THE EFFICACY OF SELECTED PLANTS LEAF EXTRACTS

(MADRE DE CACAO, LAGUNDI, LEMON GRASS AND STAR APPLE)

AS MOSQUITO LARVICIDAL

A Thesis
Presented To
The Faculty of the Graduate School
Abra State Institute Sciences and Technology
Bangued, Abra

In Partial Fulfillment
Of the Requirements for the Degree
Master of Arts in General Science

WILMA PALMOS SALVADOR

2019
 ii

CERTIFICATION

This thesis titled “THE EFFICACY OF SELECTED PLANTS LEAF

EXTRACTS (MADRE DE CACAO, LAGUNDI, LEMON GRASS AND STAR

APPLE) AS MOSQUITO LARVICIDAL”, prepared and submitted by WILMA

PALMOS SALVADOR in partial fulfilment of the requirements for the degree of

MASTER OF ARTS IN GENERAL SCIENCE (MA. GEN. SCI.) has been examined and

recommended for acceptance and approval for final defense.

ANNALIZA J. DAYAG, Ph.D.

Adviser

ELIZABETH C. FETALVERO, MA Eng. PABLO B. BOSE Jr., Ed. D.

Critic Statistician
 iii

APPROVAL SHEET

Approved by the Panel of Examiners on Oral Defense with a grade of

____________.

MARIA DIGNA T. BOSE, MST Chem. JOEY M. DELA CRUZ, Ed. D.

Member Member

NOEL B. BEGNALEN, Ph. D.

Chairman

Accepted and approved in partial fulfillment of the requirements for the degree,

Masters of Arts in General Science.

JOEY M. DELA CRUZ, Ed. D.

Dean, Graduate School

Comprehensive Examination : Passed

Date: Nov. 31, 2017 – Dec. 1, 2017
Date of Final Defense: May 07, 2019
 iv

ACKNOWLEDGEMENT

The researcher wants to express her profound gratitude to the following who

supported her to pursue this work for without them this would not be completed.

Dr. Noel B. Begnalen, Vice President for Academic Affairs, for his exceptional

ideas and suggestions to make this work meaningful and substantial.

Dr. Joey M. Dela Cruz, Dean of the Graduate School, for making himself always

available in giving insights, suggestions and comments to the researcher in making this

work a success.

Dr. Annaliza J. Dayag, the researcher’s adviser, for her understanding, having

priceless patience, valuable encouragement and untiring effort in the completion of this

study.

Dr. Pablo P. Bose Jr, the researcher’s statistician, for his humility, empathy and

tireless efforts in working the statistical part of this study, analysis and interpretation of

data despite his demanding administrative and teaching responsibilities.

Prof. Elizabeth C. Fetalvero, the researcher’s critic, for her valuable ideas and

suggestions and for sparing her time in checking the contents for the improvement of this

work.

Prof. Maria Digna T. Bose, the panel member, for her suggestions and

recommendations that contibute to make this work substantial.

Mr. Jenelyn B. Lagundino, Graduate Study clerk for his insights and pieces of

advice.

Mr. Crisostomo T. Balneg, the researcher’s school principal, for allowing the

researcher to use school science apparatus to conduct the experiments of this study.
 v

Ms. Jovelyn Pizarro, for making herself available to look for lemon grass leaves.

Colleagues (Vangie, Thelma, Lanie and Anet) and co – science teachers ( Fred,

Mical, Roda, Cy and Gillian) for their assistance, support and encouragement.

Friends, students, relatives and acquaintances for motivating and inspiring the

researcher to finish the study.

Those whose works served as the researcher’s references, her profound

commendation.

Above all to the Divine Providence for giving the researcher courage, wisdom,

good health and strength during the conduct of this study.

“To God All Be the Glory!”

WPS

DEDICATION
 vi

This work is lovingly dedicated to

Dario,

my husband who devotes his wholesome support in all my struggles

Nica, Wency, Zea

my three bubbly princesses

Nanang Tacing

my mother for her encouragement and

my brothers and sisters, nephews and nieces

You are the reason for the fulfilment of this goal.

Thank you for being an inspiration!

WILMA

ABSTRACT
 vii

THE EFFICACY OF SELECTED PLANTS LEAF EXTRACTS (MADRE DE

CACAO, LAGUNDI, LEMON GRASS AND STAR APPLE) AS MOSQUITO
LARVICIDAL, SALVADOR, Wilma P., Master of Arts in General Science (MA
Gen. Sci.), Abra State Institute of Sciences and Technology, Bangued Campus,
Bangued Abra, Philippines 78 pages.
Adviser : ANNALIZA J. DAYAG, Ph.

D._____________________________________

Mosquitoes are causative agents of dreadful diseases today such as malaria,

Japanese encephalitis, dengue and Zika virus. Annually, there are reported cases of death

not only among children but even adults and this is an alarming problem of the

community. The infestation of this insect should be addressed and such find ways to

eradicate them.

The use of commercial insecticides and pesticides are very prevalent today.

However, they impose threats not only on human being but even on the environment.

There are numerous species of plants around us. Plants may contain chemicals that could

kill pests and insects. Insecticides and pesticides that obtained from plants are more eco-

friendly and budget friendly.

This study was conducted to explore the plant diversity for a potential alternative

for synthetic insecticide to get rid of mosquito larvae. It aims further, to provide

awareness to the community with the use of cheaper but effective mosquito larvae

repellent and to help the government find solution to eradicate mosquitoes.

This study determines the efficacy of Madre de Cacao or Gliricidia sepium as

mosquito larvicidal . The leaf was extracted and the extracts were prepared to different

concentrations as 25%, 50%, 75% and 100%. The experiment was done using parallel
 viii

group design. There were five treatments on which one was set aside and served as

control. Each treatment consist of four replicates with five larvae each. The efficacy of

the concentration was tested by determining the mortality rate of the larvae. Aside from

that, madre de cacao leaf extracts was also combined to other plants such as lagundi

(Vitex negundo), lemon grass (Cymbopogon citratus), and star apple (Chrysophyllum

cainito) to further determine the efficacy of madre de cacao as mosquito larvicidal. The

efficacy of the combined leaf extracts were tested using different concentrations and by

determing the rate of mortality of the mosquito larvae.

The result of the study shows that madre de cacao leaf extract is a potential

mosquito larvicidal. The higher its concentration the more it is effective. The efficacy of

combined madre de cacao and lemon grass as larvicidal is also very effective based on

the high mortality rate of mosquito larvae at one to three hours observation. Thus, this

study therefore suggests the use of madre de cacao as insect repellent as substitute of the

commercial products, furthermore, lemon grass should be combined. In addition further

investigation or the same study should be conducted using its flowers and barks or by

using larger number of larvae to test the validity of the result of this study.
 ix

TABLE OF CONTENTS

PRELIMINARIES

Title Page..................................................................................................................i

Certification.. ..........................................................................................................ii

Approval

Sheet .......................................................................................................iii

Acknowledgement..................................................................................................iv

Dedication………………………………………………………………………...vi

Abstract……………………………………………………………………….….vii

Table of Contents………………………………………………………………....ix

List of Tables..........................................................................................................xi

List of Appendices……………………………………………………………….xii

CHAPTER

I. THE PROBLEM………………………………………………………...………1

Background of the Study.............................................................................1

Review of Related Literature………….......................................................

Statement of the Problem...........................................................................29

Hypothesis..................................................................................................30

II. THE METHODOLOGY……………………………………………………...31

Time and Place of the

Study.......................................................................31
 x

Materials…………….

……. ......................................................................31

Methods...............................................................................................…...32

Data Gathered……………….

…................................................................35

Statistical Treatment of Data.....................................................................36

III. RESULT AND

DISCUSSION……………………………………………….37

IV. CONCLUSION AND RECOMMENDATIONS……....

…………………….50

Findings….................................................................................................50

Conclusions………………………………………………………………51

Recommendations…………………………………………………..…....51

REFERENCES…..............................................................................................................52

APPENDICES……...……………………...………………………………………….....59

CURRICULUM VITAE……………………..………………...………………………..80
 xi

LIST OF TABLES
No. Title Page
1 Transformed Data on the Mortality Rate of Mosquito Larvae
On Madre de Cacao and distilled Water...................................................37

2 Transformed Data on the Mortality Rate of Mosquito Larvae

On Madre de Cacao and distilled Water...................................................39

3 Transformed Data on the Mortality Rate of Mosquito Larvae

On Madre de Cacao and distilled Water...................................................42

4 Transformed Data on the Mortality Rate of Mosquito Larvae

On Madre de Cacao and distilled Water...................................................44

5 Over – all Result of Mortality Rate of Combined Mixtures.....................47

xii
 xii

APPENDICES

No. Title Page

1 Original Values on the Mortality Rate of Mosquito

Larvae in Madre de Cacao Leaf Extract and Distilled water……………
60

1a Transformed Values on the Mortality Rate of Mosquito

Larvae in Madre de Cacao Leaf Extract and Distilled
water…………...60

1b Mean of the Transformed Data on the Mortality Rate of Mosquito

Larvae in Madre de Cacao Leaf Extract and Distilled water……….

…..61

1c Summary Mean on the Transformed Data on the Mortality Rate of

Mosquito Larvae in Madre de Cacao Leaf Extract and
Distilled water ………………………………….………………………62

1d Analysis of Variance in 3 hours……………………………………….…

62

1e Analysis of Variance in 6

hours………………………………………….62

1f Analysis of Variance in 9

hours………………………………………….62

1g Analysis of Variance in 12

hours………………………………………...62

2 Original Values on the Mortality Rate of Mosquito

Larvae in Madre de Cacao and Lagundi Leaf Extracts
…………….........63

2a Transformed Values on the Mortality Rate of Mosquito

Larvae in Madre de Cacao and Lagundi Leaf
Extracts…………………..63

2b Mean of the Transformed Data on the Mortality Rate of

 xiii

Mosquito Larvae in Madre de Cacao and

Lagundi Leaf Extracts…………………………………………………...63

2c Summary Mean on the Transformed Data of the Mortality Rate of

Mosquito Larvae in Madre de Cacao and
Lagundi Leaf Extracts …………..………………………………………64

2d Analysis of Variance in 3 hours…………………………...

……………..64

2e Analysis of Variance in 6

hours…………………………………….........64

2f Analysis of Variance in 9

hours………………………………………….65

3 Original Values on the Mortality Rate of Mosquito

Larvae in Madre de Cacao and Lemon Grass Leaf Extracts ………..
…...65

3a Transformed Values on the Mortality Rate of Mosquito

Larvae in Madre de Cacao and Lemon Grass Leaf Extracts………..……
65

3b Mean of the Transformed Data on the Mortality Rate of

Mosquito Larvae in Madre de Cacao and
Lemon Grass Leaf Extracts……………………………………………..66

3c Summary Mean on the Transformed Data of the Mortality

Rate of Mosquito Larvae in Madre de Cacao and
Lemon Grass Leaf Extracts ……………………………………….
……..66

3d Analysis of Variance in 3 hours…………………...……………………66

3e Analysis of Variance in 6

hours………………………………………….67

4 Original Values on the Mortality Rate of Mosquito

 xiv

Larvae in Madre de Cacao and Star Apple Leaf Extracts

……………….67

4a Transformed Values on the Mortality Rate of Mosquito

Larvae in Madre de Cacao and Star Apple Leaf Extracts……………..…
67

4b Mean of the Transformed Data on the Mortality Rate of

Mosquito Larvae in Madre de Cacao and
Star Apple Leaf Extracts………………………………………………..68

4c Summary Mean on the Transformed Data of the Mortality

Rate of Mosquito Larvae in Madre de Cacao and
Star Apple Leaf Extracts
…………………………………………..........69

4d Analysis of Variance in 3

hours………………………………………….69

4e Analysis of Variance in 6

hours………………………………………….69

4f Analysis of Variance in 9

hours………………………………………….69

4g Analysis of Variance in 12 hours…………………………………………

69

5 Over-all Result of Combined Mixtures………………………………….70

Documentation…………………………...……………………………………………....71
 Chapter I

THE PROBLEM

Background of the Study

Mosquitoes are notorious for their involvement in disease transmission, creating

annoyances, interfering with outdoor recreation activities, even ruining a good night sleep

and suffering infection from scratching mosquito bites when infected with bacteria. Its

size does not matter when it breaks the headline of newspapers, news on television and

radio. This blood sucking insect that grows about 5mm long is really terrifying for it

transmits tremendous diseases that are fatal or hard to cure. As years passed by there

were many cases of mosquito-borne diseases that captured lives henceforth the

eradication of these insects should be addressed and given much priority especially

during rainy seasons. In response to this need, the researcher aims to find way to help

minimize the growth of this fast breeding insect utilizing available resources in the

community in an easy and eco-friendly processes.

Mosquitoes develop long-lasting viral infections without substantial deleterious

effects, despite high viral loads. This makes mosquitoes efficient vectors for emerging

viral diseases with enormous burden on public health. (Goic et al., 2016)

According to the World Health Organization (2016), of all disease-transmitting

insects, the mosquito is the greatest menace, spreading malaria, dengue and yellow fever

which together are responsible for several million deaths and hundreds of millions of

cases every year. Mosquitoes also transmit lymphatic filariasis and Japanese encephalitis.

Dengue is one of the world’s most important mosquito-borne virus

 2

disease, with 2500 million people worldwide at risk infection and 20 million cases a year

in every in more than 100 countries (WHO, 1996)

Moreover, Asian Development Bank (ADB), reported that vector-borne diseases

such as malaria and dengue are set to become more lethal and widespread than ever and

cause far worse cases of mortality especially among children (Testa, 2017).

In the Philippines, malaria is one of the major mosquito-borne infectious diseases

affecting 14 million people who live mostly in mountainous, hard to reach areas of the

country where basic health services are relatively inaccessible. Malaria and other vector-

borne diseases brought 57 of 80 provinces are malaria endemic, and 14 million

population at risk. These endemic provinces for neglected tropical diseases are the

poorest of the poor. The parasitic diseases cause chronic disabilities and deformities

which hamper children’s growth and about 645, 232 persons are already affected with

lymphatic filariasis (WHO Philippines, 2016)

From the Philippine Star news, dated October 24, 2018, dengue cases nationwide

soared to over 138,000 including over 700 deaths in the past nine months according to the

Department of Health. The DOH Dengue Surveillance showed a 21 percent increase in

the number of potentially deadly infection recorded in various government hospitals

nationwide compared to the same period last year. From January 1 to October 6, DOH

recorded a total of 138, 444 dengue cases, up from the 114,878 cases during the same

period in 2017. Central Luzon, NCR, and Calabarzon posted the biggest number of degue

cases. Other regions include Ilocos region and Western Visayas ( Jaymalyn, 2018).

Former Health Secretary Paulyn Ubial said the public should take preventive

measures on mosquito-borne diseases like dengue. The strategy includes “getting rid of
 3

stagnant water, maintaining environmental cleanliness and eliminating potential breeding

places of mosquitoes not only within our homes but the community (Jaymalyn and

Crisostomo, 2017) .

According to Shukla et al. (2018), one of the primary reasons for the worldwide

spread of mosquitoes is due to deforestation and industrialized farming. They are causing

an alarming increase in the range mosquitoes. The World Health Organization blames

global warming for the expanding range of mosquitoes that are responsible for malaria,

yellow fever and dengue fever, causing millions at risk. It is estimated by WHO nearly

15,000 deaths per year of all ages occur only in the Indian Peninsula .Although chemical

mosquito repellents available in the market have been formulated so that they have

remarkable safety profile, but their toxicity against the skin and nervous system can cause

rashes, swelling, eye irritation and other serious problems to children. Synthetic

insecticides were introduced as an alternative but although these are effective, the insect

tends to develop resistance to such products (Jirakanjanakit et al., 2007). Aside from

being costly, the use of these repellants also generates problems such as environmental

pollution and has toxic side effect to humans. This imposes a need for other alternative

methods which are efficient, economic and environmentally safe. Botanical derivatives

materialize as a possible larvicide for the Aedes aegypti or common household mosquito

(Zhu et al., 2008).

Hence, it was felt that bio-based natural mosquito repellents would be preferred

over chemical mosquito repellents. Bio based mosquito repellent are pest management

tools that are based on safe, biologically based active ingredients derived from plants.

Benefits of bio-products include effective control of mosquito as well as human and

 4

environmental safety. These bio based products were designed to play an important role

in providing pest management tools in areas where mosquito resistance, niche markets,

and environmental concerns limit the use of products.

Ghosh (2012), also suggested to explore the floral biodiversity and enter the field

of using safer insecticides of botanical origin as a simple and sustainable method of

mosquito control. Further, unlike conventional insecticides which are based on a single

active ingredient, plant derived insecticides comprise botanical blends of chemical

compounds which act concertedly on both behavioural and physiological processes. Thus

there is very little chance of pests developing resistance to such substances. Identifying

bio-insecticides that are efficient, as well as being suitable and adaptive to ecological

conditions, is imperative for continued effective vector control management. Botanicals

have widespread insecticidal properties and will obviously work as a new weapon in the

arsenal of synthetic insecticides and in future may act as suitable alternative product to

fight against mosquito borne diseases.

Mathew et al. (2015), researched that many of the organic insecticides are based

on single ingredient, but plant products comprises of a number of chemicals which can

act on physiological process of mosquitoes. It was reported that more than two hundred

plants species belonging to different families and genera have toxic components which

are effective against insects (Govindarajan and Jebanesan, 2008 as cited by Mathew).

Extracts from various part of the plant such as leaves, roots, stem and fruits are shown

mosquitocidal (larvicidal) activities (Kalyanasundaram and Dass, 1985: Gbolade, 2001 as

cited by Mathew).

Several plant species are believed that may have chemical components
 5

that could get rid of insects. Among those plants include Madre de cacao (Gliricidia

sepium), Lagundi (Vitex negundo), Lemon grass (Cymbopogon citratus) and Star apple

(Chrysophyllum cainito)

Madre de cacao or Gliricidia sepium that is locally known as kakawate is an

effective pesticide, according to the Department of Agriculture (DA). (Calleja, 2010) It

has also same active substance that could wipe out the insects alighting on farm animals.

(Dela Cruz, 2003)

Lagundi or Vitex Negundo is believed by the old folks when leaves are burn, its

smoke is believed to drive away insects, particularly mosquitoes. (Medical health Guide,

2011)

Lemon grass (Cymbopogon citratus), a native herb from temperate and warm

regions such as India, Philippines and Malaysia, is widely used in Asian cooking and is

an ingredient in many Thai and Vietnamese foods. The aromatic and citrus scent of

lemon grass is an effective insect repellent. It helps to keep pets clean of fleas, ticks and

lice. (Medical health Guide, 2011)

Star Apple scientifically known as Chrysophyllum cainito is a tropical evergreen

tree of the family Sapotaceae. The Aytas burn dried leaves for insect repellency . (Stuart,

2018)

Due to the interest to help fight mosquitoes which are carriers of dreadful

diseases, the researcher aims to find a solution to get rid of these insects at any stage of

their life using madre de cacao (Gliricidia sepium) leaf extracts prepared in different

concentrations. The researcher also aims to determine the efficacy of madre de cacao as

mosquito larvicidal combined with other plants such as lagundi, lemon grass and star
 6

apple leaf extracts in different concentrations. It specifically, sought to determine the

efficacy of the leaf extracts to eradicate larvae of mosquitoes and to determine an

environmental friendly solution against these insects.

 7

Review of Related Literature

This part presents the life cycle of a mosquito and significant and related studies

about mosquitoes, morphological structure and characteristics of madre de cacao ,

lagundi, lemon grass and star apple plants their uses and efficacy, medicinal value and

related studies as to their larvicidal effect.

The Life Cycle of a Mosquito

Mosquitoes (Order Diptera, Family Culicidae) are some of the most adaptable and

successful insects on Earth and are found in some extraordinary places. Virtually any

natural or man-made collection of water can support mosquito production. They've been

discovered in mines nearly a mile below the surface, and on mountain peaks at 14,000

feet, and if you know where to look, there is a good possibility that there are mosquitoes

breeding just in the backyard. Not every species of mosquito causes problems for people,

but many have profoundly negative effects. Mosquitoes can be distinguished easily from

other flies by the fact that they have both a long, piercing proboscis and scales on the

veins of their wings. Approximately 176 species of mosquitoes are found in the United

States, with more than 3,000 species known throughout the world. In the United States,

only a few of these species are important as carriers of disease, but many more are

important nuisance species that dramatically affect peoples’ quality of life.

While all mosquitoes need standing water to reproduce, different mosquito species are

found in different habitats. Some mosquitoes are considered “floodwater” species that

breed in temporary water habitats, while others are considered “permanent water”

mosquitoes and breed in water sources that remain for long periods of time. Other species
 8

have evolved so specifically that they will only lay their eggs in natural or artificial

containers. (Vector Disease Control International, 2013)

The length of the mosquito life cycle varies between species and is dependent

upon environmental conditions such as temperature and moisture. However, the life cycle

of all mosquitoes is comprised of the egg, larval, pupal, and adult stages and always

aquatic.

Male mosquitoes feed on plant nectar alone, while females extract the blood of

hosts in order to develop and nourish eggs. Most mosquitoes lay their eggs directly into

water. Others lay their eggs near bodies of water but not within them.

Eggs will hatch into larvae within 24 to 48 hours. Larvae soon grow to become

approximately 5 mm in length. Most larvae breathe through air tubes. Larger larvae can

be seen floating just above the surface of infested waters. Larvae and pupae

usually cannot survive without water. If a water source evaporates before the larvae and

pupae within it transform into adult mosquitoes, those young often will die.

Within seven to ten days, larvae enter the pupal stage. Pupae are also visible upon

the surface of the breeding site. After a mosquito is fully developed, it will emerge as an

adult from its pupal case. At this time, the new adult stands upon the water and dries its

wings to prepare for flight. Adult female mosquitoes will then seek an animal on which to

feed. Females are capable of flying for miles if necessary and can lay over 100 eggs at a

time (Orkin, 2019).

The male adult mosquito will usually emerge first and will linger near the

breeding site, waiting for the females. Mating occurs quickly after emergence due to high

adult mortality rates. As much as 30% of the adult population can die per day. The
 9

females compensate for this high rate by laying large numbers of eggs to assure the

continuation of the species. Male mosquitoes will live only 6 or 7 days on average,

feeding primarily on plant nectars, and do not take blood meals. Females with an

adequate food supply can live up to 5 months or longer, with the average female life span

being about 6 weeks. To nourish and develop her eggs, the female usually must take a

blood meal in addition to plant nectars. She locates her victims by the carbon dioxide and

other trace chemicals exhaled, and the temperature patterns they produce. Mosquitoes are

highly sensitive to several chemicals including carbon dioxide, amino acids, and octenol.

The average female mosquito’s flight range is between 1 and 10 miles, but some species

can travel up to 40 miles before taking a blood meal. After each blood meal, the female

will oviposit (lay) her eggs, completing the life cycle. While some species oviposit only

once, others may lay eggs several times over the course of their lives. (Vector Disease

Control International, 2013)

Related studies about mosquitoes

The world’s deadliest animal isn’t a shark or even a human. Data shows

mosquitoes kill 250,000 animals more each year when compared to humans. Yes,

mosquitoes – the pesky bugs that suck blood and transmit viruses from person to person –

are responsible for the most animal-related deaths (830, 000 per year to be exact). For

comparison, humans are responsible for 580,000 human deaths per year, snakes account

for 60,000 deaths per year and sharks claimed just six lives per year ( Ramsey, 2018).

Mosquitoes ability to carry and spread disease to humans causes millions of

deaths every year. In 2015, malaria alone caused 438, 000 deaths. The worldwide

incidence of dengue has risen 30 – fold in the past 30 years, and more countries are
 10

reporting their first outbreaks of the disease. Zika, dengue, chikungunya, and yellow

fever are all transmitted to humans by Aedes aegypti mosquito. More than half of the

world’s population live in areas where this mosquito species is present. Sustained

mosquito control efforts are impotant to prevent outbreaks from these diseases (WHO,

1996).

World Health Organization (WHO, 2016) estimates that malaria, a mosquito

borne-infectious disease, affect 216 million people accross the world and kills about 655

000 each year.

In Indonesia, the Philippines, Thailand and Vietnam, an estimated 24,632 people

died from malaria and dengue in 1990. With global warming, deaths are expected to

increase by 18 percent in 2020, according to Asian Development Bank. The overall

burden from climate change to be expected in different Asian regions by the 2030s and

2050s is therefore projected to be much greater than reflected by the mortality estimates

shown here (Testa, 2017) .

Most of the mosquito-borne diseases have no cure. There are effective treatments

for malaria but it is a constant arm race. The parasite evolves defences; researchers and

industry race to come up with safe, effective new treatments. So, much of the focus is in

prevention (Parry, 2017).

As cited in one of the Young Naturalist Collection, throughout the world, there

are about 3,500 species of mosquitoes. The female mosquito bites people and animals

because they need the protein found in blood to help develop their eggs. Mosquitoes are

attracted to people by skin odors due to lactic acid and the carbon dioxide from breath

(Bowen 1991) and which is also present in our sweat. The chemoreceptors present in the
 11

antennae of mosquitoes perceive the smell of the sweat. The use of repellents makes a

person unattractive for feeding and therefore repels the mosquito (Maibach et al., 1966).

To prevent proliferation of mosquito borne diseases and to improve

quality of environment and public health, mosquito control is essential. The major tool in

mosquito control operation is the application of synthetic insecticides such as

organochlorine and organophosphate compounds. But this has not been very successful

due to human, technical, operational, ecological, and economic factors. In recent years,

use of many of the formal synthetic insecticides in mosquito control programme has been

limited. It is due to lack of novel insecticides, high cost of synthetic insecticides, concern

for environmental sustainability, harmful effect on human health, and other non-target

populations, their non-biodegradable nature, higher rate of biological magnification

through ecosystem, and increasing insecticide resistance on a global scale. The US

Environmental Protection Act in 1969 had rules governing the application of chemical

control agents in nature and that prompt the research to look for alternative methods of

approaching transparent mosquito control measures that focuses on public education,

monitoring and surveillance, source reduction and environment friendly least toxic larval

control. These factors have resulted in urgent search for environmental friendly, cost-

effective, biodegradable and target specific insecticides against mosquito species.

Considering these, the application of eco-friendly alternatives such as biological control

of vectors has become the central focus of the control program in lieu of chemical

insecticides (Centers for Disease Control and Prevention, 1993).

The plant world comprises a rich untapped pool of phytochemicals that

may be widely used in place of synthetic insecticides in mosquito control programme.

 12

Kishore et al. (2011), reviewed the efficacy of phytochemicals against mosquito larvae

according to their chemical nature and described the mosquito larvicidal potentiality of

several plant derived secondary materials, such as, alkanes, alkenes, alkynes and simple

aromatics, lactones, essential oils and fatty acids, terpenes, alkaloids, steroids,

isoflavonoids, pterocarpans and lignans.

Insecticidal effects of plant extracts vary not only according to plant species,

mosquito species, Insecticidal effects of plant extracts vary not only according to plant

species, mosquito species, geographical varities and parts used, but also due to extraction

methodology adopted and the polarity of the solvents used during extraction. A wide

selection of plants from herbs, shrubs and large trees was used for extraction of mosquito

toxins. Phytochemicals were extracted either from the whole body of little herbs or from

various parts like fruits, leaves, stems, barks, roots, etc., of larger plants or trees. In all

cases where the most toxic substances were concentrated upon, found and extracted for

mosquito control. (Ghosh, 2012)

Extracts from plants may be alternative sources of mosquito egg and larval

control agents, since they constitute a rich source of bioactive compounds that are

biodegradable into non-toxic products and potentially suitable for use in control of

mosquito larvae (Pushpanathan et al., 2008)

Morphological Structure of Madre de Cacao

Madre de cacao (Gliricidia sepium) is a small to medium-sized, thornless,

leguminous tree up to 10-12 m high. The bark is smooth, varying in colour from whitish

grey to deep red-brown. The trees display spreading crowns. The leaves are odd pinnate,

usually alternate, sub-opposite or opposite, to approximately 30 cm long; leaflets 5-20,

 13

ovate or elliptic, 2-7 cm long, 1-3 cm wide. The leaflet midrib and rachis are

occasionally striped red. ( Tropical Forages, 2017).

Chemical composition of Madre de cacao

The study of leaves yielded two new triterpene saponins (1 and 2), possessing

3beta

21beta, 24--trihydroxy-22-oxoolean-12-ene as aglycon, together with known aromatic

compounds. Study of bark oil by GC-MS analysis yielded 19 compounds. Leaf oil

consisted mainly of alipathics (54.9%) and terpenoids (28.1%). Major compounds from

the leaf oil were pentadecanal (18.7%), (Z)-phytol (7.8%), methyl linolenate (6.0%) and

nonanal . (Stuart, 2018)

Based from the study of Ramamoorthy et al. (1993), allelochemicals from

Gliricidia sepium were extracted, identified, and quantified using HPLC. Fifteen toxic

compounds, namely gallic acid, protocatechuic acid, p-hydroxybenzoic acid, gentisic

acid, t3-resorcyclic acid, vanillic acid, syringic acid, p-coumafic acid, m-coumaric acid,

o-coumaric acid, ferulic acid, sinapinic acid (trans and cis forms), coumarin, and

myricetin were identified and quantified.

Uses of Madre de cacao

Madre de cacao is very easy to propagate and inexpensive. The tree could re-

sprout very quickly after pruning. Many farmers plant them mainly to shade other

perennial crops like cacao, coffee and tea. Aside from this, Madre de cacao ould provide

a lot of uses to the farmers from its roots to its leaves. Its multipurpose use makes it a

good plant crop in agroforestry.

 14

Since Madre de cacao is a legume, it is useful for fixing nitrogen in the soil, thus

improving soil quality and increasing crop yields. Madre de cacao has strong roots. It

stabilizes sloping lands and reduces soil erosion. Its wood could be used as firewood,

hedges, and fencing field. The leaves are rich in nitrogen and other nutrients suitable for

green manure and fodder to farm animals.

The trees are used for intercropping in part because they fix nitrogen in the soil

and

tolerate low soil fertility, so when they are interplanted with crops they can boost crop

yields significantly, without the need of chemical fertilizers.

Madre de cacao is also used for its medicinal and insect repellent properties.

Farmers in Latin America often wash their livestock with a paste made of crushed G.

sepium leaves to ward off torsalos (botflies). In the Philippines, the extract obtained from

its leaves is made into anti-mange dog shampoo. (Wikipedia)

From the report of Dela Cruz (2003) , the bark of Madre de cacao is stripped and

cooked with grains like corn or rice and used as poisonous bait for rodents. The

advantage of Madre de cacao as bait is that the rats do not develop bait shyness that is

common in using synthetic rodenticide. The Madre de cacao has distinctive aroma that

could attract the rodents that eventually, with the right amount could terminate them.

The active substance in Madre de cacao is not a rapidly acting substance and

needs repeated doses for it to be effective. But unlike commercially prepared rat killers, it

is less lethal in case of accidents. Farmers who used Madre de cacao observed that when

rats eat it, their hands stiffen and they get bloated and die of hemorrhagic poison. After
 15

the bait was consumed it would take one to two days before the farmers could find them

dead on the fields.

The leaves of Gliricidia sepium, the multipurpose leguminous tree commonly

used in alley farming, contain a substance called coumarin. This can be converted to

dicoumerol, a substance that is chemically so similar to Vitamin K that it interferes with

Vitamin K's role in blood clotting. Warfarin, the well-known rat poison, is a similar

compound. Despite these properties Madre de cacao does not pose a threat when fed to

livestock as the coumarin is not particularly toxic. The potential risk is in feeding spoiled

leaves, in which the coumarin to dicoumerol change may have occurred. Silage should

also prove to be safe, although quality control will have to be good to avoid spoilage and

the risk of dicoumerol production.

Medicinal Value of Madre de cacao

Crude extracts have been shown to have antifungal activity. Reported to be

expectorant, sedative and suppurative. Madre de cacao is a folk remedy for alopecia,

boils, bruises, burns, colds, cough, debility, eruptions, erysipelas, fever, fractures,

gangrene, headache, itch, prickly heat, rheumatism, skin tumours, ulcers, urticaria and

wounds. (Healing Wonders of Philippine Medicinal Plant, 2015)

Related Studies About the Potentialities of Madre de cacao

Based on an article made on a survey of plants used as repellants against

hematophagous insects by the Ayta people of Porac, Pampanga, Gliricidia sepium is one

of the plants in the list as an effective insect repellent utilizing its leaves and stem.

( Obico, 2014)
 16

Madre de cacao is used by farmers in some Latin American countries to repel

insects. The leaves are ground up and combined with water. The animal is then bathed

with the resulting paste. According to some of the farmers, if this is repeated every 7-14

days, the number of torsalo (tropical warble fly) infections is decreased. No published

studies could be found to substantiate this claim. However, when interviewing farmers

and checking goats in Honduras, It was found out that the goats who the farmers claimed

to have bathed with Gliricidia had only 2-3 torsalos, while others had 10 or more. Also,

one study did indicate that the heartwood of Gliricidia contains compounds that attract

and are toxic to certain insects (e.g., southern army worm, cabbage looper, yellow woolly

bear, and Glyptotermes dilatatus, a termite. (Antonio et al., 2014)

From the report of Danny Calleja (2010), kakawate leaves can be used to

deworm pigs. Other than as pig dewormer, termite and bed bug neutralizer, anti-fungus

and bio-organic fertilizer among others, researchers have discovered the leaves as an

effective pesticide, according to the Department of Agriculture of Bicol.

A study, published in Daily Journey (2012) done by Alfredo Rabena, a full time

professor at the University or Northern Philippines in Vigan City, found that Madre de

Cacao leaves are good source of coumarins. A toxic substance that can kill almost all

types of pests and insects.

Rabena said one kilogram of kakawati leaves, soaked. In water overnight can

produce seven gallons of “botanopesticides” (botanical pesticides). He also added that

the more leaves, the pesticides will be more concentrated. Rabena said he conducted the

study in 1996 in collaboration with a Malaysian chemist Dr. Nordin Lajiz at the

University of the Philippines in Los Baños, Laguna.

 17

For three years study Dr. Rabena found that botanopesticide is really effective.

He

said, adding the technology is being used by rice and vegetable farmers in eight

municipalities in Ilocos sur. Rabena said he wants to provide Filipino farmers an

alternative to commercial pesticides to lessen the cost of production.

Field demonstrations conducted in several parts of

the Ilocos region, proved Rabena’s discovery that the kakawate leaves’ botanopesticide

effectively eliminated rice weevils, rice bugs and worms in ricefields.

Kakawate leaves are also effective anti-fungus. It can cure Tricophyton

Metagrophytes that causes skin diseases like eczema. Crumple several leaves and apply

to affected area of the skin for a salicylic acid-like effect (Tacadena, 2010).

Research has been conducted on both the antifungal and antimicrobial properties

of Madre de cacao extracts. In a brine shrimp toxicity test, a general screening method

indicative of cytotoxicity and pesticidal activity was tested. Medicarpin, one of the

compounds in the leaves and heartwood of Madre de cacao, is supposed to be antifungal.

In an antifungal study, Madre de cacao extracts inhibited the germination of Drechslera

oryzaee only 6%. However, in another study, 50 ug of stem chloroform extracts inhibited

the growth of Cladosporium cucumerinum and slightly inhibited the growth of Candida

albicans. In contrast, in another study, the antimicrobial properties of extracts from the

bark of gliricidia were tested. It was effective against bacteria causing dermatitis.

However, it was not effective against Candida albicans could be because of the quantity

of plant extract used or the type of extracts used. In another study, leaf extracts were

found to be effective against Dermatophytes microsporumcanis,

 18

Trichophytonmentagrophytes var algodonosa and T. rubrum. (Ganesan as cited in Arranguez et al.,

2019).

The researchers, Baylon et al. (2016), conclude that the 100% Neem tree

(Azadirachta indica), Madre de cacao (Gliricidia sepium), and Garlic (Allium sativum)

extract had pesticidal effect against cockroaches (Periplaneta americana).The positive

group showed more effectivity because of its shorter time compared to the plant extracts.

Based on values presented by the experiment the rest comparisons or results were

significantly differed at 0.05 level. The three extracts tested showed less effectivity

compared to the positive control Baygon. This is because of the p-value of Madre de

cacao and garlic is 0.052.

Based from the study of Mathew et al. (2015), the crude extract of Gliricidia

sepium had no effect on the life of guppy fry, the fish that commonly consumes mosquito

larvae for their food. From the control it was clear that petroleum ether had no direct

effect on these organisms. The results shows that the 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. This

indicated that the toxicity levels of the extract are dependent on time of exposure of the

larvae and the applied concentration.

Madre de cacao is well known in Central America, where the leaves or the ground

bark, mixed with cooked maize, are used as a rodenticide. This toxicity is thought to be

due to the conversion by bacteria of coumarin to dicoumerol during fermentation. May be

toxic or inhibit growth of monogastric animals such as rabbits and poultry if fed as a

sufficiently high component of the diet. Little evidence of toxic effects with ruminants
 19

fed either fresh or wilted leaves. HCN concentrations of up to 4 mg/kg and cyanogens

may be present. High levels of nitrates (during the rainy season) are suspected of causing

`cattle fall syndrome' in Colombia, but levels declined to negligible in winter. Gliricidia

may be a `nitrate accumulator'. Unidentified alkaloids and tannins have also been

reported.

Evidence of toxicity under practical feeding conditions is limited. The balance of

evidence suggests that the plant may be toxic to non-ruminants but conclusive evidence

of toxicity to ruminants under normal feeding is lacking. (Tropical forages)

According to a study reported in 1966, as cited by Torres et al. (2018), there is an

evaluated mechanism by which Gliricidia sepium exerts its rodenticidal properties. Leaf

extraction yielded the presence of coumarin asa a constituent of the phenolic fraction.

Study suggests the bacterial conversion of coumarin into the hemorrhagic agent

dicoumerol, and the pathologic evidence in rats fed on incubated leaves point to coumarin

as the basis for rodenticidal property of plant.

The phytochemical screening performed by Cruz et al. (2016), showed that the

leaves and bark extracts of Madre de cacao do contain secondary metabolites such as

alkaloids and saponin. The positive result for alkaloid was indicated by the presence of an

orange precipitate in Draggendorff's reagent and white precipitate the Mayer's reagent.

Positive result for saponin was determined by its level in the capillary tube which is half

or less than in other tube containing distilled water. Saponins have been used as

detergents, pesticides, molluscicides, apart from its industrial applications as foaming and

surface active agents which show beneficial health effects. The secondary metabolites
 20

that are not present in the leaves and bark extracts of Madre de cacao are the

anthraquinone, leucoanthocyanin, phenolic compounds, steroids, tannins, terpenoids.

Saponins were also known by their toxicity to harmful insects (anti-feeding,

disturbance of the moult, growth regulation, mortality…); the insecticidal activity of

saponins is due to their interaction with cholesterol, causing a disturbance of the synthesis

of ecdysteroids. (Chaieb, 2010)

The study conducted by Calumpang et al. (2014), proved that Madre de cacao

planted along rice fields can be promoted for insect pest management in rice production

to promote sustainability and reduce dependence on synthetic insecticides. It can be a

useful pest management strategy in organic and low-input rice production.

Related Studies About the Potentialities of Lagundi

Lagundi or Vitex negundo, commonly known as the Chinese chaste tree, five-

leaved chaste tree, or horseshoe vitex, is a large aromatic shrub with quadrangular,

densely whitish, tomentose branchlets. It is widely used in folk medicine, particularly

in South and Southeast Asia.

Leaves contain an alkaloid nishindine, flavones. Phytochemical screening of

ethanol leaf extract yielded alkaloids, iridoids, phenolic acids, flavonols and flavonoids.

(Stuart, 2018)

Based from the research conducted by Ulah et al. (2017), mosquito control is

facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of

botanical origin may serve as suitable alternative biocontrol techniques in the future. The

acetone, chloroform, ethyl acetate, hexane, methanol and petroleum ether extracts of leaf,

flower and seed of Cassia auriculata L.,Leucas aspera (Willd.), Rhinacanthus

 21

nasutus KURZ.,Solanum torvum Swartz and Vitex negundo Linn. were tested against

fourth instar larvae of malaria vector,Anopheles subpictus Grassi and Japanese

encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae).The larval

mortality was observed after 24 hour of exposure.

These results suggest that the leaf petroleum ether, flower methanol extracts of C.

auriculata, leaf and seed methanol extracts of S. torvum and leaf hexane extract of V.

negundo have the potential to be used as an ideal eco-friendly approach for the control of

the A. subpictus and C.tritaeniorhynchus. This is the first report on the mosquito

larvicidal activity of the medicinal plant extracts.

Phytochemical studies on Vitex negundo Linn revealed the presence of

volatile oil, triterpenes, diterpenes, sesquiterpenes, lignan, flavonoids, flavones,

glycosides, iridoid glycosides and stilbene derivative. (Suganthi et al., 2016)

Snake venom neutralization by the methanolic roots extracts of Vitex negundo

Linn and Emblica officinalis was performed against the Vipera russelli and Naja kauthia

venom induced lethal activity both in vivo and invitro and since no precipitating bands

were found between the plants extract and snake venom which revealed that these plants

extracts posses potent anti snake venom activity as cited by Suganthi et al. (2016), in

Alam et al. (2003), study.

The methanolic, petroleum ether and carbon tetrachloride leaf extract of Vitex

negundo Linn were screened for antibacterial and cytotoxic activity using disc diffusion

method and brine shrimp lethality bioassay respectively. Antibiotic kanamycin and anti

tumor agent vincristine sulphate were used as standard and the fractions screened for anti

bacterial activity showed prominent zone of inhibition against Bacillus megatrium,

 22

Bacillus subtilis, Salmonella typhi ,Vibrio mimicus, Aspergillus niger. When compared to

methanolic extract, the petroleum ether and carbon tetra chloride extract showed

significant cytotoxic activity (Chowdhury et al., 2009)

As cited by Suganthi et al. (2016), the larvicidal activity of flavonoid extract of

different parts of Vitex negundo Linn and Andrographis paniculata is performed against

the late III or IV instar larvae of Aedes aegypti and Anopheles stephensi and the two

plants showed good larvicidal activity and can be used to synthesis eco- friendly

insecticide.

The leaves, bark, roots, and seeds of V. negundo have long been used in

traditional medicine to heal wounds, cleanse ulcers, prevent insect bites, treat snake bites,

and relieve rheumatism. Its oil has also been used to relieve sores and sinuses. Recent

studies have proven its antiseptic, antitussive, as well as its anti-inflammatory properties.

It has been found to have antibacterial effects against Bacillus subtilis, Escherichia coli,

and Staphylococcus aureus. These properties make it an effective drug for coughs and

bronchopulmonary disorders such as bronchial asthma and acute bronchitis.

Vitex negundo Linn or lagundi is credited with innumerable medicinal activities

like analgesic, anti-inflammatory, anticonvulsant, anti oxidant, bronchial relaxant and

hepatoprotective. The plant products of Vitex negundo are variously reported to possess

insecticidal ability against stored-product pests, i.e. pantry pests include several beetles,

moths, and mite that can infest whole grains or processed foods, pests, houseflies, and

tobacco leaf eating larvae. Leaf oil of the plant is shown to have repellent action against

stored product pests. (Tandon, 2005)

 23

Amancharla et al. (1999), also tested mosquito repellent activity of aqueous

extract of Vitex negundo leaves. A new chemical ‘rotundial’ was tested for the said

activity. The chloroform fraction of the aqueous extract of the fresh leaves of Vitex

negundo by bioactivity bioactivity guided isolation yielded a pure compound rotundial

which has shown mosquito repellent activity.

A study conducted by Gokulakrishnan et al. (2015), on the larvidcidal and

ovicidal activities of Vitex negundo, they found out that the leaf extract of Vitex negundo

potential larvicidal and ovocidal agents against the dengue vector Aedes aegypti and they

have demonstrated a synergist act too.

Another study made by Rathnasagar and Thiyagaraj (2018), the larvicidal effect

of L.indica and V. negundo plant extracts showed increased mortality rate in EA and PE

extracts of V. negundo compared to L. indica other studies for larvicidal activity with

different plants also showed increased mortality with PE extracts, suggesting similar

compounds may involve in inducing mortality. The aqueous extract of both plants had

lethality only at higher concentration suggesting that compounds are more active in

organic solvents due to their chemical composition.

Related Studies About the Potentialities of Lemon Grass

Lemongrass, a common ingredient used either in dining or as an ambient scent

used in recreational spa and lifestyle activities, are also often to perceived as a pest

control solution alternative for mosquitoes.

Some would say that mosquitoes simply dislike the smell of lemongrass. Some

research has it that lemongrass contains citronella oil and musk scents that attract

mosquitoes such as carbon dioxide and lactic acid found in humans. In other words, by
 24

applying repellent with citronella oil ingredients, it blocks the scents being sensed by

mosquitoes. This makes it harder for mosquitoes to locate you. Research has shown that

citronella helps to reduce mosquito landing around 40%.

It is important to highlight that these natural repellents provide some form of

repellent against mosquitoes; however the active ingredient dosage level is not adequate

to serve as a mosquito control solution. Besides providing limited protection against

adult mosquitoes, undiluted essential oil is hazardous to health and safety. It may burn

your skin when comes into contact with, due to its high citral content. Some individuals

might be also sensitive and result in allergies, skin irritation or rashes. ( Rentokil, 2019)

Despite its ability to repel insects, its oil is commonly utilized as a “lure” to

attract honey bees. “Lemongrass works conveniently as well as the pheromone created by

the honeybee’s nasonov gland, also known as attractant pheromones. Because of this

lemon grass oil can be used as a lure when trapping swarms or attempting to draw the

attention of hived bees”.

Citronella grass (Cymbopogon nardus and Cymbopogon winterianus) grows to

about 2 meters (about 6.5 feet) and has red base stems. These species are used for the

production of citronella oil, which is used in soaps, as an insect repellent in insect sprays

and candles, and also in aromatherapy, which is famous in Bintan Island, Indonesia.

Therefore it’s assumed that its origin is from Indonesia. The principal chemical

constituents of citronella, geraniol and citronellol, are antiseptics, hence their use in

household disinfectants and soaps. Besides oil production, citronella grass is also used for

culinary purposes, in tea and as a flavoring.

 25

Lemon Grass Oil, used as a pesticide and preservative, is put on the ancient palm-

leaf manuscripts found in India as a preservative. It is used at the Oriental Research

Institute Mysore, the French Institute of Pondicherry, the Association for the Preservation

of the Saint Thomas Christian Heritage in Kerala and many other manuscript collections

in India. The lemon grass oil also injects natural fluidity into the brittle palm leaves and

the hydrophobic nature of the oil keeps the manuscripts dry so that the text is not lost to

decay due to humidity. (AgriBusiness, 2016)

The composition of lemon grass oil depends partly on geographical region or

where it is grown. Major constituents of lemon grass oil are citral, geraniol, myrcene,

citronellal and limonene. Citronella oil, which repels mosquitoes, contains large amounts

of citral. Citral gives lemon grass oil its lemony fragrance. (Blue, 2018)

Citronella candles are widely used to repel mosquitoes. Citronella oil and other

lemon grass oil extracts are used in sprays and topical applications. Studies with birds

and starved mosquitoes, using topical applications of lemon grass oil in the form of

cream or liquid paraffin solution, was effective protection against the hungry mosquitoes,

comparable to commercial mosquito repellents, according to a 2002 report in the journal

Phytomedicine as cited by Blue (2018).

Lemon grass oil is an effective insecticide against ticks, termites and dust mites,

according to study results summarized in the Sigma-Aldrich Plant Profiler. Lemon grass

also has anti-larval and anti-helmintic activity. Geraniol, a constituent of lemon grass oil,

was able to kill helminths, which are intestinal worms. A 2004 study in Brazil published

by Memórias do Instituto Oswaldo Cruz, an international journal of biological and

 26

biomedical research, showed that the essential oil of lemon grass could kill the larvae of

the mosquito that causes dengue fever.

According to Wells (2019), lemon grass is a common ingredient in cosmetic and

skin care products (lotions, soaps, tonics, deodorants, and shampoos. It deodorizes and

freshens the air, especially when you combine it with other known essential oils such as

bergamot and geranium. You can just put it in a vaporizer, oil burner, or diffuser. You

can also use it with deodorizers and air fresheners. It is an important aroma therapeutic

oil. Its aroma calms and cleans. It also relieves insomnia, stress, anxiety and irritability.

Lemongrass also prevents drowsiness. It helps relieve muscle pain, period cramps,

rheumatism, toothaches, headaches, migraines, and stomachaches. It also helps relax and

tone your muscles.

It is also an effective insect repellent. It can repel various insects such as ants and

mosquitoes. Lemongrass is a natural repellent because it has high geraniol and citral

content.

Extracts from plants may be alternative sources of mosquito egg and larval

control agents, since they constitute a rich source of bioactive compounds that are

biodegradable into non-toxic products and potentially suitable for use in control of

mosquito larvae (Pushpanathan et al., 2008). In fact, many researchers have reported on

the effectiveness of plant extracts or essential oils against mosquito larvae (Pushpanathan

et al., 2008), such as Cymbopogan citratus (lemon grass). According to an article of New

Tech Bio., lemongrass is very popular and used for medicinal, food and insect repellent

products. The lemongrass oils are used in cosmetics, soaps, perfume, dyes, and odorizes

along with thousands of other products. Lemongrass are extremely safe and is listed on
 27

the EPA’S GRAS list (Generally Regarded As Safe), unlike other insecticides containing

chemical compound like DEET (N,NDiethlymetatoluamide) used as active chemical

ingredient in insect repellent. People all over the world have come to rely on the potent

insect repellent properties of lemongrass and utilized it as a personal and areas spray. As

noted by Goddard (2002), insect repellents are important public health tools for the

prevention of vector-borne infectious diseases The actions to reduce vector-borne

diseases can result in major health gains and relieve an important constraint on

development in poor regions .Technically, an insect repellent is any chemical and natural

insecticide that causes insects or other arthropods to make directed, oriented movements

away from the source of repellent, and lemon grass can be used as natural insect

repellent.

Lemongrass is a safe and natural insect repellant that is just as effective as the

commercial chemical product and should be planted around homes, as they could have

potentials of repelling mosquitoes and reducing mosquito borne diseases. (Goselle et al.,

2017)

Based from the study of Musa et al. (2015), the larvicidal and insecticidal effect

of the oil extract of C. citratus thus shows that it is highly effective having hundred

percent mortality rate efficiency. The time taken for the mosquitoes to die decreases by

increasing the amount of oil extract; this indicates that the higher the amount of extract

the faster the mortality rate which is in line with. The grass proved to be active, that

within nineteen minutes of application all the larvae and mosquito were dead. Compared

to previous work done shows that Cymbopogon citratus and Ageratum conyzoides against
 28

Ae. aegypti have achieved 100% mortality at 120, 200 and 300 mg/L concentrations

respectively.

Related Studies About the Potentialities of Star Apple

Star apple or Chrysophyllum cainito Linn is a tree with a spreading crown,

growing to a height of 15 meters. Leaves are leathery, ovate or oblong, 7.5 to 13

centimeters long, pointed at the tip, blunt or rounded at the base and covered underneath

with silky, golden-brown, soft hairs.

Leaves contain an amorphous bitter principle, some alkaloids, and no saponin.

Phytochemical fractions of leaves yielded (% content): fats and waxes 0.934 ± 0.045,

terpenoids and phenolics 4.004 ± 0.122, alkaloids 0.166 ± 0.068, quaternary alkaloids and

n-oxides 10.678 ± 0.035, and fibers 71.122 ± 0.136.

Study evaluated the immunomodulatory effect of methanol extract of C. cainito

leaves on macrophage functions. Results showed significant dose dependent inhibition of

phagocytosis and decreased IL-6 and TNF-α as well as NO and H2O2 release by the

macrophages. Study showed an immunosuppressive effect on murine macrophages,

without effects on cell viability. GC-MS analysis showed lupeol acetate and alpha-amyrin

acetate as principal components. (Stuart, 2018)

Study evaluated C. cainito leaves for wound healing potential in male albino

Wistar rats using an excision wound model. Topical application of standardized ethanolic

extract on excision wounds showed significant faster reduction in wound area and

significant increase in tissue biochemical parameters such as hydroxyproline, hexosamine

and protein compared to untreated control.

 29

Study reports on the synthesis of environmentally friendly silver nanoparticles

using C. cainito leaves. (Stuart, 2018)

The seeds contain 1.2% of the bitter, cyanogenic glycoside, lucumin; 0.0037%

pouterin; 6.6% of a fixed oil; 0.19% saponin; 2.4% dextrose and 3.75% ash. The leaves

possess an alkaloid, also resin, resinic acid, and a bitter substance. (Morton, 1987) .

Fruits and vegetables are a good source of natural antioxidants, containing

many different antioxidant components which provide protection against harmful free

radicals which have been implicated in the aetiology of several human ailments such as

cancer, neural disorders, diabetes, arthritis, and cardiovascular disorder. An investigation

on the antioxidant and food value of Chrysophyllum showed the plant contains some

phenol, flavonoid, anthocyanin and proanthocyanidin and high antioxidant value.

According to a survey, the star apple consists of an alkaloid known as

eleagnine,

which shows a powerful antimicrobial activity. This compound has also been found to

possess antioxidant and anti-inflammatory. The star apple seeds have been used in the

preparation of ointments that are designed to fight infections. Researchers also found that

the star apple tree leaves exhibits strong antibacterial activity against common bacteria.

(Health benefits times.com)

The researcher were given wider prospective and insights to conduct this study

based from the related literature and studies.

Statement of the Problem

This study aimed to determine the efficacy of selected plants leaf extracts (Madre

de cacao, Lagundi, Lemon Grass and Star apple) as mosquito larvicidal.

 30

Specifically, the study try to search answers for the following questions:

1. What is the level of efficacy of Madre de cacao as mosquito larvicidal on

different concentrations?

a. 25 % concentration

b. 50% concentration

c. 75% concentration

d. 100% concentration

2. What is the level of efficacy of combined mixtures of Madre de cacao and

other

plants on different concentrations?

a. Madre de cacao and Lagundi

b. Madre de cacao and Lemon Grass

c. Madre de cacao and Star Apple

3. Which among the concentrations is effective in eliminating larvae?

Hypothesis

The hypothesis of the study are:

1. There is a significant difference in the level of efficacy of Madre de cacao leaf

extract in killing mosquito larvae on different concentrations.

2. There is a significant difference in the level of efficacy of combined mixture

on

different concentrations.

a. Madre de cacao and Lagundi

b. Madre de cacao and Lemon Grass

 31

c. Madre de cacao and Star Apple

Chapter II

METHODOLOGY

This chapter presents the time and place of the study, materials and methods used,

data gathering procedures and the statistical treatment of the data.

Time and Place of the Study

This study was focused on the efficacy of selected plants leaf extracts (madre de

cacao, lagundi, lemon grass and star apple) as mosquito larvicidal treated on different

concentrations. The materials were collected within Brgy. Monggoc, Pidigan, Abra. The
 32

experiment was done for almost two weeks on the residential house of the researcher at

Brgy. Monggoc, Pidigan,Abra.

Materials
The materials used in this study were:

1. Fresh and mature leaves of madre de cacao, lagundi, lemon grass and star apple

2. Common mosquito species in larvae stage

3. Other materials and equipment used were:

a. Graduated cylinder- used to measure the amount and percentage concentration

of the extract.

b. Knife – for cutting the leaves into small bits

c. Blender – an electrical appliance with whirling blades for chopping, mixing or

liquefying foods. In this study, it is used for further grinding and mixing the pounded

leaves.

d. Watch glass – used for the set up and treatment

e. Strainer covered with stockings – used to catch the larvae

f. Clear glass – used as container for mixing the blended leaf extracts

g. Spoon – used to stir and mix the blended leaf extracts

h. Dissecting tools – used to direct the larvae to the set up for observation and

used to remove the dead larvae from the set up.

i. Distilled water – used to wash the leaves and other materials and was used to

prepare solution

j. Cheese cloth/stockings- used to extract the juice of the leaves

k. Funnel – used to direct the leaf extract to the graduated cylinder

l. Mortar and pestle – used to pound the leaves

 33

m. Paper strip and pen– used to record the time it took for each larvae to die

n. Flashlight – used to illuminate dark treatments for observation

o. Cellphone with timer – used to get the time when each larvae on the set up died

Methods

The following procedures were followed in the conduct of this study:

A. Mosquito Larvae

1. Culturing the mosquito larvae.

A pail of water was placed in between banana trees. The researcher added dry

leaves and dry banana husk which served as food for the mosquito. The set up was not

disturbed for three days. On the fourth day onward, the water was checked frequently for

the presence of mosquito larvae. The larvae were very visible after five to six days.

2. Preparation of materials for collecting the larvae

The researcher used a strainer covered with white stockings to catch the larvae.

This

was made so because the stocking is softer than the strainer which was made up of metal

material. The stockings provide cushion to the larvae so that it would not get hurt or

affected after isolating it from its original habitat. Then, the larvae will be transferred into

the watch glass filled with leaf extracts solution for observation.

3. Recording the rate of larvae mortality

The researcher started to record the time when all the needed mosquito larvae

were

placed in the treatment for observation. The treatments were checked frequently to check

for the dead larvae and record the time when each larvae was died. Dead larvae were
 34

removed or isolated from the treatments so that the other larvae will not be affected. The

treatments were observed for a duration of 24 hours.

B. Preparation of the Leaf Extracts

1. Collection of leaves

The researcher collected fresh and mature leaves of madre de cacao, lagundi,

lemon grass and star apple needed for the day’s experiment. After the leaves were

removed from their twigs or from their branches they were washed with distilled water to

remove the dust and dirt.

2. Extraction

The washed leaves were cut into small bits and then it was pounded using the

mortar and pestle. After it was pounded, it was put into the blending machine for further

grinding. After the leaves were pureed, it was transferred into the cheese cloth. The

cheese cloth was squeezed for the juice of the leaves to come out.

3. Preparation of solution

After the extracts were collected, the solution needed for the set up was prepared

next. The concentration of the 100 ml solution was varied to 25%, 50%, 75% and 100%.

a. Madre de cacao and distilled water solution

To prepare a 25% concentration in a 100ml solution, the researcher mixed

25ml

of madre de cacao extract and 75 ml of distilled water, for 50 % concentration, the

researcher mixed 50 ml madre de cacao extract and 50 ml distilled water. For 75%

concentration in a 100ml solution, 75 ml madre de cacao leaf extract and 25 ml distilled

 35

water and lastly for 100% concentration this is made up of 100ml madre de cacao leaf

extract.

b. Madre de cacao and Lagundi leaf extracts

To prepare a 25% concentration in a 100ml solution, the researcher mixed

25ml

of madre de cacao extract and 75 ml of lagundi extract, for 50 % concentration, the

researcher mixed 50 ml madre de cacao extract and 50 ml lagundi extract. For 75%

concentration in a 100ml solution, 75 ml madre de cacao leaf extract and 25 ml lagundi

extract and lastly for 100% concentration this is made up of 100ml madre de cacao leaf

extract.

c. Madre de cacao and Lemon grass extracts

To obtain a 25% concentration in a 100ml solution, the researcher mixed 25ml

of madre de cacao extract and 75 ml of lemon grass extract, for 50 % concentration, the

researcher mixed 50 ml madre de cacao extract and 50 ml lemon grass extract. For 75%

concentration in a 100ml solution, 75 ml madre de cacao leaf extract and 25 ml lemon

grass and lastly for 100% concentration this is made up of 100ml madre de cacao leaf

extract.

d. Madre de cacao and Star apple extracts

To obtain a 25% concentration in a 100ml solution, the researcher mixed 25ml

of madre de cacao extract and 75 ml of star apple extract, for 50 % concentration, the

researcher mixed 50 ml madre de cacao extract and 50 star apple extract. For 75%

concentration in a 100ml solution, 75 ml madre de cacao leaf extract and 25 ml star apple
 36

extract and lastly for 100% concentration this is made up of 100ml madre de cacao leaf

extract.

Data Gathered

The researcher utilized the experimental type of research. The experiment

consisted of five treatments with four replicates for madre de cacao and distilled water

solution. There were four treatments for madre de cacao and lagundi mixture, madre de

cacao and lemon grass mixture and madre de cacao and star apple mixture. All were

arranged in a Parallel Group Design.

The set up for the following mixtures:

1. Madre de cacao and distilled water

There were twenty watch glasses and one hundred mosquito larvae needed. For

the 25% concentration, four watch glasses were used for the four replicates. Each watch

glass was filled with 25 ml of the 25% concentration and five mosquito larvae each

replicate and the control contains 25 ml and five mosquito larvae taken from the original

habitat of the larvae still in four replicates. The same procedures were done using the

50%, 75% and 100% concentrations.

2. Madre de cacao and lagundi extracts

The researcher used sixteen watch glasses and eighty mosquito larvae. For the

25% concentration, four watch glasses were used for the four replicates. Each watch glass

was filled with 25 ml of the 25% concentration and five mosquito larvae each replicate

and the control contains 25 ml of pure madre de cacao extracts with five mosquito larvae

still in four replicates. The same procedures were done using the 50%, and 75%

concentrations.
 37

3. The same procedures were followed using the mixtures of madre de

cacao and lemon grass extracts and madre de cacao combined with star apple extracts.

Statistical Treatment of Data

The following statistical tools were used in this study:

1. To determine the level of efficacy of Madre de cacao leaf extract as mosquito

larvicide, the Mean was used.

2. Analysis of Variance (ANOVA) was used to test the significant difference in the

level of effectiveness of Madre de Cacao leaf extract and the combined mixture in killing

mosquito larvae at different concentrations.

 38

Chapter III

RESULT AND DISCUSSION

This chapter presents the result and analysis of the gathered data.

Table1. Transformed data on the Mortality Rate of Mosquito Larvae on Madre de cacao
and distilled water.
Mortality Rate of Mosquito Larvae (hr)
Treatment Mean Mean Mean Mean
(3hrs) (6 hrs) (9 hrs) (12 hrs)
Control 0.71c 0.71b 0.71b 0.71b

25% 0.71c 0.97ab 1.44a 1.76 a

50% 2.17b 1.09 a 0.71b 0.71b

75% 2.17b 1.09 a 0.71b 0.71b

100% 2.35a 0.71b 0.71b 0.71b

The table presents the mortality rate of mosquito larvae on madre de cacao leaf extracts

on different concentrations. It consists of five treatments reflecting the different

concentrations of madre de cacao and four observations with three hours interval.

Looking at the means of the different concentrates, the mortality of mosquito

larvae within three hours is highest at 100% concentration and lowest at 25%

concentration. For 6 hours, it is highest on the 50% and 70 % concentrates and lowest

surprisingly at 100% concentration. Within 9 hours, the highest mortality is at 25% and

for 12 hours at 25% concentration, too.

On the analysis of variance, the first observation has F-value of 672. 69 and F –

probability of 0.00, the second observation has F-value of 4.22 and F – probability of

0.02, the third observation has F-value of 8.3.18 and F – probability of 0.002 and the last
 39

observation has F-value of 20.132 and F – probability of 0.00. At 0.05 level of

confidence, all the observations has significant difference.

Based from the data, the highest mortality rate of mosquito larvae is at the first

observation on the 100% treatment followed by the 50% and 75% concentration within 1

– 3 hours which means most of the mosquito larvae died at 50% - 100% concentration in

the duration of 3 hours. In addition, at 25% concentration all of the larvae died at the

duration of 6 – 12 hours.

Therefore, the mortality rate of mosquito larvae on madre cacao extract is higher

when concentration is higher and lower mortality rate when the concentration is low. The

higher concentration of madre cacao leaf extract the more it is effective as mosquito

larvicidal.

Larvicidal activity of extracts from leaves,flowers and roots of many plants were

reported in many studies. The phytochemical screening of those plants for saponins,

flavonoids, terpenoids, tannins, and steroids are carried out. (Joji and Beena, 2010)

Hence, based from the above findings madre de cacao has the potential as

mosquito larvicidal as stated on the study of Rabena, that madre de cacao leaves are good

source of coumarins, a toxic substance that can kill almost all types of pests and insects

cited from Daily Journey, (2012).

The phytochemical screening performed by Cruz et al. (2016), showed that the

leaves and bark extracts of Madre de cacao do contain secondary metabolites such as

alkaloids and saponin which are good as insecticide (Chaieb, 2010)

A study made by Tacadena (2010), found out that Madre de cacao leaves are

also effective anti-fungus. It can cure Tricophyton Metagrophytes that causes skin
 40

diseases like eczema. Crumple several leaves and apply to affected area of the skin for a

salicylic acid-like effect.

Furthermore, Ayta people of Porac, Pampanga uses Madre de cacao as as an

effective insect repellant based on an article made on a survey of plants used as repellants

against hematophagous insects utilizing its leaves and stem. (Obico, 2014)

Moreover, the study conducted by Krishnaveni et al. (2015), proven that the

ethanolic extract of Madre de cacao leaves was found to have an inhibitory effect on the

growth of larvae of A. aegypti. It was concluded that the ethanolic extract of Madre de

cacao was the most effective when compared to other solvent extracts by the mortality

rate of A. aegypti. In the statistical analysis, the highest significant difference was

observed between 1500 and 2000 mg/l concentration of ethanolic extract and other

extracts. However, no significant difference was observed in other concentrations. The

larvicidal activity of the plant extract may be attributed to the presence of active

compounds such as terpenoids, saponins, and steroids which were confirmed by TLC

studies and phytochemical analysis.

Table 2. Transformed data on the Mortality Rate of Mosquito Larvae on Madre de

cacao and Lagundi Leaf Extracts
Mortality Rate of Mosquito Larvae (hr)
Treatment Mean Mean Mean
(3hrs) (6 hrs) (9 hrs)
Control 0.71c 0.71b 0.71a

25% 1.77b 1.54a 0.71a

50% 1.98ab 1.35a 0.71a

75% 2.35a 0.71b 0.84a

 41

The table above shows the mortality rate of mosquito larvae on different

concentrations of Madre de cacao and Lagundi leaf extracts combined. It consist of four

treatments with different concentrations and three observations with 3 hours interval.

The result of the computed means showed that during 3 hours observation is 75 %

concentration has the highest mean and the 25% concentration has the lowest mean.

On the second observation having 6 hour duration the mortality rate of the larvae

registered highest is on the 25% concentration and the lowest registered mean is the 75%

concentration.

Lastly, on the third observation mortality rate computed values of the mean on

different treatments are very close but the 75% concentration of mortality showed to have

the highest.

This implies that the mortality rate of mosquito larvae is observed at high

concentration of madre de cacao in a three hour duration.

On the analysis of variance, the highest computed f-value is in the three hours

observation which is 35.79, followed by 6 hours observation which has 8.60 and the 9

hours observation is 1.0. When comparing their F-probability, the 3 hours observation

has 0.0 value, the 6 hours observation has 0.005 value and the 9 hours observation has

0.44 value. This means that the 3 hour observation is significantly different to the six and

9 hours observation.

Thus, using the madre de cacao and lagundi extract at any concentration

within the span of three hours, the mixture is effective in killing mosquito larvae.

Moreover, the higher the concentration of the madre de cacao extract the higher the

mortality rate as reflected on the computed mean value of the different concentrates.
 42

The result is supported with the study of Mathew et al. (2015), showing that the

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. This indicated that the toxicity levels of the extract are dependent

on time of exposure of the larvae and the applied concentration.

The leaves of Gliricidia are used in south India as a mosquito repellent, and they

have antifungal and antibacterial activity. 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. Forty two known compounds

are found in the leaves and flowers of G. sepium (Joji Reddy and Beena, 2010).

Result of Suganthi et al. (2016), also showed that the larvicidal activity of

flavonoid extract of different parts of Vitex negundo Linn showed good larvicidal effect

and can be used to synthesize eco- friendly insecticide.

Furthermore, Gokulakrishnan, et al. (2015), found that the leaf extract of Vitex

negundo has potential larvicidal and ovocidal agents against the dengue vector Aedes

aegypti and they have demonstrated a synergist act too.

Also Krishnaveni et al. (2015), said that natural insecticides may play an

important role in the 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 management.

 43

The potentiality of the madre de cacao and lagundi as insect repellent is

manifested from the above findings as claimed by other researchers.

Table 3. Transformed data on the Mortality Rate of Mosquito Larvae on Madre de cacao
and Lemon Grass Leaf Extracts
Treatment Mean Mean
(3hrs) (6 hrs)
c
Control 0.71 0.71a

25% 2.24b 0.71a

50% 2.35a 0.84a

75% 2.35a 0.84a

The table above shows the data on the mortality rate of mosquito larvae using

madre de cacao and lemon grass leaf extracts. It consists of four treatments with different

concentrations and two observations with 3 hours interval.

As registered on the table, the means of the number of dead mosquito larvae is

higher on the three hours observation compared to the six hours observation. The means

are the same in the 50% and 75% concentrations and has a higher difference compared to

the 25% concentration. Comparing the means of the 3 hours observation at any

concentration to that of the six hours observation it could be noticed that it has a great

difference.

Furthermore, there is a significant difference on the mortality rate of mosquito

larvae at 3 hours observation while on the 6 hours observation there is no significant

difference on the mortality rate as reflected on the analysis of variance using 0.05 as the

level of confidence. The 3 hours observation has F-value of 584.60 with F – probability

of 0.00 while on the 6 hours observation the F –value is 3.0 while its F – probability is

0.08.
 44

Based from the table, combined madre de cacao and lemon grass extracts

produces high mortality of mosquito. Within the 3 to 6 hours the 50% and 75%

concentrations has no significant difference thereby the efficacy of this mixture is just the

same. The efficacy of these leaf extracts mixture increases as the concentration of madre

de cacao increases.

The above result manifests that madre de cacao and lemon grass leaf extracts are

potential in killing mosquito larvae.

This is supported by a study in Brazil as cited by Blue (2010), that the essential

oil of lemon grass could kill the larvae of mosquito that causes dengue fever.

The result has proven the statement of Wells (2019), that lemon grass is an

effective insect repellent. It can repel various insects such as ants and mosquitoes.

Lemongrass is a natural repellent because it has high geraniol and citral content. You can

spray its more concentrated form around your home and the study of Mathew (2015), that

Gliricidia sepium or Madre de cacao plant have insecticidal activity, especially

mosquitocidal activity, against the larvae of the mosquitoes. It was also proven that the

extract has no lethal effect on non-targeted organisms. Therefore, it could be formulated

and used as an ecological friendly natural product for anti-mosquito activity.

Similar study was conducted by Goselle (2017), that different solvent extract of

lemon grass have significant larvicidal activity against Culex mosquitoes. Their study

further revealed that lowest mortality was recorded in wet extract of lemon grass.

Hence, extracts from plants may be alternative sources of mosquito egg and larval

control agents, since they constitute a rich source of bioactive compounds that are
 45

biodegradable into non-toxic products and potentially suitable for use in control of

mosquito larvae, (Pushpanathan et al., 2008)

Table 4. Transformed data on the Mortality Rate of Mosquito Larvae on Madre de cacao
and Star apple Leaf Extracts
Treatment Mean Mean Mean Mean
(3hrs) (6 hrs) (9 hrs) (12 hrs)
Control 0.71c 0.71b 0.71b 0.71b

25% 0.71c 0.71b 2.05a 1.27a

50% 1.85b 1.51a 0.71b 0.71b

75% 2.23a 0.93b 0.71b 0.71b

The table above shows the data on the mortality rate of mosquito larvae on madre

de cacao and star apple leaf extracts. It consist of four treatments with different

concentrations and four observations with 3 hours interval. The data reflects the means of

the dead larvae within 3 hours observation, 6 hours observation, 9 hours observation and

12 hours observation.

The highest mean in the 3 hours observation is the 75% concentration and the

lowest mean is in 25% concentration. This means that the mortality rate within 3 hours is

highest in the 75% concentration and lowest at 25% concentration.

In the 6 hours observation, the 50% concentration has the highest mean and the

25% concentration has the lowest. Thereby, data shows that mortality rate is higher in the

50% concentration than in the 25% concentration.

 46

Based from the data, in the 9 hours observation, the 25% concentration has the

highest mean and 50% and 75% concentration is the same.

On the analysis of variance, having 0.05 as level of confidence, the 3 hours

observation has an F-value of 60.70 and F-probability of 0.00, the 6 hours observation

has an F-value of 5.4 and F-probability of 0.02, the 9 hours observation has F-value of

135.448 and F- probability of 0.00 and on the 12 hours observation it has an F-value of

7.47 and F-probability of 0.008. Hence, all the observations on different treatments at a

given time intervals have significant difference.

Therefore, the data implies that at higher concentration of madre de cacao among

the treatment the higher the mortality rate and the lower the concentration of madre de

cacao in the treatment the lower its mortality rate. In addition, at 25% concentration most

of the larvae died at the span of 9 hours onwards while at 75% concentration most larvae

died after of 6 hours.

The efficacy of madre de cacao mixed with star apple extracts as mosquito

larvicidal at different concentrations is evident in this findings.

According to a survey, the star apple consists of an alkaloid known as eleagnine,

which shows a powerful antimicrobial activity. This compound has also been found to

possess antioxidant and anti-inflammatory. Researchers also found that the star apple tree

leaves exhibits strong antibacterial activity against common bacteria. (Health benefits

times.com)

This is comparable from the study of Stuart (2018), star apple leaves contain an

amorphous bitter principle, some alkaloids, and no saponin and from the research of

Bekele (2017) as cited from Secoy (1983), alkaloids are basic substances which contain
 47

one or more nitrogen atoms, usually in a heterocyclic ring. Although these chemicals are

not volatile, they may be used as repellants by burning plant material either on a fire or in

a mosquito coil to create an insecticidal smoke that repels the insects through direct

toxicity.

Madre de cacao is well known in Central America, where the leaves or the groun

d bark, mixed with cooked maize, are used as a rodenticide. This toxicity is thought to be

due to the conversion by bacteria of coumarin to dicoumerol during fermentation.

The phytochemical screening performed by Cruz et al. (2016), showed that the leaves and

bark extracts of kakawate do contain secondary metabolites such as alkaloids and

saponin. The positive result for alkaloid was indicated by the presence of an orange

precipitate in Draggendorff's reagent and white precipitate the Mayer's reagent. Positive

result for saponin was determined by its level in the capillary tube which is half or less

than in other tube containing distilled water. Saponins have been used as detergents,

pesticides, molluscicides, apart from its industrial applications as foaming and surface

active agents which show beneficial health effects. The secondary metabolites that are

not present in the leaves and bark extracts of kakawate are the anthraquinone,

leucoanthocyanin, phenolic compounds, steroids, tannins, terpenoids.

The study conducted by Calumpang et al (2014), proved that Madre de cacao

planted along rice fields can be promoted for insect pest management in rice production

to promote sustainability and reduce dependence on synthetic insecticides. It can be a

useful pest management strategy in organic and low-input rice production.

This is true when plants and their derivatives were used to kill mosquitoes and

other household and agricultural pests. These plants used to control insects contained
 48

insecticidal phytochemicals that were predominantly secondary compounds produced by

plants to protect themselves against herbivorous insects. (Shaalan et al., 2005)

Table 5. Over-all result of mortality rate of combined mixtures

Combinations Control 25% 50% 75%

Time (hrs) Time (hrs) Time (hrs) Time (hrs)

1 4 -6 7- 10 -12 1-3 4-6 7-9 10 -12 1-3 4-6 7- 10 - 1-3 4 7- 10 -

-3 9 9 12 -6 9 12

Madre de cacao - - - -
.71c 0.97b 1.44b 1.76a 2.18ab 1.09ab 2.18a 1.09a

Madre de cacao w/ lagundi - - - -

1.77b 1.54a 0.84c 0.71b 1.98ab 1.35ab 2.35a 0.71a

Madre de cacao w/ lemon - - - -

2.24a 0.97b 0.71c 0.71b 2.35a 0.71b 2.35a 0.71a
grass

Madre de cacao w/ star - - - -

.71c .71b 2.05a 1.27ab 1.85b 1.51a 2.23a 0.93a
apple

47
 48

The table above shows the mortality rate of mosquito larvae on the combined

mixtures at four observations.

As reflected in the table, the control is considered as not significant since there

were no values reflected, in the second observation , at 25% concentration the madre de

cacao with lemon grass extract has the highest mean while madre de cacao with distilled

water and madre de cacao with star apple has the lowest means. In the third observation,

at 50% concentration the madre de cacao and lemon grass mixture has the highest mean.

In the fourth observation, at 75% concentration the means of the mixture at 3 hours are

very close.

Based from the result, as the concentration increases the rate of mortality

increases. Moreover, the table evidently displays that the madre de cacao extract and

lemon grass mixture having the highest mortality rate followed by madre de cacao and

lagundi mixture.

Furthermore, the presented data in the table indicates that all the combined

mixtures of plants are indicative as larvicidal although the efficacy of each mixture

depends on their corresponding concentration.

One of the possible ways to overcome problems brought by the use of synthetic

insecticides is by the use of plant-derived insecticides as alternative for synthetic

insectides or for use in integrated management programs according to Shaalan et al.

(2005).

Ghosh et al. (2012), reviewed the mosquitocidal activities of various herbal

products from edible crops, ornamental plants, trees, shrubs, herbs, grasses, and marine

plants against different vector species.

 49

As cited by Shaalan et al. (2005), plants produce a broad range of bioactive

chemical compounds and are source of substitute agents for the control of insect vectors.

The leaves of vitex negundo contain an alkaloid nishindine, flavones.

Phytochemical screening of ethanol leaf extract yielded alkaloids, iridoids, phenolic

acids, flavonols and flavonoids, which has a larvicidal activity. (Stuart, 2018)

Lemongrass is a natural repellent because it has high geraniol and citral content

stated by Wells (2019).

According to Morton (1987), star apple leaves possess an alkaloid, also resin,

resinic acid, and a bitter substance.

The phytochemical screening performed by Cruz et al. (2016), showed that the

leaves and bark extracts of kakawate do contain secondary metabolites such as alkaloids

and saponin.

Extracts from plants may be alternative sources of mosquito egg and larval

control agents, since they constitute a rich source of bioactive compounds that are

biodegradable into non-toxic products and potentially suitable for use in control of

mosquito larvae (Pushpanathan et al., 2008)

 50

Chapter IV

CONCLUSION AND RECOMMENDATIONS

This chapter presents the findings, the conclusion and the recommendations of

this study.

Findings:

Based from the result of the study, the following are the findings:

1. The madre de cacao extract is a potent mosquito larvicidal. The efficacy of

the leaf extracts as mosquito larvicidal varies on the concentration. The 100%

concentration has higher mortality rate and the 25% concentration has lower mortality

rate.

2. The combined mixture of

a. Madre de cacao and Lagundi leaf extracts is an effective mosquito

larvicidal. The observation at 75% concentration has higher mean than at 25%

concentration. Proportionately, the higher concentration of the madre de cacao extracts in

the mixture the higher the mortality rate.

b. Madre de cacao and Lemon Grass leaf extracts is very effective as

mosquito larvicidal, too. At 50% and 100% the mortality rate is high. Proportionately, the

higher the concentration of madre de cacao extracts in the mixture the higher the

mortality rate.

c. Madre de cacao and Star Apple leaf extracts is also a potent mosquito
 51

larvicidal. The performance of the mixture varies on the concentration. In addition, the

higher the madre de cacao content the more the concentrate to be effective.

3. The mean of the combined mixture of madre de cacao and lemon grass has the

highest among the presented concentrations.

Conclusion:

Based from the findings of the study, the following conclusions were established:

1. Madre de cacao leaf extract is a potent mosquito larvicidal. The higher the

concentration of the extract the more effective it is.

2. Madre de cacao leaf extracts when combined to other plants like lagundi,

lemon

grass or star apple extracts increases the efficacy of the mixture. The higher the

concentration of madre de cacao on the mixture it become more effective as mosquito

larvicidal.

3. Madre de cacao and lemon grass mixture is the most effective concentrate.

Recommendations:

Based from the conclusion the following are recommended:

1. Madre de cacao is a potent mosquito larvicidal hence the propagation of this

plants especially on remote areas where malaria is prevalent is highly recommended.

2. Madre de cacao extracts and lemon grass combination is an effective

larvicidal

hence the propagation and use of these plants as insecticidal is highly recommended.

3. Madre de cacao and lemon grass combination should be studied on the

pharmacological and commercial viability of the extracts.

 52

4. A similar study should be conducted using other parts of the madre de cacao

such as bark and flower to verify the efficacy of this plant as mosquito larvicidal.

5. A similar study should be conducted using other methods of leaf extraction

and

may use bigger number of larvae to verify the validity of the results.

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https://drhealthbenefits.com/herbal/leaves/health-benefits-kakawate-leaves

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https://www.pinoybisnes.com/agri-business/all_about_lemongrass

American Museum of Natural History. (2009) The effectiveness of botanical extracts as

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extracts-as-repellents-against-aedes-aegypti-mosquitoes

American Museum on Natural History. (2011) Plant extracts as natural insecticides

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Insecticides

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Arranguez, M., Domino, R. & Saep M. (2019) The antifungal property of madre de cacao
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Baylon, V. et al. (2016) The pesticidal effect of Madre de cacao (Gliricidia sepium)
Neem Tree (Azadirachta indica), garlic (Allium sativum) extracts on
cockroaches (Periplaneta Americana). Retrieved from
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APPENDICES
 60

APPENDICES

Table 1. Original Values on the Mortality Rate of Mosquito Larvae on Madre de cacao
and Distilled water
Mortality Rate of Mosquito Larvae (hrs)
3 hrs 6 hrs 9 hrs 12 hrs
Treatment 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4
Control 0.0 00 0.0 0.0 0.0 0.0 0.0 0.0 0.0 00 0.0 0.0 0.0 00 0.0 0.0

25% 0.0 0.0 0.0 0.0 0.0 1.0 1.0 0.0 2.0 3.0 2.0 0.0 3.0 1.0 2.0 5.0
50% 5.0 4.0 4.0 4.0 0.0 1.0 1.0 1.0
0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
75% 5.0 4.0 4.0 4.0 0.0 1.0 1.0 1.0
0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
100% 5.0 5.0 5.0 5.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

Table 1a. Transformed Values on the Mortality Rate of Mosquito Larvae on Madre de
cacao and Distilled water
Mortality Rate of Mosquito Larvae (hrs)
3 hrs 6 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71
25% 0.71 0.71 0.71 0.71 0.71 1.22 1.22 0.71
50% 2.35 2.12 2.12 2.12 0.71 1.22 1.22 1.22
75% 2.35 2.12 2.12 2.12 0.71 1.22 1.22 1.22
100% 2.35 2.35 2.35 2.35 0.71 0.71 0.71 0.71
 61

Mortality Rate of Mosquito Larvae (hrs)

9 hrs 12 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71
25% 1.58 1.87 1.58 0.71 1.87 1.22 1.58 2.35
50% 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71
75% 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71
100% 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71

Table 1b. Mean of the Transformed data for the mortality rate of Madre de cacao and
water
Mortality Rate of Mosquito Larvae (hrs)
3 hours 6 hours
Treatment
2 3 4 Total Mea 2 3 4 Mean
1 n 1 Total
Control
0.71 0.71 0.71 0.71 2.84 0.71 0.71 0.71 0.71 0.71 2.84 0.71
25%
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1.22 1.22 0.71 3.86 0.97
50%
2.35 2.12 2.12 2.12 8.71 2.18 0.71 1.22 1.22 1.22 4.37 1.09
75%
2.35 2.12 2.12 2.12 8.71 2.18 0.71 1.22 1.22 1.22 4.37 1.09
100%
2.35 2.35 2.35 2.35 9.40 2.35 0.71 0.71 0.71 0.71 2.84 0.71
Total
8.47 8.01 8.01 8.01 32.50 8.13 3.55 5.08 5.08 4.57 18.28 4.57
Mean
1.69 1.60 1.60 1.60 6.50 1.63 0.71 1.02 1.02 0.91 3.66 0.91

Mortality Rate of Mosquito Larvae (hrs)

9 hours 12 hours
Treatment 2 3 4 Total Mea 2 3 4 Mean
1 n 1 Total
 62

Control
0.71 0.71 0.71 0.71 2.84 0.71 0.71 0.71 0.71 0.71 2.84 0.71
25%
1.58 1.87 1.58 0.71 5.74 1.44 1.87 1.22 1.58 2.35 3.39 1.76
50%
0.71 0.71 0.71 0.71 2.84 0.71 0.71 0.71 0.71 0.71 0.71 0.71
75%
0.71 0.71 0.71 0.71 2.84 0.71 0.71 0.71 0.71 0.71 0.71 0.71
100%
0.71 0.71 0.71 0.71 2.84 0.71 0.71 0.71 0.71 0.71 0.71 0.71
Total
4.42 4.71 4.42 3.55 17.10 4.28 4.71 4.06 4.42 5.19 8.36 4.60
Mean
0.88 0.94 0.88 0.71 3.42 0.86 0.94 0.81 0.88 1.04 1.67 0.92

Table 1c. Summary of the Mean on Transformed data on the Mortality Rate of Mosquito
Larvae on Madre de cacao and distilled water.
Mortality Rate of Mosquito Larvae (hrs)
Treatment Mean Mean Mean Mean
(3hrs) (6 hrs) (9 hrs) (12 hrs)
Control 0.71 c 0.71b 0.71b 0.71b
25% 0.71c 0.97ab 1.44a 1.76 a
50% 2.17 b 1.09 a 0.71b 0.71b
75% 2.17 b 1.09 a 0.71b 0.71b
100% 2.35 a 0.71b 0.71b 0.71b

Table 1d. Analysis of variance 3 hours

Source of Variation SS df MS F-value F- Prob
Replication .030 3 .010 2.439 .115
Treatment 11.053 4 2.763 672.687 .000
Error .049 12 .004
Total 64.134 20

Table 1e. Analysis of variance 6 hours

Source of Variation SS df MS F-value F- Prob
Replication .310 3 .103 3.639 .045
Treatment .479 4 .120 4.219 .023
Error .340 12 .028
Total 17.96 20

Table 1f. Analysis of variance 9 hours

 63

Source of Variation SS df MS F-value F- Prob

Replication .162 3 .054 1.087 .392
Treatment 1.650 4 .413 8.318 .002
Error .595 12 .050
Total 17.06 20

Table 1g. Analysis of variance 12 hours

Source of Variation SS df MS F-value F- Prob
Replication .156 3 .052 1.180 .358
Treatment 3.545 4 .886 20.132 .000
Error .528 12 .044
Total 21.07 20

Table 2. Original Values on the Mortality Rate of Mosquito Larvae on Madre de cacao
and Lagundi Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
3 hrs 6 hrs 9 hrs
Treatment 1 2 3 4 1 2 3 4 1 2 3 4
Control 0.0 1.0 3.0 5.0 0.0 4.0 2.0 0.0 0.0 0.0 0.0 0.0
25% 0.0 3.0 4.0 5.0 0.0 1.0 1.0 0.0 0.0 1.0 0.0 0.0
50% 0.0 3.0 2.0 5.0 0.0 2.0 3.0 0.0 0.0 0.0 0.0 0.0
75% 5.0 4.0 5.0 5.0 0.0 1.0 0.0 0.0 0.0 0.0 0.0 0.0

Table 2a. Transformed Values on the Mortality Rate of Mosquito Larvae on Madre de
cacao and Lagundi Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
3 hrs 6 hrs 9 hrs
Treatment 1 2 3 4 1 2 3 4 1 2 3 4
Control 0.71 1.22 1.87 2.35 0.71 2.12 1.58 0.71 0.71 0.7 0.71 0.71
1
25% 0.71 1.87 2.12 2.35 0.71 1.22 1.22 0.71 0.71 1.2 0.71 0.71
2
50% 0.71 1.87 1.58 2.35 0.71 1.58 1.87 0.71 0.71 0.7 0.71 0.71
1
75% 2.35 2.12 2.35 2.35 0.71 1.22 0.71 0.71 0.71 0.7 0.71 0.71
1
 64

Table 2b. Mean of Transformed Mortality Rate of Mosquito on Madre de cacao and
Lagundi Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
3 hours 6 hours
Treatment
2 3 4 Total Mea 2 3 4 Mean
1 n 1 Total
Control 0.7 0.7
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1 0.71 1 2.84 0.71
25% 1.2 1.2
1.22 1.87 1.87 2.12 7.08 1.77 2.12 2 1.58 2 6.14 1.54
50% 1.2 0.7
1.87 2.12 1.58 2.35 7.92 1.98 1.58 2 1.87 1 5.38 1.35
75% 0.7 0.7
2.35 2.35 2.35 2.35 9.40 2.35 0.71 1 0.71 1 2.84 0.71
Total 3.8 3.3
6.15 7.05 6.51 7.53 27.24 6.81 5.12 6 4.87 5 17.20 4.30
Mean 0.9 0.8
1.54 1.76 1.63 1.88 6.81 1.70 1.28 7 1.22 4 4.31 1.08

Mortality Rate of Mosquito Larvae (hrs)

9 hours
Treatment 1 2 3 4 Total Mean
Control 0.71 0.71 0.71 0.71 2.84 0.71
25% 0.71 1.22 0.71 0.71 3.35 0.84
50% 0.71 0.71 0.71 0.71 2.84 0.71
75% 0.71 0.71 0.71 0.71 2.84 0.71
Total 2.84 3.35 2.84 2.84 11.87 2.97
Mean 0.71 0.84 0.71 0.71 2.97 0.74

Table 2c. Summary of the Mean on Transformed data on the Mortality Rate of Mosquito
Larvae on Madre de cacao and Lagundi Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)

Treatment Mean Mean Mean

(3hrs) (6 hrs) (9 hrs)
Control 0.71c 0.71b 0.71a

25% 1.77b 1.54a 0.71a

 65

50% 1.98ab 1.35a 0.71a

75% 2.35a 0.71b 0.84a

Table 2d. Analysis of variance 3 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.275 3 0.092 1.658 0.244
Treatment 5.944 3 1.981 35.79 .00
Error .498 9 0.055
Total 53. 09 16

Table 2e. Analysis of variance 6 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.523 3 0.174 2,042 0.179
Treatment 2.204 3 0.735 8.597 0.005
Error .769
Total 21.99 16

Table 2f. Analysis of variance 9 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.049 3 0.016 1.0 0.436
Treatment 0.049 3 0.016 1.0 0.436
Error 0.146 9 0.016
Total 9.05 16

Table 3. Original Values on the Mortality Rate of Mosquito Larvae on Madre de cacao
and Lemon Grass Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
3 hrs 6 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.0 5.0 5.0 5.0 0.0 0.0 0.0 0.0
25% 0.0 5.0 5.0 5.0 0.0 0.0 0.0 0.0
50% 0.0 4.0 5.0 5.0 0.0 1.0 0.0 0.0
75% 0.0 4.0 5.0 5.0 0.0 1.0 0.0 0.0

Table 3a. Transformed Values on the Mortality Rate of Mosquito Larvae on Madre de
cacao and Lemon Grass Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
 66

3 hrs 6 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.71 2.35 2.35 2.35 0.71 0.71 0.71 0.71
25% 0.71 2.35 2.35 2.35 0.71 0.71 0.71 0.71
50% 0.71 2.12 2.35 2.35 0.71 1.22 0.71 0.71
75% 0.71 2.12 2.35 2.35 0.71 1.22 0.71 0.71

Table 3b. Mean of Transformed Mortality Rate of Mosquito on Madre de cacao and
Lemon Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
3 hours 6 hours
Treatment
2 3 4 Total Mea 2 3 4 Mean
1 n 1 Total
Control 0.7 0.7
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1 0.71 1 2.84 0.71
25% 0.7 1.2
2.35 2.35 2.12 2.12 8.94 2.24 0.71 1 1.22 2 3.86 0.97
50% 0.7 0.7
2.35 2.35 2.35 2.35 9.40 2.35 0.71 1 0.71 1 2.84 0.71
75% 0.7 0.7
2.35 2.35 2.35 2.35 9.40 2.35 0.71 1 0.71 1 2.84 0.71
Total 2.8 3.3
7.76 7.76 7.53 7.53 30.58 7.65 2.84 4 3.35 5 12.38 3.10
Mean 0.7 0.8
1.94 1.94 1.88 1.88 7.65 1.91 0.71 1 0.84 4 3.10 0.77

Table 3c. Summary of the Mean on Transformed data on the Mortality Rate of Mosquito
Larvae on Madre de cacao and Lemon Grass Leaf Extracts
Mortality Rate of Mosquito Larvae
(hrs)
Treatment Mean Mean
(3hrs) (6 hrs)
Control 0.71c 0.71a

25% 2.24b 0.71a

50% 2.35a 0.84a

75% 2.35a 0.84a

Table 3d. Analysis of variance 3 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.013 3 0.004 1.00 0.436
Treatment 7.731 3 2.577 584.595 0.00
Error 0.40 9 0.004
 67

Total 66.23 16

Table 3e. Analysis of variance 6 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.065 3 0.022 1.0 0.436
Treatment 0.195 3 0.065 3.00 0.88
Error 0.195 9 0.022
Total 10.03 16

Table 4. Original Values on the Mortality Rate of Mosquito Larvae on Madre de cacao
and Star apple Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
3 hrs 6 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.0 0.0 3.0 5.0 0.0 0.0 2.0 0.0
25% 0.0 0.0 2.0 3.0 0.0 0.0 3.0 2.0
50% 0.0 0.0 5.0 5.0 0.0 0.0 2.0 0.0
75% 0.0 0.0 2.0 5.0 0.0 0.0 3.0 0.0

Mortality Rate of Mosquito Larvae (hrs)

9 hrs 12 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.0 3.0 0.0 0.0 0.0 2.0 0.0 0.0
25% 0.0 4.0 0.0 0.0 0.0 1.0 0.0 0.0
50% 0.0 3.0 0.0 0.0 0.0 2.0 0.0 0.0
75% 0.0 5.0 0.0 0.0 0.0 0.0 0.0 0.0

Table 4a. Transformed Values on the Mortality Rate of Mosquito Larvae on Madre de
cacao and Star apple Leaf Extracts
Mortality Rate of Mosquito Larvae (hrs)
3 hrs 6 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71
25% 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71
50% 1.87 1.58 2.35 1.58 1.58 1.87 0.71 1.87
75% 2.35 1.87 2.35 2.35 0.71 1.58 0.71 0.71
 68

Mortality Rate of Mosquito Larvae (hrs)

9 hrs 12 hrs
Treatment 1 2 3 4 1 2 3 4
Control 0.71 0.71 0.71 0.71 0.71 1.58 0.71 0.71
25% 0.71 2.12 1.87 2.35 0.71 1.22 0.71 0.71
50% 1.87 0.71 0.71 0.71 0.71 1.58 0.71 0.71
75% 0.71 0.71 0.71 0.71 0.71 0.71 0.71 0.71

Table 4b. Mean of the Transformed data for the mortality rate of Madre de cacao and Star
apple
Mortality Rate of Mosquito Larvae (hrs)
3 hours 6 hours
Treatment
2 3 4 Total Mea 2 3 4 Mean
1 n 1 Total
Control 0.7 0.7
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1 0.71 1 2.84 0.71
25% 0.7 0.7
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1 0.71 1 2.84 0.71
50% 1.8 1.8
1.87 1.58 2.35 1.58 7.38 1.85 1.58 7 0.71 7 6.03 1.51
75% 1.5 0.7
2.35 1.87 2.35 2.35 8.92 2.23 0.71 8 0.71 1 3.71 0.93
Total 4.8 4.0
5.64 4.87 6.12 5.35 21.98 5.50 3.71 7 2.84 0 15.42 3.86
Mean 1.2 1.0
1.41 1.22 1.53 1.34 5.50 1.38 0.93 2 0.71 0 3.86 0.97
Mortality Rate of Mosquito Larvae (hrs)
9 hours 12 hours
Treatment
2 3 4 Total Mea 2 3 4 Mean
1 n 1 Total
Control 0.7 0.7
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1 0.71 1 0.71 0.71
25% 1.2 0.7
1.87 2.12 1.87 2.35 8.21 2.05 1.58 2 1.58 1 2.35 1.27
50% 0.7 0.7
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1 0.71 1 0.71 0.71
75% 0.7 0.7
0.71 0.71 0.71 0.71 2.84 0.71 0.71 1 0.71 1 0.71 0.71
Total 3.3 2.8
4.00 4.25 4.00 4.48 16.73 4.18 3.71 5 3.71 4 4.48 3.40
Mean 0.8 0.7
1.00 1.06 1.00 1.12 4.18 1.05 0.93 4 0.93 1 1.12 0.85
 69

Table 4c. Summary of the Mean on Transformed data on the Mortality Rate of Mosquito
Larvae on Madre de cacao and Star apple Leaf Extracts
Treatment Mean Mean Mean Mean
(3hrs) (6 hrs) (9 hrs) (12 hrs)
Control 0.71c 0.71b 0.71b 0.71b

25% 0.71c 0.71b 2.05a 1.27a

50% 1.85b 1.51a 0.71b 0.71b

75% 2.23a 0.93b 0.71b 0.71b

Table 4d. Analysis of variance 3 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.206 3 0.069 1.701 0.236
Treatment 7.345 3 2.448 60.694 0.0
Error 0.363 9 0.040
Total 38.11 16

Table 4e. Analysis of variance 6 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.526 3 0.175 1.667 0.243
Treatment 1.703 3 0.568 5.400 0.21
Error 0.946 9 0.105
Total 18.04 16
Table 4f. Analysis of variance 9 hours
Source of Variation SS df MS F-value F- Prob
Replication 0.040 3 0.013 1.000 0.436
Treatment 5.407 3 1.802 135.448 0.0
Error 0.120 9 0.013
Total 23.06 16

Table 4g. Analysis of variance 12 hours

Source of Variation SS df MS F-value F- Prob
Replication 0.127 3 0.042 1.0 0.436
Treatment 0.949 3 0.316 7.470 0.008
Error .381 9 0.042
Total 13.03 16
70
Table 5. Over-all result of mortality rate of combined mixtures

Combinations Control 25% 50% 75%

Time (hrs) Time (hrs) Time (hrs) Time (hrs)

1 4 -6 7- 10 - 1-3 4-6 7-9 10 -12 1-3 4-6 7- 10 - 1-3 4 7- 10 -

-3 9 12 9 12 -6 9 12

Madre de cacao - - - -
.71c 0.97b 1.44b 1.76a 2.18ab 1.09ab 2.18a 1.09a

Madre de cacao w/ lagundi - - - -

1.77b 1.54a 0.84c 0.71b 1.98ab 1.35ab 2.35a 0.71a

Madre de cacao w/ lemon - - - -

2.24a 0.97b 0.71c 0.71b 2.35a 0.71b 2.35a 0.71a
grass

Madre de cacao w/ star - - - -

.71c .71b 2.05a 1.27ab 1.85b 1.51a 2.23a 0.93a
apple

70
DOCUMENTATION
 72

THE MATERIALS USED IN THE STUDY

MaterialsUedintheSudy
 73

THE LEAVES USED IN THE STUDY

LAGUNDI
COLLECTING LEMON GRASS
WASHING

STAR APPLE MADRE DE CACAO

CUTTING
POUNDING

BLENDING EXTRACTING
 74
PROCESS OF LEAF EXTRACTION

THE LEAF EXTRACTS

STAR APPLE MADRE DE CACAO

LAGUNDI LEMON GRASS

 75

PREPARATION OF THE CONCENTRATION

MEASURING

MIXING STIRRING
 76

CULTURING MOSQUITO LARVAE

CATCHING MOSQUITO LARVAE AND CHECKING FOR DEAD LARVAE

 77

CATCHING LARVAE

CHECKING FOR DEAD MOSQUITO LARVAE

 78

CURRICULUM VITAE
 79

CURRICULUM VITAE

PERSONAL INFORMATION:

Name: Wilma P. Salvador

Sex: Female
Date of Birth: September 2, 1982
Place of Birth: Pidigan, Abra
Nationality: Filipino
Civil Status: Married
EDUCATION:
2016 - Present Master of Arts in General
Science
Major: General Science
ASIST, Bangued
Bangued, Abra

1999 – 2003 Divine Word College of Bangued CONTACT NO: 0926-586-5006

Bangued, Abra
Major : General Science
MAILING ADDRESS:
1995 – 1999 Suyo National High School
Suyo, Pidigan, Abra Monggoc, Pidigan, Abra

1989 – 1995 Pidigan Central School

Pidigan, Abra E-MAIL:
wilma.salvador@deped.gov.ph
ELIGIBILITY Board Licensure Examination for
Professional Teachers
August, 2003
WORK EXPERIENCE
SKILLS:
Current Teacher III
Gaddani National High School MS Word
Tayum, Abra 2803
MS Excel
I Year Holy Spirit Academy of Bangued MS PowerPoint
Bangued, Abra
MS Publisher
 80

2 Years Our Lady of Fatima School

Bucay, Abra
1 Year, 5 months Academy of St. Joseph
Claveria, Cagayan

4 Years St. Mary High School

Pidigan, Abra

SEMINARS ATTENDED
July 9-11, 2018 BENSS (Better, Enhanced and Nurturing Science Schoolteacher)
FOR THE LEARNERS: A Capability Building for Grade 7 and 8
Science Teachers
NMK Building, McKinley St., Zone 2, Bangued, Abra
Oct. 22-24, 2018 School-Based Training Workshop on Learning Resource
Management on Print and Non-Print Resources
Gaddani National High School
Gaddani, Tayum, Abra

May 20 – 31, 2018 Regional Training of Teachers on Science 8 Critical Content

Eurotel, Baguio City

Oct 15 - 17, 2017 Division Training-workshop for Developers on Learning Resource

Development Standards, Animation, Digitization, Manipulations,
Effective Instructional Presentations Using Powerpoint
Presentations Using Powerpoint and Hyperdoc, Instructional Short
Film Making and Basic Scripwriting
Gaddani National High School
Gaddani, Tayum, Abra

Jun 8 – 10, 2016 LRMDS Developers/Evaluators on Online Learning

Resource
Harvesting Process, Online LR QA/Evaluation, Redevelopment
Digitization and Cataloguing
Gaddani National High School
Gaddani, Tayum, Abra

Jan. 21 – 23, 2016 Division Training Workshop on the Use and Care of Science
Equipment with Content Integration
Abra High School
Bangued, Abra

Oct. 28 – 30, 2015 Training-Workshop on Improving Competencies using Varied

Strategies for Diverse Learners and Strengthening Instructional
Competence
 81

CHARACTER REFERENCES

CRISOSTOMO T. BALNEG
Poblacion, Tayum, Abra
9751897612

MARIA ANGELINA G. CRISPIN

Cabaroan, Tayum, Abra
9357883565

WILFREDO M. GANDEZA
Cabaroan, Tayum, Abra
9976316458