Professional Documents
Culture Documents
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-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
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
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Mosquitoes are considered a global treat. It is because they are vector of diseases
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.
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).
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.
symptoms of Chikungunya virus infection are fever joint pain, headache, muscle pain, joint
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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
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.
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
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
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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
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
(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
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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.
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
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
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
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
vector insect. In the publication “Some Philippine Mosquitoes,” Ladlow (2013) had
described nine (9) genera of mosquito endemic to the country: Aedeomyia, Aedes,
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Anopheles, Culex, Desvoidea, Finlaya, Mansonia, Orabhamia and Stegomyia. Contained
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
Without knowing the species of mosquito present, such exertions are unguided.
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
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).
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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.
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
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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
According to ICT Web Development Team (2019) of San Vicente, City of San
13.9 meters or 45.6 feet above mean sea level. It shares a common border with the
following barangay(s): Calendola, San Pedro, Laguna, Poblacion, San Pedro, Laguna, San
Antonio, San Pedro, Laguna, Landayan, San Pedro, Laguna, Canlalay, Biñan, Laguna,
Nueva, San Pedro, Laguna, San Francisco (Halang), Biñan, Laguna, Bagong Silang, San
City of San Pedro has a land area of 24.05 square kilometers or 9.29 square miles
which constitutes 1.25% of Laguna's total area. (ICT Web Development Team, 2019). The
city center of San Pedro is situated at approximately 14° 22' North, 121° 3' East, in the
island of Luzon. Elevation at these coordinates is estimated at 11.8 meters or 38.8 feet
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above mean sea level. Based on the great-circle distance (the shortest distance between two
points over the surface of the Earth), the cities closest to San Pedro are Biñan, Muntinlupa,
Santa Rosa, Cabuyao, Dasmariñas, and Imus. The nearest municipalities are Carmona,
General Mariano Alvarez, Silang, Binangonan, Kawit, and Pateros. Its distance from the
The nearest towns are Carmona, Cavite with 5.11 kilometers (3.18 miles) to the
South (S2°W), General Mariano Alvarez, Cavite with 8.91 kilometers (5.54 miles) to the
Southwest (S38°W), Silang, Cavite with 17.76 kilometers (11.03 miles) to the South‑
Southwest (S30°W), Binangonan, Rizal with 18.47 kilometers (11.48 miles) to the
Northeast (N52°E), Kawit, Cavite with 18.92 kilometers (11.76 miles) to the West‑
Northwest (N61°W) and Pateros with 19.95 kilometers (12.39 miles) to the North (N2°E).
San Pedro, Laguna has a total land area 9.29 square kilometers, ranked 1,546 out
of 1,634 dated January 18 1725, 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 (2019).
between Laguna and Metro Manila, so San Pedro is known as "Laguna's Gateway to Metro
Manila". San Pedro shares boundaries with Metro Manila's southernmost city, Muntinlupa
(North) bounded with Tunasan River, Biñan (South), Dasmariñas (West), Carmona and
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Gen. Mariano Alvarez (Southwest) bound with San Isidro River. Its position makes San
Pedro a popular suburban residential community, where many residents commute daily to
Epidemiology & Surveillance Unit of San Pedro, Laguna (2019) summarized that for the
year 2019, there will be 98 cases of dengue fever reported for morbidity weeks 1-38
(January to September 2019) with 1 death. The year 2019’s number of cases is 50% lower
compared to the same period last year (98 vs. 173 cases). Of the total cases reported this
period, 98 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%). Those are the primary rationale on why the researchers had chosen the locale for
their study.
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
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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
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
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. 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
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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
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
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
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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
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.
annoyance and irritation, some species can also transmit deadly diseases. Rentokil (2017)
advocates the usage of Integrated Mosquito Management (IMM) against growing mosquito
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.”
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
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The next step is called “Larviciding.” It is a treatment to control the development
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
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
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.
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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.
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).
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
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
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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.
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
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
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
Similarly, water-soluble emulsifiers stabilize vinegar as the continuous phase. In the home
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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
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.
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
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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
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.
methyl esters serve as potent oviposition stimulants for gravid Ae. aegypti.
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
Ovitraps baited with grass infusions will be evaluated in the field by Eiras et.al.
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study “Characteristics of Grass Infusions as Oviposition Attractants to Aedes (Stegomyia)
observed in areas with no vegetation, but intense human presence, whereas Aedes
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
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.
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
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
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Oviposition Attractant for Aedes aegypti Mosquitoes,” Arbaoui and Chua (2014)
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.
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
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
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Attraction of Mosquitoes to Experimental Plant Infusions,” gravid females exhibited
oak leaves, showing that attractive odorants will be produced through microbial metabolic
activity.
bamboo or white oak leaves. Plate counts of heterotrophic bacteria will be used, as well
electrophoresis (DGGE) to show that changes in the relative abundance of bacteria and the
and A. albopictus mosquitoes to infusions. DGGE profiles showed that bacterial species
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
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diversity of bacterial species, which in turn is affected by plant species, leaf biomass, and
fermentation time.
In the thesis entitled “Oviposition Responses of the Mosquitoes Aedes aegypti and
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
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
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
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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
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
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
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
Cultures will be grown for 24 h at 28 °C with constant shaking (120 rpm) and cell densities
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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
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
The isolate labels and the matching bacterium species corresponding to closest
vesicularis.
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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
foundation in the conception of their study “Effectiveness of Bamboo Leaves with Natural
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) 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
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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
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.
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
such premise that the researchers have added five (5) mL of canola oil per 300 mL of bamboo
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
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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
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).
Oils also are easy to apply with existing spray equipment and can be mixed with many other
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
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
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
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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.
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
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
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.
(Diptera: Culicidae): Efficacy, Persistence, and Elicited Oviposition Response,” and have
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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
(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.
surfactants are only active while wet and become ineffective under drying conditions.
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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
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
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
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.
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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%
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
addition, plant essential oils are natural volatile substances obtained from a variety of plants
et.al., 2014).
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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
insecticides.
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.
biodegradable leading to fewer harmful effects on human and other animals and are
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
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and are often used for surveillance of Aedes vectors. It can be modified to render it lethal
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
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
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an infected person during any of the previous bloodmeals, controlling gravid females is
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
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.
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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%.
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.
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In line with this study’s aim of determining the effectiveness of bamboo leaves with
residents of Brgy. San Vicente in San Pedro, Laguna, 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
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 &
mosquitoes. Out of those aforementioned ailment, dengue, Chikungunya and yellow fever
In addition to that Ladlow (2013), Rentokil (2017), ICT Web Development Team
(2017), and Municipal Epidemiology & Surveillance Unit of San Pedro City provided
information on Philippines and prevalent mosquito species in it. The last two sources
focused on the geography of San Pedro City and the number of dengue cases per barangay
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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
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
of the Mosquitoes Aedes aegypti and Aedes albopictus to Experimental Plant Infusions in
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Laboratory Bioassays” by Apperson et.al. (2015) and “Oviposition Responses of Aedes
(2015) have evaluated natural leaves, for examples are bamboo leaves, white oak leaves,
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
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
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performance: “Use of the CDC Autocidal Gravid Ovitrap to Control and Prevent
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
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.
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
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