Research Plan

Research Title: Larvicidal Effect of Neemtree Date to Start: Jan 08, 2023
(Azadirachta Indica) Leaves Extract
Proponents: Jed Laurenze P. Bautista Date to End: Feb 26, 2023
Khervine Chayze M. Valeros
Baby Jane E. Villanueva

Rationale
Mosquitoes are common, flying insects that live in most parts of the world. Over 3,500

types of mosquitoes can be found worldwide. Not all mosquitoes bite people or animals. When

mosquitoes bite people, the most common reactions to the bite are itching and swelling. Some

mosquitoes can be vectors. A vector is an animal, insect, or tick that spreads pathogens (germs)

to people and animals. The germs (viruses and parasites) that mosquitoes spread can make one

sick. Mosquito-borne diseases are those spread by the bite of an infected mosquito. Diseases that

are spread to people by mosquitoes include Zika virus, West Nile virus, Chikungunya virus,

dengue, and malaria. Employers should protect workers and workers should protect themselves

from diseases spread by mosquitoes. Dengue is a viral infection transmitted to humans through

the bite of infected mosquitoes. The primary vectors that transmit the disease are Aedes aegypti

mosquitoes and, to a lesser extent, Ae. Albopictus. The virus responsible for causing dengue, is

called dengue virus (DENV). Aedes aegypti is the carrier of the dengue virus. It has been

recognized be the most threatening disease to the population. The ability of this mosquito to

transmit diseases is highly dangerous, the diseases are spread to people by the infected mosquito

bite which then later causes mosquito-borne diseases like dengue. .

Dengue viral infection has become an increasing global health concern with over two-

fifths of the world’s population at risk of infection. It is the most rapidly spreading vector borne

disease, attributed to changing demographics, urbanization, environment, and global travel. It

continues to be a threat in over 100 tropical and sub-tropical countries, affecting predominantly

children. (Sarah Murrell, 2011) The global spread of dengue fever within and beyond the usual

tropical boundaries threatens a large percentage of the world's population, as human and

environmental conditions for persistence and even spread are present in all continents. (Alfonso

Guzman, 2010) In the Philippines, dengue is probably the most well-known and feared tropical

disease. The first recorded dengue epidemic in Southeast Asia occurred in Manila in 1954, and

dengue has since remained endemic. 1 In 2019, 437,563 cases were recorded in the Philippines,

contributing to the highest dengue cases ever recorded globally. (Erika P. Ong, 2022)

In order to control the dengue virus disease, many people use mosquito repellent,

larvicides, and chemicals to repel and kill mosquitoes. Nowadays, many larvicides are available

in many stores and markets. However, some larvicides that are bought in stores is not as good as

the researcher’s research findings. In addition, according to Mahmoud Osanloo (2019), frequent

use of synthetic larvicides has led to the development of resistance in many species of

mosquitoes as well as risk of environmental pollution.

In this connection, it is concerned to look on environmental and economically friendly

like plants that can be a solution to this problem. Application of active toxic agents from plant

extracts as an alternative mosquito control strategy was available from ancient times. (Anupam

Ghosh, 2012) In the Philippines, several plants have been explored and tested for its larvicidal

activity. Neemtree (Azadirachta Indica) is one of the plants that have been investigated for its

larvicidal activity, azadirachtin contents. Neemtree (Azadirachta Indica) leaves contains

azadirachtin, which is a predominant insecticidal active ingredient, having antefeedent,

ovipositional deterrence repellency, growth disruption, sterility and larvicidal action against

insects. There are various reports of control of mosquito breeding under field conditions.

(Virendra K. Dua, 2009)

 The researchers come up to the study, “Larvicidal Effect of Neemtree (Azadirachta

Indica) Leaves Extract” in order to help decrease the cases of people getting bitten by

mosquitoes causing many diseases here in our community and to test if Neemtree (Azadirachta

Indica) can be used as an effective larvicide.

Research Questions

This study aims to determine the larvicidal effect of neemtree (Azadirachta Indica) leaves

extract.

It specifically seeks to answer the following questions:

1. How effective is the neem leaves as a larvicide against mosquitoes in terms of

mortality rate?

2. Is there a significant difference between the control group and the experimental set-

ups against mosquitoes?

3. Is there a significant difference among the different concentrations of neem leaves

(Azadirachta Indica) extract against mosquitoes?

Hypothesis

 There is no significant difference between the control group and the experimental set-ups

against mosquitoes?

 There is no significant difference among the different concentrations of neem leaves

(Azadirachta Indica) extract against mosquitoes?

Expected Outcomes
 The research study “Larvicidal Effect of Neemtree (Azadirachta Indica) Leaves Extract”

is expected to have a positive results that will prove that the Neemtree (Azadirachta Indica) has

potential as an effective larvicide.

DESCRIPTION IN DETAILS

A. Preparation of Materials

One pack of neem leaves were washed then placed in a container. A (800) grams of

coconut oil were placed in a glass bowl. A glass bowl is used as the container of the coconut oil.

Tap water is also used in washing the neem leaves. A grinding jar, boiling base oil, pan, stove,

and glass jar with a lid is placed in the table for making the neem oil. A pair of scissors, 1 pack

of yeast, 1 tape, and 1 plastic bottle will be used in making a mosquito trap.

B. Procedures

One pack of neem leaves were collected then placed in a container. (800) grams of

coconut oil were also collected and placed in a glass bowl. The neem leaves were washed using

tap water. The neem leaves was then transferred into a grinding jar then added 10 drops of water

into it.

The washed neem leaves were grinded and added them to the boiling base oil. 1 cup of

coconut oil were added in a pan, then the grinded ingredients were added into it, then mixed and

cooked it at 50 °C for 15 minutes. The mixture was boiled for 15 minutes then it was removed

from the fire and allowed it to rest for 2 hours or more. The liquid were then filtered into a glass

jar and covered it with a tight lid. Then it was stored in a cool dry place where direct sunlight

cannot reach it.

Risk and Safety

In the study “Larvicidal Effect of Neemtree (Azadirachta indica) Leaves Extract,” the

researchers identified the possible risk in conducting the experiment.

Since the researchers will be handling neem oil which may cause an adverse reaction in

someone with sensitive skin or a skin disorder like eczema, it is advisable to use proper

laboratory suit.

Data Analysis

The researchers will be determining the effectiveness in mortality rate. The effectiveness

will be determined after the process of experimentation. T-test will be used to determine if there

is a significant difference between the control group and the experimental set-ups against

mosquitoes. Analysis of Variance (ANOVA) will also be used to determine if there is a

significant difference among the different concentrations of neem leaves (Azadirachta Indica)

extract against mosquitoes.

Bibliography

Sarah Murrell, Suh-Chin Wu, Michael Butler, Review of dengue virus and the development of a

vaccine, Biotechnology Advances, Volume 29, Issue 2, 2011, Pages 239-247, ISSN

0734-9750, https://doi.org/10.1016/j.biotechadv.201.11.008.

Alfonso Guzman, Raul E. Istúriz, Update on the global spread of dengue, International Journal

of Antimicrobial Agents, Volume 36, Supplement 1, 2010, Pages S40-S42, ISSN 0924-

8579, https://doi.org/10.1016/j.ijantimicag.2010.06.018.
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Perspectives and lessons from the Philippines’ decades-long battle with dengue, Volume

24, 100505, 2022, https://doi.org/10.1016/j.lanwpc.2022.100505.

Cassie C. Jansen, Nigel W. Beebe, The dengue vector Aedes aegypti: what comes next,

Microbes and Infection, Volume 12, Issue 4, 2010, Pages 272-279, ISSN 1286-4579,

https://doi.org/10.1016/j.micinf.2009.12.011.

Liu-Helmersson J, Stenlund H, Wilder-Smith A, Rocklöv J (2014) Vectorial Capacity of Aedes

aegypti: Effects of Temperature and Implications for Global Dengue Epidemic Potential.

PLoS ONE 9(3): e89783. Doi:10.1371/journal.pone.0089783,

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0089783.

Dua, V.K., Pandey, A.C., Raghavendra, K. et al. Larvicidal activity of neem oil (Azadirachta

indica) formulation against mosquitoes. Malar J 8, 124 (2009).

https://doi.org/10.1186/1475-2875-8-124.