, usage of naturally derived active ingredients as pesticide is
recommended amid increasing concerns for adverse effects from synthetic and conventional
chemicals.
Conceptual Framework
The aim of the conceptual framework is to provide a visualization of the study’s
procedure. This study’s methodology was adapted and revised from “Use of the CDC
Autocidal Gravid Ovitrap to Control and Prevent Outbreaks of Aedes aegypti (Diptera:
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Culicidae)” by Acevedo et.al. (2014). The figure that follow shows the number of
respondents, and the research material that will be used by the researchers.
Target Sample: Residents who meet the criteria (n=15)
Bamboo Leaves with Natural Oil-based

Insecticide Infusion Group
Distribution of Survey Questionnaires
Retrieval of Survey Questionnaires
Data Analysis
Outcome
Figure 1 Research Paradigm
Research Paradigm of the “Effectiveness of Bamboo Leaves with Natural Oil-based
San Gabriel GMA, Cavite"
Figure 1 simplifies this study’s paradigm. There will be an experimental group, 15
respondents under the bamboo leaves with natural oil-based insecticide infusion group.
Being previous and pioneer users of the said device, survey questionnaires will be
distributed to them. After the respondents’ completion, the survey questionnaires will be
retrieved. Consequentially, the data will be prepared for data analysis.
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Chapter 2
REVIEW OF RELATED LITERATURE
This chapter presents the citations pertinent to this study concerning the usage of
natural leaves and natural oil-based insecticide infusion as a lethal ovitrap for controlling
mosquito population. The relevant information will be used by the researchers in
developing the study.
Mosquito-borne Diseases
Almost everyone has had the unpleasant experience of being bitten by a mosquito.
Mosquito bites can cause skin irritation through an allergic reaction to the mosquito's
saliva - this is what causes the red bump and itching. But a more serious consequence of
some mosquito bites may be transmission of serious diseases and viruses such as malaria,
dengue virus, Zika and West Nile virus, which can lead to disabling and potentially
deadly effects (such as encephalitis, meningitis and microcephaly) as stated by Kavlock
(2016).
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More than its irritating bite, mosquito population must be under control out of its
ability to transmit diseases. These disorders can lead to noxious complications, which has
the potential to be a precursor to death. When unmonitored and neglected, mosquito-
borne diseases could lead to detrimental effects to a community, region or country.
Mosquitoes are considered a global treat. It is because they are vector of diseases
such as dengue, Chikungunya, Japanese encephalitis, malaria and filariasis (Gonzales,
2016). Such diseases transmissible by the said vector insect pose a threat to public and
animal health. Not only can mosquitoes carry diseases that afflict humans, but they also
can transmit several diseases and parasites that dogs and horses are very susceptible to.
These include dog heartworms, eastern equine encephalitis and West Nile virus.
According to a published guideline by the World Health Organization (2012),
incidence of dengue, a mosquito-borne disease and a leading cause of childhood
hospitalization in the Philippines, has increased 30-fold over the last 50 years, is endemic
in more than 100 countries and causes an estimated 50 million infections annually.
Dengue fever (DF) is caused by any of four closely related viruses, or serotypes: dengue
1-4. Infection with one serotype does not protect against the others, and sequential
infections put people at greater risk for dengue hemorrhagic fever (DHF) and dengue
shock syndrome (DSS).
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Dengue is transmitted between people by the mosquitoes Aedes aegypti and Aedes
albopictus, which are found throughout the world. Insects that transmit disease are
vectors. Symptoms of infection usually begin 4 – 7 days after the mosquito bite and
typically last 3 – 10 days. The indications of the said ailment can range from mild to life-
threatening.
Chikungunya virus is transmitted to people by mosquitoes. The most common
symptoms of Chikungunya virus infection are fever joint pain, headache, muscle pain,
joint swelling and rash. Outbreaks have occurred in countries in Africa, Asia, Europe,
and the Indian and Pacific Oceans. There is no vaccine to prevent or medicine to treat
Chikungunya virus infection, and most of the infected individuals feel better within a
week (Environmental Protection Agency, 2016).
Most people infected with Chikungunya virus will develop some symptoms.
Chikungunya disease does not often result in death, but the symptoms can be severe and
disabling. Most patients feel better within a week. With the absence of a cure for the
aforementioned virus, people must still avoid being bitten by the vector insect. People in
tropical and sub-tropical regions are vulnerable and must take necessary precautions.
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Centers for Disease Control and Prevention (2015) stated that Japanese
encephalitis is caused by a virus spread by infected mosquitoes in Asia and the Western
Pacific. JE virus is one of a group of mosquito-transmitted viruses that can cause
inflammation of the brain (encephalitis). It takes 5 to 15 days after the bite of an infected
mosquito to develop symptoms. In people who develop severe disease, initial symptoms
include fever, chills, headache, fatigue, nausea and vomiting. The disease can progress to
inflammation of the brain (encephalitis) and is often accompanied by seizures.
Japanese Encephalitis (JE) is a potentially severe disease. Most people who are
infected develop mild symptoms or no symptoms at all. JE occurs in Asia and parts of the
Western Pacific; it usually occurs in rural or agricultural areas, often associated with rice
farming. There is no specific treatment, but a vaccine is available.
West Nile virus is an arthropod-borne virus (arbovirus) most commonly spread by
infected mosquitoes, it can cause febrile illness, encephalitis (inflammation of the brain)
or meningitis (inflammation of the lining of the brain and spinal cord). West Nile virus
transmission has been documented in Europe, Middle East, Africa, India, parts of Asia
and Australia. Currently there is no West Nile virus vaccine available for people. The
incubation period is usually 2 to 6 days but ranges from 2 to 14 days.
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Most people (70-80%) who become infected with West Nile virus do not develop
any symptoms. In addition, about 1 in 5 people who are infected will develop a fever with
other symptoms such as headache, body aches, joint pains, vomiting, diarrhea, or rash
(Division of Vector-Borne Diseases, 2015). With that, the disease may not manifest itself
overtly.
Most people are infected with West Nile virus by the bite of an infected mosquito.
Mosquitoes become infected when they feed on infected birds; infected mosquitoes can
then spread the virus to humans and other animals. There are no symptoms in most
people. Many scientists are working on the issue of vaccine development, and there is
hope that a vaccine will be available in the future.
In an exposition, National Center for Emerging and Zoonotic Infectious Diseases
(2015) conveyed that yellow fever is a disease caused by a virus that is spread to people
by the bite of an infected mosquito. It is found in tropical and subtropical areas in South
America and Africa. The incubation period (time from infection to illness) is usually 3-6
days, initial symptoms of yellow fever include sudden onset of fever, chills, severe
headache, back pain, general body aches, nausea and vomiting, fatigue, and weakness.
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However, roughly 15% of people will have a brief period of hours to a day
without symptoms and will then develop a more severe form of yellow fever disease, in
severe cases, a person may develop high fever, jaundice (a condition that involves yellow
discoloration of the skin and the whites of the eyes), bleeding (especially from the
gastrointestinal tract), and eventually shock and failure of many organs. Roughly 20-50%
of people who develop severe illness may die. The vaccine for the virus is a live but
attenuated (less potent) strain of the virus.
Most people improve after the initial symptoms of yellow fever, still a percentage
of those severely affected may die. No specific treatments have been found to help
patients with yellow fever. If possible, patients with yellow fever should be hospitalized,
for treatment of their symptoms and to be closely observed by healthcare workers.
Yellow fever can be prevented by vaccination.
Zika virus is transmitted primarily through the bite of infected Aedes species
mosquitoes. Since 2007, Zika virus disease outbreaks have been reported in the South
Pacific, and since 2015, Zika virus has rapidly spread in the Western Hemisphere. Direct
human-to-human transmission of Zika virus can occur through sexual contact and from a
pregnant woman to her fetus, infection during pregnancy can cause severe birth defects,
including microcephaly.
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Most persons infected with Zika virus will not have symptoms; among those who
do become ill, the most common signs and symptoms are fever, rash, joint pain, and
conjunctivitis, and these usually occur within a week of infection. The illness is typically
mild, with symptoms lasting for several days to a week, and there is currently no vaccine
or specific drug to prevent or treat Zika virus infection (Department of Health & Human
Services, 2015).
Zika virus is not transmitted directly from one person to another through casual
contact. Most children and adults infected with it do not become ill enough to seek
medical care or require hospitalization, and death from Zika virus infection is rare. A
vaccine and a specific cure is still to be developed.
Philippines and Mosquito
In the Philippines, more than 300 deaths have been recorded for the year 2016 due
to dengue out of about 90, 000 cases (Geronimo, 2016). Tropical and sub-tropical regions
are areas known for wide distribution of mosquito. An international concern for it is its
capability as a vector of several diseases.
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Philippines is a tropical country, thus abounding with a plethora of species of the
vector insect. In the publication “Some Philippine Mosquitoes,” Ladlow (2013) had
described nine (9) genera of mosquito endemic to the country: Aedeomyia, Aedes,
Anopheles, Culex, Desvoidea, Finlaya, Mansonia, Orabhamia and Stegomyia. Contained
in these genera are about more than 40 species.
The above information is important for each species has its own breeding and
feeding habit. Knowing the target mosquito species is a must for control programs
regarding the said arthropod. The diversity of mosquito in the Philippines also translates
to a diversity of techniques towards addressing a specific area infested with mosquitoes.
Without knowing the species of mosquito present, such exertions are unguided.
Rentokil (2017) described the most prevalent species of mosquito in the
Philippines, those are Aedes aegypti, Anopheles spp. and Culex spp. The first described
mosquito is Aedes aegypti, its bite (responsible for spread of dengue and yellow fever)
from peaks at the change of light intensity (after sunrise and before sunset), its adult has
black and white markings, Aedes larva rests 45 degrees from the surface of the water and
the egg is black in color and is shaped like a rugby ball.
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This mosquito’s egg to adult stage takes six (6) to eight (8) days. It undergoes
complete metamorphosis (egg, larval, pupa and adult stages), its habitat is usually a
container breeder. Especially, man-made container with water, the mosquito prefers
darker colors like black and red and it can fly short distance (50 to 100m).
The Anopheles mosquito, the second mosquito described by Rentokil (2017) is
responsible for transmitting malaria, its adult has pale and dark marks on its wings and
rests on a 45-degree angle to the surface, while the larva rests parallel to surface of the
water. The egg of it is about 1mm long and has floats on its sides, additionally egg to
adult stage takes six (6) to ten (10) days that undergoes complete metamorphosis (from
egg, larval, pupa up to adult stage). The Anopheles mosquito prefers clean and unpolluted
water. Regarding its habits, it bites at night and rest indoor and outdoor, prefers darker
colors, while the female, with one blood meal, can lay about 50 to 150 eggs.
Within the same article titled “Mosquito Species,” Rentokil (2017) provided
information about Culex mosquito, the main vector for Japanese B-encephalitis. On the
appearance category, its adult’s thorax, legs and veins on the wings are always covered
with brown scales, it is dull in color, and the tip of the abdomen is always blunt.
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The larva rest 45 degrees from the surface of the water and the egg is brown, long
and cylindrical, vertical on water surface, cemented in a raft of 300 eggs [raft is usually
three (3) to four (4) mm long and two (2) to three (3) mm wide]. This mosquito’s egg to
adult stage takes six (6) to ten (10) days in a complete metamorphosis (egg, larva, pupa
up to adult stage). This kind of mosquito mainly breeds in polluted stagnant water and
drains, bites at night and usually rest indoor before and after the blood meal (sometimes
they may rest outdoor), it prefers darker colors and is a long distance flier.
Delineated above are information about the appearance, life cycle and habits of
mosquitoes commonly found in the country. It is noticeable that Aedes aegypti,
Anopheles mosquito and Culex mosquito all undergoes complete metamorphism, which
includes four (4) different phases, but its length do differs. The foresaid three (3)
classifications of mosquito prefer darker colors. Moreover, their habits do differs,
especially in the biting rhythm, preferred breeding site and distance usually flown.
General Mariano Alvarez Profile
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According to ICT Web Development Team (2017) of General Mariano Alvarez
(GMA), the municipality is geographically located at coordinates approximately 120
degree 59’15’ longitude and 14 degree 18’30’ latitude. It is at the northeastern boundaries
of Cavite Province with Laguna Province. It is bounded on the north by the Municipality
of San Pedro, Laguna on the east by the Municipality of Carmona, on the west by the
Municipality of Dasmarinas and on the south by the Municipality of Silang.
General Mariano Alvarez is approximately forty-two (42) kilometers south of the
City of Manila, the premier city of the country and seventeen (17) kilometers east of
Trece Martirez City, the capital of the province (ICT Web Development Team, 2017). It
is also approximate to the National Capital Region (NCR) of Metro Manila, the primate
region of the country. The Municipality converges with Metro Manila, through its
southern gateway of Alabang, Muntinlupa City. It is about 40 to 50 kilometers away from
said convergence points. The Aguinaldo Highway, on the west and Governor’s Drive on
the south, which connects the two major roads. It can also accessed through the Molino-
Paliparan road, which cut across the inner eastern portion of Cavite on a north-south
direction running parallel to the south Expressway and Aguinaldo Highway and joins
Governor’s Drive on the south end. Modes of transportation are land vehicles.
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General Mariano Alvarez has a total land area of 962 hectares more or less as per
“Batasang Pambansa Bilang 76” dated June 13, 1980, which created the municipality.
These areas are currently occupied by twenty-seven (27) barangays of which five (5) are
considered Poblacion barangays and twenty-two (22) others are regular barangays as
cited from the foresaid source (2017).
The municipality has elongated shape with a north, northeastern orientation. It has
a total length of about 7,040 meters and a width of about 780 meters at its smallest and
1,720 meters at its widest (ICT Web Development Team, 2017). The terrain is relatively
flat plain from the south starting at the Governor’s Drive running north covering about
one third (1/3) of the municipality’s length at Poblacion 5. From this point the terrain
gradually begin to gently slope in one direction in the middle portion of the municipality,
undulating and rolling to more than one direction towards the northeastern barangay of
Epifanio Malia, portion of Francisco de Castro, Francisco Reyes and San Jose.Inland
water bodies serve as natural boundaries with neighboring municipalities and drain its
surface waters towards Manila Bay and Laguna Bay. These are Embarcadero River on
the western boundaries with Dasmarinas and San Gabriel River on the eastern boundaries
with Carmona.
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Based on ICT Web Development Team (2017), a major tributary of San Gabriel
River traverses the inner areas of the municipality from Francisco de Castro and
Southwoods to Jacinto Lumbreras. The same tributary branches out at Inocencio Salud to
serve as boundaries between the barangay of Col. J.P Elises and Aldiano Olaes, on the
eastern side. Small streams crisscross the landscape forming natural drainage where will
bete drains from Manila Bay and Laguna de Bay.
These water bodies run and form deep gullies which gave the land a mass ridge-
like formation. Considering the height and mass of the water bodies, the ridge-like
formation is highly elevated. Thus, the terrain as described above. Man-made physical
changes, includes the presence of the Congressional Road which stretches on full length
of the municipality from the Governor’s Drive or the national highway in the south to
Francisco de Castro on the north. It runs parallel to the water bodies mentioned above on
the eastern side of the municipality. Structures such as houses, schools, commercial
establishments, institutional buildings and industrial firms are located among this road,
on gully sides of water bodies and the top flat plain areas of the ridge. Open grasslands
and agricultural areas can be found on the western near central portion of the
municipality (ICT Web Development Team, 2017).
28
The slope of the municipality ranges from 0-3% to 3-8% broad to level nearly
level is classified under 0-3% slope (ICT Web Development Team, 2017). These are
generally the flat plain level lands on the southern portion from the Governor’s Drive
extending inwards covering all Poblacion barangays, Gavino Maderan, Jacinto
Lumbreras, Ramon Cruz, San Gabriel and Severino delas Alas, 3-5% slope are gently
sloping areas with land sloping in one general direction.
These areas compress the central portion of the municipality from Barangay
Kapitan Kua, Pantaleon Granados and Marcelino Mimeje to Aldiano Olaes, Col. Jose P.
Elises and portion of Foerillo Calimag, Macario Dacon and Benjamin Tirona. As
explained by ICT Web Development Team (2017), these also include Inocencio Salud
and Bernardo Pulido.Areas on the north and northeastern side have slope of 5-8%. These
are gently undulating and rolling lands sloping in more than one general direction.
Portion of Baranay Feorillo Calimag, Macario Dacon and Benjamin Tirona and barangay
of Francisco de Castro, Francisco Reyes, Epifanio Malia, Tiniente Tiago, Nicolasa
Virata, Gregoria de Jesus and San Jose.
29
According to ICT Web Development Team (2017), Gen. Mariano Alvarez will be
originally divided into 10 barangays (more appropriately, areas) that will be parts of the
Carmona Resettlement Project. It comprised the resettlement areas in Barangays San
Jose, and San Gabriel, and a portion of Cabilang Baybay, a barangay of the municipality
of Carmona. San Jose consists of Areas A and B, while San Gabriel is composed of Areas
C, D, E, F, G, H, I, J, and K. On March 25, 1985, by virtue of legislative and
administrative acts, a referendum will be made to divide again the municipality into the
current 27 barangays.
They will be named after the revolutionary leaders of the province of Cavite and
the barangay captains and other prominent personalities who led the former 10
barangays. The first appointed officials of the barangays will be sworn into office en
masse on June 27, 1985 (ICT Web Development Team, 2017). Five of the current
barangays are classified as poblacion barangays, or those that are within the
municipality's central area. The 22 others are ordinary barangays.
The 27 barangays, are as follows (ICT Web Development Team, 2017): the first
three are the Aldiano Olaes, Barangay 1 Poblacion (Area I), and Barangay 2 Poblacion.
30
In addition are Barangay 3 Poblacion, Barangay 4 Poblacion, Barangay 5 Poblacion,
Benjamin Tirona (Area D), Bernardo Pulido (Area H), Epifanio Malia, Francisco De
Castro, Francisco Reyes, Fiorello Calimag (Area C), Gavino Maderan (Area J), Gregoria
De Jesus, Inocencio Salud (Area F), Jacinto Lumbreras (Area J), Kapitan Kua (Area F),
Koronel Jose P. Elises (Area E), Macario Dacon (Area D), Marcelino Memije (Area H),
Nicolasa Virata (San Jose), Pantaleon Granados (Area G), Ramon Cruz (Area J), San
Gabriel (Area K), San Jose, Severino De Las Alas (Area J) and Tiniente Tiago.
Regarding the number of dengue fever cases in the municipality, Municipal
Epidemiology & Surveillance Unit of General Mariano Alvarez or GMA (2016)
summarized that for the year 2016, there will be 173 cases of dengue fever reported for
morbidity weeks 1-52 (January 1 to December 30, 2016) with 1 death. The year 2016’s
number of cases is 50% lower compared to the same period last year (173 vs. 349 cases).
Of the total cases reported this period, 137 (79%) will be hospitalized and 1 (1%) will be
laboratory confirmed. Ages ranged from 0.58 to 54 years old (median is 9 years old),
with majority of the cases being among males (55%).
31
The breakdown of the dengue cases by barangay for the years 2015 and 2016 is as
follows (Municipal Epidemiology & Surveillance Unit, 2016): Bernardo Pulido=13
cases, San Jose=4 cases, Fiorello Carimag=2 cases, Tiniente Tiago=6 cases, Severino De
Las Alas=35 cases, San Gabriel=36 cases, Ramon Cruz=28 cases, Pantaleon Granados=8
cases, Nicolasa Virata=8 cases, Marcelino Memije =13 cases, Macario Dacon=11 cases,
Koronel Jose P. Elises=16 cases, Kapitan Kua =14 cases, Jacinto Lumbreras=14 cases,
Inocencio Salud=9 cases, Gregoria De Jesus=24 cases, Gavino Maderan=23 cases,
Francisco Reyes=13 cases, Francisco De Castro=26 cases, Epifanio Malia=11 cases,
Benjamin Tirona=8 cases and Aldiano Olaes = 20 cases.
For the Poblacion (Municipal Epidemiology & Surveillance Unit, 2016):
Barangay 5 Poblacion=20 cases, Barangay 4 Poblacion=10 cases, Barangay 3
Poblacion=12 cases, Barangay 2 Poblacion=21 cases, Barangay 1 Poblacion=10 cases.
As what could be inferred from the data, Brgy. San Gabriel had one of the highest
number of dengue cases (36 cases). In addition, this implies that the said barangay has a
history of mosquito infestation. Those are the primary rationale on why the researchers
had chosen the locale for their study.
32
Life Cycle of an Aedes Mosquito
It is important to know the different stages of the mosquito’s life cycle in
mitigating the adverse effects associated with it and choosing the appropriate pesticide to
be used against it. All species of the insect vector undergoes four (4) distinct stages: egg
stage, hatches when exposed to water; larva stage, or wriggler lives in water; pupa stage,
or tumbler does not feed; and adult stage (Pruitt, 2016). The adult flies shortly after
emerging and only the female ones bite and feed on blood of humans or animals.
Adult, female mosquitoes lay their eggs on the inner, wet walls of containers with
water, above the waterline. According to Division of Vector-Borne Diseases (2013),
mosquitoes generally lay 100 eggs at a time. Aedes aegypti eggs are very hardy; they
stick to the walls of a container like glue and can survive drying out for up to 8 months. It
only takes a very small amount of water to attract a female mosquito. Bowls, cups,
fountains, tires, barrels, vases and any other containers storing water make for a great
“nursery.” Its eggs hatch into larvae within 48 hours.
Larvae emerge from mosquito eggs and cause lower cuticle crack. Larvae feed on
microorganisms in the water caused by organic decomposition. After molting for three
times, the larva then becomes a pupa, and this can take up to five days in warm regions.
33
As stated by Division of Vector-Borne Diseases (2013), pupae will develop until the
body of the newly formed adult flying mosquito emerges from the pupal skin and leaves
the water. Pupae develop into adult flying mosquitoes in at least four (4) days.
After adult mosquitoes emerge, male mosquitoes feed on nectar from flowers and
female mosquitoes feed on humans and animals for blood to produce eggs (Division of
Vector-Borne Diseases, 2013). After feeding, female mosquitoes will look for water
sources to lay more eggs. Aedes aegypti only flies a few blocks during its life. Aedes
aegypti mosquitoes prefer to live near people. They can be found inside homes, buildings,
and businesses where window and door screens are not used or doors are left propped
open.
Dietary restriction extends lifespan in many organisms, but little is known about
how it affects hematophagous arthropods. Arik et.al. (2010) demonstrated that diet
restriction during either larval or adult stages extends Aedes aegypti lifespan. A. aegypti
females fed either single or no blood meals survived 30–40% longer than those given
weekly blood meals as indicated in their study “The Impact of Larval and Adult Dietary
Restriction on Lifespan, Reproduction and Growth in the Mosquito Aedes aegypti.”
34
However, mosquitoes given weekly blood meals produced far more eggs. To minimize
reproduction’s impact on lifespan, adult mosquitoes will be fed artificial blood meals
containing <10% of the protein in normal human blood, minimizing egg production. A.
aegypti fed artificial blood meals containing 25 mg/ml of BSA had significantly shorter
lifespans than those fed either 10 or 5 mg/ml.
To assess the impact of larval dietary restriction on adult lifespan, Arik et.al.
(2010) maintained larval A. aegypti on 2X, 1X (normal diet), 0.5X or 0.25X diets. Adult
mosquitoes fed 0.5X and 0.25X larval diets survived significantly longer than those fed
the 2X larval diet regardless of adult diet. In summary, dietary restriction during both
larval and adult stages extends lifespan.
Methods to Control Mosquito Population
According to Benelli (2015), the current control strategies mainly rely on
synthetic pesticides, insect growth regulators and microbial control agents. However,
synthetic chemicals lead to a number of negative implications, including high operational
costs, development of resistance and toxic effects on non-target organisms and human
health. Natural pesticides offer such benefits such as being clean, safe, eco-friendly and
cost-effective.
35
Mosquitoes have detrimental effects on human lives. Besides causing constant
annoyance and irritation, some species can also transmit deadly diseases. Rentokil (2017)
advocates the usage of Integrated Mosquito Management (IMM) against growing
mosquito population. IMM is a specially designed, holistic program proven to be an
effective approach to mosquito control in the long run. It is targeted at every stage of the
mosquito life cycle, through a simple 4-steps program which includes removing potential
mosquito breeding grounds, larviciding, thermal fogging, and using adult mosquito traps.
Based on Rentokil (2017), the first step is “Eliminate Mosquito Breeding
Grounds.” Potential breeding grounds can be eliminated by frequently checking and
removing stagnant water collected by unused items in the premises. Common breeding
grounds for Aedes mosquitoes include flower pot plate, unused container and hardened
soil of potted plant. Reducing potential breeding sites is crucial so that mosquitoes will
have less sites to breed and this will increase effectiveness of larvicide treatment
The next step is called “Larviciding.” It is a treatment to control the development
of mosquitoes' larvae (Rentokil, 2017). Larvicide treatment at mosquitoes’ hotspots can
help to reduce its infestation. A thorough inspection for any potential breeding ground
must be conducted before the larvicide treatment. After that is “Thermal Fogging.”
Thermal fogging can be carried out to eliminate breeding of adult mosquitoes.
36
It is essential to fog at the right time when the mosquitoes are active to get a more
effective control. There are 2 types of fogging, namely water-based thermal fogging and
diesel-based thermal fogging. Water-based fogging is recommended due to its properties
that are effective, environmental friendly and safe (Rentokil, 2017).
In addition is “Misting/ULV.” As described by Rentokil (2017), chemical
intervention through Misting/ULV is another way to eliminate adult mosquito, which also
break the breeding cycle. Right timing of application is essential, targeting on when the
mosquitoes are active for an effective control. And finally, “Use of Mosquito Traps.” It is
effective against mosquitoes’ species such as Aedes and Culex. Trapped adult mosquitoes
eventually die in traps, breaking the breeding life-cycle. It is also used for monitoring
activities.
Emulsion and Emulsifiers
It is common knowledge that oil and water don’t mix. If you try to mix them
together they quickly separate, with the water sinking to the bottom and the oil floating
on top. If you mix them very vigorously, one of them will break up into droplets and
disperse in the other. But even this dispersion won’t last long and the two will soon
separate as before.
37
Vigorously mixing oil and water has two possible outcomes: In one, droplets of
oil are dispersed in a continuous phase of water. In the other, droplets of water are
dispersed in a continuous phase of oil. According to Constancio (2010), the first form is
called an oil-in-water emulsion (oil droplets dispersed in water, or O/W emulsion for
short), while the second form is called a water-in-oil emulsion (water droplets dispersed
in oil, or W/O emulsion).
Distinguishing between an O/W emulsion and a W/O emulsion is very important,
because the mouth senses only the continuous phase rather than the dispersed phase.
Mayonnaise is a perfect example. Mayonnaise contains about 80% oil and 20% vinegar,
plus small amounts of egg and seasoning like mustard and salt (McKee, 2013). The
surprising thing about mayo is that even with four times more oil than vinegar, the oil is
dispersed as tiny droplets in a continuous phase of vinegar.
As a result, mayo does not feel greasy in the mouth because the mouth senses
only the continuous watery vinegar phase and not the dispersed oil droplets. As stated by
McKee (2013) in “Eggs as a Functional Emulsifier,” the vinegar becomes dispersed as
droplets in a continuous phase of oil. If the vinaigrette is prepared by slowly adding the
oil to the vinegar with very vigorous mixing, the W/O emulsion will usually stay
together.
38
Such that it stays long enough to taste, and even drizzle on some salad greens. A
W/O vinaigrette “tastes” very oily compared with mayo. Another example is butter,
which is also a W/O emulsion. Butter feels greasy in the mouth. But why does mayo exist
as an O/W emulsion while a vinaigrette, containing the same ratio of oil to vinegar, exists
as a W/O emulsion? It is because the mayo contains egg and mustard which act as
emulsifiers to stabilize the oil as droplets (Constancio, 2010). Emulsifiers are substances
that facilitate the dispersion of one phase (as tiny droplets) into another.
A simple vinaigrette does not contain emulsifiers so the smaller volume of
vinegar ends up being dispersed as droplets in a much larger continuous phase of oil.
Without an emulsifier the liquid used in excess usually forms the continuous phase.
Based on Constancio (2010), egg yolks contain lipoproteins and phospholipids, like
lecithin, that coat the surface of the oil droplets and prevent the droplets from coalescing
and forming a continuous phase even though the volume of oil is four times greater than
the volume of vinegar. But not all emulsifiers stabilize the oil as droplets. Some are better
at stabilizing vinegar as droplets. It depends on the properties of the emulsifier.
As a general rule, the continuous phase is the one in which the emulsifier is
soluble. If an emulsifier is more soluble in oil, then oil will form the continuous phase
regardless of the given proportions of the oil and that of the vinegar.
39
Similarly, water-soluble emulsifiers stabilize vinegar as the continuous phase. In the
home kitchen we have relatively few emulsifiers to pick from, namely egg yolks,
mustard, and casein in milk. Both egg yolks and mustard tend to stabilize emulsions with
oil droplets suspended in vinegar. That’s why adding a little mayonnaise, which contains
egg yolks, to a mixture of 3 parts oil to 1-part vinegar forms a fairly stable oil-in-water
emulsion.
The food processing industry has many more emulsifiers to pick from, they range
from water-soluble to oil-soluble. The solubility properties are expressed in terms of the
emulsifier’s hydrophilic-lipophilic balance or HLB (McKee, 2013). Hydrophilic
substances are water loving, while lipophilic substances are fat (oil) loving. The HLB
scale runs from 0-20. Emulsifiers with a high HLB value are hydrophilic and water-
soluble (example: sodium stearoyl lactylate). Emulsifiers with a low HLB value are oil-
soluble (example: glycerol monostearate). Emulsifiers with HLB values between 3-6
stabilize W/O emulsions, while emulsifiers with HLB values between 11-15 stabilize
O/W emulsions. Emulsifiers with intermediate values (8-10) are good wetting agents
(promote spreading of a liquid phase such as water onto a solid phase such as cocoa
powder), but relatively poor emulsifiers.
Natural Leaves as Mosquito Attractant
40
Recent studies have shown that leaves of particular plants and trees can be used to
attract gravid mosquito as indicated in the studies “Identification of Bacteria and
Bacteria-associated Chemical Cues that Mediate Oviposition Site Preferences by Aedes
aegypti” by Apperson et.al. (2008) and “Characteristics of Grass Infusions as
Oviposition Attractants to Aedes (Stegomyia) (Diptera: Culicidae)” by Eiras et.al.
(2016). The aforementioned studies proved that bamboo and white-oak leaf, along with
grass can be used a mosquito oviposition attractant.
Aedes aegypti, the global vector of dengue and yellow fever, is inexorably linked
to water-filled human-made containers for egg laying and production of progeny.
Oviposition is stimulated by cues from water containers, but the nature and origin of
these cues have not been elucidated. In the study of Apperson et.al. in 2008, it will be
shown that mosquito females directed most of their eggs to bamboo and white-oak leaf
infusions, and only a small fraction of the eggs will be laid in plain water containers.
In binary choice assays, Apperson et.al. (2008) demonstrated that microorganisms
in leaf infusions produced oviposition-stimulating kairomones, and using a combination
of bacterial culturing approaches, bioassay-guided fractionation of bacterial extracts, and
chemical analyses, demonstrated that specific bacteria-associated carboxylic acids and
methyl esters serve as potent oviposition stimulants for gravid Ae. aegypti.
41
The researchers recommended that elucidation of these compounds will improve
understanding of the chemical basis of egg laying behavior of Ae. aegypti, and the
kairomones will likely enhance the efficacy of surveillance and control programs for this
disease vector of substantial global public health importance.
Ovitraps baited with grass infusions will be evaluated in the field by Eiras et.al.
(2016) as oviposition attractants or stimulants for Aedes (Stegomyia) mosquitoes in the
study “Characteristics of Grass Infusions as Oviposition Attractants to Aedes
(Stegomyia) (Diptera: Culicidae).” Significantly higher oviposition by Aedes aegypti (L.)
will be observed in areas with no vegetation, but intense human presence, whereas Aedes
albopictus (Skuse) oviposition will be significantly higher in areas with vegetation,
regardless of human presence. Ovitraps baited with infusions of Panicum maximum
(Jacq.) collected significantly more eggs than controls containing tap water. Moreover,
the number of eggs deposited in ovitraps baited with infusion made from fresh, mature
leaves will be higher compared with dried leaves or fresh, immature leaves.
Anaerobically fermented grass infusions will be more attractive than either
aerobically fermented or sterilized infusions of P. maximum. The fermentation age of the
grass infusion also affected the oviposition response of mosquitoes. Significantly, more
eggs will be deposited in baited ovitraps containing a fresh leaf infusion of P. maximum.
42
That is, when fermented for 15- or 20-d, compared with the controls. This demonstrates
that 15- or 20-d anaerobic fermentation made of fresh, fully mature leaves of P. maximum
is the optimum infusion for ovitrap-based Aedes (Stegomyia) surveillance mosquitoes in
Brazil (Eiras et.al., 2016). Neglecting to consider these differences in infusion
attractiveness may introduce significant variation in surveillance results.
Since a safe and effective mass vaccination program against dengue fever is not
presently available, a good way to prevent and control dengue outbreaks depends mainly
on controlling the mosquito vectors. Aedes aegypti mosquito populations can be
monitored and reduced by using ovitraps baited with organic infusions. In “Bacteria as a
Source of Oviposition Attractant for Aedes aegypti Mosquitoes,” Arbaoui and Chua
(2014) conducted a series of laboratory experiments to demonstrate that the bacteria in
bamboo leaf infusion produce volatile attractants and contact chemical stimulants
attractive to the female mosquitoes. Their results showed that the female mosquitoes laid
most of their eggs (59.9 vs, 2.9 eggs) in bamboo leaf infusions when compared to
distilled water.
When the fresh infusion will be filtered with a 0.45μm filter membrane, the
female mosquitoes laid significantly more eggs (64.1 vs. 4.9 eggs) in unfiltered infusion.
43
However, when a 0.8μm filter membrane will be used, the female laid significantly more
eggs (62.0 vs. 10.1 eggs) in filtrate compared to a solution containing the residue.
Arbaoui and Chua (2014) also found that a mixture of bacteria isolated from bamboo leaf
infusion serve as potent oviposition stimulants for gravid Aedes mosquitoes. Aedes
aegypti laid significantly more eggs (63.3 vs. 3.1 eggs) in bacteria suspension compared
to sterile R2A medium. These suggest that microbial activity has a role in the production
of odorants that mediate the oviposition response of gravid mosquitoes.
In the container habitats of immature mosquitoes, catabolism of plant matter and
other organic detritus by microbial organisms produces metabolites that mediate the
oviposition behavior of Aedes aegypti and Aedes albopictus. Public health agencies
commonly use oviposition traps containing plant infusions for monitoring populations of
these mosquito species, which are global vectors of dengue viruses. In laboratory
experiments under the study “Species Composition of Bacterial Communities Influences
Attraction of Mosquitoes to Experimental Plant Infusions,” gravid females exhibited
significantly diminished responses to experimental infusions made with sterilized white
oak leaves, showing that attractive odorants will be produced through microbial
metabolic activity.
44
In his study, Arellano (2010) evaluated effects of infusion concentration and
fermentation time on attraction of gravid females to infusions made from senescent
bamboo or white oak leaves. Plate counts of heterotrophic bacteria will be used, as well
as the total counts of 4′,6-diamidino-2-phenylindole-stained bacterial cells, and 16S
ribosomal DNA (rDNA) polymerase chain reaction–denaturing gradient gel
electrophoresis (DGGE) to show that changes in the relative abundance of bacteria and
the species composition of bacterial communities influenced attraction of gravid A.
aegypti and A. albopictus mosquitoes to infusions. DGGE profiles showed that bacterial
species composition in infusions changed over time.
Principal components analysis indicated that oviposition responses to plant
infusions will be in general most affected by bacterial diversity and abundance. Analysis
of bacterial 16S rDNA sequences derived from DGGE bands revealed that Proteo-
bacteria (Alpha-, Beta-, Delta-, and Gamma-) will be the predominant bacteria detected
in both types of plant infusions (Arellano, 2010). Gravid A. aegypti will be significantly
attracted to a mix of 14 bacterial species cultured from bamboo leaf infusion. The
oviposition response of gravid mosquitoes to plant infusions is strongly influenced by
abundance and diversity of bacterial species, which in turn is affected by plant species,
leaf biomass, and fermentation time.
45
In the thesis entitled “Oviposition Responses of the Mosquitoes Aedes aegypti and
Aedes albopictus to Experimental Plant Infusions in Laboratory Bioassays,” Apperson
et.al. (2015) evaluated attraction of the mosquitoes Aedes aegypti and Ae. albopictus to
plant infusions by using a modified sticky-screen bioassay that improved the resolution of
mosquito responses to odorants. Under bioassay conditions, solid-phase micro extraction-
gas chromatographic analyses of the volatile marker chemical indole showed that
odorants diffused from bioassay cups, forming a concentration gradient. Infusions will be
prepared by separately fermenting senescent leaves of eight plant species in well water.
Plant infusions will be evaluated over an 8-fold range of leaf biomass and/or a 28d
fermentation period. The responses of gravid females of both mosquito species varied
with the plant species and biomass of plant materials used to make infusions, and with the
length of the fermentation period.
Infusions made from senescent bamboo (Arundinaria gigantea) and white oak
(Quercus alba) leaves will be significantly attractive to both mosquitoes. In general,
infusions prepared by using low biomass of plant material over a 7–14d fermentation
period will be most attractive to Ae. aegypti (Apperson et.al., 2015), a 1 g per 100 mL
concentration will be also recommended. In contrast, Ae. albopictus will be attracted to
infusions of a wide range of plant biomass over a longer fermentation period.
46
Both mosquito species will be more attracted to a non-sterile white oak leaf infusion than
to white oak leaf infusion that will be prepared using sterilized plant material and water,
thus suggesting a role for microbial activity in the production of odorants that mediate the
oviposition response of gravid mosquitoes.
The mosquitoes Aedes aegypti and Aedes albopictus are vectors of pathogenic
viruses that cause major human illnesses including dengue, yellow fever and
chikungunya. Both mosquito species are expanding their geographic distributions and
now occur worldwide in temperate and tropical climates. Collection of eggs in
oviposition traps (ovitraps) is commonly used for monitoring and surveillance of
container-inhabiting Aedes populations by public health agencies charged with managing
mosquito-transmitted illness. Addition of an organic infusion in these traps increases the
number of eggs deposited. Gravid females are guided to ovitraps by volatile chemicals
produced from the breakdown of organic matter by microbes.
On the other hand, Arellano et.al. (2015) isolated and cultured 14 species of
bacteria from attractive experimental infusions, made from the senescent leaves of
canebrake bamboo (Arundinaria gigantea) with their study entitled “Oviposition
Responses of Aedes Mosquitoes to Bacterial Isolates from Attractive Bamboo Infusions.”
47
Cultures will be grown for 24 h at 28 °C with constant shaking (120 rpm) and cell
densities will be determined with a hemocytometer. Behavioral responses to single
bacterial isolates and to a mix of isolates at different cell densities will be evaluated using
two-choice sticky-screen bioassay methods with gravid Ae. aegypti and Ae. albopictus.
The results of Arellano et.al. (2015) have shown that in behavioral assays of a mix
of 14 bacterial isolates, significantly greater attraction responses will be exhibited by Ae.
aegypti and Ae. albopictus to bacterial densities of 107 and 108 cells/mL than to the
control medium. When single bacterial isolates will be tested, seven isolates (B1, B2, B3,
B5, B12, B13 and B14) will be significantly attractive to Ae. aegypti, and six isolates
(B1, B5, B7, B10, B13 and B14) significantly attracted Ae. albopictus. Among all the
isolates tested at three different cell densities, bacterial isolates B1, B5, B13 and B14 will
be highly attractive to both Aedes species.
The isolate labels and the matching bacterium species corresponding to closest
relative are as follows (Arellano et.al., 2015): B1=Bacillus thuringiensis,
B2=Enterobacter asburiae, B3=Enterobacter cancerogenus, B4=Pseudomonas fulva,
B5=Lactococcus lactis, B6=Enterobacter gergoviae, B7=Enterobacter ludwigii, B8
translates to Klebsiella oxytoca and B9 is for the bacterial specie Klebsiella granulomatis.
48
Furthermore, B10=Pseudomonas plecoglossicida, B11=Rhizobium huautlense,
B12=Shigella dysenteriae, B13=Citrobacter freundii, and B14=Brevundimonas
vesicularis.
These only show that at specific cell densities, some bacteria significantly
influence the attraction of gravid Ae. aegypti and Ae. albopictus females to potential
oviposition sites. Attractive bacterial isolates, when formulated for sustained release of
attractants, could be coupled with an ovitrap containing a toxicant to achieve area-wide
management of Aedes mosquitoes. The researchers have considered the aforementioned
recommendation as a foundation in the conception of their study “Effectiveness of
Bamboo Leaves with Natural Oil-Based Insecticide Infusion as a Lethal Ovitrap to
Mosquito Among Selected Residents of Brgy. San Gabriel GMA, Cavite.”
Natural Oils as Insecticide
Oils have been used as pesticides for centuries and are some of the most
effective, safe alternatives to synthetic insecticides and fungicides. Safe and effective use
of any oil as a pesticide, however, requires a basic understanding of its chemical nature.
The mode of action and limitations of use must also be considered. Angelis (2016)
49
posited that boric acid, neem oil, canola oil, rotenone, cedar oil, spinosad, pyrethrum and
diatomaceous earth are among the first choice for most home and garden pest control
needs.
Regarding the usage of natural oils, Bogran et.al. (2011) have suggested that
canola oil, clove oil, cotton seed oil, garlic oil, neem oil, rosemary oil, sesame oil and
soybean oil, have the potential for acting as an insecticidal agent in their publication
“Using Oils as Pesticides.” Furthermore, regardless of the source or type, all oil-based
products have a similar mode of action. Insecticidal oils kill insects on contact by
disrupting gas exchange (respiration), cell membrane function or structure. Their toxic
action is more physical than chemical and is short-lived. Oil-based pesticides have low
residual activity and must have direct contact with the insect or mite.
Oils are most effective against soft-bodied arthropods. They are most commonly
used against mites, aphids, whiteflies, thrips, mealybugs and scale insects (Bogran et.al.,
2011). Dormant oil sprays are also used against over-wintering eggs and scales.
Horticultural and plant oils are commonly used to suppress certain fungal diseases, like
powdery mildew and black spot on rose. Stylet oils may be used to manage insect-
vectored plant viruses.
50
While oil treatments have historically targeted fruit trees and woody ornamentals,
several different types of pesticidal oils are currently marketed for house plants, flowers and
vegetables. Commercial oil products include emulsifiers to enable the oil to mix readily with
water. These emulsifiers are generally considered to be inert, but may have some insecticidal
properties. Oil formulations are generally designed to be mixed with water at concentrations
of 0.5-2.0 percent (volume/ volume), as stated by Baxendale and Cranshaw (2013). It is by
such premise that the researchers have added five (5) mL of canola oil per 300 mL of
bamboo leaves infusion for the experimental ovitrap.
In “Insect Control: Horticultural Oils,” Baxendale and Cranshaw (2013) posited that
oils have different effects on pest insects. The most important is that they block the air holes
(spiracles) through which insects breathe, causing them to die from asphyxiation. In some
cases, oils also may act as poisons, interacting with the fatty acids of the insect and
interfering with normal metabolism. Oils also may disrupt how an insect feeds, a feature that
is particularly important in the transmission of some plant viruses by aphids.
Oils pose few risks to people or to most desirable species, including beneficial natural
enemies of insect pests. This allows oils to integrate well with biological controls. Toxicity is
minimal, at least compared to alternative pesticides, and oils quickly dissipate through the
process of evaporation, leaving only little residue (Baxendale & Cranshaw, 2013).
51
Oils also are easy to apply with existing spray equipment and can be mixed with many other
pesticides to extend their performance.
According to Neudorff and Talarek (2008), canola oil is an edible vegetable oil
that can be used to control insects on a wide variety of crops, it also appears to have no
adverse effects on humans or the environment. In addition, it is an edible refined
vegetable oil obtained from the seeds of four species of rape plants, Brassica napus,
Brassica juncea, Brassica rapa and B. campestris of the family Cruciferae (mustard
family). Canola oil is considered safe for human consumption. Many types of insects are
its target pests. The products are applied either with spray or irrigation systems.
There have been numerous research studies conducted at laboratory level on plant
products as fumigants against insect pests of stored products. Besides toxicity tests,
attention has been focused to elucidate their mode of action in insects. One such study is
“Plant Products as Fumigants for Stored-product Insect Control” by Rajendran and
Sriranjini (2008) which posits that adults will be generally susceptible, whereas, eggs
will be either tolerant or highly susceptible depending on insect species and the type of
essential oil or component. The essential oils proved effective in mixture with CO2 or
ethyl formate.
52
In line with that, plant products have the potential for small-scale treatments,
space fumigations and as adjuvants for conventional fumigants. The constraints including
lack of data for single or multiple components of essential oils on sorption, tainting and
residues in food commodities, and registration protocols have been highlighted by
Rajendran and Sriranjini (2008). Unlike conventional fumigants, essential oils and their
components have certain advantages since they have been used in traditional medicine,
pharmaceutical preparations and herbal beverages and as natural flavorings.
Current measures for control of Aedes aegypti, the principal vector of dengue and
yellow fever, in many countries are based on the physical elimination of larval
development sites and the application of larvicide to domestic and urban water sources.
Specific outbreaks of vector-borne disease also are controlled by space spraying of urban
areas with insecticides. As an alternative to the use of temephos, a larvicide, biological
larvicide like the Spinosad, a mixture of two neurotoxic macrolide compounds: spinosyn
A and spinosyn D that are active mainly by ingestion, will be evaluated by Bond et.al.
(2008) in “Spinosad, a Naturally Derived Insecticide, for Control of Aedes aegypti
(Diptera: Culicidae): Efficacy, Persistence, and Elicited Oviposition Response,” and
have shown that spinosad is highly toxic to Ae. Aegypti.
53
The naturally derived insecticide spinosad represents a new generation of
biorational products developed for the agricultural industry that have a reduced spectrum
of toxicity compared with the synthetic insecticides that will be developed previously. In
assessing mortality responses, a mosquito progeny will be classified as dead if it did not
move when gently touched with the point of a toothpick by Bond et.al. (2008). The same
method will be used by the researchers of this study to determine the mortality rate of
mosquito larvae.
Oils and surfactants may provide safe alternatives to chemical insecticides and are
less prone to selection for resistance. To evaluate the insecticidal effects of some
commonly used mineral and botanical oils and some wetting and spreading surfactants
against B. argentifolii nymphs, eight surfactants, a mineral oil, Sunspray Ultra-Fine Spray
Oil, and two botanical oils, a cotton seed oil and a vegetable oil will be used, with water
as a control, for evaluation by Liu and Stansly (2010). Their study “Insecticidal Activity
of Surfactants and Oils Against Silverleaf Whitefly (Bemisia argentifolii) Nymphs
(Homoptera: Aleyrodidae) on Collards and Tomato” has demonstrated that cotton seed
oil and vegetable oil can perform as well as or better than the mineral oils and may
deserve further consideration for pest control with a recorded mortality rate between 50
and 80 percent.
54
The aforementioned study’s results demonstrate good insecticidal activity of
surfactants and oils, comparable with some conventional insecticides. In addition,
surfactants are only active while wet and become ineffective under drying conditions.
Therefore, successful use of these materials requires appropriate application methods and
environmental conditions. Water will be used for the control treatment in this study as
assimilated by this study’s researchers.
The search for new insecticides to control dengue fever, chikungunya, and Zika
vectors has gained relevance in the past decades. In line with that “Evaluation of the
Insecticidal Activity of Essential Oils and their Mixtures Against Aedes aegypti (Diptera:
Culicidae)” aimed to evaluate the larvicidal action of essential oils (EOs) from Thymus
vulgaris, Salvia officinalis, Lippia origanoides, Eucalyptus globulus, Cymbopogon
nardus, Cymbopogon martinii, Lippia alba, Pelargonium graveolens, Turnera diffusa,
and Swinglea glutinosa on Aedes aegypti. Duque et.al. (2017) determined the lethal
concentrations by probit analysis using larvae of Ae. aegypti between the third and the
fourth instars and found out that all EOs displayed insecticidal action against Ae. aegypti
larvae. Furthermore, the main compounds of the EOs with highest larvicidal activity will
be thymol (42%) and p-cymene (26.4%).
55
Due to the lack of vaccines against yellow fever, dengue fever, Chikungunya, and
Zika fever, prevention strategies are focused on the control of larvae and adult Ae.
aegypti populations. The application of synthetic insecticides is the most common
approach used worldwide (Duque et.al. (2017). Compared to synthetic products, natural
pesticides are less harmful to human health and ecosystems, and so they are widely
accepted by the general population. This is a good reason for using natural oils as source
of ingredients for design of new insecticides.
Oviposition is one of the most important events in the life cycle of mosquitoes. If
oviposition is prevented, the mosquito life cycle is disrupted and population growth
reduced. To investigate the ovicidal potential of volatile oils against mosquito,
Ignacimuthu et.al. (2014) evaluated the ovicidal and oviposition response of plant volatile
oils from aniseed, calamus, canola, cinnamon, citronella, clove, lemon, orange, thyme,
tulsi and vertiver against Culex quinquefasciatus Say. Their study “Ovicidal and
Oviposition Response Activities of Plant Volatile Oils against Culex quinquefasciatus
Say” indicates that the percent of egg hatchability will be inversely proportional to the
concentration of oils and directly proportional to the number of eggs, by such, at 200 ppm
a 90 to 100% ovicidal activity will be recorded.
56
Botanical insecticides may serve as suitable alternatives to synthetic insecticides
in future as they are relatively safe, biodegradable and readily available in many areas of
the world. Plants are considered as rich sources of bioactive chemicals and may be an
alternative source of mosquito control agents. Thus, the environmental friendly and
biodegradable natural insecticides of plants origin have been receiving attention as an
alternative green measure for controlling arthropods of public health importance. In
addition, plant essential oils are natural volatile substances obtained from a variety of
plants and have been recognized as an important natural resource of insecticides
(Ignacimuthu et.al., 2014).
In “Adulticidal Activity of Essential Oil of Lantana camara Leaves Against
Mosquitoes,” Dash et.al. (2010) have investigated the insecticidal activity of essential oil
isolated from the leaves of Lantana camara against mosquito vectors (Ae. aegypti, Cx.
quinquefasciatus, An. culicifacies, An. fluviatilis and An. Stephensi). The essential oil will
be isolated from the leaves of L. camara using hydro-distillation method. They concluded
that essential oil from the leaves of L. camara possesses adulticidal activity, with an LD90
(lethal dosage) value of 0.10 mg/cm2, against different mosquito species that could be
utilized for development of oil-based insecticide as supplementary to synthetic
insecticides.
57
Mosquitoes constitute a major public health problem as vectors of serious human
diseases like malaria, filariasis, Japanese encephalitis, dengue fever, Chikungunya and
yellow fever. Plants products are emerging as a potential source of mosquito control and
among them essential oils have special interest due to their insecticidal properties.
Furthermore, insecticides of plant origin are expected to be target selective and
biodegradable leading to fewer harmful effects on human and other animals and are
environmentally safe as compared to synthetic compound.
Lethal Ovitraps
Ovitraps or oviposition traps collect the eggs laid by the mosquitoes which
develop into larva, pupa and adult mosquitoes. The standard ovitrap utilizes water and a
container and are often used for surveillance of Aedes vectors. It can be modified to
render it lethal to immature or adult populations of Ae. aegypti. Lethal ovitraps (which
incorporate an insecticide on the oviposition substrate), autocidal ovitraps (which allow
oviposition but prevent adult emergence), and sticky ovitraps (which trap the mosquito
when it lands) have been used on a limited basis (WHO, 2017). Moreover, studies have
shown that population densities can be reduced with sufficiently large numbers of
frequently-serviced traps.
58
Populations of Aedes aegypti can be managed through reductions in adult
mosquito survival, number of offspring produced, or both. Acevedo et.al. (2014) tested
the effectiveness of the novel Centers for Disease Control and Prevention autocidal
gravid ovitrap (CDC-AGO trap) to control natural populations of Ae. aegypti under field
conditions in two isolated urban areas in southern Puerto Rico for one year. The presence
of AGO control trap per home in 81% of the houses prevented outbreaks of Ae. aegypti,
which would be expected after rains. Their study “Use of the CDC Autocidal Gravid
Ovitrap to Control and Prevent Outbreaks of Aedes aegypti (Diptera: Culicidae)”
suggests that AGO traps are useful and inexpensive mosquito surveillance devices.
Direct adult mortality can be caused by the use of space sprays or residual
insecticides to mosquito resting sites, and with a variety of residual insecticide
impregnated surfaces that are being tested, such as curtains, covers for water-storage
vessels, bed nets, and ovitraps. To be a practical tool for managing dengue vectors, a trap
must be specific, effective, inexpensive, and simple to construct and operate. Because
gravid females have fed on blood at least once to produce eggs and could have acquired
dengue viruses from an infected person during any of the previous bloodmeals,
controlling gravid females is also important to reduce dengue virus transmission.
59
Ovitraps can be used for the control of dengue vectors; conversely, a lethal
ovitrap allows egg deposition but it prevents adult emergence, thus adversely affecting
mosquito daily emergence rate. Alzona et.al. (2016) have shown that their added
Novaluron increased the attractiveness of their Mosquito Larvicidal Trap (MLT) to
female mosquitoes to lay their eggs and has killing effect on the larvae in “Mosquito
Larvicidal Trap (MLT) as Surveillance and Control Tool for Aedes Mosquitoes.” The
findings indicate that MLT is more attractive than containers or ovitraps as oviposition
medium. Moreover, MLT prevented pupal development, indicating its autocidal activity
to larvae.
To create their MLT, Alzona et.al. (2016) have used an empty evaporated milk
can, pigmented as black, that can hold approximately 370 ml of water. Lawanit board,
measuring 6 x 1 in, will be used as paddle or substrate for oviposition of Aedes
mosquitoes. The paddle will be soaked wet with water and positioned at the side of the
ovitrap can containing up to 300 ml of water with or without larvicide. These methods are
adapted by the researchers of this study, especially the container and lawanit oviposition
substrate, in making their lethal ovitrap as what could be deduced from the
instrumentation section of Chapter 3.
60
In the study “Field Testing of Ovicidal-Larvicidal Trap System with Pelletized
Extracts of Piper nigrum L. for Aedes Mosquito in Quezon City and Marikina City,”
Almanzor et.al. (2013) have had the objective of field-testing the developed ovicidal-
larvicidal admixture of Piper nigrum L. for Aedes aegypti in Quezon City and Marikina
City. In Quezon City, the OL traps with the ovicidal-larvicidal pellets will be lethal to
86.2% of the eggs or larvae. Correspondingly, 13.8% of these will be able to emerge into
adults. Similarly, in Marikina City, the % egg and larvae mortality is 86.2% while %
adult emergence is 13.8%.
An OL mosquito trap, especially when ovicide-larvicide solutions are added,
offers a simple, cheap and efficient tool for surveillance and control of mosquito. The OL
traps are effective, safe, economical and environmentally safe. Control programs can be
done using the OL trap infused with Piper nigrum L. to decrease the population of Aedes
mosquitoes and eventually reduce the incidence of Dengue. In addition, the researchers
have integrated the notions of mosquito eggs and larvae mortality rates in this study with
the formulas: mortality rate of laid mosquito eggs = no. of laid mosquito eggs - no. of
hatched mosquito eggs / no. of laid mosquito eggs and mortality rate of larvae = no. of
hatched mosquito eggs - no. of surviving pupae / no. of hatched mosquito eggs,
respectively.
61
Synthesis of the State of the Art
In line with this study’s aim of determining the effectiveness of bamboo leaves
with natural oil-based insecticide infusion as a lethal ovitrap to mosquito among selected
residents of Brgy. San Gabriel in General Mariano Alvarez (GMA), Cavite, the
researchers have located, evaluated and acknowledged a plethora of related literatures
and studies. What follows are the authors and researchers that have impacted the way this
experimental study will be conceptualized, conducted and analyzed.
Pertinent to mosquito-borne diseases, Kavlock (2016), Gonzales (2016), World
Health Organization (2012), Environmental Protection Agency (2016), Centers for
Disease Control and Prevention (2015), Division of Vector-Borne Diseases (2015),
National Center for Emerging and Zoonotic Infectious Diseases (2015), and Department
of Health & Human Services (2015) enumerated malaria, filariasis, Japanese encephalitis,
dengue fever, Chikungunya and yellow fever as serious diseases transmitted to humans
by mosquitoes. Out of those aforementioned ailment, dengue, Chikungunya and yellow
fever are spread by Aedes aegypti.
In addition to that Ladlow (2013), Rentokil (2017), ICT Web Development Team
(2017), and Municipal Epidemiology & Surveillance Unit of General Mariano Alvarez or
62
GMA (2016) provided information on Philippines and prevalent mosquito species in it.
The last two sources focused on the geography of GMA and the number of dengue cases
per barangay in the said municipality, respectively.
The life cycle of the mosquito must also be incorporated in every vector control
program targeting the foresaid arthropod. Pruitt (2016), Division of Vector-Borne
Diseases (2013), and Arik et.al. (2010) discussed the four stages by which a mosquito
egg develops into a flying adult.
For methods to control mosquito population, Benelli (2015), and Rentokil (2017)
suggested several techniques and strategies specifically calibrated for diminishing
adverse effects caused by mosquito infestation. It can be inferred that methods using
synthetic chemicals and agents are still the prevalent method for the foregoing purpose.
The researchers have used egg yolk as an emulsifier for this study’s infusion, for
the reason that it contains lecithin, an emulsifying protein. The said procedure will be
influenced by pieces of information from Constancio (2010) and McKee (2013).
The studies “Identification of Bacteria and Bacteria-associated Chemical Cues
that Mediate Oviposition Site Preferences by Aedes aegypti” by Apperson et.al. (2008),
63
“Characteristics of Grass Infusions as Oviposition Attractants to Aedes (Stegomyia)
(Diptera: Culicidae)” Eiras et.al. (2016, “Characteristics of Grass Infusions as
Oviposition Attractants to Aedes (Stegomyia) (Diptera: Culicidae)” by Eiras et.al. (2016),
“Bacteria as a Source of Oviposition Attractant for Aedes aegypti Mosquitoes” by
Arbaoui and Chua (2014), “Species Composition of Bacterial Communities Influences
Attraction of Mosquitoes to Experimental Plant Infusions” by Arellano (2010),
“Oviposition Responses of the Mosquitoes Aedes aegypti and Aedes albopictus to
Experimental Plant Infusions in Laboratory Bioassays” by Apperson et.al. (2015) and
“Oviposition Responses of Aedes Mosquitoes to Bacterial Isolates from Attractive
Bamboo Infusions” by Arellano et.al. (2015) have evaluated natural leaves, for examples
are bamboo leaves, white oak leaves, and grass, and their capability as mosquito
attractant.
To support the premise that natural oil can be used as an insecticide, the following
imparted regarding the said idea. Those are Angelis (2016), Bogran et.al. (2011),
Baxendale and Cranshaw (2013), Neudorff and Talarek (2008), and “Plant Products as
Fumigants for Stored-product Insect Control” by Rajendran and Sriranjini (2008).
Moreover, “Spinosad, a Naturally Derived Insecticide, for Control of Aedes aegypti
(Diptera: Culicidae): Efficacy, Persistence, and Elicited Oviposition Response” by Bond
64
et.al.(2008), “Insecticidal Activity of Surfactants and Oils Against Silverleaf Whitefly
(Bemisia argentifolii) Nymphs (Homoptera: Aleyrodidae) on Collards and Tomato” by
Liu and Stansly (2010), “Evaluation of the Insecticidal Activity of Essential Oils and their
Mixtures Against Aedes aegypti (Diptera: Culicidae)” by Duque et.al. (2017), “Ovicidal
and Oviposition Response Activities of Plant Volatile Oils against Culex quinquefasciatus
Say” by Ignacimuthu et.al. (2014), and “Adulticidal Activity of Essential Oil of Lantana
camara Leaves Against Mosquitoes,” by Dash et.al. (2010) have contributed, as well.
World Health Organization (2017) have defined ovitraps for mosquitoes by their
purpose and/or composition. On the other hand, the ensuing studies have assessed their
performance: “Use of the CDC Autocidal Gravid Ovitrap to Control and Prevent
Outbreaks of Aedes aegypti (Diptera: Culicidae)” by Acevedo et.al. (2014), “Mosquito
Larvicidal Trap (MLT) as Surveillance and Control Tool for Aedes Mosquitoes” by
Alzona et.al. (2016) and “Field Testing of Ovicidal-Larvicidal Trap System with
Pelletized Extracts of Piper nigrum L. for Aedes Mosquito in Quezon City and Marikina
City” by Almanzor et.al. (2013).
Gaps Bridged by the Present Study
65
With the prevalence and eminence of mosquito-borne diseases, it is paramount
and applicable to find a solution regarding the root of the problem: mosquito population.
Parallel with that, several studies have investigated the potential of a number of
alternative solutions. Most approach only target a single life stage of the mosquito, be it
in egg, larva, pupa or adult phase. In the case of this study, the targets are the Aedes
aegypti mosquito egg, larva and adult, making it a multi-stage lethal ovitrap against the
said arthropod.
The aforementioned notion will be made possible by collating and correlating
pieces of information obtained by the researchers. Through the available related literature
and studies, a lethal ovitrap, using bamboo leaves infusion as an attractant and natural oil-
based insecticide as a potent agent, will be conceptualized. This provides a conduit for
usage of natural oils as insecticide, natural leaves as insect attractant and lethal ovitraps
as mosquito control contrivance.
66
Chapter 3
RESEARCH DESIGN AND METHODOLOGY
This chapter provides the methodology of the study, which will serve as a guide
for the researchers upon conduction; it includes the Research Design, Research Locale,
Population of the Study, Data Gathering Procedure and Data Analysis Method and
Procedure.
Research Design
The researchers of this study have used a quantitative and descriptive design in
creating its scheme. According to Calderon and Gonzales (2012), quantitative research,
under the foregoing classification, has its respondents as purposely selected. It implies
that the researchers have used eligibility criteria as a guide for selecting this study’s
respondents. The quantitative design requires that the variables and/or factors under study
be converted into numerical figures.
67
In line with the purpose of this study, descriptive design will also be employed.
Descriptive research is used to obtain information concerning the current status of the
phenomena to describe "what exists" with respect to variables or conditions in a situation.
Based on (Calderon and Gonzales, 2012), the methods involved range from the survey
which describes the status quo, the correlation study which investigates the relationship
between variables, to developmental studies which seek to determine changes over time.
The group of respondents in this study have used the bamboo leaves with natural-oil
based insecticide infusion or the lethal ovitrap and their experience regarding its usage
will be described by the researchers for the ovitrap’s effectiveness assessment.
Research Locale
This study will be conducted within the municipality of General Mariano
Alvarez (GMA), Cavite, especially at Barangay (Brgy.) San Gabriel. According to ICT
Web Development Team (2017) of General Mariano Alvarez (GMA), the municipality is
geographically located at coordinates approximately 120 degree 59’15’ longitude and 14
degree 18’30’ latitude. It is at the northeastern boundaries of Cavite Province with
Laguna Province. It is bounded on the north by the Municipality of San Pedro, Laguna on
the east by the Municipality of Carmona, on the west by the Municipality of Dasmarinas
and on the south by the Municipality of Silang.
68
The breakdown of the dengue cases by GMA barangays for the years 2015 and
2016 is as follows (Municipal Epidemiology & Surveillance Unit, 2016): Bernardo
Pulido=13 cases, San Jose=4 cases, Fiorello Carimag=2 cases, Tiniente Tiago=6 cases,
Severino De Las Alas=35 cases, San Gabriel=36 cases, Ramon Cruz=28 cases, Pantaleon
Granados=8 cases, Nicolasa Virata=8 cases, Marcelino Memije =13 cases, Macario
Dacon=11 cases, Koronel Jose P. Elises=16 cases, Kapitan Kua =14 cases, Jacinto
Lumbreras=14 cases, Inocencio Salud=9 cases, Gregoria De Jesus=24 cases, Gavino
Maderan=23 cases, Francisco Reyes=13 cases, Francisco De Castro=26 cases, Epifanio
Malia=11 cases, Benjamin Tirona=8 cases and Aldiano Olaes = 20 cases.
For the Poblacion (Municipal Epidemiology & Surveillance Unit, 2016):
Barangay 5 Poblacion=20 cases, Barangay 4 Poblacion=10 cases, Barangay 3
Poblacion=12 cases, Barangay 2 Poblacion=21 cases, Barangay 1 Poblacion=10 cases.
As what could be inferred from the data, Brgy. San Gabriel had one of the highest
number of dengue cases (36 cases). In addition, this implies that the said barangay has a
history of mosquito infestation. Those are the primary rationales on why the researchers
had chosen the locale for their study.
69
Population of the Study
The population is defined as the aggregation of elements from which the sample
is actually selected. Whereas sample is the elements or the group of people who are
actually selected to participate (Calderon & Gonzales, 2012) or to be the subject of the
study. Sample size should not be too large that it may lead to difficulties in extracting
certain data and not too small, that it may make the study’s sample not representative of
the population.
Residents experiencing mosquito infestation in the Barangay San Gabriel of
General Mariano Alvarez (GMA), Cavite will be considered as this study’s population.
The researchers will employ purposive sampling technique, it is a type of sampling in
which the researchers have the ability to choose a certain sample (Calderon & Gonzales,
2012) that they deem capable in giving the sort of information they require by using a
criteria. The eligibility criteria that will be used in this study are as follows: the individual
is (1) a resident within Barangay San Gabriel of General Mariano Alvarez (GMA),
Cavite, (2) experiencing mosquito infestation in their residence, (3) have used the
bamboo leaves with natural-oil based insecticide infusion and (4) willing to participate in
the study. With the aid of the eligibility criteria, 15 respondents will be selected from the
aforementioned aggregate.
70
Data Gathering Procedure
In compliance with the regulatory and ethical concerns necessary for conducting a
study within a barangay, the researchers sought approval from Barangay (Brgy.) Captain
Concepcion D. Asilo of Brgy. San Gabriel, General Mariano Alvarez (GMA), Cavite.
Upon approval, the researchers will embark on looking for residents of the said barangay
who are qualified for their study as per the eligibility criteria. For the consent, the head of
the residence must sign the permission letter if he/she is willing to participate in the
study’s experiment.
The survey questionnaires will be given to the 15 respondents who have
previously used the bamboo leaves with natural oil-based insecticide infusion as a
contrivance made by the researchers for controlling mosquito population. Such
effectiveness assessment will be done to measure the effectiveness of the said ovitrap
upon usage by the respondents. The said questionnaires are self-made survey
questionnaires. For measuring the effectiveness of the lethal ovitrap as per the
respondents’ experience of using the lethal ovitrap, a 5-point Likert Scale technique will
be employed in the question sheets. After data gathering procedure, the data will be
tabulated and prepared for statistical treatment.
71
Data Analysis Method and Procedure
After the data was gathered, they must be statistically treated to connect it with
this study’s statement of the problem and statement of hypothesis. By such, the following
are the statistical treatments applicable concerning this study’s purpose.
1. Pearson’s R, it will be used to determine the correlation between the numbers of hour/s
upon which the bamboo leaves with natural oil-based insecticide infusion will be utilized
and its Likert Scale scoring for effectiveness by the respondents. Calderon and Gonzales
(2012) stated that in the field of statistics, this formula is often referred to as the Pearson
R test; and that it is more appropriate to conduct a Pearson correlation coefficient value to
determine how strong the relationship is between two variables. It has the formula:
𝑛(∑ 𝑥𝑦) − (∑ 𝑥)(∑ 𝑦)

𝑟=
√[𝑛 ∑ 𝑥 2 − (∑ 𝑥)2 ][𝑛 ∑ 𝑦 2 − (∑ 𝑦)2 ]
Where:
𝑟 = Pearson correlation coefficient
𝑛 = number of pairs of scores
∑ 𝑥𝑦 = sum of the products of paired scores
72
∑ 𝑥 = sum of x scores
∑ 𝑦 = sum of y scores
∑ 𝑥 2 = sum of squared x scores
∑ 𝑦 2 = sum of squared y scores
2. Chi-square, it will be used to discern if there is a significant relationship between the
numbers of hour/s upon which the bamboo leaves with natural oil-based insecticide
infusion was utilized and its Likert scale scoring for effectiveness by the respondents.
According to Calderon and Gonzales (2012), this statistical treatment is used to
determine the association between two variables; moreover, it may also be termed as chi-
square test of independence. The value of the chi-square can be solved by:
2
2
(𝑂𝑟,𝑐 − 𝐸𝑟,𝑐 )
𝑋 = ∑[ ]
𝐸𝑟,𝑐
Where:
𝑋 2 = chi-square test of independence
𝑂𝑟,𝑐 = observed frequency count at level r of Variable A and level c of Variable B
𝐸𝑟,𝑐 = expected frequency count at level r of Variable A and level c of Variable B
73

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University of Perpetual Help System GMA Campus

San Gabriel, GMA, Cavite

Senior High School Department

EFFECTIVENESS OF BAMBOO LEAVES WITH NATURAL OIL-BASED

BALDEMORO, KURT ZEDRICK M.

Senior High School Department

Background of the Study

Japanese encephalitis, malaria and filariasis (Gonzales, 2016).

Incidence of dengue, a mosquito-borne disease and a leading cause of childhood

Senior High School Department

towards the main problem, the population of mosquito.

According to Benelli (2015), the current control strategies mainly rely on

synthetic chemicals lead to a number of negative implications, including high operational

most home and garden pest control needs.

It is important to know the different stages of the mosquito’s life cycle in

Senior High School Department

attract gravid mosquito as indicated in the studies “Identification of Bacteria and

Bacteria-associated Chemical Cues that Mediate Oviposition Site Preferences by Aedes

aegypti” by Apperson et.al. (2008) and “Characteristics of Grass Infusions as

Oviposition Attractants to Aedes (Stegomyia) (Diptera: Culicidae)” by Eiras et.al.

insecticidal agent in their publication “Using Oils as Pesticides.”

With the growing concern pertinent to mosquito-borne diseases and urgency of

in General Mariano Alvarez (GMA), Cavite.

Senior High School Department

This study entitled "Effectiveness of Bamboo Leaves with Natural Oil-based

Insecticide Infusion as a Lethal Ovitrap to Mosquito Among Selected Residents of Brgy.

predicament in the country in relation to mosquito population. Because of its tropical

with the aforementioned matter.

The Philippines is an archipelagic and tropical country, with numerous bodies of

water and frequent rainfalls, plagued with mosquito-borne diseases. As a consequence,

paramount to conduct this quantitative and descriptive study.

Senior High School Department

Statement of the Problem

The general objective of this quantitative research is to determine the

effectiveness of bamboo leaves with natural oil-based insecticide infusion as a lethal

the following questions:

natural oil-based insecticide infusion was utilized?

oil-based insecticide infusion effectiveness?

Likert scale scoring for effectiveness by the respondents?

scoring for effectiveness by the respondents?

Objectives of the Study

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bamboo leaves with natural oil-based insecticide infusion effectiveness.

3. The researchers will determine if there is a significant relationship between the

Hypothesis of the Study

The hypothesis serves as an intellectual tentative answer regarding the study’s

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leaves with natural oil-based insecticide infusion effectiveness cannot be

3. The researchers believe that there is no significant relationship between the

Significance of the Study

This study must be conducted for dengue, a mosquito-borne disease, incidence

causes an estimated 50 million infections annually (World Health Organization, 2012).

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beneficiaries and donees, which are listed as the following:

natural oil infusion as a lethal ovitrap to mosquito.

Department of Health or DOH. In adherence to the national agency’s mission of

serious exertion regarding diseases transmitted and spread by mosquitoes.

Units (LGUs) fight against tropical diseases.

study regarding a related topic.

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Scope and Limitation

Insecticide Infusion as a Lethal Ovitrap to Mosquito Among Selected Residents of Brgy.

Cavite. There will only be 15 respondents from the aforementioned municipality.

To assess the effectiveness of bamboo leaves with natural oil-based insecticide