Larvicidal potency of marigold (2)

Larvicidal Potency of Marigold (Tagetes erecta) Stem and Leaf
Extract against Mosquito Larvae
A Science Investigatory Project
Presented as an Entry to the
2013-2014 Regional Science Fair
LIFE SCIENCE – GROUP
STUDENTS CATEGORY
Charlie Jhon Aspillaga
Christine Joy Arreza
Raven Daryle Mae suarez
Researchers
Jeshrel B. Plaza
Adviser
Madrid National High School
Madrid, Surigao del Sur
S.Y. 2013-2014

Larvicidal Potency of Marigold (Tagetes erecta) Stem and Leaf Extract
against Mosquito Larvae
A Science Investigatory Project 2013 – Students Category
By: Cristine JoyR. Arreza, Charlie Jhon Aspillaga, Raven Daryle Mae D. Suarez
1
Larvicidal Potency of Marigold (Tagetes erecta) Stem and Leaf
Extract against Mosquito Larvae
A Science Investigatory Project
Presented as an Entry to the
2013-2014 Regional Science Fair
LIFE SCIENCE – GROUP
STUDENTS CATEGORY
Charlie Jhon Aspillaga
Christine Joy Arreza
Raven Daryle Mae suarez
Researchers
Jeshrel B. Plaza
Adviser
S.Y. 2013-2014

2
APPROVAL SHEET
This Science Investigatory Project entitled “Larvicidal Potency of Marigold (Tagetes
erecta) Stem and Leaf Extract against Mosquito Larvae” prepared and submitted by
Charlie Jhon Aspillaga, Christine Joy Arreza, Raven Daryle Mae Suarez has been
examined and recommended for acceptance and approval for the science congress by
the school SRC.
JESHREL B. PLAZA
Research Adviser
PANEL
RUBY GRACE O. MARIBAO SULPECIA L. GUIMARY
Statistician English Teacher
LETECIA L. MOLLANIDA EVANGELINE E. CARATAO
Science Coordinator ESEP Coordinator
Francisquita J. Cosmiano
Head Teacher- IV / Chairman of SRC

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Acknowledgment
With profound gratitude and sincere appreciation, the researchers would like to
acknowledge the following persons who contributed much in the whole duration of the
Science Investigatory Project.
To Mrs. Francisquita C. Cosmiano , the Head Teacher IV, for the trust and
kindness;
To Mrs. Evangeline A. Caratao, MNHS ESEP Head, for the suggestions;
To Mrs. Sulpecia L. Guimary, for her expertise in English grammar and language;
To Mrs. Ruby Grace O. Maribao, for her expertise in statistics and for her
substantial sights;
To Mr. Jeshrel B. Plaza, II-Mendel Adviser, for the encouragement and support;
and
To GOD for the health and sound mind.
- The Researchers

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Table of Contents
Page
Title Page 1
Approval Sheet 2
Acknowledgment 3
Table of Contents 4
List of Tables 5
Abstract 6
Chapter 1 THE PROBLEM AND ITS SETTINGS
Introduction 7-8
Statement of the Problem 9
Research Objectives 9
Statement of the Hypothesis 10
Significance of the Study 10
Scope and Limitations 11
Definition of Terms 11-12
Chapter 2 REVIEW OF RELATED LITERATURE
Related Literature 13-16
Related Studies
Foreign 16-18
Local 18-20
Chapter 3 MATERIALS AND METHODS
Preparation of Marigold Leaf and Stem Extracts 21
Application of Marigold Leaf and Stem Extracts to the setups 22
Determination of effectiveness of the Marigold Extract 23
Experimental Designs 24
Schematic Diagram of the Research Methodology 25
Chapter 4 RESULTS 26
Chapter 5 DISCUSSIONS 28
Chapter 6 CONCLUSIONS AND RECOMMENDATIONS 29
Conclusion
Recommendations
BIBLIOGRAPHY 30-31
APPENDICES
Raw Data
Pictorials
Curriculum Vitae

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List of Tables
Page
Table 1 Table of Observations 24
Table 2.1 One Way Anova Test 25
Table 2.2 One Way Anova Test 25
Table 4 ANOVA Summary Table 26

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ABSTRACT
To find out the larvicidal effectiveness or potency of Marigold stem and leaf extract
against mosquito larvae, an experimental design was used. The experiment was
conducted. Ten (10) cultured mosquito larvae were placed in the five (5) set ups with
varying amount of Marigold leaf and stem extract: 10 ml, 40 ml, 70 ml, and 100 ml in 300
ml water. It was observed within 38 hours. One way anova was used as a statistical tool
to find out the Larvicidal potency of Marigold leaf and stem extract to the mortality rate of
mosquito larvae. The findings revealed that Marigold leaf and stem extract was a very
effective larvicide on mosquito larvae.

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CHAPTER 1
INTRODUCTION
A. Backgroundof the Study
Philippines is bearing the high cost brunt dealt by Aedes aegypti and Aedes
albopictus also known as the main vector of hemorrhagic dengue fever in recent years
which became a major national public health concern. The spread of both the mosquito
vectors and the viruses in the whole archipelago has led to the resurgence of epidemic
dengue fever and emergence of dengue hemorrhagic fever (dengue/DHF) in the past 25
years in many urban centers of the country.
Dengue is a mosquito-borne viral infection causing a severe flu-like illness and,
sometimes causing a potentially lethal complication called severe dengue. The incidence
of dengue has increased 30-fold over the last 50 years. Up to 50-100 million infections
are now estimated to occur annually in over 100 endemic countries, putting almost half of
the world’s population at risk because research on dengue vaccines for public health use
is still on process. http://www.iumsp.ch/Publications/pdf/infdisrev1-189.pdf
In its monthly surveillance report, the health department reported that 46,651
dengue cases, with 294 deaths, were recorded nationwide from January 1 to July 7,
2012. The figure is 13.85 percent higher than the 40,975 cases registered in the same
period last year. Official records from the Center for Health and Development of the

8
Caraga Region indicated that persons suffering with dengue fever rose by 44 percent or
678 cases from January to June 14, 2012. During the same period last year, 470 persons
were afflicted with dengue fever in the provinces of Agusan del Norte, Agusan del Sur,
Dinagat Islands, Surigao del Norte, and Surigao del Sur.Of the current 678 cases, some
160 have been recorded in Butuan City — Caraga’s regional center — while the province
of Surigao del Sur topped all Caraga provinces with 175 dengue cases.
http://www.surigaotoday.com/2012/07/dengue-cases-in-caraga-surge-bynearly.html
Since prehispanic times, Tagetes erecta, the Mexican marigold, also called Aztec
marigold, is a species of the genus Tagetes native to Mexico and Central America
reaches heights of between 50–100 cm (20–39 in) and are gathered by Aztecs for
medicinal, ceremonial and decorative purposes. It is widely cultivated commercially with
many cultivars in use as ornamental plants. This plant has been used for medicinal
purposes. Scientific study shows that thiophenes, natural phytochemicals that include
sulfur-containing rings, may be the active ingredients. They have been shown to kill gram
negative and gram positive bacteria and have the ability to kill some variety of insect
larvae. http://www.springerlink.com/content/p04q407616406618/
This study aims to know if the extract of the marigold leaves and stems is effective
of getting rid of mosquito larvae. The researchers used local methods in extracting the
leaves and stems.

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B. Statement of the Problem:
This study aims to answer the following questions:
1. Is Marigold stem and leaf extract has a potential larvicide in getting rid of mosquito
larvae?
2. Is Marigold leaf and stem extract a good substitute to commercial larvicide?
3. Is there a significant difference between the amount of marigold stem and leaf
extract in killing mosquito larvae?
C. Research Objectives
The objectives of this study are.
1. To test the Larvicidal effectiveness or potency of Marigold leaf and stem extract
against mosquito larvae.
2. To know if there is a significant difference between the amount of the marigold
stem and leaf extract in killing mosquito larvae.

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D. Statement of Hypotheses:
The study formulated the following hypotheses:
Null Hypothesis:
1. The marigold leaf and stem extract has no Larvicidal effects on the
health, behavior, and death of mosquito larvae.
E. Significance of the Study:
The results of this study will benefit the people of Surigao del Sur in the way
that this can provide a new idea on how to prevent certain mosquito transmitted
disease particularly dengue and malaria.
This study can also provide farmers a good and better stability of work for
the reason that the farmers can embed this plant and sell it for economical use that
can increase their income.

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F. Scope and Limitations
This research study is focused on the marigold stem and leaf extract as a
potential mosquito larvicide. However, this study does not aim to replace any
commercialized and synthetically produced larvicide, instead it will give idea to the
manufacturers to promote the use of non-harmful and organic substance as an
effective larvicide. Laboratory experimentation and analysis is limited only to the
Larvicidal property of this plant.
G. Definition of Terms
1. Analysis – the separation of something into its constituents in order to
find out what it contains, to examine individual parts, or to study the
structure of the whole.
2. Extract – to obtain a substance from a compound, in solid, liquid, or gas
form, by using an industrial or chemical process.
3. Extraction – the process of extracting something or being extracted.
4. Mosquito larvae – the wingless immature worm-shaped form of many
mosquitoes that develops into a pupa or chrysalis before becoming an
adult mosquito.
5. Larvicide – a chemical used to kill larvae.
6. Phytochemicals – a naturally occurring plant substance. Some
phytochemicals have been shown in research to protect against disease.
7. Potency – the state or quality of being potent.

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CHAPTER II
REVIEW OF RELATED LITERATURE
A. Related Literature
A.1 Mosquitoes/Larvae
The mosquitoes are a family of small, midge-like flies: the Culicidae. Although a
few species are harmless or even useful to humanity, most are a nuisance because they
consume blood from living vertebrates, including humans. In feeding on blood, various
species of mosquitoes transmit some of the most harmful human and livestock diseases.
Some authorities argue accordingly that mosquitoes are the most dangerous animals on
Earth.
Below is the scientific classification of a mosquito.
Kingdom: Animalia
Phylum: Arthropoda
Class: Insecta
Order: Diptera
Suborder: Nematocera
Infraorder: Culicomorpha
Superfamily: Culicoidea
Family: Culicidae

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Mosquito larvae have a well-developed head with mouth brushes used for feeding,
a large thorax with no legs, and a segmented abdomen.
Larvae breathe through spiracles located on the eighth abdominal segment, or
through a siphon, and therefore must come to the surface frequently. The larvae spend
most of their time feeding on algae, bacteria, and other microbes in the
surface microlayer. They dive below the surface only when disturbed. Larvae swim either
through propulsion with their mouth brushes, or by jerky movements of their entire bodies,
giving them the common name of "wigglers" or "wrigglers".
Larvae develop through four stages, or instars, after which
they metamorphose into pupae. At the end of each instar, the larvae molt, shedding their
skins to allow for further growth.
A.2 Marigold
Marigold or Tagetes is a genus of 56 species of annual and perennial mostly
herbaceous plants in the sunflower family (Asteraceae or Compositae). The genus is
native to North and South America, but some species have become naturalized around
the world. One species, T. erecta, is considered as a plant that has the ability to eradicate
some variety of insect larvae.
Depending on the species, marigold foliage has a musky, pungent scent, though
some varieties have been bred to be scentless. It is said to deter some

14
common insect pests, as well as nematodes. Tagetes are hence often used in companion
planting for tomato, eggplant, chili pepper, tobacco and potato. Due to
antibacterial thiophenes exuded by the roots, Tagetes should not be planted near
any legume crop.
The florets of Tagetes erecta are rich in the orange-yellow carotenoid lutein and
are used as a food color (INS-Number E161b) in the European Union for foods such as
pasta, vegetable oil, margarine, mayonnaise, salad dressing, baked goods, confectionery,
dairy products, ice cream, yogurt, citrus juice and mustard. In the United States, however,
the powders and extracts are only approved as colorants in poultry feed.
Scientific study shows that thiophenes, natural phytochemicals that include sulfur-
containing rings, may be the active ingredients. They have been shown to kill gram
negative and gram positive bacteria. Simultaneous steam distillation extractions (SSDE)
volatiles isolated from the flower of the erecta species are believed to have higher
insecticidal activity. http://www.springerlink.com/content/p04q407616406618/
Since it has an insecticidal effect, it is hypothesized that the stem and leaf extracts
can be an effective larvicide against mosquitoes.

15
Below is the scientific classification of Marigold.
Kingdom: Plantae
Division: Magnoliophyta
Class: Magnoliopsida
Order: Asrerales
Family: Calenduleae
Genus: Calendula
B. Related Studies
B.1 Foreign
Marigold, Tagetes are a useful intercrop in agriculture. Populations of
Meloidogyne nematodes have been reduced by intercropping marigolds. Other cultural
applications of Tagetes include the use of T. minuta leaves to repel mosquitoes and safari
ants (Maradufu et al. 1978) in Kenya.
Researchers have isolated several insecticidal compounds from Tagetes.
Compounds extracted from the leaves and flowers of T. minuta are toxic to Aedes aegypti
larvae (Maradufu et al. 1978 pp.24-28). Laboratory tests using polyacetylenes from
Asteraceae indicated that α-terthienyl, a phototoxic thiophene from Tagetes, had an

16
insecticidal property against A. aegypti when combined with near-UV radiation (Arnason
et. Al. 1981 pp.64-68). Morallo-Rejesus & Decena (1982) isolated a-terthienyl and 5-(3-
buten-1-ynyl)-2, 2-bithienyl as the active insecticidal components from root extracts of T.
erecta L. Thetopical LDso of partially purified T. erecta root extract was 8.1 mg/g for
Rhyzopertha dominica (F.) and 4.3 mg/g for Tribolium castaneum (Herbst) (Morallo-
Rejesus & Decena 1982 pp. 56-58). Numerous studies have evaluated the insecticidal
properties of a-terthienyl and its analogs (Philogene et al. 1985, 1986; Amason et al.
1986, 1988, 1989; Champagne et al. 1986; Evans et al. 1986; Hasspieler et al. 1988,
1990; Sen et al. 1990). The toxicokinetics of α-terthienyl were also examined for three
species of Lepidoptera (Iyengar et al. 1987 pp.65-69). The topical LDso for Manduca
sexta (L.) was 10 IJ-glg,but for Heliothis virescens (F.) it was 470 IJ-glgand for Ostrinia
nubilalis(Hiibner) the LDso was 700 IJ-g/g (Iyengar et al. 1987 pp.27-29). This difference
was likely caused by a more rapid clearance of the toxin by the more tolerant species,
which may have preadapted rapid elimination of this toxin via evolutionary associations
with the Asteraceae (Iyengar et al. 1987 pp.28-30).This elimination was facilitated by
higher levels of cytochrome P450 in those species (Iyengar et al. 1990 pp. 28-30).
Generally, the efficacy of α-terthienyl against various mosquito larvae may also be
related to the limited evolutionary association between the compound and these insects,
with the oxidative mode of action (Hasspieler et al. 1990 pp.34-36 ) being enhanced by
the lack of facile or rapid detoxification. This suggests that other naive insects, such as
stored-product pests, may be susceptible to these compounds. Many stored-product
pests were (and probably still are) either fossorial, litter-decomposing species or

17
specialists on the seeds of plants in the Leguminoseae and Graminaceae families before
synanthropic association. Thus, they could be susceptible to compounds evolved against
herbivory of tissues of other plant groups. Our objective was to evaluate tissue extracts of
T. minuta against Mexican bean weevils, Zabrotes subfasciatus (Boheman), a sexually
dimorphic bruchid of economic importance throughout the world. We are also concerned
with the production of "insecticidal crops" by developing countries as an alternative to
synthetic insecticides. Tagetes spp. has been proposed as an insecticidal crop to provide
a locally available source of α-terthienyl for mosquito control in developing countries
(Amason et al. 1981 72-73). However, in practice, potential insecticidal crops, such as T.
minuta, should be fully utilized, rather than to simply serve as a source of a single
chemical component. The efficacy of extracted material from all tissues should be
determined so that the entire plant can be used, if possible.
B. 2 Local
Marigold or more commonly known as Calendula has been used medicinally for
centuries. Traditionally, it has been used to treat conjunctivitis, blepharitis, eczema,
gastritis, minor burns including sunburns, warts, and minor injuries such as sprains and
wounds. It has also been used to treat cramps, coughs, and snake bites. Calendula has a
high content of flavonoids, chemicals that act as anti-oxidants in the body. Anti-oxidants
are thought to protect body cells from damage caused by a chemical process called
oxidation. Oxidation produces oxygen free radicals, natural chemicals that may suppress
immune function.

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Marigold has been considered beneficial in reducing inflammation and promoting
wound healing. It has been used to treat a variety of skin diseases and has been seen
effective in treatment of skin ulcerations, eczema, juvenile acne and dry phthiriasis.
Improvement has been seen in as little as 3-4 days of treatment according to the
Universitatea de Medicina si Farmacie. www.wikepedia.com/benefialeffectsofmarigold
Marigold is one of several herbs used traditionally to treat conjunctivitis and other
eye inflammations as it helps to reduce the swelling and redness of eye infections. It is
also believed that calendula may have some anti-spasmodic action, and as such, it has
been used to relieve menstrual cramps.
Marigold is used to aid the healing of wounds and internal and external ulcers. It is
an anti-septic and improves blood flow to the affected area. Some clinical studies validate
the early treatment of stomach ulcers, although further research is needed (Chakurski
1981 pp.65-66; Krivenko 1989 pp.24-25).
Marigold cream is good for acne and nappy rash. An infusion is good for digestion
and relieves colitis and symptoms of menopause. As an anti-fungal agent, it can be used
to treat athlete's foot, ringworm, and candida. The tincture applied neat to cold sores
encourages healing.
Marigold contains chemicals, which have been shown in animal studies to speed
up wound-healing by several actions that include increasing blood flow to the affected
area and promoting the production of collagen proteins. Calendula also possesses anti-

19
septic and anti-inflammatory effects due to its flavonoid content. In mouthwashes and
gargles, calendula soothes sore throat or mouth tissue; in solutions, it has been uses to
treat hemorrhoids.
Compresses of marigold blossoms are helpful for varicose veins. Results from
recent animal and laboratory studies show that calendula may also have some anti-
infective properties, particularly against fungal infections and against viruses.
Marigold's high-molecular weight polysaccharides stimulate immune system
activity (Wagner 1985 pp.88-90) and has been researched for immune system activity. It
was initially determined to have some potential therapeutic activity against the human
immunodeficiency virus (HIV): extracts significantly inhibited HIV-1 in vitro, and reduced
HIV-1 reverse transcriptase in a dose- and time-dependent manner (Kalvatchev 1997 pp.
106-108).
In one small study of about 250 women undergoing radiation therapy after surgery
for breast cancer, a commercial calendula ointment reduced the amount of skin irritation
better than another commonly-used commercial preparation. Women who used the
calendula ointment also reported less pain from the radiation. Investigations into anti-
cancer and anti-viral actions continue.

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CHAPTERIII
MATERIALS AND METHODS
A. Preparation of Marigold Leafand Stem Extracts
The plant samples of Tagetes erecta were collected. Fresh plant materials
were washed under running tap water. The collected plant materials were crushed
using mortar and pestle. The excreted juice was extracted through manual
pressing using a piece of cloth.
B. Application of Marigold leaf and stem extracts to the set-ups
Experiment 1
Five beakers of the same sizes were prepared. Each beaker contained 300
ml water and were labeled setup A for control set-up, while set-up B, set-up C,
set-up D, and set-up E for the experimental setups.
Then, approximately 50 mosquito larvae were collected and placed 10
mosquito larvae in each beaker. Marigold leaf and stem extracts were then
added to experimental set-ups at the same time. Set-up B was added with 10
ml extract, Set-up C was added with 40 ml extract, Set-up D was added with 70
ml extract, and Set-up E was added with 100 ml extract. No marigold extract
was added in Set-up A since it was the control set-up.

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The behavior and health of the mosquito larvae were observed and
recorded in tabulated form after the marigold extract was placed in the
experimental set-ups. Observations were done within 5 hours.
C. Determination of effectiveness of the marigold extract
The marigold stem and leaf extract was added at exactly 8 o’clock in the
morning. Observations shall be done within 10 hours and it was observed every
hour starting from 9:00 a.m. until 6:00 p.m. If the number of dead mosquitoes in the
treated set-ups will be greater than number of dead mosquitoes in the untreated
set-up, then the experiment is a success.
Experimental Design
Experimental Set-up Control Group
Independent
Variable
Marigold Stem
and Leaf Extract
Amount of
Extract Used
Dependent
Variable
Death of the
mosquito larvae
Untreated
Mosquito
Larvae

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Schematic Diagram ofthe Research Methodology
Preparation of Materials
EXPERIMENT 1
DIFFERENT AMOUNT OF MARIGOLD LEAF
AND STEM EXTRACTS WITH THE SAME
AMOUNT OF WATER
APPLICATION OF
MARIGOLD LEAF AND
STEM EXTRACTS TO THE
EXPERIMENTAL SETUPS
DATA GATHERING: COUNTING THE
NUMBER OF DEAD MOSQUITO LARVAE
DATA ANALYSIS

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CHAPTER IV
RESULTS
Results
The effects of Marigold leaf and stem extract to heath, behavior, and death of
mosquito larvae are shown on table 1.
Experiment1
Table 1: EFFECTS OF 10 ML, 40 ML, 70 ML, AND 100 ML MARIGOLD LEAF AND
STEM EXTRACT IN 300 ML WATER PER SET-UP TO THE HEALTH, BEHAVIOR AND
DEATH OF MOSQUITO LARVAE AFTER 38 HOURS.
Experiment 1: DIFFERENT AMOUNT OF MARIGOLD LEAF AND STEM EXTRACT
WITH THE SAME AMOUNT OF WATER.
Set-up A
(Control)
Set-up B
(10 ml
Marigold
Leaf and
Stem
Extract)
Set-up C
(40 ml
Marigold
Leaf and
Stem
Extract)
Set-up D
(70 ml
Marigold
Leaf and
Stem
Extract)
Set-up E
(100 ml
Marigold
Leaf and
Stem
Extract)
Health All mosquito
larvae are
alive
Alive but
weak
Alive but
very, very
weak
Expired,
weak
(1 remained)
Expired
Behavior Very active Very
minimal
movement
Very
minimal
movement
Not
Applicable
Not
Applicable
Dead Larvae 0 2 4 8 10

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Table 2.1 ONE WAY ANALYSIS OF VARIANCE TEST
Table 2.2 ONE WAY ANALYSIS OF VARIANCE TEST
CONTROL SET
UP
EXPERIMENTAL SET UP
Time Of Hour Set Up A2 Set Up
B2(10 Ml
Extract Of
Marigold
Leaf And
Stem)
Set Up
C2(40 Ml
Extract Of
Marigold
Leaf And
Stem)
Set Up
D2(70 Ml
Extract Of
Marigold
Leaf And
Stem)
Set Up
E2(110 Ml
Extract Of
Marigold
Leaf And
Stem)
1 0 0 0 1 4
2 0 0 0 9 16
3 0 0 1 25 36
4 0 9 9 36 64
5 0 16 36 64 100
∑A2= 0 ∑B2= 25 ∑C2= 46 ∑D2= 135 ∑E2= 216
CONTROL SET
UP
EXPERIMENTAL SET UP
Time Of Hour Set Up A
(Control)
Set Up B(10 Ml
Extract Of
Marigold Leaf
And Stem)
Set Up C(40
Ml Extract
Of Marigold
Leaf And
Stem)
Set Up D(70
Ml Extract Of
Marigold Leaf
And Stem)
Set Up E(110
Ml Extract Of
Marigold Leaf
And Stem)
1 0 0 0 1 2
2 0 0 0 3 4
3 0 0 1 5 6
4 0 3 3 6 8
5 0 4 6 8 10
∑A= 0 ∑B= 7 ∑C= 10 ∑D= 23 ∑E= 30

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Table 3: ONE WAY ANALYSIS OF VARIANCE TEST (Summary Table)
Sources Of
Variation
Sum Of
Squares
Degrees Of
Freedom
Mean Square Computed F
Between Col.
Within Col.
119.6
106.4
5-1=4
5(5-1)=20
MSB = 119.6
4
= 29.4
MSW= 106.4
20
= 5.32
F = 29.9
5.32
= 5.62
TOTAL 226 25-1=24

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Findings:
Reject the Ho and accept the alternative hypothesis since 5.62 is greater than
4.43. Therefore, there is a significant difference between the amount of leaf and stem
extract in killing mosquito larvae.

27
CHAPTER V
DISCUSSION
In set-up B (10 ml Marigold Leaf and Stem extract added in 300 ml water) only 20
percent is the mortality rate of the mosquito larvae. In set-up C with 40 ml Marigold leaf
and stem extract resulted to 40 percent mortality rate of the mosquito larvae. In Set-up D
with 70 ml Marigold leaf and stem extract it resulted to 80 percent mortality rate while in
Set-up E with 100 ml Marigold leaf and stem extract it resulted to 100 percent mortality
rate. The result shows that the Marigold Leaf and Stem extracts in 300 ml water the more
are the Larvicidal effect to the mosquito larvae. In set-up A (control set up) resulted to 0
mortality rate.

28
Conclusion
Marigold leaf and stem extract has larvicidal potency and effect in getting rid of
mosquito larvae.
Recommendation
In view of the aforementioned results, the following are recommended:
a. Further study using other plant parts of marigold as a potencial larvicide against
mosquito larvae.
b. Companies may develop mosquito larvicidal product with the use of Marigold
as main component.

29
PICTORIALS and EQUIPMENTS
Materials
Mosquito Larvae

30
Control Set up
Experimental Set up

31
Untreated Set ups
300 ml of Marigold stem and leaf
extract

32
Before the experiment
After the experiment

33
BIBLIOGRAPHY
"Genus: Tagetes L.". Germplasm Resources Information Network. United States
Department of Agriculture. 2011-01-06. Retrieved 2011-07-14.
Sunset Western Garden Book, 1995:606–607.
Soule, J. A. 1996. Infrageneric Systematics of Tagetes. Pgs. 435-443 in Compositae:
Systematics, Proceedings of the International Compositae Conference, Kew
1994, Vol. I, Eds. D.J.N. Hind & H.J. Beentje.
Everett, Thomas H. (1982). The New York Botanical Garden illustrated encyclopedia of
horticulture. Taylor & Francis. p. 3290. ISBN 978-0-8240-7240-7.
Soule, J. A. 1993. Biosystematics of Tagetes (Compositae). Ph.D. Dissertation, University
of Texas.
Soule, J. A. 1993. Tagetes minuta: A Potential New Herb from South America. Pgs. 649-
654 in New Crops, Proceedings of the New Crops Conference 1993, Eds. J.
Janick & J. E. Simon.
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you". The Tribune, Chandigarh, India (web site). Retrieved 2007-09-01.
Soule, J. A. 1996. Novel Annual and Perennial Tagetes. Pgs. 546-551 in Progress in New
Crops, Proceedings of the National New Crops Conference 1996, Ed. J.
Janick.
"Tagetes". Integrated Taxonomic Information System. Retrieved 2011-07-14.
"GRIN Species Records of Tagetes". Germplasm Resources Information Network.
United States Department of Agriculture. Retrieved 2011-07-14.

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Alameda County Mosquito Abatement District. "Biological Notes on Mosquitoes." Life
cycle of the mosquito (2011).
Centers for Disease Control and Prevention. "Dengue." Entomology and ecology (2010).
Dengue Virus Net. "Aedes aegypti." Dengue transmission by Aedes aegypti mosquito
(2011).
Department of Medical Entomology, University of Sydney and Westmead Hospital,
Australia.Aedes aegypti (2010).
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Dengue Hemorrhagic Fever, eds. D. J. Gubler & G. Kuno (Cambridge: CABI,
2001): 61–88.

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Curriculum Vitae
Name: Charlie Jhon Aspillaga
Date of Birth: June 27, 1998
Civil Status: Single
Address: Parang, Cantilan, Surigao del Sur
Parents:
Mother: Tersa T. Banguis
Father: Charlito R. Aspillaga
Educational Background
Elementary: Patong-patong Elementary School
First Honors
Secondary: Madrid National High School
Linibunan, Madrid, Surigao del Sur

36
Curriculum Vitae
Name: Christine Joy R. Arreza
Date of Birth: October 6, 1997
Address: Cancavan, Carmen , Surigao del Sur
Parents:
Mother: Rosenie C. Raz
Father: Luven P. Arreza
Elementary: Cancavan Elementary School
Cancavan, Carmen, Surigao del Sur
First Honors
Curriculum Vitae

37
Name: Raven Daryle Mae D. Suarez
Date of Birth: October 30, 1999
Address: Patongpatong,Madrid , Surigao del Sur
Parents:
Mother: Darwinda Suarez
Father: Jeome Suarez
Elementary: Patongpatong Elementary School
Patongpatong, Madrid, Surigao del Sur
First Honors

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