Chapter I

PROBLEM AND ITS SCOPE

RATIONALE OF THE STUDY

Biorational insecticides have recently become a new and

available or will likely become available source of

protection from insects in the near future. This is a

relatively, cheap and effective substitute for commercial

grade products. An example of a biorational ingredient

would be the Neem tree. It is commonly known as

“Mosquito” tree or “Bagalunga” in Filipino and is found in

several countries in Asia, it has been branded as an

inferior plant in some countries, despite that, this Tree is

found to be a good source of eco - friendly biorational

insecticides, due to the presence of Azadirachtin, the one

responsible for the growth-inhibiting properties,

meaning, this does not directly kill insects but is rather

involved in the life processing cycle of insect larvae. This

is very useful for both farmers and health consumers to

slowly and effectively eliminates a variety of insects,

however this study focuses on the comparison of

specified parts of the Neem based on the effects it has on

1
 disease harbouring insects, targeting mainly on mosquito

larvae.

The mosquito larva is a common household insect that

could be found in small contaminated or stagnant waters, the

eggs of a mosquito, are laid in empty buckets cans and hollow

trees. The eggs can survive without water up to year depending

on the type of specie it is, whenever a seasonal change occurs,

this results to the flooding of the incubating site where the eggs

will still hatch and can likely become a full grown mosquito.

With the current issues we have today, the accumulation

of insects has become a devastating problem to the masses.

The rapid growth of insects results in more discoveries and

diseases. This grace development led to the increasing of prices

in insecticide commodities. Masses would pay in large

quantities to reduce their risk of getting infected by disease

harboring insects, particularly mosquitoes.

A very serious issue regarding the rate of dengue fever

over 66,000 cases in the Philippines recently reported by the

Department of Health, and continues to rise as the population

of mosquitoes grow, the situation of communities will be drastic

2
and chaotic if not immediately taken care of directly from the

source.

The present study focuses on the selected part of the

Neem tree for it comparison in the effectiveness on producing

insect growth regulators.

STATEMENT OF THE PROBLEM

The purpose of the study is to compare the effectiveness

of the seed and leaf extract of the Neem tree (Azadirachta

indica) as biorational insecticide commodity for disease

harboring insects, it aims to:

 To ascertain and compare the time it takes for the

extracts to take effect on mosquito larvae.

 To identify which of the selected media will be more

potent in producing biorational insecticide.

 To aid in making decisions for future neem themed

products.

RESEARCH HYPOTHESIS

Ho – Both Neem specimens were equally potent and cannot

determine which of the specimens is more effective.

3
Ha – There will be a succeeding Neem specimen that is more

potent and can be determined which is more effective.

SIGNIFICANCE OF THE STUDY

This study greatly benefits the following:

Consumers. The study will benefit those who frequently deal

with insects such as agriculture and livestock because the

consumers will be more aware about a new alternative and can

choose which is the most effective in reducing the amount of

insects.

Manufacturers. The study will greatly benefit manufacturers

who are aiming for a better solution in the predicament of

agriculture and marine body poisoning since this study aims to

assist in making decisions and possible ways in reducing the

risk of poisoning.

Future Researchers. Since the study aims to compare the

specified parts of the Neem tree, future studies will know which

is more effective and focus on what kind of investigatory project

to make, future studies can also be based in the findings of this

research.

4
SCOPES AND DELIMITATIONS OF THE STUDY

This study is limited to identifying which part of the Neem

tree is the most effective in producing the best possible

products in the future, based on comparing its effectiveness on

mosquito larvae, though the researchers would like to have

more insects in this study in determining the most effective and

to ascertain which of the insects are susceptible to the extracts,

it is possible that the researchers cannot come to a conclusion

because of the unknown susceptibility of the other insects, the

researchers decided to use on one specimen in order to obtain

results. This study uses the Aqueous Neem Extraction in

removing the essentials as it is not synthesized and cannot

harm the environment.

DEFINITION OF TERMS

The following words are used in this study, their

corresponding meanings and explanations are given to make

5
this research more comprehensible to those who might come

across it.

Biorational Insecticides – are insecticides that harm the

targeted insect but does not harm natural predators of the

targeted insects.

Insect Growth Regulators (IGR) – are substances that

inhibits the life cycle of an insect from reaching maturity, these

take longer to kill than the traditional insecticides. Some IGRs

cause insects to stop feeding long before they die.

Azadirachtin – Initially found to be an active feeding inhibitor,

now known to affect 200 species of insects by acting mainly as

an antifeedant and growth disruptor, and such it possesses

considerable toxicity towards insects.

Potent – having great power, influence or effect.

Predecessor – something succeeded or replaced by

something else.

6
CONCEPTUAL FRAMEWORK

INPUT PROCESS OUTPUT

Determining
the specified
Independent Aqueous
parts of the
Neem
Variables: neem tree
Extraction
that is
Neem:
effective in
Seed producing a
Extract
biorational
Leaf Application
insectide.
on Mosquito
Water
Larvae
Temperature
Aiding in
Time making
decisions for
Dependent future neem
Variables:
themed
Mosquito products.
Larvae

7
 Chapter II

REVIEW OF RELATED LITERATURE

The neem plant is a fast growing and long living tree, native to

Burma in India. From there the neem tree has spread and is

now grown all over the world. In India the neem medicinal plant

is highly regarded because of its many uses and benefits.

However, the rest of the world is still unfamiliar with it.

Unfortunately. Neem is a fascinating and versatile plant and

using neem has many benefits.

(http://www.discoverneem.com/neem-plant.html)

Neem is POSSIBLY SAFE for most adults when taken by mouth

for up to 10 weeks, when applied inside the mouth for up to 6

weeks, or when applied to the skin for up to 2 weeks. When

neem is taken in large doses or for long periods of time, it is

POSSIBLY UNSAFE. It might harm the kidneys and liver.

8
(http://www.webmd.com/vitamins-

supplements/ingredientmono-577-

neem.aspx?activeingredientid=577&)

“Biorational” has only recently been proposed to describe those

insecticides that are efficacious against the target pest but are

less detrimental to natural enemies. The term at times has

been used to describe only those products derived from natural

sources, i.e. plant extracts, insect pathogens, etc. However, we

choose to define a biorational pesticide as “any type of

insecticide active against pest populations, but relatively

innocuous to nontarget organisms and therefore, non-

disruptive to biological control.” An insecticide can be

“innocuous” by having low or no direct toxicity, or by having

systemic or rapid translaminar activity or short field residual,

thereby minimizing exposure of natural enemies to the

insecticide.

(http://ipm.ifas.ufl.edu/pdfs/Biorational_Insecticides.pdf)

Aqueous Neem Extraction: Powdered seeds, kernels or leaves

are soaked overnight in water and the next morning the extract

9
is strained through a piece of cloth; the desired volume of

concentrated neem extract can then be used for desired

purpose. Aqueous extract is generally rich in water soluble

amino acids, pigments, soluble bitters and carbohydrates.

Some manufacturers use the process of freeze drying the

water-based neem extract producing a crystalline powder

called "neem bitters" that is also water soluble.

(http://www.neem-products.com/neem-extracts.html)

The biopesticide produced by extraction from the tree seeds

contains limonoids. Currently, the extraction process has

disadvantages such as contamination with fungi and

heterogeneity in the content of limonoids due to genetic,

climatic, and geographical variations. To overcome these

problems, production of limonoids from plant cell suspension

and hairy root cultures in bioreactors has been studied,

including the development of a two-stage bioreactor process

that enhances growth and production of limonoids with cell

suspension cultures of A. indica.

(https://en.wikipedia.org/wiki/Azadirachta_indica)

10
Mosquito larvae, commonly called "wrigglers," live in water

from 4 to 14 days depending on water temperature. Larvae of

almost all species must come to the surface at frequent

intervals to obtain oxygen through a breathing tube called a

siphon.

They hang with their heads down and the brushes by their

mouths filtering anything small enough to be eaten toward their

mouths to nourish the growing larvae.

They feed on algae, plankton, fungi and bacteria and other

microorganisms. During growth, the larva molts (sheds its skin)

four times. The stages between molts are called instars. At the

4th instar, the usual larva reaches a length of almost 1/2 inch

and toward the end of this instar ceases feeding. When the 4th

instar larva molts, it becomes a pupa.

(http://www.mosquito.org/biology)

This compound is found in the seeds (0.2 to 0.8 percent by

weight) of the neem tree, Azadirachta indica (hence the prefix

aza does not imply an aza compound, but refers to

the scientific species name). Many more compounds, related to

azadirachtin, are present in the seeds as well as in the leaves

11
and the bark of the Neem tree which also show strong biological

activities among various pest insects. Effects of these

preparations on beneficial arthropods are generally considered

to be minimal. Some laboratory and field studies have found

neem extracts to be compatible with biological control.

Because pure neem oil contains other insecticidal and fungicidal

compounds in addition to azadirachtin, it is generally mixed at

a rate of 1 ounce per gallon (7.8 ml/l) of water when used as a

pesticide.

(https://en.wikipedia.org/wiki/Azadirachtin)

A female Aedes mosquito can lay from 50 to 300 eggs in a

lifetime. There is some evidence that an infected female

mosquito can pass the dengue virus to her eggs, but

transmission by an infected mosquito to a human and from an

infected human to a mosquito remain the primary concerns.

(https://health.hawaii.gov/docd/files/2015/12/dengue-faq-

mosquitoes.pdf)

The main transmission is through mosquitoes that generally

acquire the virus while feeding on the blood of an infected

person. After virus incubation for eight to ten days, an infected

12
mosquito is capable, during probing and blood feeding, of

transmitting the virus for the rest of its life.

Infected female mosquitoes may also transmit the virus to their

offspring by transovarial (via the eggs) transmission, but the

role of this in sustaining transmission of the virus to humans

has not yet been defined.

(http://www.denguevirusnet.com/transmission.html)

Adrien-Henri de Jussieu (23 December 1797 – 29 June

1853) was a French botanist. Born in Paris as the son of

botanist Antoine Laurent de Jussieu, he received the degree

of Doctor of Medicine in 1824 with a treatise of the plant family

Euphorbiaceae. When his father retired in 1826, he succeeded

him at the Jardin des Plantes; in 1845 he became professor

of organography of plants. He was also president of the French

Academy of Sciences. De Jussieu was elected a Foreign

Honorary Member of the American Academy of Arts and

Sciences in 1850.

(https://en.wikipedia.org/wiki/Adrien-Henri_de_Jussieu)

13
Neem is considered a weed in many areas, including some

parts of the Middle East, most of Sub-Saharan Africa including

West Africa and Indian Ocean states, and some parts

of Australia.

Ecologically, it survives well in similar environments to its own,

but its weed potential has not been fully assessed.

In April 2015, A. indica was declared a class B and C weed in

the Northern Territory, Australia, meaning its growth and

spread must be controlled and plants or propagules are not

allowed to be brought into the NT. It is illegal to buy, sell, or

transport the plants or seeds. Its declaration as a weed came

in response to its invasion of waterways in the "Top End" of the

territory.

(https://en.wikipedia.org/wiki/Azadirachta_indica)

The female mosquitoes of Culex and most other species lay

their eggs in batches of 50 to 100, they make those deposits

on water that's collected in tree holes, ditches, and even your

dog's water dish. However, some species will lay mosquito eggs

on moist, often-flooded soil in anticipation of the next rise in

water. Those eggs can survive winter, waiting for spring or

14
summer rains to cover them over. Females usually deposit their

eggs at night, and can lay them about every third night, up to

three times. Entomologists at Oklahoma State University report

that the eggs are white when first deposited, then darken to

near black within a day.

They will hatch in one to three days, depending on the

temperature.

Eggs left on moist soil can last for up to a year, until the ground

is flooded again, before hatching. When the eggs hatch, the

mosquito larvae come out.

(https://www.megacatch.com/mosquito-faqs/mosquito-life-

cycle/)

15
 Chapter III

Methodology

RESEARCH DESIGN

II.
I. Gathering III. Testing
Preparation
of Materials the Set-ups
of Set-ups

Collection
Control and Control and
Experimental Experimental
set-up set-up
Gathering

RESEARCH SUBJECT

With the current issues of dengue outbreaks in the

Philippines, the researchers would like to contribute in making

a positive impact about lessening the population of mosquitoes

by regulating its growth in the society, to show that there are

more ways in killing mosquitoes than just the regular, old

fashion harmful commercial pesticides.

16
 RESEARCH LOCALE

The researchers conducted the experiment under the

jurisdiction of St. Peter’s College of Ormoc, lending the

researchers their support and apparatuses in producing the

product and experimentation in one of the labs in SPC.

RESEARCH INSTRUMENTATION

The researchers regard the Neem tree as the instrument

and a rather efficient tree specializing on warding of certain

insects that we conducted a comparative study to discern which

part of the Neem is the most effective.

RESEARCH PROCEDURE

I. Gathering of Materials

The selected parts of the Neem tree, primarily the fruit and

leaves, were gathered by the researchers in various locations

in Ormoc City. The materials were placed in separate containers

to ensure an organized and quick preparation for the Neem

extraction. The Researchers cultivated 45 individual specimens

of mosquito larva for the comparison of the experiment of the

Neem extracts.

17
Collection of Neem seeds and leaves

18
II. Preparation of Set-ups

The researchers gathered 2 sets of mortar and pestle, 2

pieces of dropper, a strainer, 8 pieces of filter paper, a funnel,

2 Florence flasks, a cloth bag and a commercial grade blender

for the apparatus in producing the Neem seed and leaf extracts.

With careful handling the samples were weighed using a normal

weighing scale and a gram scale to measure the exact weight

of the materials. The leaves weighed exactly 1 kilogram as the

water used for this extraction measured 5 liters. The seeds

weighed exactly 500 grams as the water used for this

extraction measured 500 milliliters.

Separation of Stems Separation of Branches

19
PROCESS FOR THE NEEM EXTRACTION

Procedure 1. For the Neem Seed Extraction, the researchers

did not remove the flesh of the fruit, however, removed the

stems and proceeded with pounding and grinding seeds that

measured 500 grams with a mortar and pestle. After the seeds

were properly pulverized, using a cloth, the researchers placed

this on top of the strainer to further avoid any small lumps or

excess pulp from escaping through, the researchers began

pouring 500 mL of water over the pulp to separate the Neem

seed extract.

Neem Seed Extraction

20
Procedure 2. After the extract was successfully obtained, the

researchers placed this in an air tight container, all the while

using another clean cloth to strain any further excess lumps

that may have passed through in the first procedure, ensuring

500 mL of pale, milky green liquid as the finishing product for

the Neem Seed Extract.

Thorough Filtration of the Neem Seed extract

21
Procedure 3. For the Neem Leaf Extraction, the leaves were

removed from their branches and were placed in a large

container with lid filled with 5 liters of water to soak the Neem

leaves overnight in the preparation for the Neem Leaf

Extraction.

Soaking Leaves Overnight

22
Procedure 4. After being left in an enclosed container for 2

days, the researchers placed 500 grams of Neem leaves and

500 mL of water in the commercial grade blender, the Neem

leaves were blended and pulverized properly, the leaves were

then filtered using a piece of filter paper on top of the strainer,

this procedure was done twice to further remove any clumps of

leaves that may have gone through. The given results show

500 mL of murky green liquid as the finishing product.

Neem Leaf Extraction

23
EXTRACT APPLICATION ON MOSQUITO LARVAE

Procedure 1. The researchers prepared three enclosed glass

containers, 2 containers are filled with 97 mL of water, 1

container is filled with 100 mL of water and 15 mosquito larvae

in each container, 2 containers serves as experimental setups,

the first setup, labeled setup A is added with 3 mL of Neem

seed extract, the second setup, labeled setup B is added with

3 mL of Neem leaf extract and one container serves as the

control setup and labeled as setup C where no extracts were

added. The researchers set a 4 hour interval in the inspection

of mosquito larvae.

Mosquito Larvae Set-ups

24
 CHAPTER IV

RESEARCH AND DISCUSSION

PRESENTATION

The Researchers obtained 2 distinct parts of the Neem

tree for the comparison of its effectiveness in producing a

biorational insecticide. Setup A contains Neem seed extract,

setup B contains neem leaf extract and setup C does not

contain any extract. The Aqueous Neem Extraction Process was

chosen for the extraction because of its natural properties and

does not contain any chemical in producing the Neem extracts.

Based on the table below shows the progress of the Neem seed

extract on the mosquito larvae.

Table 1. Setup A with 3mL of Neem Seed Extract

Observation Time

time 4 hrs. 8 hrs. 34 hrs. 48 hrs.

State alive Alive Alive dead

Behavior Hyper Active Weak still

Dead larvae None None 5 15

25
On the table below shows the progress of the Neem Leaf extract

on the mosquito larvae.

Table 2. Setup B with 3mL of Neem Leaf Extract

Observation Time

Time 4 hrs. 8 hrs. 34 hrs. 48 hrs.

State alive Alive Alive 5 alive

Behavior Hyper Hyper Active weak

Dead Larvae none None None 10

On the last table, it shows the progress of the Setup C where

no extracts were added.

Table 3. Setup C, no extracts were added

Observation Time

Time 4 hrs. 8 hrs. 34 hrs. 48 hrs.

State alive Alive Alive poor

Behavior Hyper Hyper active weak

Dead Larvae none None None none

26
FINDINGS

This study was to determine which part of the Neem tree

would be effective in the making of a biorational insecticide. In

the time the researchers spent observing the mosquito larvae

setups, they have found that the Neem seed extract is more

effective in biorational insecticide than the Neem leaf extract.

The Neem seed extract is more appropriate to become an

ingredient for a biorational insecticide than the Neem leaf, and

as to further conclude why the Neem seed extract was more

effective than the Neem leaf extract, it is that the seeds contain

more of the azadirachtin component in which case more potent

in its toixicity level towards mosquito larvae and other insects

compared to the Neem leaf extract.

27
DISCUSSIONS

As stated in Wikipedia on the page of Azadirachta indica,

the Neem tree can easily grow just about anywhere and does

not require any high maintenance to care for this type of tree,

branded as a class b and c weed by some parts of the world,

some of the countries do not know the full potential of the Neem

tree yet.

In addition to this, the researchers decided to plant this

tree in their respective homes to show that this tree deserves

the acknowledgment and credit than it usually has to also be

able to pass on this knowledge to people living in urban areas.

As years pass, the growth of healthy crops are

increasingly slim, resulting to risky and contaminated fruits and

vegetables because of the lack of awareness in the growth of

Biorational Insecticides, though this does little harm to full

grown insects, it will still greatly affect their offspring which will

result in suppressing the target insects’ growth and minimizing

the poison that accumulates in both terra and marine habitats.

28
 Chapter V

Summary, Conclusions and Recommendations

SUMMARY

The study entitled “Effectiveness between the Seed and

Leaf Extract of the Neem Tree (Azadirachta indica) as

Biorational Insecticide.” sought to discern which part of the

Neem tree is preferable in producing an effective and cheap

biorational insecticide in targeting mosquito larvae. The

researchers extracted the Neem seed and leaves using the

Aqueous Neem Extraction Process. After observing for

approximately 3 days, the researchers identified the Neem

seed extract as the most effective part of the Neem tree in

producing a Biorational insecticide for the targeted insect:

mosquito larvae.

29
CONCLUSION

Based on the researchers’ comparative analysis, the

following conclusions were drawn:

The Neem Seed Extract is the selected media that is more

potent in producing biorational insecticide.

It is determined to assist in making decisions for future Neem

themed products.

The Neem Seed Extract is quicker and more effective to be a

biorational insecticide than the Neem Leaf Extract.

30
RECOMMENDATIONS

Based on the garnered results of this study, the

researchers recommend the following:

Environmental Organizations should conduct activities in

promoting the unfamiliarity to the public, like the Neem Tree

that can serve as an effective substitute for manufactured

harmful commercial products of today.

Manufacturers and consumers should try producing the

extract as a DIY biorational insecticide for the protection

against particular or targeted insects and ensure the safety of

their homes from harmful insects, consumers can also easily

plant this tree without any hassle to ward off pesky insects.

Future studies could conduct a research of how many

different species of insects are susceptible to the neem seed

extract and can even create their own variety of products using

the neem seed extract as the main ingredient.

31
 BIBLIOGRAPHY

Book:

African Journal of Biotechnology, 2(9): 325-327

Internet Sources:

(http://www.discoverneem.com/neem-plant.html)
(http://www.webmd.com/vitamins-
supplements/ingredientmono-577-
neem.aspx?activeingredientid=577&
(http://www.neem-products.com/neem-extracts.html)

(http://ipm.ifas.ufl.edu/pdfs/Biorational_Insecticides.pdf

(https://en.wikipedia.org/wiki/Azadirachta_indica)

(African Journal of Biotechnology, 2(9): 325-327)

(http://www.neemfoundation.org/about-neem/neem-

iniagriculture/pest-management/)

(http://www.wisegeek.com/what-is-neem-bark.htm)

(http://www.discoverneem.com/neem-leaf.html)

(https://en.wikipedia.org/wiki/Azadirachta_indica#Tradi

tional_medicinal_use)

32
(https://en.wikipedia.org/wiki/Adrien-Henri_de_Jussieu)

(https://www.megacatch.com/mosquito-faqs/mosquito-life-

cycle/)

People: Government Official from the Department of Health

33
Name : John Gregart P. Capalla

Address : Prk.4 Brgy. Linao, Ormoc City

Birth Date : March 7, 2001

Birth Place : Ormoc City, Leyte

Experience during Experimentation :

- I really had so much fun. The best part for me was

probably the gathering of materials, most especially during the

collection of Neem Seeds. I and my team were so happy when

we got the overall outcome. All efforts did really pay off, and

we were all satisfied with the success of the study.

34
Name : Justine S. Magan

Adress : Prk. Gumamela Brgy. Punta, Ormoc City

Birth Date : October 7, 2000

Birth Place : Ormoc City, Leyte

Experience during Experimentation :

- I’ve learned a lot of new lessons in which I could

share to other people. And time spent together with my co-

researchers was a total blast. I hope our research study would

be put to great use, and would serve as an inspiration to all

those aspiring researchers.

35
Name : Shanaine Ann C. Mosende

Adress : Brgy. Don Felipe Larrazabal, Ormoc City

Birth Date : April 18, 2001

Birth Place : Ormoc City, Leyte

Experience during Experimentation :

- I experienced a lot of different things when we began to

do this research, and was shocked that the pungent odor

of the extract was especially strong because of its

concentrated juices but overall satisfied with the outcome

of the study.

36