Acknowledgements

First and foremost, I would like to thank everyone who helped and guided
me in making this project. This work would not have been possible without their
support and guidance. I would like to express my special appreciation and thanks
to my research teacher, Mr. Nestor Brito, you have been a great mentor for me. I
would like to thank you for encouraging my research and for allowing me to grow
as a researcher. Your advice on my research has been invaluable. I also want to
thank you for letting my defense be an enjoyable moment, and for your brilliant
comments and suggestions, thanks to you.

A special thanks to my family. Words can not express how grateful I am to

my mother and father for all of the help that you’ve given me. Thank you for
supporting me for everything, and especially I can’t thank you enough for
encouraging me throughout this experience. I would also like to thank my friends.
Your support, sense of humor, patience, optimism and advice was more valuable
than you could ever imagine.

Finally I thank God, for letting me through all the difficulties. I have
experienced Your guidance day by day. You are the one who let me finish my
research study. I will keep on trusting You for my future. Thank you, Lord.

i
 CHAPTER I
Background of the Study

Mosquitos are a family of small, midge-like flies. There are over 2500
different species of mosquitoes; about 200 species occur in the United States with
51 species occurring in Idaho. They have been known to cause many diseases to
people. Most mosquitoes are considered as harmful because the females
consume blood from living vertebrates including humans. Some of them acquire
or transmit harmful human livestock diseases while feeding on blood. Mosquitoes
carry diseases like dengue and malaria. Malaria is a serious and sometimes fatal
disease caused by a parasite that commonly infects a certain type of mosquito
which feeds on humans. People who get malaria are typically very sick with high
fevers, shaking chills, and flu-like illness.

According to the World Health Organization, in 2016, there were an

estimated 216 million cases of malaria, an increase of about 5 million cases over
2015. Deaths reached 445 000. Malaria may be fatal if treatment is delayed
beyond 24 hours after the onset of clinical symptoms. The incidence of dengue
has grown dramatically around the world in recent decades. Dengue is found in
tropical and sub-tropical climates worldwide, mostly in urban and semi-urban
areas. Severe dengue is a leading cause of serious illness and death among
children in some Asian and Latin American countries.

According to the World Health Organization, the actual numbers of dengue

cases are underreported and many cases are misclassified. One recent estimate
indicates 390 million dengue infections per year (95% credible interval 284–528
million), of which 96 million (67–136 million) manifest clinically (with any severity
of disease) Another study, of the prevalence of dengue, estimates that 3.9 billion
people, in 128 countries, are at risk of infection with dengue viruses.

To prevent diseases like these, using commercial mosquito repellent is the

common solution. Most of these mosquito repellents can destroy mosquitoes but
they can also do serious damage to humans as well. So I, as a researcher, decided
to come up with a study by using basil extract as an alternative mosquito
repellent which is not only affordable but also environmental friendly. It is not a
hazard to our health because the ingredients are chemical free. It is much safer to
use this alternative mosquito repellent because instead of using chemical
ingredients I used natural ones which does not contain harmful ingredients.
ii
 Statement of the Problem

The study aims to determine the potential of basil extract as mosquito

repellent. The study will specifically answer the following questions:

1. Is there any characteristic of the extract that can harm the environment?
2. What is/are the active component/s in the extract?
3. What is the mortality rate of the mosquitoes when the extract was used
on them?
4. Is there a significant difference in the mortality rate of the mosquitoes
between the usage of the extract and the commercial repellent?

Hypothesis

 Null (Ho): There is no significant difference in the mortality rate of

mosquitos between the use of the basil extract and the commercial
mosquito repellent.

 Alternative (Ha): There is a significant difference in the mortality rate of

mosquitoes between the use of the basil extract and the commercial
mosquito repellent.

 Cause and Effect: If you use the basil extract as a mosquito repellent
instead of the commercial mosquito repellent, then the population of
mosquitoes will decrease.

iii
 Significance of the Study

This study aims to explore data about the effects of basil extract when used
on mosquitoes will benefit the society, researchers, and families.

1. Society - This study will benefit the society by the reason of it is a

cheaper way of killing mosquitoes since the materials’ availability is
abundant in our country.

2. Researchers - This study will benefit researchers studying about

ocimum basilicum or the lamiaciae family. This will serve as their guide in
generating their hypothesis and results.

3. Families - This study will benefit families because it is a cheaper way

to kill mosquitoes that cause us harm and it will harmless because there is
no harmful chemicals in the extract.

Scope and Delimitation

This study will focus on the potential of basil extract as mosquito repellent.
This study will compare the effect of the extract and the effect of the commercial
repellent. The extract will only be sprayed on mosquitoes and no more other
living organisms. A certain amount of basil extract will be needed. Basil leaves are
abundant in the Philippines.

This study is not limited when it comes to the said materials since they are
sufficient and can be found in rural areas.

iv
 CHAPTER II
Review of Related Literature and Studies

This chapter includes the ideas, finished studies, generalization or

conclusions, methodologies, and others. Those that were included in this chapter
helps in formulating information that are relevant and similar to the present
study.

Related Literature

Ocimum basilicum L. (sweet basil) a member of Lamiaceae family, is native

throughout the old World and cultivated for religious and medicinal purposes.
Basil was originated in Asia and Africa. Basil is used in both Ayurvedic and Unani
system of medicine and is also popular for its culinary and ornamental uses.
Various parts of the plant of sweet basil have been widely used in traditional
medicine. The leaves and flowers of basil are used in folk medicine as a tonic and
vermifuge. Basil tea is good for treating nausea, flatulence and dysentery.

The oil of the plant has been found to be beneficial for the alleviation of
mental fatigue, colds, spasm, rhinitis, and as a first aid treatment for wasp stings
and snakebites. Studies showed that basil possesses central nervous system (CNS)
depressant, anticancer, cardiac stimulant, hepatoprotective, hypoglycemic,
hypolipidemic, immunomodulator, analgesic, anti-inflammatory, antimicrobial,
antioxidant, antiulcerogenic, chemomodulatory and larvicidal activities. The
present review article provides up-to-date information on basil chemical
properties, therapeutic benefits and pharmacological studies. (Ch et al., 2015)

Traditional agricultural systems are challenged by globally declining

resources resulting from climate change and growing population. Alternative
agricultural practices such as aquaponics (includes crop plant and aquatic species)
and hydroponics (includes crop plant only) have the potential to generate high
yield per unit area using limited land, water, and no soil. A soilless agricultural
study was conducted at the Georgia Southern University, Statesboro, GA, USA
from August to November, 2015

v
 The growth, yield, quality, and nutrition of basil (Ocimum basilicum
L.) cultivar Aroma 2, were compared between aquaponic and hydroponic systems
using crayfish (Procambarus spp.) as the aquatic species. Non-circulating floating
raft systems were designed using 95 L polyethylene tanks. Equal amounts of start-
up fertilizer dose were applied to both systems. The objective was to understand
how the additional nutritional dynamics associated with crayfish influence the
basil crop. Both fresh and dry basil plant weights were collected after harvest,
followed by leaf nutrient analysis. Leaf chlorophyll content, water pH, nitrogen
and temperature were measured periodically. Aquaponic basil (AqB) showed
14%, 56%, and 65% more height, fresh weight, and dry weight, respectively,
compared to hydroponic basil (HyB). It is logical to assume that crayfish waste
(excreta and unconsumed feed) has supplied the additional nutrients to AqB,
resulting in greater growth and yield. The chlorophyll content (plant quality) or
leaf nutrients, however, did not differ between AqB and HyB. Further research is
needed to investigate aquaponic crayfish yield, overall nutritional dynamics, cost-
benefit ratio, and other plant characteristics under soilless systems. (Day et al.,
2016)

On the other hand, mosquitoes vector many important disease pathogens.

Controls of mosquito-borne diseases often involve lowering the contact rate
between human and vector, and killing the vector. For instance, long lasting
insecticidal nets (LLINs) act on both of these levels. Unfortunately, these are at
best short-term solutions as mosquitoes are known to exhibit resistance to these
interventions. Physiological resistance to insecticides and repellents is well-
documented worldwide [1]. However, behavioural resistance (e.g. changing
encounter rates) to chemical threats and barriers is often underappreciated even
though the importance of mosquito behaviour for disease control was
emphasized as early as the 1950s [2]. For instance, it has been shown repeatedly
that an increasing proportion of mosquitoes have started blood feeding earlier,
before humans retire under their LLINs, essentially avoiding insecticides applied
to the net’s surface [2]. (Hoi, Amber Gigi and Bernard D. Roitberg, 2014)

vi
Related Studies

K. Kamsuk, W. Choochote, U. Chaithong, A. Jitpakdi, P. Tippawangkosol,

D. Riyong, B. Pitasawat. Effectiveness of Zanthoxylum piperitum-derived
essential oil as an alternative repellent under laboratory and field
applications.(2007)
Recently, there were considerable efforts made to promote the use of
environmentally friendly and biodegradable natural insecticides and repellents,
particularly from botanical sources. In this study, Zanthoxylum piperitum-derived
essential oil isolated by steam distillation was investigated and compared to the
standard synthetic repellent, N,N-diethyl-3-methylbenzamide (DEET), for
repellency against mosquitoes under laboratory and field conditions. The oil of Z.
piperitum alone and also with 5% vanillin added repelled laboratory-reared
female Aedes aegypti with the median protection times of 1.5 and 2.5 h,
respectively. These repellency times were proven to be significantly lower than
those of DEET-based products: 25% DEET and 25% DEET + 5% vanillin
demonstrating 3.5 and 5.5 h, respectively. However, when applied under field
conditions, Z. piperitum oil + 5% vanillin was found to provide better protection
against a wide range of natural mosquito populations than 25% DEET + 5%
vanillin. While the product of DEET was effective in reducing bites with 99.7%
protection, the protective effect of Z. piperitum oil product appeared complete
(100%). It had a protective effect against Aedes gardnerii, Anopheles
barbirostris, Armigeres subalbatus, Culex tritaeniorhynchus, Culex gelidus, Culex
vishnui group, and Mansonia uniformis. The better repellency against a wide
range of field mosquitoes derived from Z. piperitum oil products suggested an
advantage of efficacy by providing a broad spectrum of activity. Therefore, Z.
piperitum could have potential for use in the development of combined
repellents as a natural active ingredient, synergist, or additive to conventional
synthetic chemicals, particularly in situations when DEET is ineffective and
impractical.

Mark S. Fradin, MD. Mosquitoes and Mosquito Repellents: A Clinician's

Guide.(1998)
This paper is intended to provide the clinician with the detailed and scientific
information needed to advise patients who seek safe and effective ways of
preventing mosquito bites.For this review, clinical and analytical data were
selected from peer-reviewed research studies and review articles, case reports,
entomology texts and journals, and government and industry publications.
Relevant information was identified through a search of the MEDLINE database,
the World Wide Web, the Mosquito-L electronic mailing list, and the Extension

vii
Toxicology Network database; selected U.S. Army, U.S. Environmental Protection
Agency, and U.S. Department of Agriculture publications were also reviewed.

Evangeline T. Oparaocha, Iraneus Iwu & J.E. Ahanaku. Preliminary study on

mosquito repellent and mosquitocidal activities of Ocimum gratissimum (L.)
grown in eastern Nigeria.(2010)
The study examined the mosquito-repellent and mosquitocidal activities of the
volatile oil of Ocimum gratissimum at three different locations (World Bank
Estate, Ihitte and Umuekunne) in Imo State, eastern Nigeria, with the purpose of
sourcing for mosquito repellent that is cheap, abundant, environment and user-
friendly. Four different lotions; 20% (v/v) and 30% (v/v) concentrations each of
the extracted volatile oil in two natural oil bases (olive and palm kernel) were
made and six volunteered human baits were used to evaluate the mosquito
repellent and mosquitocidal activities of the stock materials at the three different
centres from September to November 2008. Topical application of each of the
four different lotions significantly (p <0.05) reduced the biting rate of mosquitoes
in all the three locations tested. The 30% (v/v) concentration in olive oil base
exhibiting highest average percentage repellencies of 97.2, 95.7 and 96.3% at
World Bank Estate, Ihitte and Umuekunne centres respectively while the 20%
(v/v) concentration in palm kernel oil base had the least repellency of 36.3, 41.6
and 36.3%, respectively. The other two formulations had values ranging from 67.8
to 80% in the three locations. The 30% (v/v) concentration in both olive and palm
kernel oil bases afforded all night protection against mosquito bites in all the
centres, and demonstrated fast knockdown and paralyzing effect on few
mosquitoes at the urban centre (World Bank Estate).

Marta Ferreira Maia and Sarah J Moore. Plant-based insect repellents: a review
of their efficacy, development and testing.(2011)
Plant-based repellents have been used for generations in traditional practice as a
personal protection measure against host-seeking mosquitoes. Knowledge on
traditional repellent plants obtained through ethnobotanical studies is a valuable
resource for the development of new natural products. 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 although this is sometimes a
misconception. To date insufficient studies have followed standard WHO
Pesticide Evaluation Scheme guidelines for repellent testing. There is a need for
further standardized studies in order to better evaluate repellent compounds and
develop new products that offer high repellency as well as good consumer safety.
This paper presents a summary of recent information on testing, efficacy and

viii
safety of plant-based repellents as well as promising new developments in the
field.

Dhara Shukla, Samudrika Wijayapala and Padma S Vankar. Effective mosquito

repellent from plant based formulation.(2017)
All over the developing countries in the tropical region mosquito menace has
been very acute leading to many diseases. 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 ethnobotanical studies,
development of new natural products is the need of the hour. The product
developed that offers high repellency as well as good consumer safety. Thus this
plant-based repellent made from 5 plant leaves has promising results in the field
study.

College of Chemical Sciences, Institute of Chemistry Ceylon, Rajagiriya, Sri

Lanka. Development of Herbal Mosquito Repellent Formulations.(2016)
The present study was conducted to determine the mosquito repellent activities
of some selected plant materials in order to obtain safe and efficient
herbal mosquito repellent formulations by combinations of the selected plant
materials. Azadirachta indica1 seeds were socked in hexane overnight and extract
was filtered and the filtrate was concentrated by rotary evaporator. Same
procedure was carried out for Vitex negundo2 leaves using hexane and ethanol
solvents. Hydro-distillation process was performed for Ocimum sanctum3 leaves,
Curcuma longa4 rhizomes and Citrus sinensis5 peels using Clevenger-arm
apparatus in order to obtain the essential oils. Essential oils of Cymbopogon
nardus4 leaves, Eucalyptus globulus6 leaves and Syzygium aromaticum1 buds
were purchased from a reliable source. 10% (V/V%) extract/essential oil
containing ethanol solutions were prepared using each plant extract/essential oil
and mosquito repellent activity testing was carried out using arm-in-cage method.
Volunteer’s forearm which had been rubbed with 1 ml of the test solution was
exposed to the cage where 20 blood-seeking mosquitoes had been placed and the
number of mosquitoes that aligned or biting the arm was recorded in each minute
for five minutes. Three replicates were carried out for each extract/essential oil
containing ethanol solutions. After analyzing the mosquito repellent activity of
individual extracts and essential oils, a mosquito repellent gel and a mosquito

ix
repellent spray which contained 16% (V/V%) active ingredients each were
prepared. Outdoor and indoor field trials were conducted in two days from 5 am
to 11 am by separately applying the mosquito repellent gel and the mosquito
repellent spray on volunteers’ legs. The gel and the spray showed 100% mosquito
repellency for outdoor and indoor field trials which were carried out for six hours
each day for two days.

CHAPTER III
Methodology

This chapter discusses the materials to be utilized and the sequenced

procedures on how to perform the experiment.

Materials
The study will be utilizing basil leaves (Ocimum basilicum), ethanol, mortar
and pestle, a spray bottle, a pair of rubber gloves, a container and a bottle of OFF
mosquito repellent.

Procedures
A. COLLECTION OF MATERIALS
The Basil will be collected from the localities of Tabaco. The basil will be
stored in a jar at a room temperature. The basil leaves will then be cut using a
knife while wearing rubber gloves. The mosquitoes can be found here in Tabaco.
The mosquitoes will be stored in a container where air can pass through.

x
B. EXTRACTION OF MATERIALS
After sun-drying the basil leaves, they will be pulverized using mortar and
pestle and stored in jar. The powder will be mixed with water and ethanol in
separate spray bottles.

C. REPELLENT TESTING
The mosquitoes will be divided into five which will result to 5 mosquitoes
per container. Four of the containers will be sprayed with the extract while the
last container will be sprayed with OFF mosquito repellent. Each set-up will be
compared with the reference to the mortality of mosquitoes.

Figure 1. Conceptual Framework

25% Basil Extract

And Ethanol

50% Basil Extract

Basil And Ethanol Mosquito
 
Extract 75% Basil Extract (Culicidae)
And Ethanol

100% Basil Extract

And Ethanol

xi
 CHAPTER IV
Results and Discussions

In Figure 1, the diagram of the study was presented. This explains how the
extract was used and how it was divided to compare the difference of the results.
It shows that every concentration of the basil extract will affect the mosquito.

Mosquitos Repelled
Set - up Concentration Trial 1 Trial 2 Trial 3 Mean
25% Basil Extract
1 And Ethanol 2 2 4 2.6

50% Basil Extract

2 And Ethanol 3 4 5 4

75% Basil Extract

3 And Ethanol 5 4 6 5

100% Basil Extract

4 And Ethanol
5 8 8 7

5 OFF Mosquito Repellent 6 6 9 7

Presentation of Data

Conclusion
In this study, the aim is to formulate a natural mosquito repellent based on
a mixture of basil and ethanol extract. Basil is a natural source of essential oil.
Essential oils have compounds with high repellent activity like alpha-pinene,
limonene, citronellol, citronellal, camphor and thymol. However, synthetic
chemicals are more commonly used as repellents than essential oils. These
natural products have the potential to provide efficient, safer repellents for
humans and the environment. We have 4 set-ups with different concentrations
each and 1 commercial repellent. The most effective set-up comparable to the
commercial mosquito repellent is set-up 4. Set-up 4 repelled the highest number
of mosquitoes because, compared to the concentrations of the other set-ups, set-
up 4 has the most amount of basil extract and ethanol which is why it is the one
that can be compared to the commercial mosquito repellent out of the 4 set-ups.

xii
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https://www.webmd.com/skin-problems-and-treatments/alternative-mosquito-
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https://www.npr.org/sections/goatsandsoda/2018/06/30/623865454/a-guide-to-
mosquito-repellents-from-deet-to-gin-and-tonic

https://phys.org/news/2018-08-homemade-mosquito-repellants-
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