CHAPTER I

Many chemicals have been used for the purpose of Mosquito repellency or killing,

however they are extremely harmful for human beings as well. Recently, commercial

repellent products containing plant-based ingredients have gained increasing popularity

among consumers, as these are commonly perceived as “safe” in comparison to long-

established synthetic repellents.

Our paper describes Plant-based repellents for mosquitoes for personal protection

measures. Based on the knowledge on traditional repellent plants obtained through ethno

botanical studies, development of new natural products is the need of the hour. The

product developed that offers high repellency as well as consumer safety.

The aroma components from natural products have been used for mental, spiritual

and physical healing since the beginning of recorded history. In aromatherapy, fragrance

substances (aroma/odor/scent) from various natural sources have been used for the

treatment of various disorders. The aromatherapy treatment is a natural way of healing a

person’s mind, body and soul. Many ancient civilizations, including Egypt, China and

India, have used aromatherapy as a popular complementary and alternative therapy for

more than thousands of years (Ali et al., 2015).

In recent years, various studies have revealed that olfactory stimulation through

fragrance inhalation exerts various

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psychophysiological effects on human beings. There are various methods available to

administer the fragrances in small quantities, including inhalation, massage or simple

applications on the skin surface and, sometimes, they can be taken internally (Angelucci,

2014). Many studies have suggested that the olfactory stimulation of fragrances produces

immediate changes in physiological parameters such as blood pressure, muscle tension,

pupil dilation, skin temperature, pulse rate and brain activity (Angelucci, 2014).

Mosquitoes are known vectors that transmit pathogenic microorganisms contributing

to the toll of morbidities and mortalities worldwide. These organisms proliferate in the

environment and continually affect the quality of life of each individual affected by

mosquito-borne diseases. In the Philippines, numerous mosquito-borne diseases like

dengue, malaria, and filariasis continue to affect each individual. Newer reports present

that mosquitoes likewise transmit diseases like the Chikungunya and Zika viruses. The

burden of mosquito borne diseases to the general public may bring about morbidity and

or death.Among the mosquito-borne diseases, dengue is considered to be the most

important disease in the world and in the country. Millions of lives worldwide are at risk

of the viral infection transmitted by mosquito vectors. Dengue infections are one of the

important mosquito-borne diseases, as 186,416 cases and 59 deaths were reported in

2012. In Metro Manila alone, a total of 16,046 cases and 29 deaths have been

documented.2 The same report presents that children mostly under 10 years of age are

affected. Like dengue, the Chikungunya viral infection is an emerging concern in the

country. As of 2013, a total of 4,745 cases of the viral infection were reported, and

47.03% were serologically confirmed. Of this total number, about 674 cases, with

59.40% serologically confirmed, were reported in Metro Manila.3 Both Aedes aegypti
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and Aedes albopictus have been known to transmit mosquito-borne diseases. The

occurrence of Aedes mosquitoes in the country has been documented nationwide, making

all regions of the country vulnerable to the transmission of mosquito-borne diseases

(Mistica, 2019). Mosquitoes are one of the deadliest animals in the world. Their ability to

carry and spread disease to humans causes millions of deaths every year. In 2015 malaria

alone caused 438 000 deaths. The worldwide incidence of dengue has risen 30-fold in the

past 30 years, and more countries are reporting their first outbreaks of the disease. Zika,

dengue, chikungunya, and yellow fever are all transmitted to humans by the Aedes

aegypti mosquito. More than half of the world’s population live in areas where this

mosquito species is present. Sustained mosquito control efforts are important to prevent

outbreaks from these diseases. There are several different types of mosquitoes and some

have the ability to carry many different diseases (WHO, 2020). Aedes carrier of

Chingkuya, Zika Virus, Dengue and Yellow Fever and Culex carrier of West Nile Virus

and Anopheles carrier of Malaria (WHO, 2020).

Female mosquitoes, which are the disease vectors, generally require a blood meal to

provide sufficient nutrients for egg production. (Ryelandt, 2011)

Currently, the most common method for controlling mosquitoes in people’s homes is

the use of chemical insecticides. However, the harmful effects of these chemicals on

humans, other animals, and the environment (David, 2013), as well as the development of

resistance to these chemicals by mosquitoes ( Liu, 2015).

Controlling mosquitoes is of utmost importance in the present-day scenario with

rising numbers of mosquito-borne diseases. An alarming increase in the range of

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mosquitoes is mainly due to deforestation, industrialized farming and stagnant water.

Thus, special products like mosquito repellents for combating mosquitoes are required

(Gupta, 2012).

Before the Second World War, there were only four principal repellents, namely,

Citronella oil, Dimethyl Phthalate, Indalone® and Rutgers 612. Other military-developed

mosquito repellent formulae for use on clothing were developed during the war, but they

all failed to provide the desired protection of military personnel deployed around the

world. As a result, by 1956, the United States government had screened over 20,000

potential mosquito repellent compounds. In 1953, the mosquito and other insect repellent

properties of N, Ndiethyl- 3-toluamide (DEET) were discovered and the first DEET

product was introduced in 1956. DEET spray is still the most widely used mosquito

repellent. It has generally been regarded as safe. However, toxic effects have been

recorded, including encephalopathy in children, urticarial syndrome, anaphylaxis,

hypotension and decreased heart rate. Several other compounds have been evaluated for

repellent activity, but none has had the commercial success of DEET(Adeogun, 2012).

Nowadays, there are many types of mosquito repellents which are commercially

available. Spray mosquito repellents are very common and maybe sprayed on the

clothing or skin. Some of the best sprays contain high concentrations of DEET and are

effective for several hours even when walking through deep woods where there are

thousands of mosquitoes (Adeniran, 2012). Since the discovery of DDT, mosquito

control approaches have been almost completely based on synthetic organic insecticides.

However, the extensive use of synthetic insecticides during the last five decades has

resulted in environmental hazards and also in the development of physiological resistance

 5

in major vector species (Enayati, 2012).

Even though, mosquito repellents based on chemicals have remarkable safety

profiles, they are toxic against the human skin and nervous system and may cause rashes,

swelling and eye irritations (Adeogun, 2012).

A repellent is generally defined as a substance that discourages arthropods from

landing or biting human skin (Diaz, 2016). EOs are complex mixtures of volatile organic

compounds from plants. The presence of monoterpenoids, sesquiterpenes, and alcohols

has been proven to attribute to the repellent properties of EOs (Sathantriphop et al.,

2015). In particular, citronellol, citronellal, α-pinene, and limonene are common

constituents of many EOs exhibiting repellent effects (Tong, 2013). Recent evidence has

shown that the odorant receptor neuron in a mosquito's antennal sensilla is activated by

linalool, a naturally occurring terpene alcohol found in many flowers and spice plants,

and by eucalyptol, a natural organic compound (Taylor, 2012). In general, plant EOs are

recognized as safe; however, some of them also cause side effects such as skin irritation,

mainly due to the constituents present in the plant EOs, limiting their extensive

usage ( Benelli, 2018). Natural products are not always safer than synthetic ones, and

some might cause adverse reactions. Moreover, sometimes, the available toxicity and

safety information is limited and contradictory (Dickens, 2013). At present, synthetic

repellents are more widely used than EOs, although this raises several concerns related to

environmental and human health (Lee, 2018).

Bio based mosquito repellent are pest management tools that are based on safe,

biologically based active ingredients derived from plants. Benefits of bio-products

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include effective control of mosquito as well as human and environmental safety. These

bio based products were designed to play an important role in providing pest

management tools in areas where mosquito resistance, niche markets, and environmental

concerns limit the use of products. Mosquito repellents from natural sources are not new.

Some of these mosquito repellent are derived from the following-Basil (Ocimum

basilicum), oils of Castor, Cedar, Clove, Fennel, Citronella, Eucalyptus, Neem ,

Rosemary and Catnip oil of Nepeta species having nepetalactone, Celery extract (Apium

graveolens) as well as Solanum villosum berry juice. These natural resources are good for

the environment and also have added benefit of smelling good (Shukla et al., 2017).

Lemongrass (Cymbopogon citrates. L) the leaves have a lemon like odor because

they contain an essential oil having citral as main constituent, same as what is present in

lemon peel. Leading phytochemical compounds in lemongrass leaf are terpineol,

Dipentene, Limonene, α-terpineol, citronellol, methyl heptenone, dipentene, geraniol,

limonene, nerol, farnesol, mainly (triterpenoids) (Shukla et al., 2017).

The orange (Citrus sinensus), sometimes known as sweet orange, is a valuable fruit

not only for its edibility, but for the high quantities of the substance limonene, sometimes

called d-limonene, found in its peel and zest. Limonene is a pesticide and is found in

many commercial insect repellents. Limonene an oil derived from sweet orange peel has

a 90 to 95 percent content of limonene, which is lethal to fleas, fire ants and flies.

Limonene affects insects on contact, effectively suffocating them by damaging their

respiratory systems. Many insects such as roaches, ants and silverfish do not care for the

scent of orange oil and will avoid it. Placing bits of orange peel or zest around the garden

repels flies and mosquitoes. Rubbing orange peel on the skin is a home remedy for
 7

preventing mosquito bites (Adams, 2018).

Citrus is a genus of flowering trees and shrubs that belong to the Rutaceae family.

Fruits in this genus include orange, lemon, lime, grapefruit, tangelo, pomelo, and so on.

Several types of research have proven the efficacy of citrus essential oils as mosquito

repellents. This research also shows that citrus essential oils work against both adult

mosquitoes and larvae. This is a popular and efficient way of controlling mosquitoes. If

you combine the affectivity with the fact that this is an organic approach that isn’t

hazardous to plants, animals, or the environment, you’ll agree that this is one of the best

ways to fight mosquitoes out there, Citrus fruits are used in many different ways to help

in the fight against mosquitoes. If used correctly, we might just stand a chance at winning

some of these battles. Citrus Peels contain organic chemicals that have the ability to kill

mosquito larvae. In addition, research shows that some citrus scents repel adult

mosquitoes. Unfortunately, you must extract the essential oils from the peels for this to be

effective. Simply rubbing a lemon peel your skin won’t work as a mosquito repellent.

Squirting lemon juice on yourself won’t work, either. D-Limonene; Oranges and lemons

contain the compound D-limonene. This compound is effective at chasing away

mosquitoes and some other insects. It’s also a component in many citrus essential oils

(Kripena, 2019).

Oil of lemon eucalyptus extract (also known as p-menthane-3,8-diol, or PMD) is

derived from the leaves of lemon eucalyptus trees and chemically synthesized, usually in

the form of a spray. The ingredient has earned its mosquito-repelling stripes in two

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NMSU studies led by Rodriguez. In the first, published in the Journal of Insect Science in

2015, researchers looked at eight commercially available repellents, two fragrances, and a

vitamin B patch, and noted whether each formula repelled or attracted disease-carrying

mosquitoes Aedes albopictus and Aedes aegypti when applied to participants' hands.

What they found: Cutter Lemon Eucalyptus Insect Repellent, a plant-based spray that

contains oil of lemon eucalyptus, was the only DEET-free formula to deliver strong and

long-lasting results. The second study, published in the Journal of Insect Science in 2017,

looked at how effectively wearable repellents (such as the bracelets mentioned above)

along with DEET and PMD sprays protect against female Ae. aegypti mosquitoes, the

species most likely to spread Zika. Once more, the Cutter Lemon Eucalyptus Insect

Repellent was found to be effective. Animal studies on oil of lemon eucalyptus extract

have not identified any adverse effects, although the FDA warns against using it on

children under age 3, since it can irritate the eyes. Also important: The ingredient has not

yet been studied for its ability to repel mosquitoes that carry diseases other than Zika, and

unlike DEET, PMD repellents are not recommended for protection from ticks (Felton,

2018).

1.1 Purpose

This study is to produce a low-cost, high-quality scented candle by using kalamansi

(Citrofortunella microcarpa), lemon grass (Cymbopogon citratus), and eucalyptus leaves

(Eucalyptus globulus) extracts as an anti-mosquito repellent scented candle an alternative

organic agent of mosquito repellency.

1.1.1 Description of the Problem

This research cannot be successful as well as the project of the study if there are no
 9

possible problems which will be serve as a guide and solution to make this study possible

and true. With the help of the anticipated problems of the study, the project will be more

successful.

The following are the anticipated problems of the study;

1. Duration

The problem that the researchers are facing is to produce a product with a

high amount of duration, long enough for it to do its suppose to do.

2. Effectiveness

Upon mixing the 3 main ingredients will the product be effective to repel

mosquitoes.

3. Aroma

Will the product be able to produce a wonderful and lovely aroma.

Through this study the community are able to be aware that there are natural, safe

and eco-friendly alternative anti-mosquito repellent ingredients that can be found in their

vicinity that can be turned into a multi-purpose candle. The study will help a lot to every

family unit and some associated industries. Furthermore, its preparation is easy and won’t

require much laborious.

The main concern of this study is to find out if eucalyptus leaves, lemongrass and

kalamansi extract as an effective natural alternative anti mosquito scented candle.

1.1.2 Project Scope

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The main purpose of this study is to find out if the Eucalyptus leaves, lemongrass

and kalamansi extract as an natural alternative anti mosquito scented candle. How

effective the Eucalyptus leaves, lemongrass and kalamansi extract as a main component

in making anti mosquito scented candle.

The analysis, findings and conclusions of the study under consideration were

primarily based from the procedures set and experiments done by the researchers .

1.1.3 Description of the Project

This study is to produce a low-priced, high-quality scented candle by using

kalamansi (Citrofortunella microcarpa), lemon grass (Cymbopogon citratus), and

eucalyptus leaves (Eucalyptus globulus) extracts as an anti-mosquito repellent scented

candle an alternative organic agent of mosquito eradication.

1.1.4 REVIEW OF RELATED LITERATURE

Features
Related Studies Candle/Ligh
Natural Non-Topical Repellent
t Producer
The effectiveness
of lemon grass as
√ √
natural insect
repellent
The Combo of
Lemon's
abundance and √ √ √
Candle's ability
Versus Mosquitoes
INVESTIGATIO √ √ √
N OF THE
REPELLENCE
ACTIVITY
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OF BIO-OUT, A
NATURAL
MOSQUITO
REPELLENT
Assessment of
Efficiency of Eco-
Friendly
Organic Mosquito √ √ √
Repellent
Developed using
Elephant Dung
Mosquito
Repellent Actions
of the Essential
Oils of
Cymbopogon
citratus,
Cymbopogon √ √ √
nardus and
Eucalyptus
citriodora:
Evaluation and
Formulation
Studies
EUCALEM-O;
Anti-Mosquito
√ √ √ √
Scented Candle

1.1.4.1 The Effectiveness of Lemon Grass as Natural Insect Repellent

The researchers wants to determine the effectiveness of lemon grass as a natural

insect repellent. Their objective is to produce a natural insect repellent, a topical repellent

using only lemon grass extract. About 20 pieces of lemongrass was collected near the

house of the researcher. Cleaned then ready for the extraction. The extraction process was

done through Steam Distillation Technique. The researchers use candies as a test subject

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for their study. One candy was treated (applied) with the extract while the other was not.

After (14) minutes, the candy without the lemon grass were approach by insects, while

the other candy that was treated with lemongrass was not approach by insects.

1.1.4.2 The Combo of Lemon's abundance and Candle's ability Versus Mosquitoes

The first goal of the study was to compare the degree of personal protection

against bitinginsects provided by geraniol, linalool, and citronella candle (5%) vapors

outdoors, where such products arecommonly used. At a distance of 1.0 m, citronella

candles reduced the number of female mosquitoes caughtin Centers for Disease Control

and Prevention traps by 35.4% and sand flies by 15.4%, linalool candlesreduced female

mosquitoes by 64.9% and sand flies by 48.5%, while geraniol candles reduced

femalemosquitoes by 81.5% and sand flies by 69.8%. By increasing the distance to 2 m

and 3 m, the repellencydropped significantly. The second goal was to compare the degree

of personal protection provided by the bestperforming candle, geraniol, under conditions

of high and low biting pressure. The introduction of geraniolcandles to protect volunteers

in a high biting pressure environment reduced the mosquito pressure by anaverage of

56% and the sand fly pressure by 62% over a distance of 1.0 m. In the low biting

pressureenvironment, geraniol reduced the mosquito pressure by an average of 62%. No

sand flies were present atthis site.

1.1.4.3 INVESTIGATION OF THE REPELLENCE ACTIVITY OF BIO-OUT, A

NATURAL MOSQUITO REPELLENT

Mosquito repellents are important tools for prevention of dreadful diseases as well as

painful mosquito bites. Lemongrass essential oil (Cymbopogon citratus) has been
 13

investigated for its repellent activity against mosquitoes respectively. Tests were

performed without the contribution of human volunteers. Number of visible mosquitoes

was noticed. Approximately 80% repellence activity has been observed. Lemongrass oil

exhibited an average of 30 min protection time against mosquitoes to make them away

from the observed area, All-out (50-80%) protection time in comparison to the activity of

the best known chemical insect repellent, N, N-diethyl-mtoluamide (DEET). Determining

the effectiveness of lemon grass (Cymbopogon citratus) as mosquito repellent is the main

goal of the study. Specifically, it intends to apply the extraction procedure techniques to

extract the essential oils of lemon grass. It can be concluded that lemon grass oil is a

promising natural repellent due to its safety advantage over chemical repellents. The

study aims to make a cream formula made from natural ingredients.

1.1.4.4 Assessment of Efficiency of Eco-Friendly Organic Mosquito Repellent

Developed using Elephant Dung

Natural ingredients ike Nerium flowers, Tulsi, Neem leaves, Lemon grass oil and

Natural loban were used in combination with the elephant dung to reinforce the potency

of the mosquito repellent. Smoke toxicity of organic mosquito repellent was evaluated.

Emission test for commercial and developed repellent was also compared and it is

evident that there is 57% reduction in the carbon-di-oxide emission from developed

repellent than commercial repellent and there are no traces of emission of carbon

monoxide from the developed repellent. Also, cage test proves that the developed

mosquito repellent is highly efficient against mosquitoes and hence can be more

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effectively used for control of mosquitoes. According to the study Usage of elephant

dung for production of mosquito repellant has not been reported before. The developed

organic herbal was quite efficient in repelling and killing the mosquitoes without harming

the human health. This study resulted in development of eco-friendly organic herbal

repellent with long lasting protection, safe for human life, animal skin and humans with

no side effect, as an alternative to commercially available synthetic chemical repellents.

The results of this investigation indicated that the elephant dung could be beneficial for

the control of mosquito borne diseases.

1.1.4.5 Mosquito Repellent Actions of the Essential Oils of Cymbopogon citratus,

Cymbopogon nardus and Eucalyptus citriodora: Evaluation and Formulation Studies

According to the study essential oils extracted by steam distillation

from Cymbopogon citratus (DC.) Stapf, Cymbopogon nardus (L.) Rendle and Eucalyptus

citriodora Hook were evaluated for their repellent effects against Anopheles

arabiensis mosquitoes under laboratory conditions. Blended oils, ointments and cream

formulations of the oil of C. nardus in different bases were also evaluated. At 10 % and

20 % concentrations, all the oils showed a minimum of 90 % and 95 % relative

protection, respectively, soon after application. These were not significantly different in

efficacy from NN-diethyl-meta-toluamide (DEET), P<0.05. Afterwards however, only C.

nardus oil produced more than 70 % protection for 3 h and 4 h at a concentration of 10 %

and 20 %, respectively. Combinations of 10 % or 20 % of each of the oils of C.

citratus and C. nardus gave better protection than their individual effects. In the presence

of the oil of E. citriodora, the effects of C. citratus and C. nardus were significantly

reduced at both concentrations. The oil of C. nardus in oleaginous ointment (i.e.
 15

Vaseline) provided above 80 % relative protection for as long as 4 h which was better

than the effect of the 20 % oil, P<0.05. Similarly, the oil in hydrophilic ointment, 1:1 w/w

blend of PEG 4000 and PEG 600, gave better repellent effect than the 20 % oil (P<0.05).

However, the effect of the o/w cream was not statistically different from the 20 % oil,

P>0.05. The blends of C. citratus and C. nardus oils and the ointment formulations of the

oil of C. nardus showed improvement on the percentage of protection of the oil of C.

nardus. However, both the blending and the formulations did not improve the duration of

protection. In addition, both the percentage and duration of protection were not as good

as DEET, P<0.05. This study demonstrated the potential of essential oil of C. nardus as

topical repellent against An. arabiensis.

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 CHAPTER II

PRODUCT DESCRIPTION

This chapter discusses the functionality and capability of the product. It describes the

specification of design of the product which affect the product limitations and issues.

2.1 Product Perspective and General Features

The product will repel mosquitoes and produce nice aroma. Thus, to materialize the

different expectation, the following features being realized:

2.1.1 Use

The flicker of the candlelight creates a soothing ambiance that can add effect and

create a cozy and lovely atmospheres in homes. It can also be a source of light in the dark

when there is no electricity, can also be a stylish way to add touch to any part of the

house. Apart from that its therapeutic scent have a capability to help relax, feel calm and

help the brain get into a peaceful mindset. Its aroma also can kill mosquitoes.

2.1.2 Ingredients

The ingredients of the product are purely from nature without adding harmful chemical

ingredients and since its from nature anyone can make it.

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Budget Friendly

Environmental Friendly

EUCALEM-C
Usable

User Friendly

Anti Mosquito
 CHAPTER IV

PRODUCT PROTOTYPE

This section provides detailed information on the application to undertake during the

product use. It comprises of the different product prototypes that shows its control and

functionalities of the product. It defines the aspect of the product that contributes to its

user friendliness and environmental friendliness.

4.1 User-Friendliness

4.1.1 Light

Candles have been used as a source of light to illuminate. Candles

were used to aid travelers at night and light homes, especially from families that

lived from far flung areas unreachable to electricity.

4.1.2 Aroma

Can also be a therapy as the user inhales the aroma promotes

concentration and has calming and clarifying properties that are helpful when you’re

feeling angry, anxious, run-down and help control emotional stress or just want to feel

relaxed.

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4.1.3 Anti-Mosquito

With its aroma molecules, stops the mosquitoes from biting and

transmitting Dengue and other diseases harmful to human being.

4.2 Environmental-Friendliness

4.2.1 Anti-Mosquito

The product only targets mosquitoes and not other insects which stabilize the

ecosystem