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).

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.

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 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).

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

2
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.

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.

3
 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.

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

4
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

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.

5
 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.

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,

6
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.

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).

7
 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

rest outdoor), it prefers darker colors and is a long distance flier.

8
 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.

San Pedro, Laguna Profile

According to ICT Web Development Team (2019) of San Vicente, City of San

Pedro in Laguna, the municipality is geographically located at coordinates approximately

14.3502, 121.0557, in the island of Luzon. Elevation at these coordinates is estimated at

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

Pedro, Laguna, United Bayanihan, San Pedro, Laguna

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

9
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

national capital is 26.64 kilometers (16.55 miles).

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).

San Pedro is located in Region 4A or Calabarzon. San Pedro is the boundary

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

Metro Manila for work.

Regarding the number of dengue fever cases in the municipality, Municipal

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.

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

11
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. 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

Restriction on Lifespan, Reproduction and Growth in the Mosquito Aedes aegypti.”

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.

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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

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.

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

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 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.

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

15
 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).

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

16
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

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.

Similarly, water-soluble emulsifiers stabilize vinegar as the continuous phase. In the home

17
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

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

18
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.

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

19
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.

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

20
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.

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

21
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.

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

22
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

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.

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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

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.”

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

24
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.

Furthermore, B10=Pseudomonas plecoglossicida, B11=Rhizobium huautlense,

B12=Shigella dysenteriae, B13=Citrobacter freundii, and B14=Brevundimonas

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

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) 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

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.

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

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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

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).

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

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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

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.

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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

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.

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.

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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

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%).

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%

ovicidal activity will be recorded.

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).

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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

utilized for development of oil-based insecticide as supplementary to synthetic

insecticides.

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

33
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.

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

34
an infected person during any of the previous bloodmeals, controlling gravid females is

also important to reduce dengue virus transmission.

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.

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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%.

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.

Synthesis of the State of the Art

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 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 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

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 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

in the said municipality, respectively.

37
 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),

“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

38
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

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

39
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

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.

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