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 Joy R. 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






3

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






4

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






5

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






6

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.






7

CHAPTER 1

INTRODUCTION

A. Background of 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

http://www.iumsp.ch/Publications/pdf/infdisrev1-189.pdf






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.

http://www.surigaotoday.com/2012/07/dengue-cases-in-caraga-surge-bynearly.html

http://en.wikipedia.org/wiki/Prehispanic

http://en.wikipedia.org/wiki/Tagetes

http://en.wikipedia.org/wiki/Mexico

http://en.wikipedia.org/wiki/Central_America

http://en.wikipedia.org/wiki/Cultivar

http://en.wikipedia.org/wiki/Ornamental_plant

http://en.wikipedia.org/wiki/Thiophene

http://en.wikipedia.org/wiki/Gram_negative


http://en.wikipedia.org/wiki/Gram_positive

http://en.wikipedia.org/wiki/Bacteria

http://www.springerlink.com/content/p04q407616406618/






9

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.






10

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.






11

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.






12

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

http://en.wikipedia.org/wiki/Family_(biology)

http://en.wikipedia.org/wiki/Midge

http://en.wikipedia.org/wiki/Fly

http://en.wikipedia.org/wiki/Vertebrates

http://en.wikipedia.org/wiki/Humans

http://en.wikipedia.org/wiki/Livestock






13

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

http://en.wikipedia.org/wiki/Thorax_(insect_anatomy)

http://en.wikipedia.org/wiki/Abdomen

http://en.wikipedia.org/wiki/Spiracle

http://en.wikipedia.org/wiki/Algae


http://en.wikipedia.org/wiki/Pleuston

http://en.wikipedia.org/wiki/Marine_propulsion

http://en.wikipedia.org/wiki/Instar

http://en.wikipedia.org/wiki/Metamorphosis

http://en.wikipedia.org/wiki/Pupae

http://en.wikipedia.org/wiki/Asteraceae

http://en.wikipedia.org/wiki/Compositae

http://en.wikipedia.org/wiki/North_America

http://en.wikipedia.org/wiki/South_America

http://en.wikipedia.org/wiki/Tagetes_minuta






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.

http://en.wikipedia.org/wiki/Insect

http://en.wikipedia.org/wiki/Nematode

http://en.wikipedia.org/wiki/Companion_planting

http://en.wikipedia.org/wiki/Companion_planting

http://en.wikipedia.org/wiki/Tomato

http://en.wikipedia.org/wiki/Eggplant

http://en.wikipedia.org/wiki/Chili_pepper

http://en.wikipedia.org/wiki/Tobacco

http://en.wikipedia.org/wiki/Thiophenes

http://en.wikipedia.org/wiki/Legume

http://en.wikipedia.org/wiki/Tagetes_erecta

http://en.wikipedia.org/wiki/Carotenoid

http://en.wikipedia.org/wiki/Lutein

http://en.wikipedia.org/wiki/Food_coloring

http://en.wikipedia.org/wiki/Poultry

http://en.wikipedia.org/wiki/Thiophene



http://en.wikipedia.org/wiki/Gram_positive


http://www.springerlink.com/content/p04q407616406618/






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.






18

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 -

http://www.wikepedia.com/benefialeffectsofmarigold






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.






20

CHAPTER III

MATERIALS AND METHODS

A. Preparation of Marigold Leaf and 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.






21

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






22

Schematic Diagram of the 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






23

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.

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






24

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






25

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






26

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






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BIBLIOGRAPHY

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Janick & J. E. Simon.

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you". The Tribune, Chandigarh, India (web site). Retrieved 2007-09-01.

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35

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






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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






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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



Larvicidal potency of marigold (2)

Education

Transcript of Larvicidal potency of marigold (2)