INSECTICIDAL ACTIVITY OF PATCHOULI (Pogostemon heyneanus) AND

KAKAWATE (Gliricidia sepium) LEAF EXTRACTS AGAINST MOSQUITO

A Research Project

Presented to the

Faculty of Senior High School

INFANT JESUS SCHOOL

Kalibo, Aklan

In Partial Fulfillment

Of the Requirements for the Subject

CAPSTONE PROJECT

Bianca R. Nemis

Christine Grace N. Popes

Fhenzie Dhana C. Retoriano

March 30, 2019

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AKNOWLEDGEMENT

The framework of success in the pursuit of this study lies behind the guidance and

participation of the following persons. The researchers would like to express their sincere and

heartfelt gratitude and immeasurable appreciation to the following people whose services,

support and shared knowledge at every point turned the study to a success.

Great Almighty, the Beloved Father, for the source of life and strength, of knowledge

and wisdom that enable the researchers to perform well and use their sterling capacity to the

fullest. Also, for His guidance and mercy through every circumstance encountered that hindered

the researchers for the final completion of the study. His endless love indeed makes all things

possible.

Sir Jimmy Boy Estrellas, the school’s director/principal, for his valuable comments and

constructive suggestions that immensely improved the researchers’ study.

Ms. Kareen Retoriano, the research adviser, for her supervision, patient guidance, time

and immense knowledge that assisted the researchers in many ways right through course of this

work. Furthermore, for offering her valuable effort and never-ending support during hard times

of this study, her creative advices and suggestions helped the researchers surpassed the

challenges encountered. It is with her supervision from start to end that this work came into

existence.

Ms. Jyanee Yecla, the researchers’ mentor, who provided insight and expertise that

greatly, assisted the researchers during the planning and development of the product. The

knowledge she shared and useful critiques she contributed notably helped the researchers.

Ms. Jale Ann Pearl Yecla, for assisting the researchers in doing the Capstone Product,

her critiques and useful suggestions helped the development of the product.
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 3

Mrs. Miralyn Mationg, the vice principal/registrar, whose reminders and constant

motivation encouraged the researcher to meet the deadlines and progress on schedule; for her

approval for the permission that were needed for the completion of research project.

Ms. Ma. Rosalie Zonio, for her act of generosity and willingness to thoroughly scan the

research paper for possible grammatical errors.

Mr. Eleandro Dominguez, school’s mathematics coordinator, for the help he rendered

toward the statistical analysis of data.

Mrs. Jasmin Nicolas, the researchers’ class adviser, for her vital support and enthusiastic

encouragement for this study.

Mrs. Mary Joy Fuentes, for her moral support and invaluable time extended with

unconditional love in assisting the researchers in their Capstone Project, her words of wisdom

immensely influenced the researchers to strive harder for the completion of the project.

Mr. Paterno Luciano, the school’s laboratory technician, for his generosity and

willingness to offer his valuable time in guiding the researchers during experimentation. Further,

for his effort to respond with the researchers’ request in preparing the equipments needed for the

study.

To the researchers’ parents, Mr. & Mrs. Joel Nemis, Mr. & Mrs. Donato Popes, Mr. &

Mrs. Zalde Retoriano, for the unceasing love, prayers, and sacrifices for the completion of the

study and throughout this academic endeavor. They not only assisted the researchers financially

but also extended their support morally and emotionally. This research is heartily dedicated for

all their unwavering sacrifices.

Lastly, the researchers are immensely grateful to each and every one who did not hesitate

to offer their helping hands all through this research.

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ABSTRACT

Leaf extracts of patchouli (Pogostemon heyneanus) and kakawate (Gliricidia sepium)

were evaluated for their insecticidal activity against adult mosquito. There were 12 cages filled

with 10 mosquitoes each. These insects were exposed to four different treatments namely;

Treatment A (75% Patchouli / 25% Kakawate), Treatment B (50% Patchouli / 50% Kakawate),

Treatment C (25% Patchouli / 75% Kakawate), and Treatment D (No Treatment Received)

which served as a control. The treatments made had three replications each. Insecticidal effect

was observed after two hours of exposure to the formulated leaf extracts. Highest mortality was

recorded on mosquito treated with 50% patchouli (P. heyneanus) and 50% Kakawate (G.

sepium). The leaf extracts showed a low insecticidal activity against mosquito in Treatment A.

All treatments tested showed various level of insecticidal activity based on the mortality rate.

The analysis of variance showed that there was a difference among the three different treatments

and the control (p > 0.05). This study presented that the insecticidal activity of patchouli

(Pogostemon heyneanus) and kakawate (Gliricidia sepium) against mosquito might be useful as

botanical insecticide and safer alternatives than other chemically-based insecticide.

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TABLE OF CONTENTS

TITLE PAGE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

ACKNOWLEDGEMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

ABSTRACT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

TABLE OF CONTENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

CHAPTER

I INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Background of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Statement of the Problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Hypotheses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Conceptual Framework . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Scope and Limitations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Significance of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Definition of Terms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

II REVIEW OF RELATED LITERATURE AND STUDIES . . . . . . . . . . . 14

Mosquito . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Three Genera of Mosquito . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Chemical Insecticide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

Human & Animal Threat, Environmental Contamination and

Resistance of Insecticide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

Patchouli (Pogostemon heyneanus) . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Kakawate (Gliricidia sepium) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

III METHODOLOGY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
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Materials and Apparatus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

Experimental Design and Treatments . . . . . . . . . . . . . . . . . . . . . . . . 23

Preparation of Plant Extracts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Collection of Mosquito Specimens . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Insecticidal Bioassay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Locale of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Statistical Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

IV RESULTS AND CONCLUSION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Data Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Summary of Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Recommendation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

V PROJECT DESCRIPTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
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CHAPTER I

INTRODUCTION

Background of the Study

Mosquito is one of the most abundant insect in urban and rural environment and has been

associated in the transmission of numerous deadly diseases to humans. Philippines, a tropical

country and a biodiversity hotspot, is highly prone to the incidence of mosquito-borne diseases.

Dengue had become one of the most alarming public health issues in recent years and had pose

danger to the lives of many Filipinos. To get rid of the diseases caused by these blood-sucking

insects, most people utilize high amounts of commercially available insecticides. These products

are chemical-based which are known to contain compounds that are harmful to human health,

environment and toxicity to non-target organisms (Mandavgane et al., 2005). The negative

effects associated with the use of these products have driven the researchers to formulate organic

insecticides which are effective, safer, and affordable than currently available products (Gillij,

2007).

Patchouli (Pogostemon heyneanus) is a green and tropical herbaceous plant belonging in

the family Lamiaceae. It is known for its expensive essential oil, used as a source in perfume and

healthcare industries. Traditionally, the boiled root of the plant is used in curing cough and

asthma while the leaf is used for headache, jaundice, etc. (Dharmadasa et al., 2014). On the other

hand, kakawate (Gliricidia sepium) is a leguminous tree belonging to the family Fabaceae. The

leaves contain antifungal and antibacterial properties (Jose & Reddy, 2010). In the Philippines,

kakawate is usually used by the farmers along rice paddies to reduce rice insect pests in the fields

(Navasero et al., 2014).

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This study sought to scientifically determine the insecticidal activity of patchouli

(Pogostemon heyneanus) and kakawate (Gliricidia sepium) leaf extracts against mosquito.

Although there were studies conducted to assess the insecticidal activity of kakawate, there were

no previous studies of its insecticidal effect when combined to patchouli thus, this is the aim of

the study.

Statement of the Problem

Generally, the study aimed to scientifically determine the “Insecticidal Activity of

Patchouli (Pogostemon heyneanus) and Kakawate (Gliricidia sepium) Leaf Extracts against

Mosquito.”

Specifically, this study was conducted to answer the following questions:

1. What are the percent mortality rates of mosquito based on different treatments of

patchouli (Pogostemon heyneanus) and kakawate (Gliricidia sepium) leaf extracts?

2. What concentration of patchouli (Pogostemon heyneanus) and kakawate (Gliricidia

sepium) leaf extracts provided the highest insecticidal activity against mosquito?

3. Is there is a significant difference on the insecticidal activity among different treatments

of patchouli (Pogostemon heyneanus) and kakawate (Gliricidia sepium) leaf extracts?

Hypotheses of the Study

Based on the problems presented, this study derived with the hypotheses that state:

Ha: There is a significant difference on the insecticidal activity against mosquitoes of different

formulations of patchouli (P. heyneanus) and kakawate (G. sepium) leaf extracts.
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 9

Ho: There is no significant difference on the insecticidal activity against mosquitoes of different

formulations of patchouli (P. heyneanus) and kakawate (G. sepium) leaf extracts.

Conceptual Framework

The conceptual framework of the study is shown in Figure 1 below.

Independent Variables Dependent Variables

Concentration of Patchouli Insecticidal Activity against

(Pogostemon heyneanus) and Kakawate Mosquito
(Gliricidia sepium)

Figure 1. Research paradigm showing the relationship of independent and dependent variables.

Scope and Limitations

The study covered a concise introduction about mosquito-borne diseases, chemical

insecticide and nature of patchouli (Pogostemon heyneanus) and kakawate (Gliricidia sepium). It

focused on the insecticidal activity of patchouli (Pogostemon heyneanus) and kakawate

(Gliricidia sepium) against mosquito. The researchers utilized experimental design in testing the

insecticidal activity of both plants using different concentrations. The study is only limited to the

leaf extracts of patchouli (P. heyneanus) and kakawate (G. sepium) plants. The mixtures of two

botanical leaf extracts were only investigated and did not include the insecticidal activity of each

pure extracts. The insecticidal properties of patchouli (P. heyneanus) and kakawate (G. sepium)

were not also investigated in this study. The researchers used natural extraction to obtain the leaf
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 10

extracts of patchouli (Pogostemon heyneanus) and kakawate (Gliricidia sepium). Different

concentrations (Treatment A with 75% patchouli / 25 kakawate, Treatment B with 50% patchouli

/ 50 kakawate, Treatment C with 25% patchouli / 75 kakawate and Treatment D as a control)

with three replicates were made to determine the most effective among the treatments based on

mortality rate of mosquito.

Significance of the Study

The study assessed the insecticidal activity of patchouli (Pogostemon heyneanus) and

kakawate (Gliricidia sepium) against mosquito. The importance of this study is to produce cost

efficient, environmental friendly and toxic-free insecticide by the use of all-natural components

specifically patchouli (P. heyneamus) and kakawate (G. Sepium). The results derived from the

study will be beneficial to the following:

Community. The finding of the study will help the community to produce their own

organic mosquito sprays and prevent mosquito-borne diseases, instead of purchasing

commercially available products.

Industry. The results of the study will prompt the pharmaceutical industry to use natural

materials from patchouli (P. heyneanus) and kakawate (G. sepium) in producing

insecticidal products which are toxic-free, cost effective, and environment friendly. It can

be an alternative to chlorinated hydrocarbons as an active ingredient.

Environment. The findings of this study will be beneficial to the environment in

becoming more sustainable in a way of reducing toxic chemicals released by a

chemically-made mosquito sprays. This will also reduce the use of the chemical-based

products that bring harmful effect to both humans and environment.

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Future Researchers. The information derived from the study will serve as underpinnings

for researches relevant to this. Moreover, the study will impact literature reviews relevant

to the insecticidal activity of patchouli (P. heyneanus) and kakawate (G. sepium)

especially against mosquito.

Definition of Terms

The following terms are defined in order to provide understanding on how the terms were

used in the study:

Concentration. The amount of a component in a given area or volume. (www.merriam-

webster.com/)

In the study, it referred to the percentage by volume of patchouli (Pogostemon

heyneanus) and kakawate (Gliricidia sepium) leaf extracts in a solution and is described as 75%

patchouli / 25% kakawate, 50% patchouli / 50% kakawate, and 25% patchouli / 75% kakawate.

Exposure. The condition of being presented to view or made known. (www.merriam-

webster.com/)

In the study, it referred to the length of time in which the mosquitoes were exposed in

different treatments.

Insecticide. Any toxic substance that is used to kill insects and primarily to control pests

that infest cultivated plants or to eliminate disease-carrying insects in specific areas.

(https://www.britannica.com/)
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 12

In the study, it referred to the mixture of two organic leaf extracts of patchouli

(Pogostemon heyneanus) and kakawate (Gliricidia sepium) that is used to kill insects specifically

mosquitoes.

Kakawate (Gliricidia sepium). It is a perennial, medium-sized (2-15 m high) legume tree

which is mostly deciduous during the dry season but is reported to remain evergreen in humid

areas. (www.feedipedia.org/)

In the study, it pertained to one of two green organic plant used as major component in

creating insecticide by extracting the leaf extracts needed to formulate different concentrations of

the said insecticide. This plant is also investigated combined with patchouli to determine its

insecticidal activity.

Mosquito. It is a slender long-legged fly with aquatic larvae.

(https://en.oxforddictionaries.com/)

In the study, it referred to the subject in the investigation of the insecticidal activity of

patchouli (P. heyneanus) and kakawate (G. sepium). Each treatment with three replications

consisted of 10 adult mosquitoes.

Patchouli (Pogostemon heyneanus). It is species of plant from the family Lamiaceae

which is cultivated for its fragrant leaves in South and South-East Asia and also outside this

region (uses.plantnet-project.org/)
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In the study, it referred to plant that was tested for its suitability and its insecticidal

activity. It was also pounded to obtain the leaves extracts that was used together with kakawate

(Gliricidia sepium) in formulating insecticide.

Replicate. It is a nature of a repetition of a scientific experiment or trial.

(https://en.oxforddictionaries.com/)

In the study, it pertained to every treatment that appears more than once in the experiment

to provide means for estimating experimental error. It is also referred to the number of trials in

each treatment.

Treatment. The techniques or actions customarily applied in a specified situation

(www.merriam-webster.com/)

In the study, it is referred to the different concentration of patchouli (Pogostemon

heyneanus) and kakawate (Gliricidia sepium) leaf extracts in the experimental group and is

described as Treatment A, Treatment B, Treatment C and Treatment D.

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CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

This chapter presents literatures and studies of other related researches which the

researchers reviewed and found significant to the present study. This section cites the literature

references about mosquito, chemical insecticide, patchouli (Pogostemon heyneanus) and

kakawate (Gliricidia sepium).

Mosquito

Mosquitoes are kind of insects which belong to the order Diptera, true flies and have

been around for over 30 million years. Mosquitoes are said to be like flies because of their wings

with scales, long legs and the females which consist of long mouth called proboscis in piercing

skin. There are approximately 2,700 species of mosquitoes with the majority belonging to three

major genera namely, Aedes, Anopheles, and Culex. Mosquitoes are attracted to heat, light,

perspiration, body odor, lactic acid and carbon dioxide. They use chemical, visual and heat

sensors to locate their prey. The chemical sensors of mosquitoes are used to detect carbon

dioxide and lactic acid from up to 100 feet away while heat sensors to detect warm-blooded

mammals and birds in their vicinity (Freudenrich, 2001). According to National Pest

Management Association, mosquitoes in general are vectors of numerous diseases including

malaria, yellow fever, dengue, encephalitis, Chikungunya.

Three Genera of Mosquito

Three important genera of mosquito were described by Rogers (2019) namely:

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 15

Anopheles Mosquitoes

Anopheles is the only known carrier of malaria, which also transmits filariasis

and encephalitis. Anopheles mosquitoes are easily recognized in their resting position, in which

the proboscis, head, and body are held on a straight line to each other but at an angle to the

surface. The life cycle is from 18 days to several weeks.

Culex Mosquitoes

The genus Culex is a carrier of viral encephalitis and, in tropical and subtropical climates,

of filariasis. It holds its body parallel to the resting surface and its proboscis is bent downward

relative to the surface. The life cycle, usually 10 to 14 days, may be longer in cold weather. The

northern house mosquito (C. pipiens) is the most abundant species in northern regions, while the

southern house mosquito (C. quinquefasciatus) is abundant in southern regions, namely the

tropics and subtropics.

Aedes Mosquitoes

The genus Aedes carries the pathogens that cause yellow fever, dengue, Zika fever, and

encephalitis. Like Culex, it holds its body parallel to the surface with the proboscis bent down.

The wings are uniformly coloured. The life cycle may be as short as 10 days or, in cool weather,

as long as several months.

Chemical Insecticide

The use of synthetic insecticide originated in the mid-20th century, it is a powerful tool

and primary agents to control insects all over the area. In general they penetrate insects readily
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 16

and are toxic to a wide range of species. They have also been essential in improving the health of

both humans and domestic animals because through their use and controls the cases of malaria,

yellow fever, and typhus, among other infectious diseases reduced in many areas of the world.

The main synthetic groups that mostly contain the insecticide are the chlorinated hydrocarbons,

organic phosphates (organophosphates), and carbamates. With the dangerous compounds that

constitute its formulation, there are possibilities that it may pose serious threats misapplied,

spilled or misused. But harmful effects of these chemicals remain essential in modern agriculture

despite their environmental drawbacks.

Human and Animal Threat, Environmental Contamination and Resistance of Insecticides

The use of synthetic insecticides has also resulted to several serious issues. The chief

issue is the contamination of the environment and the species develop resistance. Since

insecticides are poisonous compounds, the accumulation of some in the environment can affect

and pose a serious threat to both wildlife and humans besides of harmful insects. For instance, it

contains chlorinated hydrocarbons such as DDT, aldrin, dieldrin, heptachlor, and BHC which

contaminate the soil and water supplies. The stability of DDT and its relatives leads to their

accumulation in the bodily tissues of insects that constitute the diet of other animals higher up

the food chain, with toxic effects on the latter. Thus, studies found that the effects of DDT and its

relative caused a decrease of population in birds of prey such as eagles, hawks, and falcons.

Also, the contaminated water can lead to wildlife poisoning or fish kills.

A person who drinks water contaminated with insecticide from runoff can experience

acute to chronic poisoning effects. Insecticide runoff can also negatively impact surrounding
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 17

wildlife by killing or poisoning food supplies such as insects or plants. Excess runoff from

insecticides may also kill fish.

Another problem with insecticides is the tendency of some target insect populations to

develop resistance as their susceptible members are killed off and those resistant strains that

survive multiply, eventually perhaps to form a majority of the population. Resistance denotes a

formerly susceptible insect population that can no longer be controlled by a pesticide at normally

recommended rates. Furthermore, the World Health Organization (WHO) defines resistance

to insecticides as "changes in an insect that increase its ability to withstand or overcome the

effects of one or more insecticides.” Insects that developed resistance to first insecticide may

also resistant to a second which contains a similar mode of action to the first. Insecticides may

also encourage the growth of harmful insect populations by eliminating the natural enemies that

previously held them in check.

In addition, animals are not target of these harmful compounds, but also to humans. The

use of one common organophosphate, parathion, was drastically curtailed in the United States in

1991 owing to its toxic effects on farm laborers who were directly exposed to it. According to

the University of Missouri Extension's website, 24 hours of exposure may lead to acute allergic

reactions such as difficulty in breathing, skin and nose irritation. Furthermore, insecticides can

cause chronic poisoning even in small amount when a person is exposed in long period. A

person can experience chronic poising when drinking contaminated water with insecticide. It can

lead to nervousness, slow movement, twitching and a decline in good health.

Because of the problems associated with the heavy use of some chemical insecticides,

current insect-control practice combines their use with biological methods in an approach

called integrated control. In this approach, a minimal use of insecticide may be combined with
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 18

the use of pest-resistant crop varieties; the use of crop-raising methods that inhibit pest

proliferation; the release of organisms that are predators or parasites of the pest species; and the

disruption of the pest’s reproduction by the release of sterilized pests (Rogers, 2018).

Patchouli (Pogostemon heyneanus)

Patchouli (Pogostemon heyneanus) is a tropical herbaceous aromatic plant, commonly

called as ‘patchouli’. Patchouli (P. heyneanus), a native of the Indo-Malaysian region, is a highly

aromatic herb widely distributed and frequently cultivated in India, Philippines, Malaysia, China

and South America. It is a large, straggling, evergreen perennial plant with stems that are woody

at the base. It can be erect, but is often semi-scandent, producing long horizontal branches up to

1.5 metres long. The plant is an important source of pogostemonine, patchouli resinoid, and

trimethylamine, and the oil extracted from the leaves contains patchoulene, patchouli alcohol,

and eugenol as the major components. Moreover, the plant is known for its essential oil, used as

a source in perfume as well as in healthcare industries. Traditionally, the root decoction of the

plant is used in cases of cough and asthma. The leaf poultice is used for boils, headache,

jaundice, etc. (Dharmadasa et al., 2014). The patchouli leaves are used along with clothes stored

in the wardrobe since they deter insects like flies, ants, moths, gnats and mosquitoes. They are

also used as a leech repellent and remedy for poisonous insect bites. Cultivation of this plant

outside houses also helps to keep mosquitoes at bay.

As reported by Wong (2018), patchouli essential oil is a type of essential oil commonly

used in aromatherapy. Oil from patchouli offers a variety of health benefits to people. It is used

as an ingredient producing incense and perfume, skin-care products, acne treatment, anxiety,

constipation, dandruff, eczema, indigestion and insomnia. In addition, patchouli essential oil is
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 19

said to act as an insect repellent. It contains antibacterial and antifungal properties that promote

burns and wounds to heal and reduce inflammation.

According to Das et al., (2015) in their study “Synergistic mosquito-repellent activity

of Curcuma longa, Pogostemon heyneanus and Zanthoxylum limonella essential oils,” the

complete protection times (CPTs) produced by P. heyneanus were 23.2 (5%), 49.2 (10%), 123.4

(20%) and 125.2 (30%) minutes in the laboratories trials of repellent efficacy. In fact, patchouli

(P. heyneanus) essential oil provided slightly greater complete protection times (CPTs)

compared with the other essential oils used namely, C. longa and Z. limonella. Furthermore,

among the tested mixtures of essential oils, the C. longa – P. heyneanus mixture (1:2) provided

48.4 (5%), 133.4 (10%), 244.6 (20%) and 246.6 (30%) minutes of CPT and the Z. limonella – P.

heyneanus mixture (1:2) produced 46.6, 120.2, 228.4 and 243.4 min of CPT at the tested

concentrations (5–30%). Further evaluation of the mixtures of C. longa, Z. limonella and P.

heyneanus indicated that the mixture in a ratio of 1:1:2 offered the longest duration of protection.

This mixture provided 117.2, 219.2, 329.4 and 344.8 min of CPT at the 5, 10, 20 and 30%

concentrations, respectively.

In the study of Murugan et al., (2010) entitled “Volatile oil composition of Pogostemon

heyneanus and comparison of its composition with patchouli oil,” the researchers investigated

the components of volatile oils of patchouli. In 96.0% of oil, there are 26 components identified

in the leaves of Pogostemon heyneanus. The researchers stated that acetophenone (51.0%), beta-

pinene (5.3%), (E)-nerolidol (5.4%), and patchouli alcohol (14.0%) are the four major

constituents of the patchouli oil.

Furthermore, Filho et al. (2009) stated the essential oil components of leaves and twigs of

Pogostemon heyneanus in their study entitled “Essential Oil Components of Pogostemon

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 20

heyneanus Benth, Piper hispidinervum C. DC. And Ocimum americanum L. Obtained in the

Amazon.” The researchers claimed and identified that P. heyneanus is composed of 21.9%

patchouli alcohol as a main component. This result was obtained through the identification of

essential oils using hydrodistillation and analyzed by gas chromatography coupled to a mass

spectrometer.

The study aimed to characterize the essential oil components of leaves and twigs

of Pogostemon heyneanus Benth, Piper hispidinervum C. DC. and Ocimum americanum L.

collected around the city of Belém, Pará. The essential oils were obtained by hydrodistillation

and analyzed by gas chromatography coupled to a mass spectrometer. The main components

identified in the essential oils were patchouli alcohol 21.9 % (Pogostemon heyneanus Benth),

safrole 64.3 % (Piper hispidinervum C. DC.) and limonene 24 % (Ocimum americanum L.).

Kakawate (Gliricidia Sepium)

Gliricidia sepium is a medium sized leguminous tree introduced into India from the

American continent. This tree is used in Mexico as shade for cocoa and coffee plantations and

for this reason it is called ‘madrecacao’ which means mother of cocoa. Gliricidia is a Latin name

which means rodent poison which in some country it is also utilized as a poison for rodents (Jose

& Reddy, 2010). ). Gliricidia sepium often simply referred to as kakawate is a medium size

leguminous tree belonging to the family Fabaceae. This plant was introduced in Philippines in

1600s and 1800s to provide shade to tea plants (Nazli et al., 2008). The active medical

compounds present in kakawate are afrormosin, medicarpin, tannin, and some isoflavins. Tannin

is reported to have antidiarrheic, antidysentric, antinephritic, antiviral, bacterial, pesticide,

psycotrophic, and viricide activities (Akharay et al., 2012). The leaves of kakawate are used in
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 21

south India as a mosquito repellent, and they have antifungal and antibacterial activity (Jose

&Reddy, 2010). In the Philippines, kakawate is usually found along rice paddies and some

farmers believe that it can reduce rice insect pests in their fields (Navasero et al., 2014).

Morphological and biochemical features of kakawate (G. sepium) leaves contain 2.4%

nitrogen, 0.1% phosphorus, 1.8% potassium, calcium and magnesium. It grows fast and is

tolerant to pruning and it can thrive in dry, moist, acidic soils or even poor degraded, infertile

soils under rainfed conditions. The foliage can be used as green manure. Gliricidia is a root

nodulating, N-fixing and multipurpose legume (Kumar & Mishra 2013). Various phytochemicals

such as flavonoids, triterpenoid, coumarin, coumaric acid, melilotic acid, and stigmastanol

glucoside have been identified and isolated from various parts of this plant (Jose & Reddy,

2010).

According to Navasero et al., (2014) in their study entitled “Repellency of Volatile

Organic Chemicals of Kakawate, Gliricidia sepium to Rice Green Leaf Hopper,” kakawate is

believe by some farmers to reduce rice insect pests in Philippine’s field. The use of kakawate for

insect pest management in rice production can be promoted to reduce dependence on synthetic

insecticides. It can also be a useful pest management strategy in organic rice production

(Navasero et al., 2014).

In addition to that, Obico & Ragragio (2014) study entitled “A survey of plants used as

repellents against hematophagous insects by the Ayta people of Porac, Pampanga province,

Philippines” highlighted that gliricidia sepium is one of the 7 species of plant that is most

important in repelling insects, The study lists 54 species of plants used by the Ayta people of

Porac, Pampanga with insect-repellent property which includes the G. sepium which is usually

used by the people as hedge around the houses. The Ayta people utilized mainly a combination
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 22

of the dried stem and leaf of the plant, which is burned to elicit its repellent activity against

hematophagous insects.

Meanwhile, the repellent activity of G. sepium has been studied by testing it against adult

Mosquito Aedes aegypti (L.). The maximum repellency percentage afforded by the repellent G.

sepiumwas 78%, whereas with citronella oil, the repellency percentage was 74%. It can be

concluded that the ethanolic leaves extract of kakawate is non-irritating and longer lasting than

the standard repellent (Nazli et al., 2008).

Furthermore, as reported by Krishnaveni et al., (2015), the ethanolic extract of G. sepium

leaves was found to have an inhibitory effect on the growth of larvae of A. aegypti. The

researchers concluded that the ethanolic extract of G. sepium was the most effective when

compared to other solvent extracts by the mortality rate of A. aegypti. The larvicidal activity of

the plant may be attributed to the presence of active compounds such as saponins, flavonoids,

terpenoids, triterpenoids, and alkaloids which were confirmed by TLC studies and

phytochemical analysis. Natural insecticides may play an important role in future regarding the

control of mosquitoes and may thus contribute for the control of vector transmitted diseases such

as malaria, dengue fever, and many others. Thus, this work is contributing evidence for the

potentiality of botanicals in the public health integrated management (Krishnaveni et al., 2015).

In addition, according to Mathew et al., (2014) in their study “Larvicidal Activity of

Gliricidia Sepium Leaf Extracts on Mosquito Larvae,” the crude extract of the leaves of

Gliricidia sepium could be used as one of the source of finding chemical substances to design

insecticidal agents, especially anti-mosquito agents. Crude extract of the leaves of the Gliricidia

sepium plant has activity against the larvae of mosquitoes. The toxic effects of the extract on the

larvae depend on the concentration and duration of exposure.

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 23

CHAPTER III

METHODOLOGY

This chapter presents the procedure, experimental design and treatment, and the materials

that were utilized to derive valid and relevant results of the study.

Materials and Apparatus

Table 1. Materials and Methods Used in the Study

Materials

100 mL Graduated cylinder (Pyrex ™)

250 mL Erlenmeyer Flask (Pyrex™)

75 mm Separatory Funnel (Pyrex ™)

Stirring rod

Vials with cork

100 mL Spray bottles

Mortar and Pestle

Cheese cloth (30cm x 30 cm)

Cage (20cm x 20 cm)

Sieve net

Cotton balls

Experimental Design and Treatments

The study made use of the experimental method of research, a systematic and scientific

approach to research in which the researcher manipulates one or more variables, and controls and
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 24

measures any change in other variables (Blakstab, 2008). In this study, the experiment consisted

of four treatments with three replicates arranged in a Single-Factor Completely Randomized

Design (CRD). The researchers used the method of lottery sampling to derive corresponding

cages for each treatment and replication. The target population was 10 adult mosquitoes in each

treatment (Kamaraj et al. 2010). Since there were four treatments which was replicated three

times, 12 testing cages and a total 30 adult mosquito were used for each treatment. The

mosquitoes were treated with different concentrations of patchouli (P. heyneanus) and kakawate

(G. sepium) extracts mixture to determine its insecticidal activity against mosquito. A total of

120 adult mosquitoes were exposed to four (4) different treatments in three (3) replicates. Each

cage contained 10 adult mosquitoes which were applied with different preparations namely:

Treatment A (75ml Patchouli/25 ml Kakawate, Treatment B (50ml Patchouli/50 ml Kakawate),

Treatment C (25ml Patchouli/75 ml Kakawate) and Treatment D which is the control (Table 2).

Table 2. Preparation of Treatments

Treatments

Treatment A (75mL Patchouli/25mL Kakawate)

Treatment B (50mL Patchouli/50mL Kakawate)

Treatment C (25mL Patchouli/75mL Kakawate)

Treatment D Received no treatment

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 25

Figure 2. Layout of experimental with treatment arranged in a Single-Factor Completely

Randomized Design

TB R2 TD R2 TA R2

TB R1 TC R2 TD R3

TA R3 TA R1 TC R3

TD R1 TB R3 TC R1

Legend: A – 75ml Patchouli / 25ml Kakawate

B – 50ml Patchouli / 25ml Kakawate

C – 25ml Patchouli / 75ml Kakawate

D – No Treatment
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 26

Figure 3. Flow of process conducted throughout the study in determining the insecticidal activity

of patchouli (P. heyneanus) and kakawate (G. sepium)

Preparation of Materials

Collection of Patchouli
and Kakawate Leaves

Collection of Adults
Pounding of Patchouli Male and Female
and Kakawate Leaves Mosquitoes

Extraction of Patchouli
and Kakawate Leaves

Testing the Prepared

Extracts on the Adult
Mosquitoes

Observation

Times of Replication
Data Gathering: Counting
of Dead Mosquitoes

Analysis, Interpretation, and Conclusion

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 27

Preparation of Plant Extracts

The following were the procedures done in order to obtain leaf extracts of patchouli

(Pogostemon heyneanus) and kakawate (Gliricidia sepium) that was utilized as insecticide

against mosquitoes.

1. Fresh young leaves of patchouli (P. heyneanus) and kakawate (G. sepium) were gathered

in Banga, Aklan.

Figure 4. Gathering of Patchouli (P. heyneanus) and Kakawate (G. sepium) leaves

2. Dry and damaged part of the leaves were discarded. Healthy leaves of the plants were

selected and rinsed with water to remove unnecessary substances. The leaves were

wrapped in a plain paper to prevent it from desiccating. While it is still fresh, it was

brought to the laboratory for extraction process.

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 28

Figure 5. Screening of Patchouli (P. heyneanus) and Kakawate (G. sepium) Leaves

3. The researchers pounded the patchouli and kakawate leaves separately using mortar and

pestle. The pounded leaves were placed inside a cheese cloth and were squeezed to obtain

the pure extracts of the plant leaves.

Figure 6. Extraction of Patchouli (P. heyneanus) and Kakawate (G. sepium) Leaf Extracts
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 29

4. The extracts were subjected to filtration using separatory funnel and filter paper to

separate the residue from the filtrate. The residue was discarded while the filtrate

from the Erlenmeyer flask was decanted into vials for treatment preparation.

Figure 7. Filtration of Patchouli (P. heyneanus) and Kakawate (G. sepium) Leaf Extracts

5. The extracts from the vials were decanted into graduated cylinder to obtain the desired

preparations: Treatment A (75ml Patchouli/25ml Kakawate), Treatment B (50ml

Patchouli/50ml Kakawate), and Treatment C (25ml Patchouli/75 ml Kakawate). The

extracts of both plants with specific volume were mixed inside the beaker. The treatments

were then transferred to spraying bottles (100ml).

Figure 8. Formulation of Various Patchouli (P. heyneanus) and Kakawate (G. sepium)
Leaf Extracts Treatments
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 30

Collection of Mosquito Specimens

Mosquito served as the test insect in determining the insecticidal activity of patchouli (P.

heyneanus) and kakawate (G. sepium). These mosquitoes were collected in an open area using

sieve nets. This method was repeatedly used to obtain the desired number of mosquito samples

totaling to 120 mosquitoes (10 mosquitoes in each cage) and was placed inside the insect-rearing

cage inside the laboratory to prevent them from escaping.

Figure 9. Collection of Mosquito Specimen

Insecticidal Bioassay

Mosquitoes were selected from the insect-rearing cage for the testing of insecticidal

activities. They were gently collected from the cage and were carefully transferred into the

testing cages (10 mosquitoes for each cage). The mosquitoes were allowed to acclimatize in the
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 31

cages for an hour and were then exposed to the different treatments through spraying the entire

sides of each cage. A regular-sized cotton ball was sprayed within corresponding treatments just

enough to wet the whole cotton ball and was also placed inside corresponding testing cages to

prolong the effect of the treatments inside the cage. All sides of the cage were covered with an

observable white net to allow viewing. A fabric sleeve was added to the front side of the test

cage to allow access by a human forearm (Yoon, 2015). The number of dead mosquitoes in each

cage was recorded every 30 minutes within two (2) hours. The mean of the number of dead

mosquitoes was used to determine the insecticidal effect of patchouli (P. heyneanus) and

kakawate (G. sepium) (Antonio et al., 2014).

Locale of the Study

This study was conducted in the Chemistry Laboratory of Infant Jesus School, Mabini

St., Extension, Kalibo, Aklan from the month of January to February 2019.

Statistical Tools

The data obtained from the test was analyzed using the following statistical tools:

To determine the insecticidal effect of patchouli (P. heyneanus) and kakawate (G.

sepium) on mosquitoes, the mean was used. The mean was obtained by getting the sum of

mosquitoes that died in three replicates and was divided by three. The higher the obtained mean,

the more effective the treatment is.

On the other hand, to determine whether there is significant difference in the insecticidal

effect of patchouli (P. heyneanus) and kakawate (G. sepium) on mosquitoes as to different

concentrations, the results were statistically analyzed by mean of two-way Analysis of Variance
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 32

(ANOVA) and results were statistically significant at p = 0.05, Tukey test was used. The data

were analyzed using the statistical computer program MiniTab 2018.

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 33

CHAPTER IV

RESULTS AND CONCLUSION

This chapter describes the analysis of data followed by a discussion of the research

findings. This chapter also contains the presentation of data in figural and tabular form with its

corresponding interpretation. The summary of findings, conclusion derived from the findings and

the recommendations are also presented.

Data Analysis

Table 3. Mortality rate of mosquito species exposed to Treatment A (75ml Patchouli / 25ml

Kakawate) extract mixture across different length of the exposure

Mortality rate

Replication Length of Exposure (%) per

Replication

Less than 1 hr 1 hr 1 hr 30 min 2 hrs (f) (%)

(30 min)

1 0 3 3 1 7 70

2 0 1 2 3 6 60

3 1 5 2 0 8 80

(f) 1 9 6 4 21 70

(%) (3.33%) (30%) (23.33%) (13.33%) (70%)

Table 3 presents the mortality rate of mosquito when exposed to Treatment A

concentrated with 75% patchouli (P. heyneanus) and 25% kakawate (G. sepium). There are three

replications, and each has 10 adult mosquitoes exposed to patchouli (P. heyneanus) and
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 34

kakawate (G. sepium) for two (2) hours. The findings showed that in Treatment A the mortality

rate is seven (7) or 70% in Replication 1, six (6) or 60% in Replication 2, and eight (8) or 80% in

Replication 3. It is also shown in the results that the reduction of mosquito started at 1 hour in

Replication 1 and 2, but earlier in Replication 3 at 30 minutes. Treatment A gave a mean

mortality rate of 7 mosquitoes or exactly 70%.

Table 4. Mortality rate of mosquito species exposed to Treatment B (50ml Patchouli / 50ml

Kakawate) extract mixture across different length of the exposure

Mortality rate

Replication Length of Exposure (%) per

Replication

Less than 1 hr 1 hr 1 hr & 30 2 hrs (f) (%)

(30 min) min

1 1 2 5 0 8 80

2 0 1 5 3 9 90

3 3 1 5 1 10 100

(f) 4 4 15 4 27 90

(%) (13.33%) (13.33%) (50%) (13.33%) (90%)

Table 4 shows the mortality rate of mosquito when exposed to Treatment. There are

three replications, and each has 10 adult mosquitoes exposed to 50% patchouli (P. heyneanus)

and 50% kakawate (G. sepium) for two (2) hours. There are eight (8) or 80% mortality rate in

Replication 1, nine (9) or 90% in Replication 2, and all mosquitoes died or 100% mortality rate

in Replication 3. It is also shown in the results that the reduction of mosquito started at less than
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 35

1 hour (30 minutes) in Replication 1 and 3 but at 1 hour in Replication 3. Treatment B gave an

average of mortality rate values of 9 or 90%.

Table 5. Mortality rate of mosquito species exposed to Treatment C (25ml Patchouli / 75ml

Kakawate) extract mixture across different length of the exposure

Mortality rate (%)

Replication Length of Exposure per Replication

Less than 1 hr 1 hr 1 hr & 30 2 hrs (f) (%)

(30 min) min

1 4 4 1 0 9 90

2 0 2 4 3 9 90

3 1 5 1 1 8 80

(f) 5 11 6 4 26 86.67

(%) (16.67%) (36.67%) (20%) (13.33%) (86.67%)

Table 5 presents the mortality rate of mosquito species when exposed to Treatment C.

There are three replications, and each has 10 adult mosquitoes exposed to patchouli (P.

heyneanus) and kakawate (G. sepium) for two (2) hours. The findings showed that in Treatment

C nine (9) or 90% of the mosquitoes died in Replication 1, same data observed in Replication 2

with nine (9) or 90%, and the mortality rate in Replication 3 is eight (8) or 80%. It is also shown

in the results that the reduction of mosquito started at less than 1 hr (30 min) in Replication 1 and

3, but at 1 hr in Replication 2. The results of analysis showed that the mean mortality rate of
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 36

Treatment C treated with 25% patchouli (P. heyneanus) and 75% kakawate (G. sepium) is 8. 67

of 86.67%.

Table 6. Mortality rate of mosquito species exposed to Treatment D (Received No Treatment as

a Control) extract mixture across different length of the exposure

Mortality rate (%)

Replication Length of Exposure per Replication

Less than 1 hr 1 hr 1 hr & 30 2 hrs (f) (%)

(30 min) min

1 0 0 0 0 0 0

2 0 0 0 0 0 0

3 0 0 0 0 0 0

(f) 0 0 0 0 0 0

(%) (0%) (0%) (0%) (0%) (0%)

Table 6 presents the mortality rate of mosquito when exposed to Treatment D (Received

No Treatment) as a control. There are three replications, and each contains 10 adult mosquitoes

without patchouli (P. heyneanus) and kakawate (G. sepium) for two (2) hours. The findings

showed that in Treatment D no mortality was detected in Replication 1, same data observed in

Replication 2 and Replication 3. Thus, there was 0% mortality rate in Treatment D.

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 37

Figure 10. Percent mortality of the mosquito after exposure to four different treatments

Insecticidal Activity of Patchouli (Pogostemon heyneanus) and Kakawate

(Gliricidia Sepium) against Mosquito

100

90

80
Treatment A
70
Treatment B
Mortality Rate (%)

60

50 Treatment C

40 Treatment D
(Control)
30

20

10

The graph displays the average percent mortality of mosquito exposed in four

concentrations. The mean percent mortality rate was derived from the three replications of each

treatment. The findings show that Treatment B treated with 50% patchouli (Pogostemon

heyneanus) and 50% kakawate (Gliricidia sepium) has the highest mortality rate among the four,

having 90%. Treatment C contained with 25% patchouli (Pogostemon heyneanus) and 75%

kakawate (Gliricidia sepium) has 86.67% mortality rate. Treatment A treated with 75% patchouli

(Pogostemon heyneanus) and 25% kakawate (Gliricidia sepium) has 70% mortality rate.

Treatment D which served as the control had no mortality at all.

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 38

Table 7. Significant difference of insecticidal activity of patchouli (Pogostemon heyneanus) and

kakawate (Gliricidia sepium) among treatments

Source DF Adj SS Adj MS F-Value P-Value

Treatments 3 39.75 13.250 5.39 0.003

Error 44 108.17 2.458

Total 47 147.92

The p-value in columns lesser than 0.05 is significantly different.

Table 7 shows the analysis of significant difference among four treatments of (P.

heyneanus) and kakawate (G. sepium). The computed p-value of treatments was 0.003 which is

lesser than 0.05. The insecticidal activity provided by three treatments with a control one of

patchouli (P. heyneanus) and kakawate (G. sepium) was significantly different with each other

(p > 0.05).

Table 8. Summarized comparisons of significant difference of treatment level

Treatments N Mean Grouping

B 12 2.25000 A

C 12 2.16667 A

A 12 1.75000 A

D 12 0.00000 B

ANOVA followed by Tukey's HSD. The values in columns followed by same letters are not

significantly different (p > 0.05).

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 39

Table 8 presents the Tukey pairwise comparisons among treatments with 95% confidence

level. Among the tested treatments of patchouli (P. heyneanus) and kakawate (G. sepium) against

mosquito, Treatment A, B, and C had no significant difference. The findings also clearly

indicated that the insecticidal activity provided by Treatment D was significantly lesser at all

concentrations with a mean value of 0.25000. Treatment D is significantly different from

Treatment A, B and C.

Table 9. Tukey Simultaneous Tests for Significant Differences of Four Treatments

Difference of Difference SE of Simultaneous T-Value Adjusted

Treatments of Means Difference 95% CI P-Value
Levels

B-A 0.500 0.640 (-1.211, 2.211) 0.78 0.863

C-A 0.417 0.640 (-1.294, 2.128) 0.65 0.915

D-A -1.750 0.640 (-3.461, -0.039) -2.73 0.043

C-B -0.083 0.640 (-1.794, 1.628) -0.13 0.999

D-B -2.250 0.640 (-3.961, -0.539) -3.52 0.006

D-C -2.167 0.640 (-3.878, -0.456) -3.38 0.008

The values in adjusted p-value lesser than 0.05 is significantly different.

Table 9 shows the analysis between the differences of insecticidal activity of each

treatment of patchouli (P. heyneanus) and kakawate (G. sepium) from one another. It is shown in

the results that Treatment D and B are significantly different for having the lowest p-value of

0.006. Treatment D and C, and Treatment D and A is significantly different with 0.008 and 0.043

p-values, respectively. On the other hand, the finding also shown that there is no significant
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 40

difference between Treatment C and B for having the highest p-value of 0.999. Treatment C and

A is not significantly different with p-value of 0.915. Furthermore, Treatment B and A has no

significant difference, having p-value of 0.863.

Summary of Findings

This study scientifically determined the insecticidal activity of patchouli (Pogostemon

heyneanus) and kakawate (Gliricidia sepium) against mosquito. The study consisted of four

treatments having four different concentrations Treatment A is treated with 75% patchouli (P.

heyneanus) and 25% kakawate (G. sepium); Treatment B with 50% patchouli (P. heyneanus) and

50% kakawate (G. sepium); Treatment C with 25% patchouli (P. heyneanus) and 75% kakawate

(G. sepium); Treatment D with no treatment at all. The treatments were investigated to analyze

the insecticidal activity of leaf extracts against mosquito.

The results indicated the following:

1. Significant insecticidal effect was noted for the leaf extracts of patchouli (Pogostemon

heyneanus) and kakawate (Gliricidia sepium) against mosquito. Four treatments were

tested and the mortality rate of mosquito was analyzed. The rate of adult mosquito

mortality was 90% in Treatment B. Moreover, Treatment C produced an average

mortality of 86.67%. Mortality rate of mosquito decreased to 70% in Treatment A.

Treatment D which acted as a control variable of this study had 0% percent mortality.

2. Among the treatments, the cage treated with 50% patchouli (P. heyneanus) and 50%

kakawate (G. sepium) provided the highest insecticidal activity against mosquito with

90% mortality rate or nine (9) out of ten (10) mosquitoes died in this treatment.
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 41

3. In the statistical analysis, there are significant differences between Treatment D-B and

Treatment D-C, with p-values of 0.006 and 0.008, respectively. Treatment D-A also

showed significant difference of 0.043. However, no significant differences were

observed in Treatment C-B, Treatment C-A, Treatment B-A with p-values of 0.999,

0.915, and 0.863 respectively.

Conclusion

Based on the findings derived from the study, the following conclusions are drawn:

1. Among the tested treatments, the highest mean percentage mortality rate of mosquito

after 2 hours of exposure was obtained in Treatment B. Treatment C yielded the second

highest insecticidal effect against mosquito. However, the mortality effect of Treatment

A was lower than those of the other treatments. Treatment D yielded no mortality rate.

2. Treatment B concentrated with 50% patchouli (Pogostemon heyneanus) and 50%

kakawate (Gliricidia sepium) leaf extracts was the most effective and provided the

highest insecticidal activity when compared to other three treatments by means of

mortality rate of mosquito.

3. The mortality rates that were measured in four different treatments including the control

were analyzed. The analysis of variance among treatments resulted to p-value of 0.003.

Therefore, there is a significant difference on the insecticidal activity of different

treatments of insecticide made mainly of patchouli (P. heyneanus) and kakawate (G.

sepium) leaf extracts against mosquito.

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 42

Recommendations

The foregoing findings and conclusions lead to the following recommendations:

1. Further studies on the larvacidal and repellent activity of patchouli (Pogostemon

heyneanus) and kakawate (Gliricidia sepium) against mosquito

2. Additional scientific approach is needed to better understand how the differences

between treatments occur, including studies on the chemical volatilization.

3. Conduct studies about the insecticidal activity of patchouli (Pogostemon heyneanus) and

kakawate (Gliricidia sepium) against other kind of insect.

4. Conduct similar studies by utilizing an identified mosquito specimen.

5. Addition of natural preservatives and fragrance suitable for the product to make it long-

lasting and to mask the unpleasant smell.

6. Comparison of formulated natural insecticide to the commercially available chemical-

based insecticide.

7. Utilization of patchouli (Pogostemon heyneanus) and kakawate (Gliricidia sepium)

extracts’ residue (derived from filtered patchouli and kakawate) as mosquito repellent
 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 43

CHAPTER V

PROJECT DESCRIPTION

Insecticide against mosquito plays an important role in preventing or reducing the

incidence of mosquito-borne diseases. However, the continued use of chemical insecticide leads

to adverse effects in humans, wildlife and environments. A plant-derived insecticide made out of

patchouli (Pogostemon heyneanus) and kakawate (Gliricidia sepium) were used to minimize the

harmful effects brought by chemical-made insecticides. The materials that were used in this

project are all natural and available at no-cost. Both botanical plants are accessible anywhere but

the researchers gathered these plant specimen in Banga, Aklan. The researchers used natural

extraction method in order to obtain the leaf extracts needed to formulate different treatments

with various extracts’ concentration. Three concentrations were formulated with the control and

each insecticidal activity was investigated using captured mosquitoes. These mosquitoes were

collected in Banga, Aklan using sieve net. There are three (3) cages for each treatment with 10

mosquitoes each. Three replications were done in this experimental study. Moreover, the number

of mortality in each cages were counted every 30 minutes within 2 hours. The percent mortality

rate of mosquito was calculated and the highest one was considered as the most effective among

treatments.

The present study was done in Chemistry Laboratory in Infant Jesus School with the

supervision of their capstone adviser. The researchers used personal protective equipments such

as laboratory gowns, gloves, and mask in order to protect themselves in possible incidences.

The present study revealed the insecticidal activity of leaf extracts of patchouli

(Pogostemon heyneanus) and kakawate (Gliricidia sepium) against mosquito.

 INSECTICIDAL ACTIVITY AGAINST MOSQUITO 44

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