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Chapter 1

The Problem and Its Background

This chapter presents the background of the study, statement of the problem, hypotheses
of the study, review of related literature, review of related studies, scope and delimitations,
significance of the study, definition of terms and overall view of the study.

Introduction

Dengue is a mosquito-borne viral disease that has rapidly spread in all regions of WHO in
recent years. Dengue virus is transmitted by female mosquitoes mainly of the species Aedes
aegypti. This mosquito also transmits chikungunya, yellow fever and Zika infection. Dengue is
widespread throughout the tropics, with local variations in risk influenced by rainfall, temperature
and unplanned rapid urbanization (World Health Organization, 2018).

Dengue viruses are primarily maintained in a human-to-mosquito-to-human cycle. The


primary vector is the Aedes aegypti mosquito, which is highly adapted to human habitations.
Aedes albopictus can also sustain dengue virus transmission in humans. Other species maintain a
monkey-mosquito cycle in south-east Asia and western Africa. Dengue virus transmission from
non-human primates to humans appears to be rare. The spread of vectors following urbanization
and the decline in vector-control efforts has partially contributed to the increased incidence of
dengue virus infections. However, dengue is not confined to urban settings and is increasingly
reported from rural areas. Additionally, factors such as population growth, globalization and travel,
and climate change facilitate increased transmission of dengue viruses. Dengue exhibits substantial
temporal and geographic variability (World Health Organization, 2016).

Dengue cases and even malaria diseases are one of most common disease we get from
mosquito bites. The number of dengue cases reported increased from 2.2 million in 2010 to 3.2
million in 2015. Although the full global burden of the disease is uncertain, the initiation of
activities to record all dengue cases partly explains the sharp increase in the number of cases
reported in recent years. Philippines health officials have reported 33,748 suspected cases of
dengue through the first three months of 2016, up from 24,927 cases reported during the same
period in 2015. Consider also the bites we get from ants and other insects are one of the problems
we face today. A better way to prevent this is to use insect or mosquito repellent, but since most

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of the factory-made repellents causes other side effects, most of the Filipino citizens prefer to use
organic repellents (Cogan, 2016).

Camansi seeds are one of the nutritious seeds eaten by mankind, due to its high content of
Niacin (B3) which is greater than those of almonds, peanuts and other nuts. It also contains both
Vitamins B1 and B2 and other minerals found such as phosphorus, potassium, sodium etc. proves
that camansi seed is one of the edible and nutritious nuts. Camansi seeds have been analyzed for
its moisture, ash, crude protein and fat content (Orwa et al., 2009).

The research on efficacy of Artocarpus Camansi seed extract aims to develop an


insecticidal agent as a first prevention step against dengue and malaria diseases.

Statement of the Problem

The study aims to the determine the insecticidal capacity of Camansi Seed Extract against
mosquito larvae. Specifically, it aims to find answers to the following:

1. What is the effect of the larvicidal activity of Camansi seeds extract against mosquito
larvae?

2. What is the level of Effectiveness of Camansi seeds extract in terms of:


2.1 Color
2.2 Odor
2.3 Effectiveness (time)
3. Is there a significant difference in the larvicidal activity between Mosquito Larvae with
treatment and with no treatment?

Hypotheses of the study

The researcher formulated the following hypothesis:

Ho: There’s no significant difference in the larvicidal activity between Mosquito Larvae with
treatment and with no treatment.

Ha: There’s a significant difference in the larvicidal activity between Mosquito Larvae with
treatment and with no treatment.

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Review of Related Literature

Breadnut (Artocarpus Camansi)


According to Tilburt (2008), herbal plants are the objects of interest in the field of medicine.
Politically speaking, the Philippine government officials seek to find ways as to how medical drugs
could be easily availed by the Filipinos remarkably those who are poor and also solutions for
fungal growth in agricultural lands which could heavily affect the production of food and etc. thus,
having an impact on the economy.

The Philippines has bountiful forest resources especially in Mindanao. About 8,000 species
of plants in these forests have been known to possess certain medicinal properties (Kumar &
Jnanesha, 2016).

One of these is Artocarpus Camansi, commonly known as breadnut or Kamansi. It is a


member of the family Moraceae (mulberry). Flowering is monoecious with male and female
flowers on the same tree at the ends of branches, with the male inflorescence appearing first. Male
flowers are club-shaped, up to 3 cm (1.2 in) in diameter and 25–35 cm (10–14 in) long or longer.
Thousands of tiny flowers with two anthers are attached to a central spongy core. Female
inflorescences consist of 1500–2000 reduced flowers attached to a spongy core. Unlike breadfruit,
the individual flowers do not fuse together along their length. Leaves are alternately arranged, with
length 40–60 cm (16–24 in), moderately dissected with 4–6 pairs of lobes and sinuses cut half way
to the midrib. New leaves on young trees can be 76 or more cm (30 in) long. They are densely
pubescent, with many white or reddish-white hairs on upper and lower veins, lower leaf surface,
and petiole. Blade is dull green with green veins. Two large green stipules enclose the bud, turning
yellow before dehiscing. Kamansi trees are now found in cultivation in the Philippines, where it is
typically grown as backyard tree (Ragone, 2009). Kamansi trees are trees with single trunk and
have spreading canopies. They grow in tropical regions. Trees can grow 10-15 m with a trunk 1 m
in diameter or larger. Latex can be found in every part of the tree.

Analysis of Camansi seeds samples were analyzed for moisture, ash, crude protein, crude
fiber and fat content using the methods described by 1274 Afr. J. Agric. Res. AOAC in 1990. The
mineral contents of phosphorus, potassium, sodium, calcium, iron, manganese, copper and
magnesium were determined using the method of Ilelaboye and Pikuda in 2009. Ten amino acids
(arginine, histidine, isolueucine, leucine, lysine, phenylalamime, tyrosine, cystine, tryptophan and

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methionine) were estimated using the method of Ketiku in 1973. The predominant acid in Camansi
seeds are lactic acid (0.317 mg/kg) and citric acid (0.185 mg/kg). The seeds exhibited trace
amounts of acetic acid (0.050 mg/kg), butyric acid (0.012 mg/kg) and malic acid (0.012 mg/kg).
The organic acids evaluated (malic and citric acid) were lower than the values obtained for Syrian
sumac and Chinese sumac fruits (1568.04 mg/kg for malic and 56.93 mg/kg citric acid) by Kossah
in 2009. The results indicate that breadnut seeds are low in organic acid content and the fresh
breadnut seeds are susceptible to spoilage (Adeleki & Abiodun, 2010).

Artocarpus species are rich in phenolic compounds including flavonoids, stilbenoids,


arylbenzofurons and Jacalin, a lectin. Oil is rich in unsaturated fatty acid comparing well with
melon seeds, soybean and groundnut oil. Artocarpus camansi seeds are a good source of minerals
and contains more niacin than other nuts. A 100 g edible portion yields amino acids--methionine
3.2 g, leucine 2.6 g, isoleucine 2.4 g, and serine 2.1 g, comprising 50% of 14 amino acids analyzed
(Stuart, 2016).

According to Lewis (2012), methionine is needed in the human diet for many reasons,
including protein-building and metabolism. It is environmentally safe and harmless to citrus plants,
mammals and birds. It was first discovered the pesticide properties of methionine while cloning
genes that regulate amino acid metabolism in 1998. They later found this amino acid to be effective
against yellow fever mosquito larvae, tomato hornworm and Colorado potato beetle. Methionine
disrupts an ion channel that controls nutrient absorption in larvae with an alkaline intestine. The
human nutrient amino acid methionine has been shown by us to disrupt amino acid-modulated ion
transport systems in caterpillars and other insect larvae that possess an alkaline midgut. methionine
kills larvae of mosquitos and caterpillars.
A lectin was isolated and purified from the seeds of Artocarpus camansi. It was a complete
lectin since it agglutinated erythrocytes without trypsin treatment and addition of metal ions
(Lewis, 2010).

Lectins have deleterious effects against larvae, developing stages and mature forms of
insects from orders Coleoptera, Diptera, Hemiptera, Homoptera, Hymenoptera, Isoptera,
Lepidoptera and Neuroptera. Insecticide activity of lectin is generally evaluated by bioassays that
incorporate the lectin into artificial diets offered to insects, with insects dying from nutritional
deprivation. It has been shown that lectins are resistant to proteases present in the insect gut, (a

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property responsible for their active presence in the digestive tract). Lectin may also cross the
midgut epithelial barrier by transcytosis, entering the insect circulatory system and resulting in a
toxic action against endogenous lectins involved in haemolymph self-defense mechanisms
(Macedo et al., 2011).

According to Bonotano (2015), niacin will change the odor of your perspiration which for
some people repels mosquitoes. Some insect repellent use niacin as active ingredients. Thiamine
produces a foul smell that repels female mosquitos. Since these vitamins are commonly present in
our bloodstreams, it will take longer time for these vitamins to be effective. It is required to drink
supplements contain B1, B2 and B3 to 3-4 weeks.

Citric acid relieves mosquito bites and reduces itchiness, it has great ability to relinquish
such bites from mosquitos. Lactic acids attract mosquitos, causing the person to get more bites
(Palermo, 2018).

Review of Related Studies

A recent study by Agricultural Research Service (ARS) scientists and their collaborators
at the University of British Columbia in Okanagan, Canada, identified three breadnut
compounds—capric, undecanoic and lauric acids that act as insect repellents. ARS is the chief
intramural scientific research agency of USDA. In the study, chemist Charles Cantrell and his
colleagues at the ARS Natural Products Utilization Research Unit (NPURU) in Oxford, Miss., and
the University of British Columbia scientists collected smoke extracts by burning sun-dried
clusters of flowers in the traditional method used by people in Pacific regions. Capric, undecanoic
and lauric acids, which are saturated fatty acids, were found to be significantly more effective at
repelling mosquitoes than DEET, the primary insect repellent used against biting insects (Avant,
2013).

For the first time, breadnut was shown to work as a repellent, confirming it as a valid folk
remedy, according to Cantrell. These same compounds found in breadfruit and other folk remedies
were shown to be highly active and the most repelling in a different study that examined a variety
of saturated and unsaturated fatty acids. Cantrell teamed with Uli Bernier, a chemist in the
Mosquito and Fly Research Unit at the ARS Center for Medical, Agricultural and Veterinary
Entomology in Gainesville, Fla., and scientists at the University of Mississippi to evaluate the

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compounds. The test involved cloth treated with different concentrations of compounds and worn
by volunteers. These compounds were shown to provide effective protection against mosquitoes
(Avant, 2013).

The use of plant extracts in insect repellent and insecticide use in insect pests such as adult
mosquitoes has been widely practiced by many indigenous cultures around the planet for thousands
of years. This can be accomplished by volatilizing plant essential oil or by burning plant tissue to
yield smoke.

A research study done by Matthew Aaron Glover that seeds generated from Artocarpus
Camansi were all effective at preventing landings of both Aedes aegypti and Anopheles gambiae
being comparable to commercially available 10% N,Ndiethyl-meta-toluamide (DEET). It was also
shown that the seeds of breadnut have strong ovicidal effects whereas Cinnamomum zelyanicum,
Cuminum cyminum, and Rosmarinus officinalis possess strong repellent effects against adults of
Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. Traditional indigenous
knowledge was used to identify the male inflorescences of Artocarpus Camansi as a potential
source of insect repellent phytochemistry and ethanol extracts effectively repelled mosquitoes in
a bioassay (Jones et al., 2012).

In a study conducted by Krishnappa (2013), B. rubra L. and Artocarpus Camansi seeds


extracts showed ovicidal activity on twenty-five early third instar larvae of A. aegypti. The results
have shown that Artocarpus Camansi exhibited greater ovicidal property providing 100 %
mortality when compared to B, rubra L. The seeds extracts obtained from the two-plant were an
excellent potential for controlling A. aegypti mosquitoes.

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Conceptual Framework

The study was guided by a conceptual framework which postulates that the extraction of
Breadnut (Artocarpus camansi) seeds can be used to develop an insecticidal agent as a first
prevention step against dengue and malaria disease.

Independent Variable Intervening Variable Dependent Variable

Testing of Mortality
Artocarpus Larvacidal Activity rate of the
Camansi dead
seeds (with and without mosquito
treatment) larvae

Figure 1. The research variable in the study

Figure 1: A conceptual framework presents the Independent Variable (Artocarpus Camansi


seeds), Intervening Variable (Testing of Larvicidal Activity (with and without treatment) and
Dependent Variable (Mortality rate of the dead mosquito larvae).

Scope and Delimitation

This research determined Camansi seed extract as effective and safe natural larvicidal agent
against mosquitoe. The assay was done simultaneously with having a treatments and without
treatments to determine the efficacy of the formulation against mosquito larvae. Since testing was
conducted at the specific period of time, the data obtained in this research were true only at the
time larvae samples were taken. However, the data obtained may serve as baseline for further
comprehensive researches that may be conducted.

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Significance of the Study
This study aims to develop an insecticidal agent as a first prevention step against dengue
and malaria diseases. The outcome of this study could bring great contribution to the following:

Department of Health. This study may be beneficial to the Department of Health for the possible
production of alternative medicines that might contribute to the prevention or delay of some types
of cell damage.

Community. This will inform the community about the benefits of seeds extract against dengue
and malaria diseases and will gain knowledge as to what components of seeds extract are
responsible for giving such effects on their health.

Researchers. The researchers and other medical courses would benefit from this study since they
will be able to know the first prevention step against dengue and malaria diseases. Also to prevent
other lung and skin diseases caused by factory-made repellents. This may also be beneficial to the
future researchers for the formulation of related studies. As for the benefit of the general public,
this serves as the cheapest yet effective way to prevent mosquito-caused diseases. The results of
this study can be used as reference for their advanced studies.

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Definition of Terms

In order to understand fully some of the technical terms to be used in the study, they are
defined exactly according to how they were used and are arranged alphabetically as follow:

Artocarpus camansi. It refers to the breadnut, is a medium-sized tree found in the mulberry family
Moraceae. Native to Papua New Guinea, it is a relative of the breadfruit and is commonly used as
a staple crop.

Chemical. It refers to a substance produced by or used in a chemical process.

Dengue. It refers to a mosquito-borne tropical diseases caused by the dengue virus.

Larva. It refers to a distinct juvenile form many animals undergo before metamorphosis into
adults.

Malaria. It refers a mosquito-borne infectious disease affecting humans and other animals caused
by single-celled microorganisms belonging to the Plasmodium group.

Mosquito. It refers to a family of small, midge-like flies (Culicidae). In feeding on blood, some
of them transmit extremely harmful human and livestock diseases, such as malaria, and yellow
fever.

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Chapter 2
Methods
This chapter presents the discussion and presentation on the research design, preparation
of materials, data gathering procedure and data analysis tool.

Research Design

This research used the posttest true experimental research design. In this type of research
design, which uses two groups. Experimental group is given the treatment (Mosquito Larvae)
while the other group (control) will be used through the observation. In this process, mosquito
larvae are allowed to submerge into these solutions.

Preparation of Materials

The materials used in the study were the following: 100 g of breadnut seeds, ethanol, knife,
cheesecloth, beakers, graduated cylinders, spoon, jar, water and weighing scale

Data Gathering Procedure

Phase I - Collection

Figure 1. Breadnut seeds


The plant material was collected at Kidapawan.
Phase II- Prior to extraction

Figure 2. Peeling of camansi seeds Figure 3. Weighing the sample

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Figure 4. Extracting the Camansi seeds in a blender Figure 5. The extracts filtered
through the use of cheesecloth and the
camansi seeds extract was put in a jar.
Phase III – Measuring the camansi seeds extract
The camansi seeds extract was measured in beaker and graduated Cylinder to measure the
right amount of liquid extraction.

Phase IV - Concentrations of Camansi seeds extract.


From the original concentrated extract, an aliquot of 2, 4, 6, 8 and 10ml of the extract were
taken and diluted to 20 mL to make series of concentrations of 10.0, 20.0, 30.0, 40.0 and 50.0%
of camansi seed extract. These concentrations were tested for its insecticidal activity towards the
mosquito larvae.

Phase V - Screening of Camansi seed extract with and without treatment for Larvicidal
activity

Figure 6. Camansi seeds extract without treatment (left) and Ethanolic camansi seeds extract
with mosquito larvae (right)
Phase VI – Computation
The data collected from Phase V were computed using the statistical program
which is the Microsoft Excel and the transformation of percentage to probit table from the study
of Finney (1952).

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Figure 7. Probit Transformation Table

Data Analysis Tool

To compute for the LC50 of the Camansi seeds extract against mosquito Larvae, probit
analysis was conducted. In this type of test, the researchers aimed to find the effective
concentration that has median lethal concentration. By plotting the response of the larvae to various
concentrations of Camansi Seed Extract, the researchers may be able to compute the LC50.

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