Professional Documents
Culture Documents
Larvicidalpotencyofmarigold2 141217080205 Conversion Gate01
Larvicidalpotencyofmarigold2 141217080205 Conversion Gate01
Larvicidalpotencyofmarigold2 141217080205 Conversion Gate01
Jeshrel B. Plaza
Adviser
S.Y. 2013-2014
Madrid National High School
Madrid, Surigao del Sur
Jeshrel B. Plaza
Adviser
S.Y. 2013-2014
1
Madrid National High School
Madrid, Surigao del Sur
APPROVAL SHEET
JESHREL B. PLAZA
Research Adviser
PANEL
Francisquita J. Cosmiano
Head Teacher- IV / Chairman of SRC
2
Madrid National High School
Madrid, Surigao del Sur
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
To Mrs. Francisquita C. Cosmiano , the Head Teacher IV, for the trust and
kindness;
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
- The Researchers
3
Madrid National High School
Madrid, Surigao del Sur
Table of Contents
Page
Title Page 1
Approval Sheet 2
Acknowledgment 3
Table of Contents 4
List of Tables 5
Abstract 6
4
Madrid National High School
Madrid, Surigao del Sur
List of Tables
Page
5
Madrid National High School
Madrid, Surigao del Sur
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.
6
Madrid National High School
Madrid, Surigao del Sur
CHAPTER 1
INTRODUCTION
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
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
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
7
Madrid National High School
Madrid, Surigao del Sur
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
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
8
Madrid National High School
Madrid, Surigao del Sur
1. Is Marigold stem and leaf extract has a potential larvicide in getting rid of mosquito
larvae?
3. Is there a significant difference between the amount of marigold stem and leaf
C. Research Objectives
1. To test the Larvicidal effectiveness or potency of Marigold leaf and stem extract
9
Madrid National High School
Madrid, Surigao del Sur
D. Statement of Hypotheses:
Null Hypothesis:
1. The marigold leaf and stem extract has no Larvicidal effects on the
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
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
10
Madrid National High School
Madrid, Surigao del Sur
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
G. Definition of Terms
adult mosquito.
11
Madrid National High School
Madrid, Surigao del Sur
CHAPTER II
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.
Kingdom: Animalia
Phylum: Arthropoda
Class: Insecta
Order: Diptera
Suborder: Nematocera
Infraorder: Culicomorpha
Superfamily: Culicoidea
Family: Culicidae
12
Madrid National High School
Madrid, Surigao del Sur
Mosquito larvae have a well-developed head with mouth brushes used for feeding,
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,
they metamorphose into pupae. At the end of each instar, the larvae molt, shedding their
A.2 Marigold
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
Depending on the species, marigold foliage has a musky, pungent scent, though
13
Madrid National High School
Madrid, Surigao del Sur
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
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
Since it has an insecticidal effect, it is hypothesized that the stem and leaf extracts
14
Madrid National High School
Madrid, Surigao del Sur
Kingdom: Plantae
Division: Magnoliophyta
Class: Magnoliopsida
Order: Asrerales
Family: Calenduleae
Genus: Calendula
B. Related Studies
B.1 Foreign
applications of Tagetes include the use of T. minuta leaves to repel mosquitoes and safari
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
15
Madrid National High School
Madrid, Surigao del Sur
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-
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
pests were (and probably still are) either fossorial, litter-decomposing species or
16
Madrid National High School
Madrid, Surigao del Sur
specialists on the seeds of plants in the Leguminoseae and Graminaceae families before
herbivory of tissues of other plant groups. Our objective was to evaluate tissue extracts of
dimorphic bruchid of economic importance throughout the world. We are also concerned
synthetic insecticides. Tagetes spp. has been proposed as an insecticidal crop to provide
(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
B. 2 Local
Marigold or more commonly known as Calendula has been used medicinally for
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.
17
Madrid National High School
Madrid, Surigao del Sur
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
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
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
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 -
18
Madrid National High School
Madrid, Surigao del Sur
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-
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 -
19
Madrid National High School
Madrid, Surigao del Sur
CHAPTER III
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
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,
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
20
Madrid National High School
Madrid, Surigao del Sur
The behavior and health of the mosquito larvae were observed and
recorded in tabulated form after the marigold extract was placed in the
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
Experimental Design
Independent Dependent
Variable Variable
Amount of Untreated
Marigold Stem Extract Used Death of the Mosquito
and Leaf Extract mosquito larvae Larvae
21
Madrid National High School
Madrid, Surigao del Sur
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 ANALYSIS
22
Madrid National High School
Madrid, Surigao del Sur
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.
Experiment 1
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.
23
Madrid National High School
Madrid, Surigao del Sur
24
Madrid National High School
Madrid, Surigao del Sur
25
Madrid National High School
Madrid, Surigao del Sur
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.
26
Madrid National High School
Madrid, Surigao del Sur
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.
27
Madrid National High School
Madrid, Surigao del Sur
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.
28
Madrid National High School
Madrid, Surigao del Sur
Materials
Mosquito Larvae
29
Madrid National High School
Madrid, Surigao del Sur
Control Set up
Experimental Set up
30
Madrid National High School
Madrid, Surigao del Sur
31
Madrid National High School
Madrid, Surigao del Sur
32
Madrid National High School
Madrid, Surigao del Sur
BIBLIOGRAPHY
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. 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.
Gupta, Y.C.; Y. D. Sharma and N.S. Pathania (2002-09-09). "Let the flower of gods bless
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.
33
Madrid National High School
Madrid, Surigao del Sur
Centers for Disease Control and Prevention. "Dengue." Entomology and ecology (2010).
Dengue Virus Net. "Aedes aegypti." Dengue transmission by Aedes aegypti mosquito
(2011).
34
Madrid National High School
Madrid, Surigao del Sur
Curriculum Vitae
Parents:
Educational Background
35
Madrid National High School
Madrid, Surigao del Sur
Curriculum Vitae
Parents:
Educational Background
Curriculum Vitae
36
Madrid National High School
Madrid, Surigao del Sur
Parents:
Educational Background
37