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Chapter I

INTRODUCTION

Background of the Study

Mosquitoes are among the well-known group of insect vectors that

transmit deleterious human diseases, such as malaria, yellow fever, encephalitis

and dengue which pose as the major public health challenges eroding

development in the poorest countries of the world. Dengue fever and dengue

hemorrhagic fever are vector-borne viral diseases on the society today. Some 2.5

billion people are at risk around the world and up to 100 million cases occur each

year (Wettstein et al., 2012).

The medical importance of mosquitoes as vectors for the transmission of

serious diseases that cause morbidity, mortality, economical loss, and social

disruption such as malaria, lymphatic filariasis, and viral diseases is well

documented. The best way of reducing the incidence of these diseases is by

mosquito control, which is frequently dependent on applications of conventional

synthetic insecticides (Wettstein et al., 2012).

Mosquitoes can have both positive and negative impacts on the

ecosystem. As part of their useful role, the larvae of mosquitoes live in water and

provide food for fish and other wildlife, including larger larvae of other species

such as dragonflies. The larvae themselves eat microscopic organic matter in the

water, helping to recycle it. Adult mosquitoes make up part of the diet of some
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insect-eating animals, such as birds, bats, adult dragonflies and spiders. They

also help pollinate some flowers, when they consume nectar.

Eradication or reduction of disease-carrying mosquitoes can help prevent

the spread of different diseases particularly dengue, hence, pesticide spraying

and fumigation have been practiced. However, environmental and health hazard

issues that arise from the use of these methods are well documented (US EPA,

2012).

The use of natural products poses an alternative method of control and/or

eradication of disease-carrying mosquitoes. It has been customarily considered

to be one of the safest ways of controlling pests. To find effective and affordable

ways to control the mosquito and prevent the spread of dengue, several plants

have been tested (Cavalcanti et al. 2004).

Ampalaya is a year-round vegetable, extensively cultivated in the

Philippines for its bitter edible fruit. In the Philippines, juice expressed from the

infusion of leaves or leaf juice can be drink for fevers.

In the past, synthetic organic chemical insecticides-based intervention

measures for the control of insect pests and disease vectors have resulted in

development of insecticide resistance in some medically important vectors of

malaria, filariasis and dengue fever.

The frequent use of chemical insecticides to manage insect pests’ leads to

a destabilization of the ecosystem and enhanced resistance to insecticides in

pests suggest a clear need for alternatives. Plants are the chemical factories of

nature, producing many chemicals, some of which have medicinal and pesticidal
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properties. The insecticidal and acaricidal properties of a number of plants have

been discovered a long ago, and some of the plants can compete with synthetic

means of control.

Higher air temperature can facilitate the transmission of dengue by

increasing the replication rate of the dengue virus in disease carrying mosquitos

such as Aedes aegypti, Anopheles stephensi, Culex quinquefaciatus and other

species (Tipayamongkholgul, 2005). Thus researchers look for convenient, safe,

and efficient herbal alternatives which can be easily obtained from the

environment to prevent and control dengue outbreaks.

Since dengue is a major public health problem, and Momoridca charantia

abundant in the Philippines, this study is therefore designed to determine the

larvicidal effect of Ampalaya (Momoridca charantia) leaf extract against mosquito

larvae.

Statement of the Problem

This study aims to determine the Larvicidal Effect of Ampalaya

(Momoridca charantia) Leaf Extract against mosquito larvae.

Specifically, this study seeks to answer the following questions:

1. Which concentration of Ampalaya (Momoridca charantia) Leaf Extract is

most effective against mosquito larvae?

1.1. 50% Ampalaya aqueous leaf extract

1.2. 75% Ampalaya aqueous leaf extract

1.3. 100% Ampalaya aqueous leaf extract


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2. Is there a significant difference between the effects of the different

concentrations of Ampalaya (Momoridca charantia) Leaf Extract against

mosquito larvae in terms of:

2.1. mortality rate

2.2. time of effectiveness

Hypotheses

1. Ampalaya (Momoridca charantia) leaf extract had a larvicidal effect

against mosquito larvae.

2. There is an effective concentration of Ampalaya (Momoridca charantia)

Leaf Extract against Mosquito larvae.

3. There is a significant difference between the effects of the different

Concentrations of Ampalaya (Momoridca charantia) Leaf Extract against

Mosquito larvae in terms of:

3.1. mortality rate

3.2. time effectiveness

Theoretical Framework

According to Bayer Environmental Sciences and Reinert, there are about

3500 species of mosquitoes, grouped into 3 sub families, Toxorhnychitinae,

Anophelinae, and Culicinae. Although there are over 3500 species of mosquito

through the world, all of which live in specific habitats, exhibit unique behavior

and bite various animals.


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According to Lim (2008), Momordica charantia is tropical vine of the family

Cucurbitaceae widely grown for edible fruit, which is among the most better of all

vegetables. In the Philippines it is commonly known as Ampalaya. It grows in

areas where annual precipitation ranges from 480 mm to 4100 mm. All parts of

the plant are equally important as fruits of Ampalaya (Momordica charantia)

contain Charantin, Polypeptide p as its constituent elements and also the leaves

of the plant contains Charantin, Polypeptide p as the main component.

The plant contains also several bioactive glycosides (including momordin,

charatine, charnantitosides, Goya glycosides, and momordicosides) and other

terpenoids compounds (including momordicin 28, momordicinin, momordicilin,

momordenol and momordol). It also contains cytotoxic (Ribosome inactivity)

proteins such as momorcharin and momordin. Also the aerial parts of the Bitter

gourd / Ampalya possess a number of active ingredients including tannins,

flavonoids, alkaloid’s, quinines and phenol. The juice extracted from the leaf of

Momordica charantia that will be used in this study.


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Conceptual Framework

INPUT PROCESS OUTPUT

To determine the Culturing of Larvicidal effect of


mosquitoes
larvicidal effect of Ampalaya (M.
Collection of Ampalaya
Ampalaya (M. charantia) leaf extract
(M. charantia) leaf
charantia) leaf extract on mosquitolarvae is
Ampalaya (M.
determined
charantia) leaf
extraction by pounding

Preparation of
Replicates for each
concentration

Preparation of
Ampalaya (M.
charantia) extract
concentrations

Testing for results

Figure 1. Scheme for the Conceptualization of the study


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The aim of the study is showed in the first box. To determine the larvicidal

effect of Ampalaya (M. charantia) leaf extract in mosquito larvae. Through the

process the researchers could achieve the objective. The process involves both

the independent (concentrations) and dependent (larvae) variables. The process

involves culturing of mosquitoes, collection of Ampalaya (M. charantia) leaf,

Ampalaya (M. charantia) leaf extraction by pounding, Preparation of Replicates

for each concentration, Preparation of Ampalaya (M. charantia) extract

concentrations, and testing for results. The researchers expect to found out if

Larvicidal effect of Ampalaya (M. charantia) leaf extract on mosquito larvae is

determined

Significance of the Study

This study is considered beneficial and relevant to the following

stakeholders:

Community. People who suffer from the dengue that caused by the

Mosquitoes, may found an abundant supply of Larvicide for mosquito larvae that

can be seen commonly just in their backyard.

Manufacturers. They could make this effective product that could be

available in the market and would again earn money from it.

Future researchers. This study may take as their take off for the further

investigation regarding the Larvicidal Effect of Ampalaya (Momordica charantia)

Leaf Extract against Mosquito larvae as Mosquito killer or repellent.


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Scope and Delimitations

The coverage of this study will determine the effectiveness of Ampalaya

leaf extracts as mosquito larvicide. The said experiment will be conducted within

Tacloban City area only. The study will be conducted in two weeks which includes

the culturing of mosquito larvae and collection of Ampalaya leaf. The materials to

be used will be gathered around the researchers’ school laboratory. The study

will limit only on the larvicidal property of Momordica charantia and other

chemically bounded larvicidals will not be used in this experiment.

Definition of Terms

The following terms are derived to convey their actual meaning as

employed in the textual presentation of the study.

Ampalaya (Momordica charantia). Also known as bitter melon, bitter

gourd, bitter squash or balsam-pear in English, has many other local names (An,

2016). In this study, this material will serve as the extracted Momordica charantia

leaf to be used as larvicide against mosquito larvae.

ANOVA. According to Oxford Dictionaries, ANOVA is the analysis of

variance, a statistical method in which the variation in a set of observations is

divided into distinct components. In this study, it will serve as the instrument in

order to analyze the results of the study.

Concentration. According to Oxford Dictionaries, concentration is the

amount of dissolved substance in a given volume of solvent. Obtained by dividing

the volume of the pure extract by the volume of the distilled water and multiplying
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the quotient by 100. Using the research instrument is is used to determine with

the concentration of Ampalaya leaf extract has the most effective larvicide

Extract. According to Webster’s, extract is a decoction, a solution, or

infusion made by drawing out from any substance that which gives its essential

essence. In this study, refers to the extraction of Ampalaya (Momordica

charantia) leaf that will serves as a larvicide against mosquito larvae.

Homogenous. According to Collins dictionaries, homogenous describes a

group or thing which has members or parts that are all the same. In this study, it

refers to the type of the sample which is the mosquito larvae.

Larvae. According to Cambridge dictionaries, it is an active immature form

of an insect, especially one that differs greatly from the adult and forms the stage

between egg and pupa, e.g., a caterpillar or grub. In this study, larvae will serve

as the sample of the experiment.

Leaf. According to oxford dictionaries, leaf is a flattened structure of a

higher plant, typically green and blade-like, that is attached to a stem directly or

via a stalk. Leaves are the main organs of photosynthesis and transpiration. In

this study, it refers to the part of Ampalaya (Momordica charantia) plant that will

be extracted to be used as larvicide against mosquito larvae.


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Chapter II

REVIEW OF RELATED LITERATURE

This chapter presents related theories and information related to the study

at hand.

Related Literature

In the past years, the plant kingdom has been of great interest as a

potential source of insecticidal products. Many species in the plant kingdom

synthesize a variety of secondary metabolites which play a vital role in defense of

plants against insects/mosquitoes. Plants may be alternative source for mosquito

repellent agents since they constitute a rich source of bioactive chemicals

(Shankar, Saravanan, & Ragavi, 2014).

Plant products can be used, either as an insecticide for killing larvae or

adult mosquitoes or as repellents for protection against mosquito bites,

depending on the type of activity they possess (Tekie, 2013).

Products of secondary plant metabolisms may be responsible for the chemical

communication between plants and insects. Allelochemicals have been

considered as potential natural insecticides and can be used for insect/mosquito

management in integrated control (Su, n.d.).

Repellents have an important part in protecting man from the bites of

insect pests. An effective repellent will be useful in reducing man vector contact

and in the interruption of disease transmission. Mosquito repellents may be one


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of the most effective tools for protecting human from vector-borne diseases and

nuisance caused by mosquitoes (Barnard, 2000).

Repellent compounds should be nontoxic, non-irritating and long lasting.

Repellents are substances that act locally or at a distance, deterring an arthropod

(insect/mosquito) from flying to, landing on or biting human or animal skin.

Usually, insect repellents work by providing a vapor barrier deterring the

arthropod (insect/mosquito) from coming into contact with the surface and

sometimes, applied on to the skin for protection. Repellents of plant origin do not

pose hazards of toxicity to humans and domestic animals and are easily

biodegradable. Natural products are safe for humans when compared to that

synthetic compound (Perry, 2014).

Mosquito repellent chemicals present in the plant contain phytochemicals

like, methone, limonene, beta pinene, alpha pinene and linaliol (Eun, 2001).

Burning wood and adding repellent plants to it probably works in several ways.

First, the smoke may disguise human kairomones and disrupts convention

currents essential in mosquito host location. Secondly, burning may, release

repellent irritant molecules and the molecules released by the plants also may be

insecticidal.

Therefore, the use of plants in mosquito control is an alternative pest

control method for minimizing the noxious effects of some pesticides compounds

on the environment (Fatope, 2001). Botanicals have widespread insecticidal

properties and will obviously work as a new weapon, and in future may act as

suitable alternative product to fight against vector mosquitoes (Ghosh, 2012).


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It may be concluded that natural products from plants of insecticidal and

medicinal values have higher efficiency in reducing mosquito menace due to their

repellent toxicity.

Related Studies

Mosquito-borne diseases, such as filariasis, malaria, dengue, yellow fever,

and Japanese encephalitis, contribute significantly to disease burden, death,

poverty, and social debility in tropical countries. The direct and indirect

contributions of such effects to treatment efficacy through reduced larval feeding

and fitness need to be properly understood in order to improve the use of

botanical insecticides for of A. stephensi. These and other naturally occurring

insecticides may play a more prominent role in mosquito control programs in the

future (Wandscheer et al., 2004).

The recently increased interest in developing plant-based insecticides as

an alternative to chemical insecticides, it was undertaken to assess the larvicidal

potential of the various fruit wall extracts of M. charantia against two species of

mosquito vectors, An. stephensi and Cx. quinquefasciatus. Among the extracts

tested, petroleum ether extract was found more effective than carbon

tetrachloride and methanol extracts towards anopheline and culicine larvae after

24 and 48 hrs of exposure respectively. Thus, all fruit wall extracts of M.

charantia are toxic to both the larval species. M. charantia may, therefore, act as

an effective biolarvicide against mosquitoes in the future (Maurya et al., 2009).


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Rahuman and Venkatesan (2008) have reported that the Larvicidal activity

of crude hexane, ethyl acetate, petroleum ether, acetone, and methanol extracts

of the leaf of five species of cucurbitaceous plants, Citrullus colocynthis, Coccinia

indica, Cucumis sativus, Momordica charantia, and Trichosanthes anguina, were

tested against the early fourth instar larvae of Aedes aegypti L. and Cx.

quinquefasciatus .

The larval mortality was observed after 24 h of exposure. The petroleum

ether extract of C. colocynthis and methanol extract of M. charantia were more

effective than the other extracts. Phytoextracts are emerging as potential

mosquito control agents, with low-cost, easy-to-administer, and risk-free

properties as compared to isolated or synthesized biopesticides and can be used

successfully in mosquito management.

Kamaraj et al. (2009) have reported that the highest mortality was found in

leaf petroleum ether and flower methanol extracts of Cassia auriculata against

the larvae of An. subpictus and Cx. tritaeniorhynchus. Elango et al. (2009) have

reported that the maximum repellent activity was observed at 500 ppm in

methanol extracts of Aegle marmelos and A. lineata and ethyl acetate extract of

Cocculus hirsutus, and the mean complete protection time ranged from 90 to 120

min with the different extracts tested against A. subpictus; no egg hatchability

was observed with ethyl acetate extract of A. marmelos; methanol extracts A.

marmelos, A. lineata, and C. hirsutus were exerted.


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M. charantia exhibited encouraging larvicidal effects against An. stephensi

and Cx. quinquefasciatus. Toxicological studies have shown that M.charantia is

safe for human health, and there are no toxic effects (Chopra, 2004).

Cucurbitaceous plants contain many compounds and are commonly used

in traditional medicines and have been reported to possess various biological

activities. M. charantia is widely used as a vegetable, an antidiabetic, and for

other common oilments. Moreover, its insecticidal activity was confirmed by

Kumar et al. (1979). The larvicidal effect of A. indica and M. charantia have

significant larvicidal potential against Cx. quinquefasciatus Further, the extracts

are eco-friendly larvicides as well as safe for use, as evidenced by use of plant

extracts as ingredients in oral medicines and ointments (Grover and Yadav,

2004).

The addition of B. thuringiensis var. israelensis with plant extracts caused

a significant mortality due to the avoidance of treated diet and may be due to

increased toxicity. It can therefore be concluded that B. thuringiensis var.

israelensis and plant compounds caused swelling of the gut epithelial cells. At

naturally occurring concentrations, allelochemicals produce midgut lesions,

reduce feeding and growth and increase mortality (Lindroth et al., 2008).

A study that the present results of M. charantia against the first-to fourth

instars larvae and pupae was recorded to produce a considerable mortality and

bacterial insecticide, B. thuringiensis against the first-to fourth instars larvae and

pupae also evidenced a considerable mortality. An attempt has been made to

evaluate the role of medicinal plant to control mosquitoes. Natural products are
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generally preferred in vector control measures due to their less deleterious effect

on non-target organisms and their innate biodegradability. In the context of

resistance developed by the mosquito larvae against chemical insecticides, it is

worthwhile to identify new active compounds from natural products against

mosquitoes. The findings of the present investigation revealed that the leaf crude

extract of M. charantia.
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Chapter III

METHODOLOGY

This chapter contains the research design and the methodology used in

the study. It incorporated the research materials and instruments, data gathering

procedure, and statistical treatment. Also, this chapter shows how the

researchers came up with the necessary data for this study, and how these data

were analyzed, interpreted and presented in easiest possible way.

Research Design

The study was conducted to determine the effectiveness of Ampalaya leaf

extract as larvicide against mosquito larvae. This study will utilize the

experimental design, specifically Completely Randomized Design (CRD) since

the sample (10 mosquito larvae per set-up) is homogenous, and the objects are

randomly assigned to the treatment.

Preparation of Ampalaya (Momordica charantia) Leaf Extract

A 2 kg Ampalaya (Momordica charantia) leaves was collected from Brgy.

91 Abucay, Tacloban City. It was gently washed with running water to be freed

from dust and contaminant, and will be air dried for an hour.

The collected Ampalaya leaves were cut into pieces and grind using

mortar and plastic cups to produce an extract. Then, the extract was strained

using a piece of cloth making 50 mL extract to obtain the sufficient amount of

pure extract for the different solutions.


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Preparation of the Different Treatments

Three solutions was prepared: 50% (5ml), 75% (7.5ml) and 100% (10ml).

The extracts were added with controlled amount of water to complete the 10 ml

solution; where in 5ml of the 50% concentration with water and 2.5ml of water for

the 75% concentration. While the last treatment remained as pure Ampalaya leaf

extract which is the 100% concentration.

Culturing the Mosquito Larvae and Containing in Experimental Set-up

A container with rainwater was set in an area where mosquitoes are

abundant inside the residential premises. After 14 days, the container was

observed for the presence of mosquito larvae. Using a medicine dropper, the

larvae were put in different glass containing a total number of 10 larvae in each

set up.

Application of Treatments to Mosquito Larvae

Five treatments were prepared namely: the three extracts, the positive

control with pepper and the negative control with water. The three solutions were

measured using a graduated cylinder to ensure the accuracy of the 10 mL

solution of the 50%, 75% and 100% concentrations with Amplaya aqueous leaf

extract then applied into the set-up. After 2 hours, observation was able to

determine the mortality rate and time of effectiveness. If the larvae showed no
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response to the agitation of the glass, the mosquito larvae is considered dead.

An LED light will be used to confirm the state of the larvae.

Statistical Analysis

The gathered data was analyzed through One-Way Analysis of

Variance (ANOVA) in order to determine with concentration of ampalaya leaf

extract has the most effective larvicide in terms of mortality rate and time of

effectiveness. Hence, the ampalaya leaf extract as larvicide against mosquito

was effective. In line with this, Microsoft® Excel 2013 was used to compute the

data that was gathered. The resulted data was subject to attest the significant

difference between the effects of the different concentrations of Ampalaya

(Momoridca charantia) leaf extract against mosquito larvae in terms of mortality

and time effectiveness.


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Chapter IV

RESULTS AND DISCUSSION

This chapter presents the data generated, its evaluation, interpretation

and discussion of the results. Each set of data gathered was analyzed and

interpreted to shed light on the problems that the study was ought to investigate.

The tables below show the data obtained from the experiments performed.

Interpretation of Data

Treatment Mortality
50% Ampalaya aqueous leaf extract 100%
75% Ampalaya aqueous leaf extract 100%
100% Ampalaya aqueous leaf extract 100%
Positive Control (Pepper) 100%
Negative Control (Water) 0

Table I. Mean Mortality of Ampalaya (Momoridca charantia) Leaf Extract

The mean mortality of the different Ampalaya (Momoridca charantia) leaf

extract concentrations are shown in the table above. These were then compared

to the mean mortality of the positive control which is the pepper and the negative

control which is the water. According to the data, it was found out that all the

concentrations and the positive control had a mean mortality of 10 while the

negative control showed the least mean mortality among the treatments which is

0.

Time of Effectiveness
Treatment
(sec.)
Set-up A 50% Ampalaya aqueous leaf extract 3,060
Set-up B 75% Ampalaya aqueous leaf extract 2,160
Set-up C 100% Ampalaya aqueous leaf extract 1,260
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Set-up D Positive Control (Pepper) 1,800


Set-up E Negative Control (Water) 0

Table 2. Duration of Effectiveness of the Different concentration on Mosquito

Larvae

Immediately after the application of the Ampalaya (Momoridca charantia)

leaf extract on mosquito larvae, the time was determines through the use of a

stopwatch. Results showed that the most effective treatment was the 100% seed

extract with 1,260s. The 75% seed extract also had a positive effect having a

time 2,160s while the 50% seed extract had the longest time of effectiveness with

3,060s. The positive control showed the least time of effectiveness with 1800s

while the negative control showed no larvicidal effect. According to the data, the

researchers can conclude that the relationship between the time of effectiveness

and the concentration of the extracts are inversely proportional because the time

of effectiveness decreases as the concentration increases.

Source of Sum of Degrees of Mean


Fcalc Ftab
Variation Squares Freedom Square
Between
300 2 150 36.05 3.40
treatments
Within
100 24 4.16
Treatments
TOTAL 400 26

Table 3. ANOVA Summary Table


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The ANOVA table above, shows that the calculated value of F is 36.05 and

the table value of F is 3.40, since Fcalc > Ftab, reject H˳. There is significant

difference in the effect of the different concentrations of Ampalaya leaf extract on

mosquito larvae in terms of mortality rate and the time of effectiveness.


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Chapter V

SUMMARY OF FINDINGS, CONCLUSION AND RECOMMENDATIONS

This chapter presents the summary, conclusions, and recommendations

drawn out of the data gathered by the researchers.

Summary

Three concentrations namely the 50%, 75% and 100% were obtained

from the Ampalaya (Momoridca charantia) leaf extracts and were tested for

larvicidal property against mosquito larvae. The following data are the summary

drawn from the conducted study:

a. Based on the results, the different concentrations of Ampalaya

(Momoridca charantia) leaf extracts showed a larvicidal property against

mosquito larvae. The three concentrations exhibited little to no difference

in terms of mortality rate but differed in terms of time of effectiveness.

b. Based on the ANOVA result, there is a significant difference between the

effects of the different concentrations in terms of time effectiveness and

mean mortality rate.

c. Since all the concentrations have the same mean mortality rate, which is

10, they are all considered to be effective in killing mosquito larvae. The

positive control exhibited similar mean mortality rate with the three

concentrations, which is 10, while the least mean mortality among the 5

treatments was the negative control, which is 0.


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d. The results showed a significant difference in terms of time effectiveness.

The 50% leaf extract had longest time in taking effect against mosquito

larvae at 3,060s. The 75% seed extract also showed larvicidal property

having a time of effectiveness at 2,160s while the 100% seed extract had

the fastest rate of killing mosquito larvae at 1,260s among the three

concentrations. The positive control had a time of effectiveness at 1800s

while the negative control showed no larvicidal effect on mosquito larvae.

Conclusion

The Ampalaya (Momoridca charantia) leaf extract as a larvicide against

mosquito larvae was proven effective. All the 50%, 75% and 100%

concentrations were found to exhibit the same mean of mortality but they differ in

time of effectiveness. The 100% concentration was found most effective having

the shortest time of killing mosquito larvae.

Recommendations

The researchers managed to use the Ampalaya leaf extract as larvicide

against mosquito. However, there is still room for improvements in making a

larvicide.

The researchers recommend the following:

1. To look for another plant that contains lectin that can be used in

conducting the same study.


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2. Other plant parts of Ampalaya specifically the pericarp as mosquito

larvicide.

3. To use other solvent such as ethanol for the preparation of the extract.
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APPENDIX A

Preparation of Ampalaya Leaf

Culturing of mosquito larvae Preparation of Different Treatments

Transferring of the Mosquito Larvae Application of Ampalaya Aqueous Leaf


Extract
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APPENDIX B

ANOVA SUMMARY TABLE

Replicates
1 2 3
Treatment Row Total (Row Total)^2
50% 10 10 10 30 900

75% 10 10 10 30 900

100% 10 10 10 30 900

Positive control 10 10 10 30 900


(Pepper)
Negative control 0 0 0 0 0
(Water)

c
( X a 2 + X b 2 + X c 2) To find the sum of squares
SSBT =∑ ¿
i=a r
between treatments, first, using the
c 2 2 2
( 30 + 30 + 30 )
¿∑ ¿ treatments from the raw data
i=a 9
c
( 900+900+900 ) gathered, square the value of Xa,
¿∑ ¿
i=a 9
Xb, Xc, Xd and Xe then add
c
2700
¿∑ ¿ together. After finding the sum of the
i=a 9
c squares, divide it with the number of
¿ ∑ ¿ 300
i=a
replicates which is 9.
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c
(x 2 a+ x2 b+ x 2 c) treatments from the raw data
SSWT =∑ ¿
i=a r
gathered, square the value of Xa,
c
(300+ 300+300 )
¿∑ ¿ Xb, Xc, Xd and Xe then add
i=a 9
c
900 together. After finding the sum of the
¿∑ ¿
i=a 9
squares, divide it with the number of
c
¿ ∑ ¿ 100 replicates which is 9.
i=a

To find the sum of squares

between treatments, first, using the

To find the total sum of the


SST = SSBT + SSWT
= 300 + 100 squares, add the sum of the squares
= 400
between the treatments and the sum

of the squares within treatments.

DFBT = t – 1 one from the number of treatments


=3–1
=2 which is 3.

To find the degree of freedom

between the treatments, subtract


To find the degree of freedom
DFWT= t (r- 1)
= 3 (9-1) within the treatments, multiply the
= 3 (8)
= 24 number of treatments to the

difference of the number of replicates

and one.
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DFTOTAL = DFBT + DFWT To find the total degree of


= 2 + 24
= 26 freedom, add the difference from

DFBT and the product from DFWT.

MSBT = SSBT ÷ DFBT To find the mean squares


= 300 ÷ 2
= 150 between the treatments, divide DFBT

fromSSBT.

Source of Sum of Degrees of Mean


Fcalc Ftab
Variation Squares Freedom Square
Between
300 2 150 36.05 3.40
treatments
Within
100 24 4.16
Treatments
TOTAL 400 26

MSWT = SSWT ÷ DFWT


= 100 ÷ 24 To find the mean square within the
= 4.16
treatments, divide SSWT from DFWT.

The ANOVA Summary Table


APPENDIX C

STATISTICAL ANALYSIS

The statistical analysis used in the study is the One-Way Analysis of

Variance (ANOVA). This statistical test of significance is used in accordance with

the following procedures:

1. There is no significant difference in the effect of the different

concentrations of ampalaya leaf extract on mosquito larvae in terms of:

2. The level of significance used is ɑ – 0.05

3. Raw Data Recorded:

Replicates
Treatment Row Total (Row Total)^2
1 2 3

50% 10 10 10 30 900

75% 10 10 10 30 900

100% 10 10 10 30 900
Positive control
10 10 10 30 900
(Pepper)
Negative control
0 0 0 0 0
(Water)

4. ANOVA Summary Table

Source of Sum of Degrees of Mean


Fcalc Ftab
Variation Squares Freedom Square
Between
300 2 150 36.05 3.40
treatments
Within
100 24 4.16
Treatments
TOTAL 400 26
APPENDIX D
SET-UP CHARTS

Set – Up A

Trials
Mortality Count Mean of Mortality
1 2 3

10 10 10 10

Average Time of
Time of Effectiveness 3210 3060 2910 Effectiveness
(sec.) 3060

The researchers use three trials for the 50% concentration of ampalaya

leaf extract. The results for each trial in terms of mortality count are the same

while the time of effectiveness was different. Referring to the specific objectives,

the mean mortality and the average time of effectiveness were the only needed

results for Table 1.

Set – Up B

Trials
Mortality Count Mean of Mortality
1 2 3

10 10 10 10

Average Time of
Time of Effectiveness 2060 2360 2060 Effectiveness
(sec.) 2160

The 75% concentration of ampalaya leaf extract showed greater effect

than the 50% concentration in Set-up A in terms of time of effectiveness. Both


concentrations displayed the same mortality count. The mean mortality and the

average time of effectiveness, referring to the specific objectives, were the only

needed results for the Table 1.

Set – Up C

Trials
Mortality Count Mean of Mortality
1 2 3

10 10 10 10

Average Time of
Time of Effectiveness 1360 1260 1160 Effectiveness
(sec.) 1260

The 100% concentration of the ampalaya leaf extract is the most effective

concentration. It showed the same mortality count but fastest time of

effectiveness among the other concentrations. The mean mortality and the

average time of effectiveness were the only needed results for Table 1 referring

to the specific objectives.

Set – Up D

Trials
Mortality Count 1 2 3 Mean of Mortality
10 10 10 10

Average Time of
Time of Effectiveness 1800 1800 1800 Effectiveness
(sec.) 1800

The positive control which is the pepper showed the great effect on the

sample with the fastest time of effectiveness but the same mortality rate with the

other concentrations. Because it is commonly used by DOH and DOST, it was

expected that the positive control will show a great effect.

Set – Up E

Trials
Mortality Count 1 2 3 Mean of Mortality

0 0 0 0

Average Time of
Time of Effectiveness Effectiveness
(sec.)

Water was used as the negative control. Since it is the normal habitat of

mosquito larvae, it was expected that the results would show 0 mortality count

and time of effectiveness is not applicable.

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