Chapter 1

THE PROBLEM AND ITS SETTING

Background of the Study

Mosquitoes are the carriers of severe and well-known illnesses such as

malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus

and yellow fever. These diseases produce significant morbidity and mortality in

humans and livestock around the world (Murugen, et.al, 2012). And mosquitoes

are common flying insects that are found around the world.

Repellents are expensive nowadays, and the researcher wanted to see if

this repellent would be a cheaper alternative. Repellents are tools for prevention

of insect-borne diseases as well as painful or uncomfortable insect bites. The 2

most effective and widely used products are N,N-diethyl-3-methylbenzamide

(deet), which is applied to exposed skin, and permethrin, which is applied to

clothes.

Biasong is characterized by small flowers (thus the “small-flowered”

moniker) with fewer stamens than other papedas and oblong-obovate, few-loculed

fruits. They have numerous flat, pointed, reticulate seeds. The fruits were not used

only for food but for hair-washing, and it had little economic importance.

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 The peels of the Biasong are just thrown away and they don’t have any

other uses. And it will be treated as a garbage. In view of an increasing interest in

developing peel origin repellants as an alternative to chemical repellants, thus

feasibility of Biasong peels extract as repellent against Aedes aegypti was made

and conducted by the researcher.

Statement of the Problem

The main problem of the study is to determine the effectiveness of the

Biasong peels extract as repellent against Aedes aegypti

Specifically, the study sought to answer the following questions:

1.Is the repellent from Biasong peels extract effective against Aedes

aegypti?

2. Does the Biasong peels extract have the potential as an effective

repellent against Aedes aegypti?

3. Is there a significant difference between the alternative repellent

compared to the commercial chemical repellent?

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Hypotheses

From the problems stated, the following hypotheses were formulated:

H1:

1. The repellent from Biasong peels extract is effective against Aedes

aegypti.

2. The Biasong peels extract have the potential as an effective repellent

against Aedes aegypti.

3. There is a significant difference between the alternative repellent

compared to the commercial chemical repellent.

H0:

1. There is no significant difference between the alternative repellent

compared to the commercial chemical repellent.

Objectives of the Study

1. To identify the repellent from Biasong peels extract effect against

Aedes aegypti.

2. To evaluate the effectiveness of the repellent from Biasong peels extract

against Aedes aegypti.

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 3. To produce an effective and low-cost repellent from Biasong peels

extract alternative to chemical repellents.

Conceptual/Theoretical Framework

To provide a working framework for this study, the following factors are

considered to the success of the production of the repellent from Biasong peels

extract against Aedes aegypti:

1. Production the repellent from Biasong peels extract against Aedes

aegypti.

2. The use of bioassay to evaluate the effectiveness of the alternative

repellent.

3. Application of the repellent from Biasong peels extract against Aedes

aegypti.

The following diagram shows the schematic relationship of the variable

involve in the theoretical base of this study.

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 Production the repellent from Biasong peels extract against Aedes
aegypti.

Application of the
The use of bioassay to evaluate repellent from
the effectiveness of the Biasong peels
alternative repellent. extract against
Aedes aegypti.

Figure 1. Schematic diagram of the Study

Significance of the Study

The study the feasibility of Biasong peels extract as a repellent against

Aedes aegypti will be a great benefit to the people of Magallanes and its

environment. It is because it can be offered as a solution for the making of an

alternative repellent instead of using chemical repellent. The researcher desire to

assess the repellent against Aedes aegypti. The study principally aims to

determine the effectiveness of feasibility of Biasong peels extract as a repellent

against Aedes aegypti.

This study was concerned on converting Biasong peels extract as the

larvicidal repellent against Aedes aegypti. The Biasong peels are utilized for the

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repellent preparation. The study needs to be pursued for it can help the

community of Magallanes, Agusan del Norte. It can help as a reference for other

researches, students and future researchers.

Scope and Limitation of the Study

This study focused on the feasibility of Biasong extract as a repellent

against Aedes aegypti was characterized through drying and making a solution.

Then, the researcher used bioassay to evaluate the effectiveness of the Biasong

peels extract feasiblity of Biasong peels extract as a repellent against Aedes

aegypti against Aedes aegypti.

The materials were collected in the municipality of Magallanes, Agusan

del Norte. The duration of the study will be from January to March 2018 and

conducted at Magallanes National High School, town of Magallanes, province of

Agusan del Norte, Philippines.

Definition of Terms

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 Repellent – a substance that keeps away insects.

Mosquitoes – any of a family (Culicidae) of dipteran flies with females

that have a set of slender organs in the proboscis adapted to puncture the skin of

animals and to suck their blood and that are in some cases vectors of serious

diseases.

Biasong – is a species of wild citrus among those called papedas native to

southern Philippines, particularly islands of Cebu and Bohol.

Aedes aegypti – the yellow fever mosquito, is a mosquito that can spread

dengue fever, chikungunya, Zika fever, Mayaro and yellow fever viruses, and

other diseases.

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 Chapter 2

REVIEW OF RELATED LITERATURE

A. Biasong

Figure 2. Citrus micarantha

The Biasong fruit is also known as Citrus micrantha var. micrantha. The

aroma of the fruit is similar to that of the samuyao. The tree reaches 7.5 to 9

meters in height. Leaves are 9-12 cm long, 2.7- 4.0 cm wide, broadly elliptical to

ovate, crenate, thin with base rounded or broadly acute; apex acutely blunt

pointed. Petioles are 3.5-6 cm long, broadly winged, up to 4 cm wide, with wings

(phyllodes) sometimes larger than the leaf. Flowers are small, four-petaled, white

with thin purple edge, 12-13 mm in diameter, forming cymes of two to five. There

are 15-17 equal stamens. The ovary is obovoid, with 6-8 slender, distinct locules.

Fruits are obovate to oblong obovate, 5-7 cm long, with diameter of 3-4 cm,

averaging 26 grams in weight. Their skin is rather thick, fairly smooth or with

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transverse corrugations. The pulp is juicy, grayish and acid, while juice cells are

short and blunt to long, slender and pointed. Sometimes containing a minute,

greenish nucleus. They have numerous, flat, pointed, reticulate seeds. Citrus

micrantha, the small-flowered Papeda or Biasong, is one of the parents of the

common lime (Key lime), Citrus aurantifolia (Wikipedia, 2018).

B. Aedes aegypti Mosquitoes

Figure 3. Aedes aegypti mosquito

Aedes aegypti, the yellow fever mosquito fever, is a mosquito that can

spread dengue fever, chikungunya, Zika fever, Mayaro and yellow fever viruses

and other diseases. The mosquito can be recognized by white markings on its legs

and a marking in the form of a lyre on the upper surface of its thorax. This

mosquito originated in Africa, but it is now found in tropical and subtropical

regions throughout the world. The yellow fever mosquito belongs to the tribe

Aedini of the dipteran family Culicidae and to the genus Aedes and subgenus

Stegomyia (Wikipedia, 2018).

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C. Other related studies and its application

Another related researches that, the efficacy of prepared citronella leaf

cakes was evaluated on three different parameters such as flammability, burning

time and mosquito repellency test. Also, the cakes were sprayed with different

concentrations of citronella oil. Based on the results obtained from these

parameters, the residual percentage of each combination of cakes was calculated

and it suggested that Neem powder cake has the most effective repellency activity

when impregnated with 10% citronella oil (Rani, Nandini,et. al, 2013).

The results of the repellent activity of plant extract of A. nilagirica plants

at five different concentrations of 50,150, 250, 350, and 450 ppm were applied on

skin of fore arm in man and exposed against adult female mosquitoes. In this

observation, the plant crude extract gave protection against mosquito bites

without any allergic reaction to the test person, and also, the repellent activity is

dependent on the strength of the plant extracts. The adult mortality was found in

methanol extract of A. nilagirica, with the LC50 and LC90 values of 205.78 and

459.51 ppm for A.stephensi, and 242.52 and 523.73 ppm for A. aegypti,

respectively. This result suggests that the leaf extract have the potential to be an

ideal eco-friendly approach for the control of vector mosquito as target species

(Chellasamy,P. et.al ,2012).

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 In this study, various Citrus grandis’s fruit peel extracts (hexane, ethyl

acetate, methanol and essential oil) have been tested to determine its repellency

effect against ovipositing female and blood hungry Aedes aegypti mosquitoes.

The findings showed that oviposition activity by gravid female mosquitoes were

successfully deterred by ethyl acetate at 10 ppm. In another study, pure essential

oil effectively repelled female Ae.aegypti mosquitoes from attacking the bait in

modified tunnel test. In addition, animal model applied with 20% essential oil

showed good protection rate (94.7%). These results indicated that Citrus

grandis’s fruit peel extracts have repellency effect on female Ae. Aegypti

mosquitoes (Jamil, Mailina, et.al, 2017).

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 Chapter 3

METHODOLOGY

A. Collection and Preparation of Materials

The biasong peels were collected from the town of Magallanes, Agusan

del Norte. These biasong peels were removed from dirt and then dried at room

temperature. The peels were then powdered coarsely. The fruit peels have been

extracted (crude methanol extract, hexane fraction, ethyl acetate fraction, queous

fraction and crude water and essential oil) as described.

Figure 4. Biasong peels. (Source: from www.google.com)

B. Preparation of Stock Solutions

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 One gram of crude methanol extract, hexane fraction, ethyl acetate

fraction, queous fraction and crude water extracts was first dissolved in 100 ml of

respective solvent (stock solution). Similarly, 1 ml of essential oil was dissolved

in 100 ml of acetone (stock solution). From the stock solution, serial dilution was

prepared.

C. Repellency Activity

Repellency effects of different concentration of Biasong essential oil

against female Aedes aedypti mosquitioes. Forty 5-7 days old blood, starved

female Ae. Aegypti mosquitoes were released into a mosquito cage measuring 30

cm x 30 cm x 30 cm. Rats (weighed 100) abdomen were shaved. Treatment of 1

ml of acetone containing 5%, 10%, and 20% of extract were applied on its

exposed abdomen. DEET and acetone was used as the positive and negative

control, respectively. At the start of the experiment, the forty mosquitoes were

released into the mosquito cage. The stopwatch was immediately started as soon

as the rat was placed on the top of the mosquito cage where the treated surface

was exposed to the mosquitoes. The total number of bites was recorded for 3

minutes at each interval. The control and treatment tests were replicated 3 times.

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