University Research Proposal: Essential Oils as Mosquito Repellents

This research proposal investigates the potential of essential oils as mosquito repellents, addressing the growing concerns of diseases spread by mosquitoes. The study aims to examine the synergistic activity of essential oils, such as Litsea and peppermint oil, against mosquitoes using olfactometers. The research will focus on the behavioral changes of mosquitoes when exposed to essential oils, utilizing a quantitative experimental design with a randomized controlled trial method. A sample size of 100 participants will be divided into groups to assess the efficacy of different essential oils. Statistical analyses, including T-tests and ANOVA, will be used to evaluate the repellent behavior and mean values. This study seeks to provide safer and environmentally friendly alternatives to synthetic insecticides, highlighting the significance of plant-based repellents in vector control. The project will use an olfactometer to measure the repellent behavior of mosquitoes exposed to the essential oils.

Running head:RESEARCH PROPOSAL
RESEARCH PROPOSAL
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Abstract
Of the insects, mosquito species are the most important; since they transmit major diseases
viz. Malaria, dengue, zika, chikungunya, encephalitis and filariasis of different forms; pose
immense threats to both humans and animals. Such diseases leads to significant deaths and
morbidity worldwide. The key technique for controlling these diseases is the protection of the
vectors and the use of adult mosquito repellent. The commercial products are being used that
leads to many skin related problems. Various studies have indicated the use of essential oils
as a mosquito repellent. Keeping that in mind, this proposal aims to examine the synergistic
activity of essential oil against mosquitoes using olfactometers. Statistical analysis revealed
that essential oils like Litsea oil, peppermint oil are potent repellant against mosquitoes. The
essential oils are natural plant products which may be useful in designing effective mosquito
repellents based on safer and newer herbal ones.
Specific aims
This study aims to inquire the impact of essential oils on the behavior of the mosquito.
The research objectives are:
 To examine the importance of essential oil to repel mosquitoes
 To examine the behavioral change of mosquitoes on use of potential essential oil
 To examine the use of olfactometer to know the efficacy of the essential oil
Introduction
Mosquitoes historically were believed to cause malaria. It has been known over the
last few decades that mosquitoes spread not only malaria but also a large number of other
significant pathological complications such as dengue, chikungunya, yellow fever, Japanese
Encephalitis (JE), filariasis, and so on. Multiple strategies to controlling the mosquitoes were
established. The latest methods for managing mosquitoes are based on synthetic insecticides.
Because of their swift action, synthetic insecticides are the first line of action, but their

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continuous use can lead to resistance growth and adverse environmental effects. These
factors have created a need to look for alternative insecticides that are easily biodegradable
(Navayan et al., 2017). The use of vector control plant extracts has many attractive features,
since they are biodegradable, less dangerous and abundant stock of chemicals of diverse
biological activity (Markus & Linder, 2015). Essential oils have been used by entomologists
as mosquito larvicides, or several researchers are employed in the vector control system for
repelling insect pests (El-Seedi et al., 2017)
Essential oils are chemicals derived from plants and are "responsible for the
distinctive odor or taste of the plant from which they originate," according to the EPA (PDF).
Essential oils can offer many benefits (Conti et al., 2014). Users of essential oils have found a
number of uses for these naturally derived oils, from stress relief to restful sleep. While
disease-ridden mosquitoes have plagued tropical regions throughout the past, in the United
States they are increasingly appearing. So, use of an essential oil can be of good help. It is
important to remember that essential oils can be used topically on the skin (but not sensitive
skin near the eyes, etc.), spread with a diffuser, or make a spray by diluting the oil with water
and either witch hazel toner (found in the beauty aisle) or vodka. Essential oils can be very
strong, although they are natural and plant-based. There are certain smells that mosquitoes
love, so it is an opportunity to get such smells in your yard for these irritating pests (Tisgratog
et al., 2016). But just like there are smells that mosquitoes enjoy, so there are others they hate
and make sure they avoid. Citronella, Basil, Peppermint, Garlic, Rosemary, Lemon Balm,
Lemongrass, Lavender, Neem, Marigold, Catnip, Bergamot, Eucalyptus, are some scents that
repels mosquitoes (Ojewumi, Adeyemi & Ojewumi, 2018). Mosquitoes hate wearing scented
oils and this may be one way to keep them away from skin (and blood). Citronella is one of
the most common scents which the mosquitoes dislike. All-natural treatment of Mosquito
Squad consists of essential oils and kills mosquitoes when in contact, and then continues as a

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repellent for up to 14 days. This interrupts the mosquitoes 'life cycle and face trouble in
surviving on skin. An olfactometer is a tool used to detect and measure dilution of odours.
Olfactometers are used in laboratory environments, most often in market research, in
combination with human subjects to measure and assess human smell (Uniyal et al., 2016). It
is used in this study to measure the efficacy of essential oils that can be used as a repellant.
This study will draw attention on the behavior of mosquitoes on using essential oils.
Research designs
Naturally, occurring botanical compounds include a wide variety of chemical active
ingredients that can interact with all of the mosquito's biological processes, thus interrupting
its life cycle and dispersal and reducing harm to both humans and animals. Some reports of
serious side effects caused by commercial product such as DEET include long-term
application of the repellent, strong, repeated or whole-body. Yet for certain individuals,
DEET can lead to adverse reactions to the skin, seizures and brain disturbances, exhaustion,
respiratory problems and possibly even cancer. Whereas the use of essential oils have high
beneficial factors such as some essential oils like citronella contains antibacterial effects that
treats minor cuts and scrapes and also control body odors caused by bacteria. Some essential
oils participates in aromatherapy along with controlling mosquitoes (Ali et al., 2015).
Research Method
A quantitative study will be done to check the importance of essential oil and examine
the behavioral change of mosquitoes on application of essential oils. An experimental design
will be set up which will follow descriptive cohort study. Randomized controlled trial method
will be used that will assign participants into experimental group or control group. When the
analysis will be performed, the only potential difference in a randomized controlled trial
(RCT) between the control and the experimental groups is the outcome variable being tested.
The olfactometer will be used to observe the repellent behavior of the mosquito. The

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olfactometer will be kept on the table with pressurized flow of air. One arm will be exposed o
essential oils and other arm will be exposed to isopropanol treated paper which is used as
control. Then the mosquitoes will be exposed and mosquitoes count will be taken from both
the arms.
Sample size
A number of 100 random people will be chosen irrespective of age and gender. The
group will be divided into two groups. One will be provided with different essential oils or
mixture of essential oils and other group will be not be provided with any repellants. A trial
period of two month will be taken to know the efficacy.
To examine the use of olfactometer to know the use of essential oils , 20 essential oils that
will be evaluated effective against mosquitoes from above method will be chosen. The test
mosquitoes will be reared in laboratory condition with proper nutritional condition. Then the
repellant behavior of the mosquitoes exposed to 20 essential oils will be studied using Y-
maze olfactometer.
Statistical analysis
Statistical method such as T-test and Anova will be used to analyze the efficacy. The
repellence and mean values will be evaluated using ANOVA one way for variance ratio
calculation and the least significant difference (LSD) To evaluate the different combinations
of essential oils K and D module will be used.
Significance
Essential oils are gaining significance over commercial products like DEET. The
essential oils like litsea oil, pepper mint oil, tea tree oil, lavender oil and many more. There
are many different strategies in use for the control of mosquito vectors such as attractants,
insecticides, repellents, control of the biological environment and genetics, etc. which have
their own merits and demerits. It is therefore important to develop safe and environmentally

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friendly techniques for controlling vector-borne diseases, especially in the mosquito region.
Plant-based repellents are also widely used in rural communities

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References
Ali, B., Al-Wabel, N. A., Shams, S., Ahamad, A., Khan, S. A., & Anwar, F. (2015). Essential
oils used in aromatherapy: A systemic review. Asian Pacific Journal of Tropical
Biomedicine, 5(8), 601-611.
Conti, B., Flamini, G., Cioni, P. L., Ceccarini, L., Macchia, M., & Benelli, G. (2014).
Mosquitocidal essential oils: are they safe against non-target aquatic
organisms?. Parasitology research, 113(1), 251-259.
El-Seedi, H. R., Azeem, M., Khalil, N. S., Sakr, H. H., Khalifa, S. A., Awang, K., ... & Borg-
Karlson, A. K. (2017). Essential oils of aromatic Egyptian plants repel nymphs of the
tick Ixodes ricinus (Acari: Ixodidae). Experimental and Applied Acarology, 73(1),
139-157.
Markus, A., & Linder, C. (2015). U.S. Patent No. 9,079,152. Washington, DC: U.S. Patent
and Trademark Office.
Navayan, A., Moghimipour, E., Khodayar, M. J., Vazirianzadeh, B., Siahpoosh, A.,
Valizadeh, M., & Mansourzadeh, Z. (2017). Evaluation of the mosquito repellent
activity of nano-sized microemulsion of Eucalyptus globulus essential oil against
Culicinae. Jundishapur J. Nat. Pharm. Prod, 12.
Ojewumi, M. E., Adeyemi, A. O., & Ojewumi, E. O. (2018). Oil extract from local leaves-an
alternative to synthetic mosquito repellants. Pharmacophore, 9(2), 1-6.
Tisgratog, R., Sanguanpong, U., Grieco, J. P., Ngoen-Kluan, R., & Chareonviriyaphap, T.
(2016). Plants traditionally used as mosquito repellents and the implication for their
use in vector control. Acta tropica, 157, 136-144.
Uniyal, A., Tikar, S. N., Mendki, M. J., Singh, R., Shukla, S. V., Agrawal, O. P., ... &
Sukumaran, D. (2016). Behavioral response of Aedes aegypti mosquito towards
essential oils using olfactometer. Journal of arthropod-borne diseases, 10(3), 370.a

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