FEASIBILITY OF USING POWDERED OREGANO (Origanum vulgare),

LEMONGRASS (Cymbopogon citratus) AND MARIGOLD (Tagetes erecta) AS

MOSQUITO LARVICIDE

A Research Paper Submitted to

Davao Oriental Regional Science High School

In Partial Fulfillment of the Requirements in Practical Research 2

(Quantitative Research)

PRINCE JOHN G. CAPAROS

KHYLA METZI A. DEBUTIACO
ELYZA PATRICE C. GORDULA
MARC LESTER B. OBATONON
ALLENA JYN O. RIMANDO
ERIKKA M. SULATRE
Researchers

KRISTAL G. ENTRINO
Researcher Adviser

January 2023
 DEPARTMENT OF EDUCATION
Region XI
Schools Division of the City of Mati
Davao Oriental Regional Science High School

APPROVAL SHEET

This Research/Capstone Project entitled “FEASIBILITY OF USING

POWDERED OREGANO (Origanum vulgare), LEMONGRASS (Cymbopogon
citratus) AND MARIGOLD (Tagetes erecta) AS MOSQUITO LARVICIDE” has
been prepared by ALLENA JYN O. RIMANDO, ELYZA PATRICE C. GORDULA,
ERIKKA M. SULATRE, KHYLA METZI A. DEBUTIACO, MARC LESTER B.
OBATONON, PRINCE JOHN G. CAPAROS, is submitted to the School Scientific
Review Committee for acceptance and approval for Research Proposal Defense.

KRISTAL G. ENTRINO
Research Adviser

SCHOOL RESEARCH REVIEW COMMITTEE

Accepted and approved by the committee on Oral Examination with a grade of _______.

GENE PEARL A. LUNA

Chairperson

LLOYD U. ANDRES LORIE MAE A. BABIERA MARIA FE M. DUMARAN

Member Member Member

MELBERT I. FLORES RODRIGO A. SALIMACO, JR. HAIDEE M. SIASON

Member Member Member

JAIME S. YU
Member

Accepted and approved by the School Scientific Review Committee.

JESSICA M. LUMAPAS
Secondary School Principal I
 TABLE OF CONTENTS

TITLE PAGE

Cover Page………………………………………………………………………………………………………i

Approval Sheet……………………………………………………………………………………………….ii

Table of Contents…………………………………………………………………………………………..iii

List of Figures………………………………………………………………………………………………...v

Chapter I – INTRODUCTION

Background of the Study…………………………………………………………………………1

Statement of the Problem……………………………………………………………………….3

Objectives……………………………………………………………………………………………..4

Hypotheses……………………………………………………………………………………………4

Conceptual Framework………………………………………………………………………….4

Significance of the Study………………………………………………………………………..5

Scope and Limitation…………………………………………………………………………….6

Definition of Terms……………………………………………………………………………….7

Chapter II – REVIEW OF RELATED LITERATURE

Oregano……………………………………………………………………………………………….9

Lemongrass Oil…………………………………………………………………………………….10

Marigold………………………………………………………………………………………………11

Larvicide…...…………………………………………………………………………………………13

Mosquito……………..………………………………………………………………………………14

Mosquito Larvae…....…………………………………………………………………………….15

Summary…………………………………………………………………………………………....16

Chapter III – METHODOLOGY

Research Design………………………………………………………………………………….19

Locale and Duration………………………………………………………………..…………..19

Materials…………………………………………………………………………………………….20
 Instrumentation…………………………………………………………………………..……..21

Experimental Setup…………………………………………………………………….....…..22

Data Gathering Procedures………………………………………………………………….22

Data Analysis………………………………………………………………………………….....23

REFERENCES

APPENDICES

A - Materials

B – Instrumentation

C – Experimental Setup

LIST OF FIGURES
FIGURE TITLE PAGE

1 Conceptual Framework of the Study 4

2 Satellite view of the Location 18

Chapter I
 INTRODUCTION

Background of the Study

Mosquitoes are a primary public health concern worldwide. They significantly impact

local economies and are responsible for the spread of illnesses (La Quarta, et al., 2022).

Mosquito-borne illnesses are a dangerous concern for human and animal health in all

tropical and subtropical areas (Erasga, 2018). Malaria, filariasis, chikungunya, yellow fever,

dengue fever, and Japanese encephalitis are just a few of the diseases transmitted by

mosquitoes that cause significant morbidity and mortality in humans and animals worldwide

(Sharma et al., 2019). Moreover, the past three decades have seen a tremendous global

growth of numerous mosquito-borne diseases despite centuries of control efforts (Vega Rúa

& Okech, 2019).

The second most lethal mosquito-borne illness is dengue fever. It is a viral illness

brought on by one of the four serotypes of the dengue virus. In the Philippines, the dengue

virus outbreak, spread by mosquitoes, is currently affecting the whole country (Dumilon, et

al., 2020). Regarding dengue fever cases, the Philippines had the seventh-highest rate in the

world from 2004 to 2010 (Lozano, 2018). In addition, the country recorded the most cases in

August 2019, with about 150,000, and more than 400,000 by the end of the year (Ligue &

Ligue, 2022).

Locally, 38,169 dengue cases were reported in the Department of Health-Davao

Region between 2011 and 2015. Dengue cases increased during the study period from 4115

cases in 2011 to 9507 cases in 2012, peaking at 10,762 cases in 2013, the year with the

highest number of reported dengue infections during the study period (Iguchi, et al., 2018).

On the other hand, dengue cases in the area are most prevalent in Davao City. The city

recorded 4,495 cases of dengue in 2019, which is 50% of the total occurrences in the region.

In the first half of 2021, the city had already recorded 1,431 dengue cases, a 66% rise from
the same time the previous year (Ligue & Ligue, 2022). Thus, dengue is still endemic in every

region, with hundreds of thousands of cases reported yearly.

One method of controlling mosquito-borne diseases is to reduce the population by

reducing the availability of oviposition sites and/or reducing the likelihood that the resulting

larvae will develop into adulthood. Larval source management (LSM) aims to reduce the

number of adult mosquitoes by concentrating on the mosquito's aquatic, immature stages

(Stanton, et al., 2021). Egg, larva, pupa, and adult are the four stages of the mosquito's life

cycle. One of the aspects of mosquito life cycles is the aquatic stage of immaturity. In a

larvicidal mosquito control strategy, this phase is crucial (Mariam, et al., 2021).

With the possible illnesses caused by mosquitoes, it is indeed inevitable to eliminate

them in their early stage using natural ingredients abundant in a specific locality. In

Giatropolous, et al., (2022) study, the use of plant essential oils (EOs) and their constituents,

primarily terpenes and a variety of aromatic and aliphatic constituents, is currently one of

the most promising alternative low risk strategies aimed at mosquito control. Numerous EOs

are recognized as toxic to various mosquito species' larvae. Some EOs are also identified

against mosquitoes as ovicides, oviposition deterrents, pupicides, adulticides, antifeedants,

growth and/or reproduction inhibitors.

The use of larvicides and repellents, especially in endemic areas, is an effective way to

lessen the burden of the disease. Larvicides are applied to breeding habitats to help reduce

the adult mosquito population in nearby areas. However frequent use of chemical

insecticides like larvicides and repellents has harmed the environment and made mosquitoes

more resistant (Sanei-Dehkordi, et al., 2021).

Additionally, larviciding is one of the most significant malaria and dengue prevention

strategies involving 55 nations. Nevertheless, microbial infections because of overuse of

chemical larvicides are another health issue that has jeopardized both human and

environmental health as well as produced vector resistance. Gram-positive Staphylococcus

2
aureus and gram-negative Pseudomonas aeruginosa are two frequent opportunistic bacteria

that cause a variety of infections like skin ailments like discomfort, swelling, and changes in

skin color in humans. Other recently emerging concerns for the health systems include

microbial drug resistance and adverse effects of chemical medications. Therefore, it is crucial

to create new medications and insecticides with fewer negative effects (Osanloo, et al., 2022).

These studies focused on the formulation of liquid larvicides made of herbs.

However, little attention is given on using and combining two or more plant components in

creating a more effective herbal powdered larvicide. Furthermore, more general studies

address reducing mosquito larvae rather than targeting fully developed mosquitoes to

decrease their likelihood of maturing. Thus, the researchers distinguished this study of using

three plants in eliminating mosquito larvae in a form of herbal powdered larvicide with no

negative effects on humans and on the environment.

According to World Health Organization (2022) in their Dengue Situation Update on

deaths, dengue-related deaths in 2022 are higher compared to the cases reported in 2021. To

avoid the continuous increase in Dengue cases and other mosquito diseases, this powdered

larvicide can be an effective tool in preventing the disease from spreading and controlling the

mosquito larvae population in areas. Also, this may reduce the number of cases of mosquito-

related diseases and the chances of getting the infections.

Statement of the Problem

This research study aims to determine the feasibility of using powdered oregano

(Origanum vulgare), lemongrass (Cymbopogon citratus), and marigold (Tagetes erecta) as

efficient herbal mosquito larvicide.

Specifically, it seeks to answer the following questions:

1. What is the performance of each powdered herb as mosquito larvicide?

 2. Is there a significant difference among the treatments in eliminating mosquito

larvae?

Objectives

This study aims to: 3

1. Determine the performance of the herbal mosquito as larvicide.

2. Determine if there is significant difference among the treatments in eliminating

mosquito larvae.

Hypotheses

Alternative

Ha: The herbal powdered treatments are efficient as herbal mosquito larvicide.

Ha: There is a significant difference among the treatments in eliminating mosquito

larvae.

Null

Ho: The herbal powdered treatments are not efficient as herbal mosquito larvicide.

Ho: There is no significant difference among the treatments in eliminating mosquito

larvae.

Conceptual Framework

Independent Variable Dependent Variable

Powdered Oregano Numbers of

(Origanum vulgare), mosquito larvae
Lemongrass
eliminated
(Cymbopogon citratus),
and Marigold (Tagetes
erecta)
 Figure 1: Conceptual Framework of the Study

Figure 1 illustrates the conceptual framework of the study. It consists of three

4
variables, powdered oregano (Origanum vulgare), lemongrass (Cymbopogon citratus), and

marigold (Tagetes erecta) as the independent variable and number of mosquito larvae

eliminated as the dependent variable.

The diseases mosquitoes can bring are prevalent problems to society. It requires

action to create alternatives in preventing such causable illness. Thus, this study intends to

investigate the effectiveness of powdered oregano (Origanum vulgare), lemongrass

(Cymbopogon citratus), and marigold (Tagetes erecta) as an effective larvicide. Oregano

(Origanum vulgare) has active ingredients such as carvacrol, thymol and a-terpinene which

are effective in repelling mosquitoes (Sharififard et al., 2018). Also, lemongrass

(Cymbopogon citratus) contains natural insect repellent compounds such as citral and

geranyl acetate (Grifford, 2022). Marigolds (Tagetes erecta) contain a natural compound

pyrethrum used in many insect repellents and they have a distinctive smell that repels

mosquitoes (Boehme, 2021).

In conclusion, making productive powdered oregano oregano (Origanum vulgare),

lemongrass (Cymbopogon citratus), and marigold (Tagetes erecta) can be effective in

eliminating mosquito larvae which can lessen the dilemma mosquitoes can bring.

Significance of the Study

This study of powdered oregano (Origanum vulgare), lemongrass (Cymbopogon

citratus), and marigold (Tagetes erecta) will hold significant impacts to the community as an

efficient herbal mosquito larvicide. It will also provide crucial information and knowledge

that is beneficial to different individuals and future studies.

 Moreover, the study is beneficial to the following:

Households. This study will provide an effective mosquito larvicide, chemical free,

which will make it safe for humans and the environment. Also, the said product of this study

will benefit humans to avoid mosquito-borne diseases.

Health Organizations. This study will provide assistance in uncovering new

5
solutions in fighting life-threatening diseases like dengue, yellow fever, and other illnesses

caused by mosquitoes.

Field of Science. This study will aid the said field in determining the efficiency of

powdered oregano (Origanum vulgare), lemongrass (Cymbopogon citratus), and marigold

(Tagetes erecta) as herbal mosquito larvicide. Moreover, it will provide additional

knowledge on existing researches.

Future Researchers. This study will open doors for future researchers to refine

and expand their study. This may serve as a source of information or reference for their

research.

Scope and Limitations

This study aims and focuses on determining the effectiveness of powdered oregano

(Origanum vulgare) leaves, lemongrass (Cymbopogon citratus), and marigold (Tagetes

erecta) in eliminating mosquito larvae. The researchers will use cultured mosquito larvae as

the samples in this study and the herbal powdered treatments as the eliminating compound.

Additionally, it will highlight the benefits of the finished product and its quality. Also, this

study will be conducted within March to April 2023.

In making this study, there will be multiple processes to be followed and to be done.

At the end of this research study, there will be an expected outcome of a new and efficient

way in killing mosquito larvae.

Definition of Terms

6
This section presents words that will be used in the study. These terms will be

conceptually and operationally defined.

Larvicide Larvicide refers to the regular application of

microbial or chemical insecticides to water bodies or

water containers to kill the aquatic immature forms of

the mosquito (the larvae and pupae) (Wilson & Choi,

2019).

Lemongrass Lemongrass, (Cymbopogon citratus), also

called sweet rush, a species of oil grass in the family

Poaceae, often used in cooking (Hill, 2022). In this

study, the extract of this herb will be used as the

treatment.

Marigold Marigold (Tagetes erecta) is a flowering,

herbaceous annual plant that is known for its large

flowerheads (Sharma, 2022). In this study, this herb

will be used as an ingredient for the treatment.

Mosquito Mosquitoes are disease-causing vectors that are

major causative agents of human filariasis, malaria, and

many other viral diseases like dengue, Japanese

 encephalitis, Zika, and West Nile virus (Mondal, et al.,

2021).

Mosquito Larvae Mosquito Larvae are ferocious insect-killers

with cannibalistic tendencies. But the larvae hunt so

quickly, their attack appears only as a blur to the

human eye (Osborne, 2022). In this study, these will be

the sample objects.

Oregano Oregano (Origanum vulgare) is grown in the

7
Mediterranean, Euro-Siberian and Iran-Siberian

regions. Essential oils of oregano are widely known for

their antipathogenic activity demonstrated in cellular

and animal studies (Oh, et al., 2022). In this study, this

is referred to as the primary ingredient in the herbal

treatment.
 Chapter II

REVIEW OF RELATED LITERATURE

This chapter presents the related literature and studies after the thorough and in-

depth search done by the researchers. This will also present literature about Oregano

(Origanum vulgare), Lemongrass (Cymbopogon citratus), and Marigold (Tagetes erecta),

Mosquito, Mosquito Larvae and Larvicide.

Plant Components Efficacy

Oregano

Oregano (Origanum Vulgare) is a common species of the mint family Lamiaceae's

Origanum genus (Hassaballa & Alsiddig, 2022). There are six different subspecies Oregano,

but the most valuable among them is Origanum vulgare, also known as Greek oregano, a

native to the Mediterranean region and is grown almost everywhere in the world (Węglarz, et

al., 2020). According to Crawford (2022) there are many different types of oregano around

the world that can help people, not only in the culinary field but also by killing mosquitoes

that carry different types of diseases such as dengue.

Apart from this, according to Hassaballa and Alsiddig (2022), many medical uses for

oregano (Origanum vulgare) exist but are not widely known. Oregano (Origanum vulgare)

works well as a parasite repellent, anti-inflammatory, antioxidant, and antibacterial and

antifungal agent. As an essential oil or as a supplement are the two (2) most popular uses.

These benefits of oregano essential oil may be valuable to the culinary, cosmetics, and

pharmaceutical industries.

Moreover, oregano (Origanum vulgare) is regarded as a good antiseptic and insect

repellent. Due to the capacity to spread a variety of infections, such as bacteria, viruses, and

eucaryotes, mosquitoes pose a serious threat to worldwide public health (Schrieke, et al.,

2022). Oregano (Origanum vulgare) may be one of the most efficient methods for

preventing mosquitoes because it is a significant problem in the researchers’ neighborhood.

An Italian field study found that Origanum vulgare essential oils, both emulsified and crude,

were effective larvicidal agents against Culex pipiens (Giatropoulos & Bellini., 2022).

Oregano (Origanum vulgare) is usually regarded as nontoxic for humans and contains

chemicals that are known to repel mosquitoes and is regarded as a powerful antiseptic and

insect repellant. It contains various active chemicals that have been shown to be quite

efficient at keeping mosquitoes away, including carvacrol, thymol, and -terpinene

(Sharififard, et al., 2018).

In this analysis, the leaves of the oregano (Origanum vulgare) will be used by the

researchers as one of the variables in making the powdered mosquito larvicide because

oregano (Origanum vulgare) is abundant and is acquainted with the researchers.

Lemongrass

Lemongrass, or Cymbopogon citratus, is a fragrant and perennial herb (Sousa, et. al.,

2022). Cymbopogon citratus and Cymbopogon nardus are grasses that produce citronella.

Common names include lemon grass, barbed wire grass, silky heads, citronella grass, fever

grass, tanglad, or serai (Toungos, 2019).

Equally, citronella is an essential oil derived from the stems and leaves of many

lemongrass (Cymbopogon spp.) species. Citronella from lemongrass (Cymbopogon citratus)

has a repellency effect against Anopheles culicifacies for 11 hours, Anopheles stephensi and

Anopheles dirus for six to eight hours (Kumar, et. al., 2022). The extract has the potential to

repel mosquitoes of the Aedes aegypti species by as much as 90% (Yanti & Sari, 2019). A

study on mosquitoes’ repellency found that citronella obtained from lemongrass

(Cymbopogon citratus) has a 100% repellency effect against Anopheles culicifacies for 11

hours (Wangai, et al., 2022).

Furthermore, citronella (Cymbopogon citratus) or lemongrass oil can be used for

children in tropical insect repellency. Citronella can deter mosquito biting for two hours,

showing lesser effectiveness than DEET (N, N-diethyl-3-methyl benzamide) in terms of

10
duration of protection against mosquito bite (Lee, 2018). On the other hand, long-term

usage of DEET, a relatively toxic synthetic chemical, can result in harmful reactions in the

body (Handayani, et al., 2022).

Lemongrass (Cymbopogon citratus) is utilized as a natural insect repellent and aids

in the prevention of insect-borne diseases such as malaria, dengue fever, and probably Lyme

disease (Toungos, 2019).

Citronella leaves are utilized in various cultures for their aromatic and therapeutic

properties, including the treatment of fever, digestive and menstrual issues, and as an insect

stimulant and repellent. It is non-toxic and non-irritating, however in some people it can

induce dermatitis (Sousa, et.al., 2022).

In this study, the lemongrass (Cymbopogon citratus) the researchers will be using is

processed into powder. Lemongrass (Cymbopogon citratus) is very accessible and is known

to repel mosquitoes. The powdered larvicide that the researchers will be making will be

crushed from the stems and leaves of lemongrass (Cymbopogon citratus) along with the

other variables.

Marigold

The Asteraceae family includes the significant genus Tagetes (marigold), which has

at least 56 species (Salehi, et al., 2018). According to Bakshi and Ghosh (2022), the

traditional medicinal plant known as marigold (Tagetes species) has great therapeutic

efficacy and is used to cure a variety of illnesses. In particular, the role of Tagetes as an

insecticide, mosquitocide, nematicide, bactericide, and fungicide is emphasized in this study

(Sachin & Homraj, 2021). Kour and Riat (2021) also said that essential oil from Tagetes

plants can be used as a natural pesticide.

As stated by Salehi et al. (2018), phenylpropanoids, carotenoids, flavonoids,

thiophenes, and other varied chemicals with unique characteristics are present in marigold

extracts. In addition, Iamba and Yaubi (2021) proclaimed that benzaldehyde, linalool,

11
monoxide, piperidone, limonene, ocimene, Lage tone, and valeric acid are among the volatile

chemicals generated by marigold (Tagetes erecta), which attract both pests and their natural

enemies. As well to a lesser quantity of sesquiterpene and oxygenated chemicals, the

essential oils extracted from Tagetes' aerial component primarily comprise monoterpene

hydrocarbons (ocimenes, limonene, terpinene, etc.) and acyclic monoterpene ketones

(tagetones, dihydrotagetone, and tagetenone) (Bakshi & Ghosh, 2022).

The African marigold, or Tagetes erecta, which is a common medicinal plant grown

in Africa (Chauhan, et al., 2022). In Dr. Kanthethi et al. (2020) analysis remarks that 90% of

mosquitoes were repelled by materials that had been treated with Tagetes patula flower

extracts. In prior research of Salehi, et al. (2018), Tagetes patula had antioxidant and

cytotoxic activity, however the extract did not show promising antibacterial activity against

fungus (Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis,

Microsporum anisopliae, and Beauveria bassiana).

In this situation, a mixture of plant extracts from Tagetes minuta and other herbs

has shown promise and may soon be used as a powerful bed bug control agent. Further

research is needed to clarify the bioactive principles, mechanism, and safety of this result,

nevertheless. According to the study's findings, a combination of the potential plants'

extracts has exceptional insecticidal activity (Walukhua & Nyukurib, 2020).

Stated by Ponkiya, et al. (2018), a widely known plant named marigold has a natural

insecticidal property caused by a chemical known as “a-terthienyl”. Other hazardous

substances found in all of the constituents that are harmful to human health include

alkaloid, papain, terpenes, and cyanogenic glycosides. It also contains pyrethrin, a natural

substance that works well as a mosquito repellent. Marigold (Tagetes erecta) is known to

repel some common insect pests and nematodes (Agrawal, et al., 2018).

In this research, the flower of the marigold (Tagetes erecta) will be crushed to be

used by the researchers in the mosquito larvicide. The mosquito larvicide is expected to be

12
effective in eliminating mosquitoes as it will be mixed with two herbal plants while having no

negative effects on people’s health.

Larvicide

Mosquito control is the primary method of preventing the spread of mosquito-borne

diseases. According to Wong et al. (2019), killing larvae is a more effective method of

reducing mosquito populations because larvae live in water and are not as mobile as adults.

Mosquitoes are primarily controlled by using insecticides on their breeding sites, which

include poor drainage systems, tires, and garbage in urban areas. However, this pollutes the

environment and harms non-targeted organisms. One method of controlling mosquito-borne

diseases is, Larval source management (LSM) is the environmental, biological, or chemical

manipulation of a mosquito-infested environment with the goal of reducing the adult

mosquito population by targeting the immature, aquatic stages of the mosquito. (Stanton, et

al., 2021)

Larviciding, along with habitat modification, habitat manipulation, and biological

control, is a method of larval source management (LSM) that involves the use of biological or

chemical insecticides to kill the immature stages of mosquitoes. (Runge, et al., 2021).

Larviciding was widely used in the first half of the twentieth century, with the greatest

success outside of Sub-Saharan Africa, but it eventually fell out of favor. LSM, particularly

larviciding, has been reconsidered within an integrated vector management approach over

the last decade. (Runge, et al., 2021)

As Zhou et al. (2020) proclaimed, larval control and environmental management

played important roles in malaria elimination, it has been shown to effectively kill mosquito

larvae and reduce adult abundance. However, environmental concerns have been raised

about the potential toxicity of larvicides and insecticides to non-target organisms, including

humans. As a result, the World Health Organization (WHO) and the Pan American Health

Organization (PAHO) jointly promote and encourage the use of non-chemical vector control

strategies, emphasizing the importance of biological control (Arias-Castro, et al., 2020).

 Furthermore, a larval control strategy extends the useful life of adult mosquito

insecticides by reducing the size of the population being selected for resistance, and the

strategy is equally effective in controlling both indoor and outdoor biting mosquitoes (Derua,

et al., 2019). 13

As larvicides are widely popularized, there are different types of larvicides whether

powdered or liquid larvicides. First, microbial larvicides target mosquito larvae in confined

breeding habitats and its larval control reduces both the number of mosquitos that enter

houses and the number of mosquitos that bite and rest outside (Zhou, et al., 2020).

EOs also have strong mosquito larvicidal properties. Many of the essential parts of

EOs have been shown to have larvicidal effects via at least three different mechanisms:

neurotoxicity, growth inhibition, and metabolic pathway disruption (Kelly, et al., 2022).

Wong et al. (2019) stated that EO can be extracted from the entire plant or a specific part of

the plant that contains a high concentration of active ingredients. Lastly, Giatropoulos et al.

(2022) said that the continued use of synthetic chemical larvicides which contains toxic

chemicals raises concerns about the environment, non-target organisms, and humans, as

well as the development of resistant mosquito populations.

Mosquito

Mosquito populations are growing exponentially and this is a major problem for

many countries as mosquitoes transmit various diseases such as filariasis, encephalitis, Lyme

disease, yellow fever, malaria, chikungunya fever, dengue fever, and epidemic polyarthritis

(Haria, 2021). Mosquito-borne diseases are major human and animal health problems in all

tropical and subtropical countries. For centuries, people have explored various ways to

combat the threat of mosquito-borne diseases (Erasga, 2019).

Even so, mosquitoes locate and distinguish between vertebrate hosts for blood meals

via their sense of smell. Disrupting a mosquito's sense of smell can reduce host-seeking

behavior. Insect repellent use alters mosquito olfactory responses and is a personal
protection strategy against host mosquitoes (Afify & Potter, 2020). Mosquito control and

personal protection from mosquito bites are the most meaningful measures to control

several life- threatening diseases that are transmitted exclusively by the bite of blood-sucking

mosquitoes (Khater, et al., 2019).

Undoubtedly, mosquitoes are one of the most significant threats to human and

veterinary health worldwide. Their ability to carry and transmit disease to humans kills

millions of people every year (da Silva & Ricci - Junior, 2020). Mosquitoes are, by far, the

world’s deadliest animal for humans: an estimated 780,000 died from the transmission of

disease from mosquitoes in 2016 (Ritchie, et al., 2018). Around 120 million people

worldwide suffer from mosquito bites and nearly 4 million people become seriously ill from

mosquitoes, especially in Tanzania (Tariq, et al., 2022).

The most important reasons for the rise in dengue are increased breeding sites for

Aedes mosquitoes, ineffective mosquito control, increased urbanization and accelerated

population growth (Haria, 2021). Hence, the distribution of dengue virus has increased

dramatically, although the actual number of dengue cases has been underestimated due to

many misclassifications. A recent estimate puts 390 million people infected with dengue

(Lopez, et al., 2019).

Since, Aedes aegypti is the mosquito responsible for the transmission of dengue and

yellow fever arboviruses, the two most prevalent human diseases (Tariq, et al., 2022). In

2017, about 700,000 people died from mosquito bites from three different species of

mosquitoes (Anopheles, Aedes and Culex) (Afify & Potter., 2020).

Mosquito Larvae

The mosquito's life cycle consists of four stages: egg, larva, pupa, and adult, and it

takes a month to complete. Furthermore, the larvae of A. aegypti are known to be larger than

others within 7-10 days of the larval stage entering the pupal stage (Setlur, et al., 2023).
 In addition, Stanton et al. (2021) also stated that mosquito larvae are fixed in space

throughout their development to adulthood. Typically, eggs hatch into larvae within 2-3 days

of oviposition and pupae take 5-10 days to metamorphosize, though the rate of this process is

highly dependent on temperature and nutrient availability. The presence of specific types of
15
vegetation, microbiota, predators, algal density, shade, and water depth all influence larval

development.

A series of life stages are experienced by mosquito eggs as they develop in standing

water where mosquitoes also lay their eggs. Depending on the species, ovipositing females

will choose either natural or artificial, temporary or permanent, standing water. Anopheles

stephensi prefers containers such as water tanks, some species prefer brackish habitats

(Anopheles aquasalis), while others prefer Riceland habitats (Anopheles arabiensis) (Choi

and Wilson, 2019)

In Lutz et al. (2019) study, aquatic detritivores, Aedes aegypti larvae inhabit small

spaces like tin cans and vases. The adult Ae Mosquitoes from the aegypti species have

evolved numerous behavioral adaptations to live around people. However, the knowledge of

larval adaptations is fairly limited. The acknowledged mosquito breeding sites as ornamental

concrete and tiled bowls, basins, pools, and other cavities filled with stagnant water that are

surrounded by lush vegetation and are an important part of the architectural design of

various tropical cities. Such ornamental basins can be found in a variety of public

places where people congregate (Arias-Castro et. al, 2020).

In a range of aquatic habitats, from small, transitory sites to large, permanent ones,

mosquito larvae and their predators coexist. Anopheles larvae were found less frequently and

in lower abundance in habitats with a wide variety of predators and competitors, according

to a study by Eba et al. (2020).

Summary
 Mosquitoes are nuisances that produce ectoparasites and transmit conditions similar

to malaria, dengue fever, yellow fever, responsible for thousands of deaths. Aedes aegypti is

the mosquito responsible for the transmission of dengue and unheroic fever arboviruses, the

two most current mortal conditions. Additionally, mosquitoes lay their eggs, more

specifically, they go through different life stages. Temperature and the availability of

nutrients have a significant impact on how they develop. Ornamental concrete with standing16

water is known to be a mosquito larvae breeding ground.

In order to help the spread of mosquito- borne conditions multiple ways for guarding

oneself against mosquito mouthfuls are developed. Similar system is larvicide, a chemical

that prohibits mosquito larvae from developing into adult mosquitoes. Nevertheless,

larvicides play an essential role in the increasing rate of mosquito-borne illnesses. There are

two types of larvicides; synthetic and organic in a form of essential oils, still synthetic has a

problem of being dangerous to humans, particularly children and domestic creatures,

because they beget vexation, rashes or disinclination.

The Origanum genus of the mint family Lamiaceae includes several species of

oregano, including Origanum vulgare. Oregano (Origanum vulgare) comes in a variety of

forms that are beneficial to individuals both in the kitchen and in terms of warding off

insects that spread diseases like dengue. Mosquito-repelling compounds are found in

oregano, which is typically thought to be safe to humans. It contains a number of active

compounds, such as carvacrol, thymol, and terpinene, which have been demonstrated to be

highly effective at repelling mosquitoes.

Cymbopogon citratus, also known as lemongrass, is a fragrant perennial herb. It is

possible to extract citronella oil/lemongrass oil from the plant's stem and leaves.

Lemongrass (Cymbopogon citratus) extract offers additional advantages, especially in the

stem and leaves, which can be used to ward against mosquitoes. The citronella larvicide is

more successful at killing and developing mosquitoes when compared to other mosquito
repellants, candles, and other goods available for purchase in markets. Processing

lemongrass (Cymbopogon citratus) into powder has no detrimental effects on one's health.

The major genus Tagetes (marigold), which has at least 56 species, is a member of

the Asteraceae family. In so far, as humans have long used Tagetes for medicinal purposes,

including as analgesics, antiseptics, carminatives, diuretics, antispasmodics, anthelmintics,

stimulants, vermin repellents, and the treatment of stomach and intestinal diseases, analysis17

notes that 90% of mosquitoes were repelled by materials that had been treated with Tagetes

flower extracts.
 Chapter III 18

METHODOLOGY

In this chapter, the methodology that will be used in the research study will be

discussed by the researchers. Details about the research design, locale and duration,

materials, instruments, experimental setup, research procedures and data analysis are

provided in the following paragraphs.

Research Design

The study is experimental in nature. According to Nair (2021), experimental research

is a scientific method of conducting research using two variables: independent and

dependent. Independent variables can be manipulated to apply to dependent variables and

the effect is measured. The different treatments of powdered oregano (Origanum vulgare),

lemongrass (Cymbopogon citratus), and marigold (Tagetes erecta), which are the

independent variables, will be modified and be utilized as the powdered treatments to

determine the level of effectiveness as mosquito larvicide, which is the dependent variable.

The study intends to learn about the effects of the powdered oregano (Origanum vulgare),

lemongrass (Cymbopogon citratus), and marigold (Tagetes erecta) on mosquito (Culicidae)

larvae based on the research problem.

Locale and Duration

This experimental research will be conducted at Purok Panaghiusa 2, Capitol Hill,

Barangay Central, Mati City, Davao Oriental since the location is big enough for the cages

that will be used for the experiment. The location is chosen since it is accessible and
convenient for the researchers. The experimentation will last for six (6) weeks, starting on

March 2023 until April 2023.

Fig. 2 Satellite view of the location.

Materials

The researchers will use the following materials in this experimental study: Oregano

leaves, lemongrass, and marigold flowers will be used in making the mosquito larvicide

Moreover, food processor, strainer, jar, funnel, and weighing scale will also be needed.

Lastly, plastic bottles and basins will be needed for raising mosquito larvae. Photos of these

materials are found in Appendix A.

First among the materials are the Oregano (Origanum vulgare). They will be used as

one of the variables in the study. The leaves will be dried and pulverized.
 Second among the materials is the lemongrass (Cymbopogon citratus). They will be

used as one of the variables in the study. It will be dried and pulverized.

Marigold (Tagetes erecta) comes in second place among the materials. The flower of

marigold (Tagetes erecta) will be used in the study since it contains a natural compound

used in many insect repellents.

Food processor comes next in the list of materials. This will serve as a tool in order to
20
crush the sun-dried oregano (Origanum vulgare), lemongrass (Cymbopogon citratus), and

marigold (Tagetes erecta).

Next among the materials are plastic bottles. The researchers will need 5 plastic

bottles and it will serve as a container for the mosquito larvae.

Basin is the next material on the list. It will be used in gathering mosquito larvae. The

researchers will use 1 large basin for the gathering of mosquito larvae, and 5 small basins will

be needed as a container for the application of treatments.

Weighing scale follows in the materials. Weighing scale will be used in measuring the

variables to keep the measurement accurate.

Next is the strainer. The researchers will use a strainer to remove the mosquito larvae

in the basin.

Funnel is the next material on the list. This will be used for pouring the powdered

treatments into the jar.

Next among the list is jar. This will serve as the container of the powdered

treatments. The researchers will need 5 jars and will make sure that the jar is clean and not

contaminated.

Sumilarv larvicide is the final component of the materials. The researcher will use

this powdered larvicide as a control in the study.

Instrumentation
 The research instrument is essential for the duration of the study and it will be used

in this research experiment as a tool to gather data. Furthermore, the researchers will

conduct an observation to collect the data. Referring to Appendix B, the table consists of the

quantity of eliminated mosquito larvae and the specific timeline of its death that are needed

in the study. Thus, this table will help the researchers record the presentation of data before,

during and after the experiment.

Experimental Setup
21

There will be five (5) treatments in this study, each with 5 replicates. The treatments

in this setup will demonstrate the feasibility of using powdered oregano (Origanum

vulgare), lemongrass (Cymbopogon citratus), and marigold (Tagetes erecta) as a mosquito

larvicide.

The following components that make up the treatments are:

Treatment 1 – powdered oregano (Origanum vulgare)

Treatment 2 –powdered lemongrass (Cymbopogon citratus)

Treatment 3 –powdered marigold (Tagetes erecta)

Treatment 4 – 33.33 % powdered oregano (Origanum vulgare), 33.33 % powdered

lemongrass (Cymbopogon citratus), and 33.33 % powdered marigold

(Tagetes erecta)

Treatment 5 – commercial mosquito larvicide, control

Data Gathering Procedures

The procedure will cover gathering of materials, powdered oregano (Origanum

vulgare) making, powdered lemongrass (Cymbopogon citratus) making, powdered marigold

(Tagetes erecta) making, gathering of mosquito larvae, and recording.

Gathering of Materials
 To begin, the researchers shall gather all of the needed essential materials to be

utilized in the study. This includes various materials such as oregano (Origanum vulgare),

lemongrass (Cymbopogon citratus), and marigold (Tagetes erecta), food processor, plastic

bottles, basin, jar, funnel, strainer and weighing scale.

Powdered Oregano (Origanum vulgare) Making

After the gathering of materials, the researchers will proceed in making powdered

oregano (Origanum vulgare). It will be then washed with tap water, which will then be sun-

dried for three days until the leaves turn dry, then pulverized using a food processor to
22
become oregano (Origanum vulgare) powder.

Powdered Lemongrass (Cymbopogon citratus) Making

To create the powdered lemongrass (Cymbopogon citratus) the researchers will wash

the lemongrass (Cymbopogon citratus) with tap water and will be sun-dried for five days.

After the leaves turn dry, it will be then crushed using a food processor.

Powdered Marigold (Tagetes erecta) Making

Moreover, the procedures in making powdered marigold (Tagetes erecta) will be the

same in making powdered oregano (Origanum vulgare) and powdered lemongrass

(Cymbopogon citratus).

Gathering of Mosquito Larvae

The researchers will gather clean empty plastic bottles and will be cut into half. Also,

a basin with water will be used to gather mosquito larvae, then transfer it into the plastic

bottles and will be placed into the small basins respectively.

Recording

After applying the treatments to the mosquito larvae, the number of eliminated

mosquito larvae will be recorded. To record and monitor the significant information, tables

and graphs shall be utilized. And at the end of the experiment process, the researchers will
provide data about the feasibility of using powdered oregano (Origanum vulgare) leaves,

lemongrass (Cymbopogon citratus), and marigold (Tagetes erecta) as mosquito larvicide.

Data Analysis

To identify and analyze the outcomes, the data to be gathered will be presented in a

one-way analysis of variance (one-way ANOVA). According to Bevans (2022), the data from

one categorical independent variable and one quantitative dependent variable are the subject

of one-way analysis of variance (one-way ANOVA). At least three levels should be present in

the independent variable. In the study, one-way analysis of variance will be used as it
23
contains five (5) treatments with 5 replicates. The researchers will be able to compare and

evaluate the efficacy of powdered oregano (Origanum vulgare), lemongrass (Cymbopogon

citratus), and marigold (Tagetes erecta) as a mosquito larvicide using various measurements

as the herbal powdered treatment.

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020-00767-3.
 APPENDICES

APPENDIX A - MATERIALS

https://www.realliving.com.ph/home-improvement/gardening/5-reasons-why-you-should-grow-

oregano-at-home

Figure 3. Oregano (Origanum vulgare)

 https://ttseeds.com/product/lemongrass/

Figure 4. Lemongrass (Cymbopogon citratus)

https://www.facebook.com/ transitionph/posts/marigoldit-is-being-

grown-as-trap-crop-in-agriculture- against-nematodesmosquitos/

3167072003324944/

Figure 5. Marigold (Tagetes erecta)

https://shopee.com.my/Electric-Household-Meat-Slicer-Mixing-Food-Processor-for-Kitchen-

i.7737591.1286165177

Figure 6. Food Processor

 https://www.pinterest.ph/pin/ 675610381577614930/

Figure 7. Jar

https://www.indiamart.com/ proddetail/plastic-drinking-bottle-

18358355597.html

Figure 8. Plastic Bottle

https://www.alatoneplastics.com.ph/ 24-inches-diameter-basin

Figure 9. Basin
https://www.excell-scale.com/en/product-274820/IP68-Waterproof-Weighing-Scale-ESW-Plus-ESW-E-

Plus.html

Figure 10. Weighing scale

https://planmystudy.co.in/image/cache/catalog/funnel%2065mm-500x500.jpg

Figure 11. Funnel

https://ae01.alicdn.com/kf/HTB19cOmSFXXXXcoXVXXq6xXFXXXa/

UESH-Kitchen-Flour-Colander-Sifter-Sieve-Oil-Mesh-Wire-Strainer-16cm-Dia.jpg

Figure 12. Strainer

 https://shopee.ph/1-Box-Sumitomo-Chemical-Sumilarv-0.5G-Pyriproxyfen-Larvicide-for-Mosquito-

Control-Anti-Dengue-i.150987898.3572428392

Figure 13. Control (Sumilarv)

APPENDIX B – INSTRUMENTATION

Table 2: Mosquitoes Death Index per treatment

Treatment #

Quantity
APPENDIX C – EXPERIMENTAL SETUP

Treatment 1 – powdered oregano (Origanum vulgare)

Treatment 2 –powdered lemongrass (Cymbopogon

citratus)

Treatment 3 –powdered marigold (Tagetes

erecta)
 Treatment 4 – 33.33 % powdered oregano
(Origanum vulgare), 33.33 % powdered lemongrass
(Cymbopogon citratus), and 33.33 % powdered
marigold (Tagetes erecta)

Treatment 5 – commercial mosquito larvicide, control