larvicidal effect of ampalaya (momordica charantia) fruit juice on aedes mosquito larvae

LARVICIDAL EFFECT OF AMPALAYA (Momordica charantia)

FRUIT JUICE ON Aedes MOSQUITO LARVAE

A Research Paper Presented to

the Faculty of the College of Arts and Sciences

Cebu Doctors University

Mandaue City, Cebu

In Partial Fulfillment of

the Course Requirements for the Degree

Bachelor of Science in Biology

Submitted By:

Amtul. Noor

Asunala. Rajesh Kumar

Chevagoni. Suresh

Doddaka. Ghnapti Krishna Sayogi

Gollapally. Vikram Goud

Kosaraju. Narendra kumar

Koutharapu. Deepak Chandran

Pagadala. Devi Vara Prasad

Rama. Mounika

Rama. Annapurna

Rama. Mounika

Rama. Nikhil Raj

Sah. Sitesh Kumar

Senguttuvan. Rahul Anand

Tamang. Nima

Uppugalla. Ruzu Rohith Reddy

Mary Grace G. Villaflor, M.S.

Research Mentor

September 30, 2013

ENDORSEMENT

This research proposal entitled Larvicidal Effect of Ampalaya (Momordica charantia)

Fruit Juice on Aedes Mosquito larvae was prepared and submitted by Amtul. Noor,

Asunala. Rajesh Kumar, Chevagoni. Suresh, Doddaka. Ghnapti Krishna Sayogi, Golapally.

Vikram Goud, Kosaraju. Narendra Kumar, Koutharapu. Deepak Chandran, Pagadala. Devi Vara

Prasad, Rama. Annapurna, Rama. Mounika, Rama. Nikhil Raj, Sah. Sitesh kumar, Senguttuvan.

Rahul Anand, Tamang. Nima, Uppugalla. Ruzu Rohit Reddy to the Proposal Hearing

Committee as endorsed by:

Mrs. Mary Grace G. Villaflor, M.S. Mr. Vitaliano V. Fernandez, M.S Research Mentor Research Coordinator

PROPOSAL HEARING COMMITTEE

Mr. Vitaliano V. Fernandez, M.S Chair, Technical

Ms. Cathy C. Bangos Ms. Marose Y. Abello Panelist, Statistician Panelist, Member

Leticia G. Cabrera, Ph.D. Panelist, IERC

ACCEPTANCE OF APPROVAL

This research proposal entitled Larvicidal Effect of Ampalaya (Momordica charantia)

Fruit Juice on Aedes Mosquito larvae upon the unanimous approval of the PROPOSAL

HEARING COMMITTEE with the grade of PASSED is hereby accepted for implementation

in partial fulfillment of the requirements for the degree for Bachelor of Science in Biology.

Atty. Roel S. Hortelano College Dean

Noted by: Leticia G. Cabrera, Ph.D. Director of Research

March 26, 2013 Date of Proposal Hearing

ENDORSEMENT

This research paper entitled Larvicidal Effect of Ampalaya (Momordica charantia)

Fruit Juice on Aedes Mosquito larvae was prepared and submitted by Amtul. Noor,

Asunala. Rajesh Kumar, Chevagoni. Suresh, Doddaka. Ghnapti Krishna Sayogi, Golapally.

Vikram Goud, Kosaraju. Narendra Kumar, Koutharapu. Deepak Chandran, Pagadala. Devi Vara

Prasad, Rama. Annapurna, Rama. Mounika, Rama. Nikhil Raj, Sah. Sitesh kumar, Senguttuvan.

Rahul Anand, Tamang. Nima, Uppugalla. Ruzu Rohit Reddy to the Oral Examination

Committee as endorsed by:

Mrs. Mary Grace G. Villaflor, M.S. Mr. Vitaliano V. Fernandez,M.S Research Mentor Research Coordinator

ORAL EXAMINATION COMMITTEE

Mr. Vitaliano V. Fernandez, M.S Chair, Technical

Ms. Cathy C. Bangos Ms. Marose Y. Abello Panelist, Statistician Panelist, Member

Leticia G. Cabrera, Ph.D. Panelist, IERC

ACCEPTANCE

This research paper entitled Larvicidal Effect of Ampalaya (Momordica charantia)

Fruit Juice on Aedes Mosquito larvae upon the unanimous approval of the ORAL

EXAMINATION COMMITTEE with the grade of PASSED is hereby accepted in partial

fulfillment of the requirements for the degree for Bachelor of Science in Biology.

Atty. Roel S. Hortelano College Dean

Noted by: Leticia G. Cabrera, Ph.D. Director of Research

September 30, 2013 Date of Oral Defense

Abstract

Objective: To determine the larvicidal effect of Ampalaya (Momordica charantia) fruit

juice on Aedes mosquito larvae.

Research design: The study was experimental in nature, using completely randomized

design.

Research locale: Aedes mosquito eggs were collected from one of the researchers

residence in Mandaue city, breeding and hatching was done in the zoology laboratory of

CDU.

Research subject: Two (2) to four (4) days old Aedes mosquito larvae.

Research instrument: Beaker, Measuring Cylinder, Plastic Containers, Spoon.

Data collection procedure: Aedes larvae were first treated with water, then with

different concentrations of Ampalaya fruit juice and finally with commercial larvicide.

During each trial the numbers of dead larvae were counted after time interval of 20

minutes, 40 minutes and one (1) hour. The larvae that floated and did not show any

movement or even did not respond to very slow stirring of the plastic spoon were

considered dead larvae and the one which showed movement were counted as alive

larvae.

Result: The larvicidal effect was not superior to ABATE 1SG.

Conclusion: In conclusion Ampalaya fruit juice had a larvicidal effect, but it was

inferior to the positive control ABATE 1SG.

ACKNOWLEDGMENT

The group would like to express their gratitude to several personalities, without

whom it would not have been possible to complete their research. The proper guidance,

support and precious help of these individuals who, in one way or another contributed to

the completion of this paper making the final product truly a team effort.

To Mrs. Mary Grace G. Villaflor, thesis adviser for her hours of work, advice on all

information, suggestions and patience. She always guided the group in the right tract

through her knowledge, insights and excellent constructive suggestions.

To Mr. Vitaliano Fernandez, research coordinator and chairman of the department

of Biology for providing the group with the required supervision, direction and the

invaluable information.

To Ms. Cathy Bangos for her advice and supervision in the statistical analysis.

To Ms. Marose Y. Abello for her valuable time, encouragement and input in this

research.

To Mr. Luvin C. Comendador, the laboratory technician for providing the necessary

equipment to perform the experiment.

To the parents, for their continuous support and unwavering encouragement.

To the almighty God, for the hope and guidance.

Table of Contents

Title i

Endorsement ii

Abstract iv

Acknowledgment v

Table of contents vi

INTRODUCTION

Background of study 1

Theoretical framework 2

Review of Related Literature 4

Significance of Study 5

Objective 6

General Objective 6

Specific Objective 6

Scope and Limitation 7

Hypothesis 8

MATERIAL AND METHODS

Research Design 9

Research Location 9

Research Subject 9

Collection of Sample 10

Mosquito larvae collection 10

Hatching of eggs 10

Procedure of Extraction 10

Data Collection 11

Definition of Terms 12

RESULTS AND DISCUSSION

Result 13

Discussion 17

CONCLUSION 18

RECOMMENDATION 19

BIBLIOGRAPHY 20

APPENDIX A Certification of Aedes larvae 23

APPENDIX B Certification of Ampalaya 24

APPENDIX C Data sheet 25

APPENDIX D Time Table of Activities 26

APPENDIX E Research Documentation 27

APPENDIX F Research Budget 28

RESEARCHERS BIODATA 29

INTRODUCTION

Background of study

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

deleterious human diseases, which pose as the major public health challenges eroding

development in the poorest countries of the world (Awad and Shimaila, 9 (4) : 637). Their

medical importance as vectors for the transmission of serious diseases that cause

morbidity, mortality, economic loss and social disruption such as malaria, filariasis, yellow

fever, dengue and other viral diseases as well (Becker et al. 325). The species of Aedes are

important vectors in the transmission of dengue, yellow fevers and chikungunya fevers,

and the like. Dengue is a serious and ancient problem spread by the bite of the infected

Aedes mosquitoes and its complications affects the kidney, liver, brain and blood which

could be very fatal. Dengue fever is an actual frequent epidemic viral disease transmitted

by the mosquito Aedes aegyptii. These are the burning issue now-a-days. These health

conditions are endemic in the developing countries of Africa, Latin America and Asia.

World Health Organization (88), Brown (2 : 123- 140) and Lee et al. (44: 105 -

112), reported about the development of physiological resistance to the chemicals by

mosquito species. Some mosquito species developed resistance to malathion (Guneady et

al. 50 (1) : 45- 54), a conventional pesticide, and to deltamethrin (Chen wen-mei 33,(1) :

14 - 20) like the adult Culex pipiens. These mosquitoes have developed resistance against

currently used insecticides, thereby hindering their effective control and possibility of

eradicating them in areas where they are prevalent. Furthermore, the adverse effects of

conventional insecticides on the environment and animals, including humans, and the

limited number of available insecticides compel continued search for safer plant

insecticides and larvicides that could be used in the control of these vectors. Medicinal

plant extracts and their constituents have proved to be biodegradable, have low

mammalian toxicity and induction of resistance while their activities were similar to those

of the standard drugs, such as temephos and methoprene.

These viral diseases and their resistance to currently used insecticides were the

reasons that researchers decided to study whether Ampalaya fruit extract could also be a

possible mosquito larvicide.

Theoretical framework

According to Bayer Environmental Sciences and Reinert (16: 175- 188), 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. Despite these different behaviors, all mosquitoes share some common traits, such

as four stage life cycle. After a female mosquito bites the animal and obtains the blood, it

lays eggs on the surface of stagnant water, in a depression, or on the edge of a container

where rain water may collect and flood the eggs. The eggs hatch in two to three days and

mosquito larvae or wriggler emerges. Mosquitoes are most efficiently and economically

destroyed when they are in the larval stage and are concentrated in their breeding site (Ellis

9). The larvae of Aedes mosquito were unable to survive in water of a high degree of

salinity due to exosmosis. (Wigglesworth, V.B.).

The larvae live in the water, feed and develop into the third stage of the life cycle

which takes four to ten days and is called a pupa or tumbler. The pupa also lives in the

water but do not feed. Finally, the mosquito emerges from the pupal case and it is ready to

bite as a fully developed mosquito (Evelyn, 6).

Aedes bites only human beings. Twenty-four to 48 hours following the emergence

of the adult mosquito from the pupa and after mating has taken place, the female seeks out

the blood for egg production in which it has two peaks for the biting time, one at the dawn

just after sunrise and the second at the dusk before the sunset. After female mosquito feeds

on a person whose blood contains virus, it can transmit dengue either immediately or after

an incubation period of 8 10 days, during which the virus multiplies (Evelyn, 7)

According to T. K. Lim (344-348), Momordica charantia is tropical vine of the

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

vegetables. An English name for the fruit is bitter melon or bitter gourd. In the Philippines

it is commonly known as Ampalaya, Hindi karela; Nepali- Tite Karela; Tamil pakal,

pavakka; Marathikarke. The fruit has a distinct warty looking exterior oblong shape. It is

hollow in cross section with relatively thin layer of flesh surrounding a central seed cavity

filled with large flat seeds and pith. 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 whereas SEEDS contains Charantin , Polypeptide-p along with Vicine as its

constituents.

Along with these components the plant contains several biologically active group,

chiefly Momordican I, Momordican II and Cucurbitacian B. The plants 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, alkaloids, quinines and phenol. Fruit consist of amino acids such as Beta

alanine, Gamma Alanine, Glutamic acid, Tryptamine and Gama amino-butyric acid among

others. Seeds also contain amino acids and include Arginine, Aspartic acid, Glycine,

Leucine, Lysine and Histidine (T. K. Lim, 347-353). The juice extracted from the fresh

fruit of Momordica charantia has been used in our research.

Review of related literature

Larvicides are chemical substances or group of insecticides used to stop mosquito

larvae from maturing into biting adults that transmit various diseases. Dichloro-diphenyl-

trichloroethane (DDT), pyrethruim, heptachlor, diedrin and lindane were the most

commonly used larvicides in the past to achieve these control measures. The commonly

and repeatedly used larvicides are fuel oils, kerosene and insecticide formulations (Truman

et al., 207). Synthetic organic larvicides, although very efficacious to target species such

as mosquitoes could be detrimental to a variety of animal life including man.

NICC reported that organophosphate temphos which is a Larvicide usually used in

breeding site, though slightly toxic to target organism, may cause headache, loss of

memory and irritability to man. Besides, the incessant use of chemical insecticides had

often led to the disruption of natural biological control system, and outbreak of insect

species as noted by (Chaithong et al., 31(1): 138- 144). Moreover, these chemicals could

be carcinogenic, mutagenic and teratogenic.

Many plant juices could be used to drive mosquitos away, eg. Eucalyptu

citriodora, Pterigeron odorus, Santalum lanceolatum, Momordica charantia. (Zohara and

Uriel, 183). Larvae can be killed by keeping it into hypertonic solution as due to the

difference in pressure gradient and concentration gradient osmosis can take place and then

cell loose its water shrink and dies. (Biology, p.180). Plants that were termite or insect

resistant, were used as fish poison and folkloric ally in treating malaria and fever as well

as those with reported insecticidal, mosquito repellent and mosquitocidal activities had

demonstrated promising larvicidal activities . However, no co-relation had been

established between larvicidal activities and these properties but an earlier study with a

common medicinal and vegetable plant of Momordica charantia Linn (Family:

Cucurbitaceae), had shown the insecticidal activity of this plant against mustard saw fly.

(T.K. Lim, 356)

The larvicidal nature of fleshy fruit wall of Momordica charantica Linn (Family:

Cucurbitaceae) was found first time, in the management of mosquitos done in the year

2009 November by Batabya et al. ( P.1205-10) in view of the recently increased interest in

developing plant based insecticides as an alternative to chemical insecticides. This study

was undertaken to know the larvicidal potential of the various fruit juices of Momordica

charantia (Cucurbitaceae) against culex species of mosquito vector.

Significance of study

This research could help the Rural and Community Health Workers to enhance

rural based projects promoting locally grown plant for medical purpose which could serve

as the cheaper alternative to expensive drugs available in the market. This research would

also help in finding the active ingredient in Ampalaya fruit and potentially spearhead the

manufacturing of commercial larvicides that would benefit the people. The farmers could

gain profit by cultivating this plant as cash crops. Industries could probably get benefitted

from this study since they could produce new larvicides from local resources. The

botanists could also be benefitted as they would get new information about this plant. This

would create an interest towards further research and exploration. Government would also

be equally benefitted by exporting larvicides and nature would also be free and fresh since

this juice is natural and biodegradable unlike some other commercial larvicides which are

non-biodegradable.

Objectives

General objectives

The study aimed to determine the larvicidal effect of Ampalaya (Momordica

charantia) fruit juice on Aedes mosquito larvae and also compare its larvicidal nature with

commercial larvicide (ABATE 1SG).

Specific objective

This study aimed to:

1. keep a tally of number of Aedes mosquito larvae killed after 20, 40 and 60 minutes of

exposure to

A) Ampalaya fruit juice in tap water

a.1) 25%

a.2) 50%

a.3) 75%

B) Commercial larvicide (ABATE 1SG)

2. Compare the number of Aedes mosquito larvae killed by the administration of Ampalaya

fruit juice and commercial larvicide (ABATE 1SG).

3. Determine the significant difference in the proportion of dead wrigglers between

Ampalaya fruit juice and commercial larvicide (ABATE 1SG).

Scope and limitation

This experimental study involved the determination of the larvicidal effect of

various concentrations of juice extracted from the fruit of Ampalaya on the two (2) four

(4) days old larvae of Aedes. The determinations of chemical composition of the juice

were not included in the study. The concentrations of Ampalaya fruit juice below 25% and

above 75% were not considered in the experiment. The collected eggs were of Aedes

mosquito as the eggs were laid separately, were elongated and black in color. But, the

species of the Aedes mosquito were unidentified.

Hypothesis

Statement of Null Hypothesis (Ho)

There was no significant difference in the proportion of dead wrigglers between

Ampalaya fruit juice as Aedes mosquito larvicide and a commercial larvicide.

Statement of Alternative Hypothesis (Ha)

There was a significant difference in the proportion of dead wrigglers between

Ampalaya fruit juice as a mosquito larvicide and a commercial larvicide.

MATERIALS AND METHODS

Research design

The study was experimental in nature, using Single Variable completely

randomized design to determine the larvicidal effect of Momordica charantia fruit juice

(variable) on Aedes mosquito larvae (constant), which were selected and assigned

randomly. Also it was compared with commercial larvicide (ABATE 1SG) as positive

control and water as negative control.

Research locale

The preparation of the different concentrations of Momordica charantia fruit juice

was conducted in Cebu Doctors University research laboratory.

The collection of mosquito eggs was conducted at one of the researchers residence

located near CDU, Mandaue City. The breeding and hatching of the eggs were conducted

at Cebu Doctors University, zoology laboratory. This area was suitable for conducting

experiment as it is well lighted, well-furnished and well equipped with the materials

needed for experiment.

Research subject

The research subjects were two (2) to four (4) days old Aedes larvae i.e. 2nd -3rd

instar larvae which were randomly selected from the basin with naturally bred and hatched

larvae.

Collection of sample

Mosquito larvae collection

White plastic containers coated in black were used as breeding site of Aedes

mosquito. The containers were lined with filter papers and half filled with clean water. The

plastic containers with filter paper were placed in the area where Aedes mosquitoes

naturally bred such as human surrounding. The containers were checked every morning

for the presence of mosquito eggs. When the tiny black patches were seen on the filter

papers, the filter papers were removed from the black container, air dried and were stored

into a clean and dry container. The larvae were then brought to Dr. Emmanuel Pardian

for authentication of identification (Appendix E ).

Hatching of Aedes Eggs

The transparent plastic containers were filled with clean water. The filter paper

with eggs were cut into pieces and then placed in the plastic containers to allow the

hatching of eggs. The set ups were checked daily for the hatching of the eggs and once the

larvae were seen, they were transferred into another plastic container and labeled

accordingly as to how many days old. After the collection of desired amount of larvae

(between one (1) to four (4) days), they were transferred to nine (9) different cups for

experimental setups (three (3) for each concentration), three (3) for negative control and

three (3) for positive control. Using a plastic spoon, six larvae were randomly selected and

assigned in each of the fifteen (15) cups.

Procedure of extraction

The selected, fresh and clean fruits of Momordica charantia were bought from the

mall and brought to Mr. Vitaliano Fernandez for authentication of identification (Appendix

F). After confirming that they were really Momordica charantia, they were then brought

into the research laboratory for the extraction process. Nearly three and half (3.5)

kilograms of fresh fruits of Momordica charantia were washed and chopped into small

pieces. Then, they were crushed in a mixer. The juice was then taken out and filtered using

a clean cloth. Then three (3) different concentrations of juice were prepared by mixing it

with proper amount of water. For the 25% concentration, 25 ml of pure juice and 75 ml of

water were mixed, for 50% concentration, 50 ml pure juice and 50 ml were mixed and for

the 75% concentration, 75 ml pure juice and 25 ml water were mixed. The prepared

different concentrations of juice were used in the experiment along with water as negative

control and larvicide (ABATE 1SG) as positive control.

Data collection

The experiment was conducted on the 6th of December 2013 in the zoology

laboratory in CDU. The three plastic cups were taken and filled with 25 ml, 50 ml and 75

ml of pure water. The three different concentrations were obtained by adding 75 ml, 50 ml

and 25 ml of pure Momordica charantia fruit juice respectively. As for the negative

control, 100 ml of pure water was kept in one cup. Then six (6) naturally bred larvae were

placed in each cup and the cups were kept on the table. The set ups were checked every 20

minutes until one (1) hour. A stop watch was used to note down the duration of time. Same

steps were followed for three (3) trials along with positive and negative control set ups.

At the end of each trial, the numbers of dead and alive larvae were noted. The

larvae that floated and did not show any movement or even did not respond to very slow

stirring of the plastic spoon were considered dead. The number of dead and alive larvae

were counted and recorded in the tabulated data sheet.

Definition of terms

Aedes Mosquito- Aedes mosquitoes are typically small mosquitoes. They usually have

black and white stripes marking on the body and legs. They usually bite only during day

time.

Larvae - larvae are the distinct juvenile forms of many insects which undergo complete

metamorphosis. It is the feeding stage of the insect development. It is constant.

Larvicide - An agent for killing larvae, in this experiment, it is the Ampalaya juice.

Commercial larvicide - Insecticide that is specially targeted against larval stage of any

insects, in this experiment, ABATE 1SG.

Ampalaya fruit juice - It is the juice extracted from the fresh Ampalaya fruit with the

help of grinder. After the extracted of juice, desired amount of water is added to make

different concentration (25%, 50% and 75%). It is variable.

RESULTS AND DISCUSSIONS

Result

The tables below showed the summary of the data gathered from the research and the

Mean of the different concentrations.

Table 1

Percentage (%) of dead Wrigglers

A) After 20 minutes

Substance used

No. of Trials

ABATE 1SG

25% of Ampalaya juice



Trial 1 50.00 0.00 0.00 16.67

Trial 2 66.67 0.00 16.67 16.67

Trial 3 33.33 0.00 0.00 0.00

MEAN 50.00 0.00 5.56 11.11

Based on the result, it can be seen that the ABATE 1SG is 50% effective after 20 minutes

whereas highest concentration of Ampalaya fruit juice is only 11.11%.

B) After 40 minutes

Substance used

No. of Trials

ABATE 1SG




Trial 1 66.67 16.67 33.33 33.33

Trial 2 66.67 0.00 16.67 33.33

Trial 3 66.67 0.00 16.67 16.67

MEAN 66.67 5.56 22.22 27.78

Based on the result, ABATE 1SG was 66.67% effective after 40 minutes whereas 25%,

50% and 75% concentrations of Ampalaya fruit juice were comparatively less effective and

showed 5.56%, 22.22% and 27.78% effectiveness respectively.

C) After 1 hour

Substance used

No. of Trials

ABATE

1SG

25% of

Ampalaya

juice

50% of

Ampalaya

juice

75% of

Ampalaya

juice

Trial 1 83.33 33.33 50.00 66.67

Trial 2 83.33 16.67 33.33 50.00

Trial 3 100.00 16.67 33.33 50.00

MEAN 88.89 22.22 38.89 55.56

As shown in the above table, the ABATE 1SG was more effective after 1 hour as compared

to 25%, 50% and 75% concentrations of Ampalaya fruit juice.

Table 2

Comparison of Results

A) After 20 min

Group No. of Trials

Average % of dead Wrigglers

P value

Remarks

With ABATE 1SG 3 50.00 0.038 Since the P < 0.05 level of significance, the null hypothesis (Ho) was rejected.

With 25% of Ampalaya fruit juice

3 0.00


3 5.56


3 11.11

With ABATE 1SG 3 50.00 0.000 The Ho was rejected because the P value is < 0.05 level of significance.


3 11.11

Based on the given results, the P value (0.038) was less than 0.05 level of significance.

Hence, the null hypothesis (Ho) was rejected. It implies that the number of dead wrigglers

using Ampalaya fruit juice in different concentrations and using ABATE 1SG were

significantly different. More dead wriggler were found when ABATE 1SG was used.

Hence, Ampalaya fruit juice is not an effective mosquito larvicide.

B) After 40 min

Group No. of Trials Average % of dead Wrigglers

P value

Remarks



3 5.56


3 22.22


3 27.78



3 27.78

In this table it can be seen that the percentage (%) of dead larvae was only 27.78% for the

75% conc. of Ampalaya fruit juice. The P value at 40 minute is 0.027, which is less than

0.05 level of significance. Hence, the null hypothesis (Ho) was rejected. The results also

show that the number of dead wrigglers using Ampalaya fruit juice in different

concentrations and ABATE 1SG differed significantly even after 40 minutes.

C) After one (1) hour

Group No. of Trials Average % of dead Wrigglers

P value

Remarks



3 22.22


3 38.89


3 55.56



3 55.56

Based on the above results, the percentage (%) of dead larvae was only 55.56% for the

75% conc. of Ampalaya fruit juice. Here, the P value after one (1) hour was 0.020 which is

less than 0.05 level of significance. Therefore, the null hypothesis (Ho) was rejected. More

dead wrigglers were found by treating with ABATE 1SG (88.89). Hence, the number of

dead wrigglers using Ampalaya fruit juice in different concentrations (even at 75%

concentration) and ABATE 1SG were significantly different after one (1) hour.

Discussion

Based on the experiment, it was seen that more number of larvae died in the highest

concentration of Ampalaya fruit juice probably because of hyper tonicity (Biology, p.180)

of the solution as the tonicity of Ampalaya fruit juice was more compared to cells of the

larvae, exosmosis took place eventually resulting in their death.

An experiment performed on Aedes mosquito larvae showed that the larvae of

Aedes mosquito were unable to survive in water of a high degree of salinity due to

exosmosis. (Wigglesworth, V.B.)

However, few larvae died in the lowest concentration of Ampalaya fruit juice. This

showed the presence of certain chemicals in the Ampalaya fruit juice that had some

larvicidal property. Therefore Ampalaya fruit juice had larvicidal effect on Aedes larvae

even though it is minimal. Some researchers have shown that many plant juice like

Eucalyptus citriodora, Pterigeron odorus, Santalum lanceolatum, Momordica charantia

were used to drive mosquitoes away (Zohara and Uriel, 183) but Ampalaya fruit juice did

not prove to be as effective as them.

Some researches (Maurya Para et al., 2009) have also shown the larvicidal nature

of fleshy fruit wall of Momordica charantia to Culex mosquito larvae.

Thus, it can be said that the larvicidal property of Ampalaya fruit juice is selective

to Culex mosquito larvae as proven by the work of (Batabyal L et al. para.1) while not so

effective in the case of Aedes mosquito larvae.

CONCLUSION

In conclusion, the Ampalaya fruit juice had a larvicidal effect, but it was inferior to

the positive control ABATE 1 SG.

RECOMMENDATION

From this very research, it was found out that the larvicidal property of

Momordica charantia fruit juice was minimal. Even the 75% concentration solution was

not very effective in killing Aedes mosquito larvae. However, the other parts of

Momordica charantia plant such as leaves, root, stem etc. might be useful in killing the

Aedes mosquito larvae. Further research using other parts of Ampalaya was therefore

recommended for further study if they have larvicidal effect.

As mentioned before, the Ampalaya fruit juice has minimum larvicidal effect when

it comes to Aedes mosquito larvae. But, it might be effective in killing mosquito larvae of

other genera such as Anopheles, Culex. It can also be tried for different species of

mosquitoes.

BIBLIOGRAPHY

Books

Becker, et al. Mosquitoes and their Control. New York: Kluwer Academic/Plenum

Publishers, 2003.

Biology. New Delhi: National Council of Educational Research & Training. 2006.

T.K. Lim. Edible Medicinal and Non-Medicinal Plants. Springer Dordrecht Heidelberg

London New York, 2012.

Truman, et al. Scientific Guide to Pest Control Operations 3rd ed. Cleveland Ohio:

Harvest Publishing Company, 1976.

Zohara Xaniv & Uriel Bachrach. Handbook of Medicinal Plants. New York: Food

Products Press, Haworth Medical Press, 2005.

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APPENDIX A

Certification of Aedes Larvae

APPENDIX B

Certification of Ampalaya Fruit

APPENDIX C

Data sheet

Group:

Total No. of Larvae

Trials

Times

No. of dead larvae No. of alive larvae

20 min 40 min 1 hrs 20 min 40 min

1 hrs

6 T1

T2

T3

Mean

APPENDIX D

Time Table of Activities

Date Activity

January 14, 2013 Writing the research title

February 4, 2013 Approval of research title

February 15-25, 2013 Writing and submission of Rationale, Significance

of study and Statement of problem

February 28 - March 5,

2013

Writing and submission of Theoretic Background,

Review of related Literature, scope and limitation

March 6- 9, 2013 Writing and submission of Methodology and

Bibliography

March 26, 2013 Proposal Hearing and Approval of the Study

November 3- 14, 2013 Mosquito larvae collection and storage

December 4, 2013 Buying of the equipment necessary for the research

December 2, 2013 Hatching of the mosquito eggs

December 6, 2013 Transferring of the mosquito larvae

December 6 , 2013 Buying of the Mimordica charantia fruit

December 6, 2013 Treatment of different age of larvae with juice

December 6, 2013 Data analysis

December 7, 2013 Submission of data to the advisor and statistician for

Statistical analysis

September 28, 2013 Preparation of oral defense

September 30, 2013 Oral defense

December 9, 2013 Revision

December 9, 2013 Signature

APPENDIX E

Research Budget

Transportation:

Jeepney fare : Php 10 x 10 Php 250.00

Supplies:

Box : Php 45 x 6 Php 270.00

Spoon : Php 20 x 2 Php 40.00

Glass : Php 30 x 2 Php 60.00

Ampalya : Php 70 x 3.5 Php 245.00

Xerox : Php 2 x 1000 Php 2000.00

Extra

Lunch Php 1300.00

Breakfast Php 300.00

---------------------------------

Total : Php 4465.00

APPENDIX F

Research Documentation

Collection of Ampalaya fruit and Commercial larvicide ABATE 1SG

Performing of experiment in the lab with various concentration of fruit juice

Performing of experiment in the lab with commercial larvicide ABATE 1SG

RESEARCHERS BIODATA

PERSONAL INFORMATION:

NAME : AMTUL. NOOR

DATE OF BIRTH : 06-01-1994

HOME ADDRESS : HYDERABAD, ANDHRA PRADESH, INDIA

CURRENT ADDRESS : LAHUG, PHILIPPINES

CONTACT NUMBER : +63 9339929582

CIVIL STATUS : SINGLE

FAMILY BACKGROUND

FATHERS NAME : MOHD. RAHMATULLAH

OCCUPATION : ENGINEER

MOTHERS NAME : AMTUL. SAMI

OCCUPATION : HOUSE WIFE

EDUCATIONAL BACKGROUND

ELEMENTARY SCHOOL : ST. ANNS HIGH SCHOOL TARNAKA

HIGHER SECONDARY SCHOOL : SRI CHAITANYA JUNIOR COLLEGE

PRESENT SCHOOL : CEBU DOCTORS UNIVERSITY

EXPERIENCE IN RESEARCH : LARVICIDAL EFFECT OF AMPALAYA

(Momordica charantia) FRUIT JUICE ON

Aedes MOSQUITO LARVAE


NAME : ASUNALA. RAJESH KUMAR


HOME ADDRESS : WARANGAL, ANDHRAPRADESH, INDIA




FAMILY BACKGROUND

FATHERS NAME : ASUNALA. SATHISH

OCCUPATION : FARMER

MOTHERS NAME : ASUNALA. LEELA

OCCUPATION : HOUSEWIFE


ELEMENTARY SCHOOL : NAVABHARATHI HIGH SCHOOL

HIGHER SECONDARY SCHOOL : NARAYANA JUNIOR COLLEGE






NAME : CHEVAGONI. SURESH


HOME ADDRESS : POCHAMPALLY, ANDHRAPRADESH,

INDIA




FAMILY BACKGROUND

FATHERS NAME : CHEVAGONI. VENKATESH

OCCUPATION : BUSINESSMAN

MOTHERS NAME : CHEVAGONI. VENAMMA



ELEMENTARY SCHOOL : BHASHYAM PUBLIC SCHOOL

HIGHER SECONDARY SCHOOL : SRICHAITHANYA JUNIOR COLLEGE





PERSONAL INFORMATION

NAME : DODDAKA. GHNAPTHI KRISHNA SAYOJI


HOME ADDRESS : MACHILIPATNAM, ANDHRAPRADESH,

INDIA




FAMILY BACKGROUND

FATHERS NAME : D.SAYOJI RAO

OCCUPATION : JOURNALIST

MOTHERS NAME : D.RAMANI



ELEMENTARY SCHOOL : A LIITLE FLOWER THE LEADER

HIGHER SECONDARY SCHOOL : SRI CHAITHANYA JUNIOR COLLEGE






NAME : GOLLAPALLY. VIKRAM GOUD


HOME ADDRESS : HYDERABAD, ANDHRAPRADESH, INDIA




FAMILY BACKGROUND

FATHERS NAME : GOLLAPALLY. MADHUKAR GOUD

OCCUPATION : BUSINESS

MOTHERS NAME : GOLLAPALLY. ASHA JYOTHI



ELEMENTARY SCHOOL : SR. NATIONAL HIGH SCHOOL






PERSONAL INFORMATION

NAME : KOSARAJU. NARENDRA KUMAR


HOME ADDRESS : GUNTUR, ANDHRAPRADESH, INDIA




FAMILY BACKGROUND

FATHERS NAME : KOSARAJU. SAMBASIVA RAO


MOTHERS NAME : KOSARAJU. SUNITHA



ELEMENTARY SCHOOL : ST.ANNSS SCHOOL

HIGHER SECONDARY SCHOOL : SRICHAITANYA JUNIOR COLLEGE






NAME : KOUTHARAPU. DEEPAK CHANDRAN


HOME ADDRESS : TENALI, ANDHRAPRADESH, INDIA




FAMILY BACKGROUND

FATHERS NAME : K. RAMA MOHANA RAO

OCCUPATION : RETD. EMPLOYEE

MOTHERS NAME : ABBURI. NAGENDRAM

OCCUPATION : RAILWAY EMPLOYEE


ELEMENTARY SCHOOL : MONTESSORI PUBLIC SCHOOL, TENALI

HIGHER SECONDARY SCHOOL : SRI MARUTHI VIDYA NIKETAN






NAME : PAGADALA. DEVI VARAPRASAD


HOME ADDRESS : KHAMMAM, ANDHRAPRADESH, INDIA




FAMILY BACKGROUND

FATHERS NAME : PAGADALA. MOHAN RAO

OCCUPATION : BUSINESS MAN

MOTHERS NAME : PAGADALA. SRI LATHA



ELEMENTARY SCHOOL : GEETHANJALI VIDYANIKETHAN







NAME : RAMA. ANNAPURNA



CURRENT ADDRESS : CEBU CITY, PHILIPPINES



FAMILY BACKGROUND

FATHERS NAME : RAMA. PRAKASH


MOTHERS NAME : RAMA. SRIDEVI










NAME : RAMA. MOUNIKA






FAMILY BACKGROUND

FATHERS NAME : RAMA. PRABHAKAR


MOTHERS NAME : RAMA. RAJITHA




HIGHER SECONDARY SCHOOL : SR. JUNIOR COLLEGE FOR GIRLS






NAME : RAMA. NIKHIL RAJ






FAMILY BACKGROUND

FATHERS NAME : RAMA. PRABHAKAR


MOTHERS NAME : RAMA. RAJITHA



ELEMENTARY SCHOOL : NALANDA HIGH SCHOOL







NAME : SAH. SITESH KUMAR


HOME ADDRESS : SIRAHA, NEPAL

CURRENT ADDRESS : MANDAUE CITY, PHILLIPINES



FAMILY BACKGROUND

FATHERS NAME : SAH. SURYA NARAYAN

OCCUPATION : FARMER

MOTHERS NAME : SAH. BINDU DEVI

OCCUPATION : TEACHER


ELEMENTARY SCHOOL : NEW ENGLISH SECONDARY BOARDING

SCHOOL

HIGHER SECONDARY SCHOOL : PRASADI HIGHER SECONDARY SCHOOL






NAME : SENGUTTUVAN. RAHUL ANAND


HOME ADDRESS : VALAPADY, SALEM, TAMILNADU, INDIA

CURRENT ADDRESS : SAMBAG-1, URGELLO, CEBU,

PHILIPPINES



FAMILY BACKGROUND

FATHERS NAME : SENGUTTUVAN

OCCUPATION : VETERINARY DOCTOR

MOTHERS NAME : RANI



ELEMENTARY SCHOOL : GREEN PARK SCHOOL

HIGHER SECONDARY SCHOOL : SKV HIGH SCHOOL






NAME : TAMANG. NIMA


HOME ADDRESS : KATHMANDU, NEPAL

CURRENT ADDRESS : SAMBAG, CEBU, PHILLIPINES



FAMILY BACKGROUND

FATHERS NAME : TAMANG. PANCHA BAHADUR


MOTHERS NAME : TAMANG. MANJU



ELEMENTARY SCHOOL : BHANU BHAKTA MEMORIAL H.S.

HIGHER SECONDARY SCHOOL : CAPITAL COLLEGE AND RESEARCH

CENTER






NAME : UPUGALLA. RUZU ROHITH REDDY


HOME ADDRESS : HANAMKONDA, ANDHRAPRADESH,

INDIA




FAMILY BACKGROUND

FATHERS NAME : UPUGALLA. RAMCHANDRA REDDY


MOTHERS NAME : UPUGALLA. RAJITHA



ELEMENTARY SCHOOL : VRS & VJR SCHOOL

HIGHER SECONDARY SCHOOL : SRI CHAITHANYA JUNIOR COLLEGE





larvicidal effect of ampalaya (momordica charantia) fruit juice on aedes mosquito larvae

Documents

research proposal

research paper

mounika rama

annapurna rama

director of research

aedes mosquito larvae

rajesh kumar chevagoni

narendra kumar koutharapu