Kappaphycus alvarezii (Red Seaweed) extract as an attractant for Aedes

aegypti mosquito

Ma. Angelica G. Dela Cruz

Evan Nathaniel T. Gumapac
Charmane Lizette S. Tarcena

September 2014

Table of Contents

Abstract

Chapter 1: Introduction

1-6

Introduction

1-2

Background of the Study

2-3

Statement of the Problem

Significance of the Study

4-5

Scope and Delimitations

Chapter 2: Review of Related Literature and Related Studies
Introduction

6
7-11
7

Life Cycle
7-9
Egg

7-8

Larvae

8-9

Pupae

Adult

Appearance

9-10

Behavior

10-11

Attractant

11

Chapter 3: Methodology

12-14

Introduction

12

Research method used

12

Procedures used
Experimental Design

12-14
14

Chapter 4: Presentation, Analysis, and Interpretation of data

15-19

Chapter 5: Summary, Conclusion, and Recommendation

20-21

Summary of Findings
Conclusion
Recommendation
Acknowledgement

20
20-21
21
ii

Bibliography

iii

Appendices

iv

The extract of Kappaphycus alvarezii, a species of red algae

seaweed, is largely cultivated here in the Philippines and carrageenan
is extracted from it for commercial uses. The waste products from the
processed seaweed has no use and could pollute the environment if
not properly disposed, but there might be another use for this
extracted waste product. The extract has shown nutrients and minerals
that are important for biological growth. This makes the extract
favourable for bacteria to grow and produce gases such as carbon
dioxide and ammonia. Because of these variables present, the extract
could be a possible attractant for Aedes aegypti, a mosquito that
spreads

diseases

such

as

dengue

and

yellow

fever.

For

the

experimentation, the researchers have prepared three setups and each

setup has three replicates. Setup A consists of 100% water, Setup B
consists of 75% solution and 25% water, and Setup C consists of 50%
solution and 50% water. The nine containers were placed in the same
location and were observed for six days. On the sixth day, Setup A has
an average of 31 eggs, Setup B has an average of 48 eggs and Setup C
has an average of 46 eggs. Based on the observations of the
experimentation of the researchers, it was concluded that 75% solution
of Kappaphycus alvarezii extract is the most favourable nesting place
for Aedes aegypti compared to 100% water and 50% solution.

CHAPTER I
INTRODUCTION

The Philippines is an archipelago that is located just above the equator; this
makes the Philippines a tropical country. It is surrounded by vast bodies of water
and has plenty of rivers and streams. The rivers and streams come from heavy
rains that are very abundant throughout the year, especially during the wet season.
With all of these factors present in our country, this makes the Philippines a
tropical paradise, perfect for many plant and animal life-forms to thrive
throughout the islands. Animal life-forms consist of a variety of species of
mammals, birds, fishes and insects. Insects usually favor the climate in our
country because it provides them with abundant food and warm temperature helps
them in their life-cycle.
Although the Philippines have a large biodiversity, not all of the species are
actually benevolent and some could cause harm and even deaths. One of the
deadliest species present and is abundant in the Philippines is surprisingly not a
giant monstrous beast, but instead is a small flying insect. This insect is known as
the Aedes aegypti, more commonly known as the dengue mosquito or the yellow
fever mosquito. This species of mosquito has been reported to cause 42,500 cases
of dengue fever and 200 deaths as of June 2013. The number of cases here in the
Philippines is relatively high compared to other Southeast Asian countries. After
typhoon Haiyan strokes the Visayas and southern Luzon region, epidemics have
been reported around these areas due to stagnant waters present in the leftovers of

the typhoon. The dengue mosquito is relatively more intelligent and adapted
compared to other species of mosquito. You could not get easily rid of the eggs
because it could survive for one year without water and could still survive when
washed up. The eggs are also laid separately, unlike other species of mosquitoes
that lays its eggs in one bundle. It also has a strategy of attacking from behind and
could lessen its humming sound in order to avoid it from being noticed.
The climate of the Philippines is perfect for the life cycle of the dengue
mosquito. It could just take more or less one and a half weeks for its life-cycle to
be completed, unlike other regions of the world that would take weeks or even
months for the dengue mosquito to complete its life cycle. Because of the
perfectly adapted traits of the mosquito and the favorable tropical climate of the
Philippines, it is very difficult to control the population of Aedes aegypti. The
objective of this research is to find another way to control the population of Aedes
aegypti. It aims to use the extract of the red seaweed Kappaphycus alvarezzii to
attract the mosquitoes and lay their eggs in the attractant. In this way, it would be
much easier to get rid of the eggs of the mosquito, therefore making it easier to
control the population of the dengue mosquito.

Background of the Study

Mosquitoes are tiny fly-like insects which are present in almost all
households. Species of this insect are mostly harmful. Similar to other files, a
mosquitos life cycle undergoes four stages. The cycle may start at the eggs being
laid mostly in aquatic or damp areas. However, some adult mosquitoes lay eggs

varying from one another. Each of the species considers the waters situation.
When the eggs hatch they will turn into larvae and when it matures, it will
become a pupa. Some species are harmless but most are not. One of the most
harmful and known specie of mosquitoes is the Aedes aegypti mosquito. It could
be distinguished easily due to the white markings that could be found in its legs
and body. This mosquito came from Africa. However, it could now be mostly
found in tropical and subtropical areas around the world. This specie served as a
vector to most killer diseases nowadays. One of the most serious cases is the
dengue fever. This is a mosquito-borne type of disease, caused by a dengue virus,
that is present in most tropical regions like the Philippines. Aedes aegypti is
considered as the principal transmitter of the said disease. According to the
Department of Health of the Philippines, as of July 2, 2014 more or less 31,088
dengue cases have been reported. Although there was a huge decrease in
comparison to the number of cases in 2013, the figures are still big to be not taken
seriously. A lot of preventions and remedies are being measured and considered
regarding this matter. One of the most ways to ensure effectivity in preventing the
spreading of mosquitoes is with the use of an attractant. An attractant works like a
magnet to mosquitoes for them to lay eggs. Mosquitoes have a characteristic
where they do not lay all their eggs in one placethis is called as selective
oviposition. By the use of an attractant, one could be able to control the
environments which are mostly aquatic that mosquitoes breed in. An attract could
be any solution or liquid that has bacteria which is capable for larvae to feed in. in
this case, it is best to use something that is an excess or a left-over for practicality.

The researchers made use of Kappaphycus alvarezii which is a species of red alga
that is mostly yellow or green in color. This is considered as one of the main
sources of carrageenan which is commercially necessary for applications such as
in cosmetics and industrial applications. There for this particular species of red
alga is commonly used. Most of the time, the seaweed undergoes a drying process
to be able to extract carrageenan wherein the residue or excess is filtered. This
excess is the drippings which the researchers utilize to be able to achieve a
solution which served as an attractant.

Statement of the problem

The study Kappaphycus alvarezii extract as an attractant for Aedes aegypti
mosquitoes primarily aims to develop an attractant utilizing Kappaphycus
alvarezii drippings to control the environment of Aedes aegypti mosquitoes when
breeding.
Specifically, the researchers seek to determine the answers to the questions as
follow:
1. Is seaweed extract capable to magnet or attract Aedes aegypti
mosquitoes?
2. Will there be any significant difference of having an attractant in the
breeding of Aedes aegypti mosquitoes in comparison to normal water?
3. What would be the most responsive concentration of solution?
100% water
75% seaweed solution

 50% seaweed solution

Significance of the study

The study Kappaphycus alvarezii extract as an attractant for Aedes
aegypti mosquitoes will benefit biologisst, entomologists, those who are inclined
with pest control, people from the seaweed industry and ordinary people in the
researchers locale since it includes the trapping of Aedes aegypti mosquitoes that
could be done in a simple setup but still could attain good results. Moreover, the
attractant can be a great controlled environment for Aedes aegypti mosquitoes.
Considering the selective ovipostion characteristic of Aedes aegypti mosquitoes
wherein they lay their eggs in different areas for the eggs to spread. Bearing that
in mind, there is a good possibility that the process of curing the spread of the
mosquitoes will be much harder to handle. By attracting the mosquitoes, one
could easily limit the reproduction of adult mosquitoes by killing it by the time it
reaches the larvae stage. Dictating the area where the Aedes aegypti would attach
their eggs will make it effortless to prevent the range of the regions to increase.
Scope and Delimitation
The research will utilize the following materials: 9 pieces of large
microwavable containers, 9 pieces of filter paper cut into strips 62 x 4 cm, 17 mL
KD Fertilizer, and 2L of Wilkins distilled water. The microwavable containers and
filter papers were borrowed from the UP Los Baos crop protection laboratory.
The KD fertilizer was shipped from Davao City and the Wilkins distilled water
was bought from 7eleven convenient store.

In preparation for the setup, the extract was diluted with the ratio of 17 mL
of the extract / 2L of distilled water. Afterwards, the diluted solution was poured
into the microwavable containers measuring 500 mL per each treatment. The
containers had the strips of filter paper around the inside. The treatments were
then divided into three groups which are replicated thrice. Setup A contained
100% water, Setup B contained 75% solution and 25% water and Setup C
contained 50% solution and 50% water. Then, the treatments were placed in the
same environment and were observed for 6 days.
The research entitled Kappaphycus alvarezii extract as an attractant for
Aedes aegypti mosquitoes will only test the feasibility of the extract from
Kappaphycus alvarezii as an attractant to Aedes aegypti mosquitoes. At the same
time, only concerned with the extract of one specific species of red alga-Kappaphycus alvarezii. In addition, the study will be concerned with one specific
type of subject, Aedes aegypti mosquito.
The researchers will no longer be concerning with the side effects of the
seaweed extract to the mosquitoes and is critically focused in the attractant
property of the solution only. At the same time, the researchers are no longer
responsible for the other formulations of the extract. The researchers do not claim
application of the attractant property of the solution to other species of
mosquitoes. Likewise, the researchers do not claim the same results in all or
different kind of environment. Lastly, the researchers had depended on the
accommodation of the findings only.

CHAPTER II
Review of Related Literature and Studies

Introduction
Aedes aegypti, commonly known as the yellow mosquito or the dengue
mosquito, is a disease-causing ectoparasite that could transmit viruses and
microorganisms into a host and cause deadly diseases (i.e. yellow fever, dengue
fever, chikungunya, etc). This species is observed to inhabit every continent
excluding Antarctica. It most likely originated from Africa and is most common in
tropical regions. Aedes aegypti is an agent that transmits several tropical diseases.
The female is the only one that bites mammals, including humans, and suck out
blood that is needed for its eggs to mature. The female Aedes aegypti could find a
host by detecting several compounds that are released by mammals such as lactic
acid, carbon dioxide, ammonia and octenol. While the female mosquito feeds off
the blood of the host, there is a large risk that several viruses could enter the body.
Life Cycle
Egg
After the female consumes the nutritious blood of the host, The females
would lay eggs in clear, stagnant water usually found in flower vases, uncovered
buckets or barrels, a small clear pond, discarded tires, pots, tanks, saucers etc. The
female dengue mosquito is usually more motivated to lay their eggs in stagnant
bodies of water or containers that have sufficient amounts of a specific type of
fatty acids that is associated with several species of bacteria that degrades leaves

or organic matter in water. On average, the female Aedes aegypti could lay 100 to
200 eggs per batch, dependent on the amount of blood it had consumed. A female
mosquito has the capacity to lay up to 5 batches of eggs in her lifetime. Unlike
most species of the family Culicidae that usually lays its eggs in clustered batches,
the Aedes aegypti lays its eggs separately. Instead of laying the entire clutch of
eggs in just one location, the female mosquito would lay the eggs in several
different locations. The number of locations depends on the quality and
availability of water. The shape of the eggs is long and ovular. They are white in
appearance when laid but would turn black after a few minutes. The eggs could
develop within two days in warm, tropical areas but would take more or less than
a week in less temperate regions. If the supply of water dried out, the eggs could
survive for a year. Once the egg is submerged in water, it would immediately
hatch. This unique survival mechanism of the dengue mosqito makes the control
of its population difficult.
Larvae
Within 2 days or a week, given that there is no disturbance of the medium
where the eggs are laid, the larvae of the mosquito, called wrigglers or kiti kiti
in tagalog, would hatch from the egg. It feeds off the organic particles in the
water. For most of its larval stage, the wriggler spends within the surface of the
water but it would swim to the bottom of the body of water when it feeds or when
it is disturbed. The development of the larvae is dependent on its gender and the
temperature of the environment. Male wrigglers usually develop faster than
female wrigglers; therefore it would go through the pupae stage first. The warmer

the temperature, the faster the wriggler would develop. If the temperature is
warm, it would take up to 3-4 days for the wriggler to develop into its pupae
stage. But if the temperature is cool, it would take weeks or even months for the
wrigglers to develop as long as water supply is sufficient. The larvae have four
instars; if it went through its fourth instar, which would take three days, it could
measure up to eight millimeters.
Pupae
After the fourth instar of the wriggler, it is time for the larvae to develop
into its pupae stage. The larvae, located on the surface of the water, would shed its
skin before curling itself and turning into pupae. The pupae responds to stimuli,
therefore is mobile. If disturbed, it would swim to the bottom of the body of water
or container. Although it is mobile, the pupae cannot feed. It would take
approximately two days for the pupae to develop if the temperature is warm and if
there would be no disturbance.

Adult
After two days, the adult mosquito would emerge from the cocoon. In
order to hatch, the adult would take air in its abdomen so it would expand. As its
abdomen continues to expand, the cocoon would break and the head would come
out first. After half an hour or so, the adult mosquito has fully emerged from the
cocoon but is not ready to fly yet. It would rest on top of the stagnant water for its
exoskeleton to harden. After this, it would find a safe place for its wings to dry.

Finally, the adult Aedes aegypti is ready for flight and would find a host (if it is a
female) for the cycle to start anew.

Appearance
Adult Aedes aegypti is relatively small compared to other
species in the family Culicidae, having a size of three to four
millimetres in length excluding the legs. Aedes aegypti could be
distinguished from other mosquitoes by the appearance of white scales
that appears like white stripes on the joints and white spots on the
thorax, abdomen and head of the mosquito. Male and female Aedes
aegypti mosquito could be distinguished by its size, the antennae,
palps and the patterns of the white stripes and spots. Female Aedes
aegypti are larger in size compared to the male. The antennae of the
male are attractive compared to that of the females antennae. The
palps of the females are white tipped and the proboscis is all black.
The wings of the dengue mosquito is translucent. Its legs are curved at
rest, just like other mosquitoes.

Behavior
The Aedes aegypti is relatively more intelligent and well adapted
compared to other species of mosquito. It has learned to live in the
environment of humans and has adapted to lay its eggs in man-made
containers. Dengue mosquitoes does not live far away from human
environments, only going as far as 90 meters. Aedes aegypti prefer
human blood and would not consume animal blood unless human

10

blood is not present. Unlike other mosquitoes that would attack on the
head or on the arm and be easily noticed, dengue mosquitoes attack
from behind and would bite on the ankles, elbows or other regions of
the body where it would not be easily noticed. It has also developed to
decrease its humming sound, unlike other mosquitoes that have an
irritating buzz. The manner of the dengue mosquito laying its eggs is
also different. Most mosquitoes would prefer to lay their eggs in
stagnant polluted waters, but Aedes aegypti would lay its eggs in clear
water in order to avoid other organisms in the body of water that might
prey on the mosquito or the eggs. Instead of laying its egg in closelybundled clusters, it lays its eggs in separately and in different
locations.
Kappaphycus alvarezii
Kappaphycus alvarezii is a species of red algae seaweed that is
important for many commercial products because of the cell wall
polysaccharide

and

the

carrageenan

extracted

from

it.

The

carrageenan of the red seaweed could be used products such as

chocolate, frozen desserts, cheese, pharmaceutical products etc. The
species of Kappaphycus are the largest among the red algae and has a
very fast growth rate that could increase its biomass twice in just as
less as fifteen days. Kappaphycus alvarezii is native here in the
Philippines and is largely cultivated in seaweed farms. The seaweed is
used in many commercial products and as it is processed in factories, it
results in waste products that is just thrown away.

11

Attractant
In order to understand how mosquito attractants work, it must
be first understood on how mosquitoes behave, how they bite and
what compounds they are attracted to. Mosquitoes use complex set of
cues in order to detect humans. These are short-range, mid-range and
long-range. The main diet of most mosquitoes are usually fruit and
nectar, but females in most species of mosquitoes feed on the blood of
mammals in order for the eggs to mature and be nourished.
Mosquitoes usually rest in discreet areas or underside of foliage. Flight
is activated when a mosquito receives a cue from a host. A common
example is carbon dioxide, which hosts usually release. A human
breath consists of 4% carbon dioxide and humans could breathe up to
250ml of carbon dioxide per minute. Carbon dioxide could be a midrange or a long-range cue. As soon as the mosquito detected the cue,
it slowly flies upwind and heads towards host which serves as the
source of carbon dioxide. Common household mosquito attractants
could emit 350ml to 450ml of carbon dioxide per minute. The fouler
the attractant, the greater chance it could attract more mosquitoes.
Fouler attractants usually mean more carbon dioxide to be released.

Related Studies
Because of the progressively increasing population of mosquitoes here in
the Philippines, the Department of Science and Technology has made several
effective mosquito attractants. One of these attractants is the Ovicical-Larvical
Mosquito Trap. A whole set of this mosquito trap consists of a piece of plywood

12

stick similar to that of a wooden ice cream stick, A black cup or a black container
and four packs of pellets. The black color of the container could attract some
species of disease-carrying mosquitoes such as the dengue virus. The OL solution
could also attract disease-carrying mosquitoes etc.

There have also been several studies about different plant species and their
effects to mosquitoes, such as the study of Geraldine Marcos and Denise Pacol
which states the larvicidal effect of Lantana camara leaf extract on mosquito. It is
stated in the data analysis that there is a significant difference among the extract
that is formulated and percentage of mortality on mosquito. The researchers of
this study have concluded that Lantana camara extract could be used as a larvicide
for mosquitoes.
There has also been another study regarding the properties of Azadirachta
indica A. Juss as a mosquito repellant. The neem leaaf has always been
used as a treatment for malaria through oral use. The researchers of
this study have concluded that the neem extract has no side effects on
the skin of pigs. The researchers of this study have also concluded that
the effectiveness of the extract decreases as time passes.
There have also been many studies about decaying leaves as
an attractant to mosquitoes and this is also considered as a common
mosquito attractant. The fallen leaves are exposed in water and are
left there for several days in order for bacteria that produce carbon

13

dioxide which is a natural attractant to mosquitoes. The bacteria is also

favorable to the mosquitoes because it serves as food for the larvae.
There are also studies regarding the different compounds that
the mammals release. Examples of these compounds are lactic acid,
sweat and carbon dioxide. These serves an cues to mosquitoes which
helps them find mammals

14

CHAPTER III
RESEARCH METHODOLOGY

This chapter outlines the procedures and experimental design used in the
study to determine the significant relationship between the amounts of Red
Seaweed extract (Kappaphycus alvarezii) and the number of eggs to be laid off by
Yellow Fever mosquito (Aedes aegypti).

Materials
9 pieces of large microwavable containers
9 pieces of filter paper cut into strips 62 x 4 cm
17 mL KD Fertilizer
2L of Wilkins distilled water

Research Method used

This study used the qualitative and quantitative methods of research. The
qualitative method includes the description of the physical and chemical
properties of red seaweed extract before the experiment. This study emphasizes
the importance of the physical properties of an attractant for mosquitos.
Quantitative method focused on the dilution of red seaweed extract in water,

15

amount of water and red seaweed extract in a container, number of replicates for
each treatment and the number of eggs that are observed on the filter paper.

Procedures Used
The study involves the extraction of red seaweed extract from the red
seaweed, dilution of the extract, preparation of the attractant and the analysis of
the experimentation.
1. Processing of Kappaphycus alvarezii Extract
The K. Alvarezii extract used in the study were obtained from the
Province of Davao. The extract was purchased from a professor in a
University in Davao. The professor used the extract as a fertilizer for her
research. The extract was extracted manually. The K. Alvarezii was placed
on a net and there was a container below the net. This set up was placed
outside to be exposed under the sun. The seaweed was exposed under the
sun for three days for it to dry. The drippings were collected by the
container under the net. After three days, the drippings were gathered and
was treated with different chemicals. It was tested in a laboratory for
chemical analysis.
2. Dilution of the Red Seaweed Extract
According from the review of related literature, mosquitos tend to lay
off on transparent solutions like stagnant waters. The physical properties of
the extract was considered in the study. The extract has a translucent feature. It

16

was diluted in water to be an effective attractant. The concentration was in the

ratio of 17 ml of extract in 2000 ml or 2 liters of distilled water.

3. Preparation of the Experiment

Nine sterilized containers were used in the study. For each container
has one filter paper measuring 62 x 4 cm. The amount of varying solutions
to be placed inside the container is 500 ml. The study used three set ups
which there are one controlled and two experimental group. Each treatment
has three replicates.
4. Preparation of the Set up
There are different concentration for each set up. The first set up was
100% distilled water solution. The second set up was 75% diluted seaweed
extract solution and 25% distilled water solution. The third set up was 50%
diluted seaweed extract solution and 50% distilled water solution. Each set
up has three replicates. The experimentation was conducted for six days
under room temperature and was placed inside a room .The number of eggs
present on the filter paper was recorded on the 3rd and 6th day of the
experimentation. The recorded data will be used in the study to determine
the effectiveness of the attractant.

5. Disposal of Solution

17

After the sixth day of observation and experimentation, the filter

papers with mosquito eggs were collected. The collected filter papers were
dried off. An alcohol solution was placed on the attractant solution to kill the
mosquitoes in larva stage before it turns into adult. The researchers will get
the mean of the number of eggs for each solution on the 3 rd and 6th day. Using
this formula:

Where: = mean
fx = amount of eggs for each replicate in the same solution
f = the number of replicates in a solution

Experimental Design
The red seaweed extract attractant for mosquito experiment was conducted
using three different set up. 17 ml of the seaweed extract was added to 2000 ml of
distilled water for dilution. For repeatability, each experimental set-up was
repeated 3 times. Each set-up contains a 500 ml solution. A filter paper was placed
inside each container. The three conditions were 100% distilled water solution,
75% diluted extract and 25% distilled water solution, and 50% diluted extract and
50% distilled water solution. The experiment were observed for six days and the
recording of the data was on the third and sixth day of the experiment. Filter
papers were collected and dried off. An alcohol solution was placed in every
attractant solution for proper disposal.

18

CHAPTER IV
DISCUSSION OF RESULTS

Introduction

The purpose of this study is to determine the feasibility of Kappaphycus

alvarezii extract as an attractant for Aedes aegypti mosquitoes. The results
regarding the number of eggs present in the samples will be discussed. The
data collected as well as the average of each sample and the significant
difference are present in this chapter.

The data has been collected by counting the eggs that were visible in the
filter paper. Aedes aegypti mosquitoes could be distinguished because the
eggs are black when they are attached to the filter paper. The extract is in a
liquid state and is dark brown in color. The extract has been diluted since the
transparency is one of the requirements of being an attractant. The
measurement for each sample is 500 ml. There has been three setups that were
replicated thrice. The solution used has been diluted and has a ratio of 17 mL
of extract/ 2L of H2O. The solution has been diluted since the physical
appearance of the water was considered.

19

Table 1a. Number of eggs of laid by dengue mosquito, Aedes aegypti in

seaweed extracts

and water at 3 days after trapping.

Treatment/Attracta

R E P L I C A T I O N

nt

Seaweed Extract 38

Mean

II

III

37

41

38.67 a

20

14

20.33 b

10

4.33 c

50%
Seaweed Extract - 27
75%
Control (Water)

ANALYSIS OF VARIANCE (Completely Randomized Design)

Source
Variation

of Degrees

F-

F-Tabular

of

Compute

5%

1%

Freedom

d
5.14

10.93

Treatment

Error

SS

MS

1770.89
146.0
0

Total

885.44

36.39**

24.3
3

1916.8
9

20

Computation based on transformed values,

** - highly significant difference
Table 1b. Number of eggs of laid by dengue mosquito, Aedes aegypti in seaweed
extracts

and water at 6 days after trapping.

Treatment/Attracta

R E P L I C A T I O N

Mean

I
nt
Seaweed Extract 44

II
44

III
49

45.67

50%
Seaweed Extract - 80

46

17

47.67

75%
Control (Water)

20

74

31.33

ANALYSIS OF VARIANCE (Completely Randomized Design)

Source

of Degrees

Variation

of

SS

MS

FComputed

Freedom
Treatment
2
476.22
238.11
0.29 ns
Error
6
4936.00
822.67
Total
8
5412.22
CV (%) Computation based on transformed values,

F-Tabular
5%
1%

5.14

10.93

ns Not significant
Table 1a.
Significantly highest number of eggs were laid by adult dengue mosquito
in 50% seaweed extract which constitute about 61.06% (38.67 / 63.33 x 100) of
the total number of eggs laid at 3 days after trapping (Table 1a). This was
followed by 75% seaweed extract where about 32.10% (20.33 / 63.33 x 100) of

21

the eggs were laid. This indicates that it is better to use 50% seaweed extract since
aside from being more economical, significantly higher number of eggs were laid
by the dengue adult mosquito. In contrast, only about 6.84% (4.33 eggs) of the
eggs were laid in water alone. This suggests that in the presence of oviposition
attractant, the dengue mosquito adults will lay majority of their eggs in water with
seaweed extract as attractant.
Table 1b.
At 6 days after trapping, significantly the same number of eggs were laid
in the different concentrations of seaweed extracts and water alone (36.61% of
eggs were laid in 50% Seaweed extract, 38.26% eggs in 75% extract and 25.13
in water alone) which means that the three treatments were equally attractive to
the adult dengue mosquito. However, the results for water for the different
replications is quite inconsistent wherein in Replication I, no eggs were laid
while in Replication III, 74 eggs were counted. Since the containers are placed
very close among each other, there is dilution of odor hence the adult mosquito
could not delineate which containers had the attractant. In order to remedy this
problem, there is a need to place the containers at least 1 meter apart and the
number of replications could be increased to as many as 10.

This experiment should have been conducted in combination with the

release of water bug (Diplonychus rusticus), a voracious feeder of mosquito
wrigglers. However, due to time constraint, this was not undertaken. Results of
the experiment indicate that instead of using plants to kill the dengue mosquito, it

22

is possible to attract them and release predator like water bug which could
consume almost all wrigglers present in the container. The technique of attracting
dengue mosquito adults to lay majority of her eggs in the oviposition attractant
and subsequent release of water bug could be a novel approach in significantly
reducing the population of dengue mosquito since it is environment friendly and
safe to humans.

Chapter V

23

SUMMARY, CONCLUSION, AND RECOMMENDATION

This chapter states the summary procedure, the conclusion and the
recommendation. The summary procedure is a short description of the
methodology of the research study. The conclusion is the answer to the question
Could K. alvarezii extract be used as an attractant to A. aegypti mosquito and
What solution is most effective in attracting A. aeypti mosquito. The
recommendation states the suggestions of the researchers on the continuation and
the further use of the study. It also states other possible uses of the K. alvarezii
extract.
Summary Procedure
First off, the researchers would collect the seaweed, dry it, and collect its
drippings. Then, chemical compounds would be added to the drippings to make
the full chemical composition of the extract. The researchers would dilute the
extract in distilled water with the measurement of 17ml of seaweed extract to 2
liters of distilled water. The researchers would make 3 setups each having 3
replicates. Filter paper would be suspended in the containers. Setup A contains
100% distilled water, Setup B contains 75% solution and Setup C contains 50%
solution. These Setups would be placed in a similar area. The number of mosquito
eggs would be recorded at the third and sixth day of the experimentation. After
the sixth day, the mosquito eggs on the filter paper would be preserved and the
larvae would be killed by the use of 70% ethyl alcohol.

24

Conclusions
The researchers have concluded that Kappaphycus alvarezii extract is an
effective mosquito attractant due to the results. The results show that the seaweed
extract solution contains more eggs in the third and sixth day of the
experimentation compared to that of the 100% distilled water.
The researchers have also concluded that 75% solution of K. alvarezii
extract is the most effective solution that is able to attract A. aegypti mosquito
based on the computed mean as of the sixth day.
It is concluded by the researchers that the number of eggs as of third day
has a significant value of the number of eggs because the number of eggs in each
replicate in a setup is far from the other replicates of different setups. The number
of eggs as of the sixth day has an insignificant value because the number of eggs
in each replicate in a setup is not far from the other replicates of different setups.
The researchers have concluded that the different scent of each extract are mixed
up because each setup did not have appropriate distant from the other setups,
resulting to the insignificant value as of the sixth day.

Recommendation

25

The researchers recommendations for the continuation and the further use of this
study are:

The use of water bugs or other predaors that could kill the Aedes aegypti

in the controlled container as long as it does not emerge to the adult stage.
The use of chemicals that could kill the Aedes aegypti in the controlled

container as long as it does not emerge to the adult stage.

The further study and specification of the compounds or elements within

the extract that attracts adult Aedes aegypti.

The further study of the behavior of Aedes aegypti within the controlled

container.
Other compounds that might be added to the extract that could attract
more mosquitoes.

The researchers also recommend other possible uses of the Kappaphycus alvarezii
extract:

The use of K. alvarezii extract as an attractant to other species of

mosquito.
The use of K. alvarezii extract as a growth enhancer.
The use of K. alvarezii extract as a pesticide.
The use of K. alvarezii extract as a medium for bacterial growth.

Take note of the following:

If ever the experiment is to be re-conducted, there must be a distance of

one meter for each setup. This is done in order to avoid the mixing of the
scent of the solutions, which could affect the selective ovi-position of the
A. aegypti mosquito.

26

Bibliography
Johnson, J. (1998). Biology Visualizing Life. Florida: Holt, Rinehart, and Winston
Klemm, B., Reed, A., Pottenger, F., Porter, C. & Speitel, T. (1999). The Living
Ocean: Biology and Technology of the

Marine Environment.

Honolulu, Hawaii: University of Hawaii

27

Lning, K., Yarish, C., Kirkman, H. (1990). Seaweeds: Their Environment,

Biogeography, and Ecophysiology. New York : Wiley
Christophers, R. (1960). Aedes Aegypti: the Yellow Fever Mosquito: Its life
history, bionomics, and structure.. Cambridge University Press,
New York,

Webliography:
http://en.wikipedia.org/wiki/Aedes_aegypti
http://www.denguevirusnet.com/life-cycle-of-aedes-aegypti.html
http://en.wikipedia.org/wiki/Kappaphycus_alvarezii
http://www.beyondpesticides.org/mosquito/documents/mosq_traps.PDF

28

http://mda.maryland.gov/plants-pests/Pages/mosquitoes_host_attractants.aspx
http://www.microscopyuk.org.uk/mag/art98/aedrol.html
http://www.dengue.gov.sg/subject.asp?id=12
http://www.cdc.gov/dengue/resources/30jan2012/aegyptifactsheet.pdf
http://www.microscopyuk.org.uk/mag/indexmag.html?
http://www.ecdc.europa.eu/en/healthtopics/vectors/mosquitoes/Pages/aedesaegypti.aspx
http://medent.usyd.edu.au/photos/aedes%20aegypti.htm
https://www.google.com.ph/webhp?sourceid=chromeinstant&ion=1&espv=2&ie=UTF8#q=aedes%20aegypti%20male%20appearance

Acknowledgement

Our research topic could not be possible without the help and support
of our beloved authority figures of our school, universities, government agencies

29

and other people who cared enough to help us. The researchers would like to
acknowledge the following people and authority figures with utmost gratitude:
Our research adviser,
Mrs. Donabel G. Simporios, who supported and helped us throughout our
research project; she always came with us when we have to go to different
universities and government agencies to conduct our project. She also helped us
in coming up with the ideas for our project.
Our beloved school principal,
Sir Jay Mathias A. Arellano, for supporting and allowing us to go to
different government agencies and universities.
Our science coordinator,
Ms. Ma. Allen B. Secretario, for supporting us and giving us permission
to go to different government agencies and universities.
Biologist in Bureau of Fisheries and Aquatic Resources,
Ms. Daisy F. Ladra, for greatly helping us by consulting and suggesting
that we could use seaweed extract for our research project.
A professor from a university in Davao,
Ms. Graciella A. Caballero, for the K. alvarezii extract that she offered to
us and for the suggestion and consultancy of ideas.
Dean of the University of the Philippines, Los Baos,
Sir Domingo E. Angeles, for giving us permission for consultancy and
suggesting us people to whom we could consult.

30

Research professor and Deputy Director of the University of the Philippines, Los
Baos,
Sir Pio A. Javier, for suggesting us the idea of K. alvarezii extract as an
attractant to A. aegypti and for helping us in our research paper. He is one of the
most important people who helped us because he came up with the main idea of
our research.
Our class adviser and a research teacher,
Sir Alexander A. Vivar, for the support that he has given to us and for
the help in making letters for government agencies and universities.
To all of our subject teachers, for understanding our hard work and
excusing us from his/her class whenever we are out of the school for our research
project.
To all of our classmates and friends who supported and encouraged
us in our research project
To our parents and relatives for understanding our hard work and for
supporting us throughout our research project. They were always there when we
needed help, guidance and comfort. They also helped us financially in our project
and gave us permission to go to different universities and government agencies.
And last but not least, to our dear and loving God the father because
all of this would not be possible without His help. He guided us through all of our
trips and gave us knowledge and wisdom so we could conduct our project well.

31

Appendices
Appendix A
Table a1: The raw data
SAMPLES

DAY 3

DAY 6

32

A1 (100% H20)

A2 (100% H20)

10

20

A3 (100% H20)

74

B1 (75% SOLUTION)

27

80

B2 (75% SOLUTION)

20

46

B3 (75% SOLUTION)

14

17

C1 (50% SOLUTION)

38

44

C2 (50% SOLUTION)

37

44

C3 (50% SOLUTION)

41

49

Appendix B

33

Consultancy of the research project.

Appendix C

34

Laboratory in University of the Philippines, Los Baos where they breed larvae
Aedes aegypti and waterbugs for research purposes.

Appendix D
The materials needed for the experimentation.

35

Appendix E
The dilution of K. alvarezii extract in distilled water

36

Appendix F
The process of making the solutions and preparation of the setups
37

Appendix G
The setups

38

Appendix H

39

The Aedes aegypti eggs collected on the filter papers as of day si

40