Surigao del Sur State University

Cantilan, Surigao del Sur

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ACKNOWLEDGEMENT

Foremost, we would like to express our sincere gratitude to our advisor Mrs. Maria

Lourdes Sering for the continuous support of our study and research, for her patience,

motivation, enthusiasm, and immense knowledge. Her guidance helped us in all the time of

research and writing of this thesis. We could not have imagined having a better advisor and

mentor for our study.

Besides our advisor, we would like to thank the rest of our thesis committee: Mrs.

Anamarie Acevedo, Mrs. Gemma Gruyal, and Mr. Junielito Cortes, for their exemplary

guidance, valuable feedback and constant encouragement throughout the duration of our

research study. Their valuable suggestions were of immense help throughout our work. Their

perceptive criticism kept us working to make this study in a much better way.

We would like to give our sincere gratitude to our families who provide the advice and

financial support. To our friends Jordan Arpilleda, Tristan Charlie Cago, Joseph Condontol, and

Jeruel Mahinay for the stimulating discussions, for the sleepless nights we were working

together, and for all the fun we have had. The product of this research paper would not be

possible without all of them.

Above all, to our Almighty God for the health, and sound mind.
 Surigao del Sur State University
Cantilan, Surigao del Sur
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ABSTRACT

To find out the larvicidal potency of Marigold Leaf and Stem Extract against Mosquito

Larvae, an experimental design was used. The experiment was conducted. The mosquito larvae

were cultured with the use of Mosquito OL Trap, it was covered with a mosquito net and the

eggs became mosquito larvae for 5 days. 10 cultured mosquito larvae were placed in 3 set-ups

with varying amount of Marigold Leaf and Stem Extract: 50ml, 75ml, and 100ml; Synthetic

Pesticide; and a control set-up without treatment. It was observed within 2 hours. It can be seen

in each trial that the mortality rate of the mosquito larvae are in an increasing trend as the level

of concentration increases. Therefore, it can be seen also that the mortality rate is directly

proportional to the level of concentration.

One way ANOVA was used as a statistical tool to find out the Larvicidal Potency of

Marigold Leaf and Stem Extract and LC50 to count the mortality rate of Mosquito Larvae. The

findings revealed that Marigold Leaf and Stem Extract was a very effective larvicide of Mosquito

Larvae.
 Surigao del Sur State University
Cantilan, Surigao del Sur
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CHAPTER 1

INTRODUCTION

Philippines is bearing the high cost brunts 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. Capeding, M.R. (2012, May 30) Dengue in the Philippines. Retrieved from

http://www.denguematters.info

Dengue is the most common arthropod-borne viral (arboviral) illness in humans. It is

transmitted by mosquitoes of the genus Aedes, which are widely distributed in subtropical and

tropical areas of the world. Shepherd, S. M. (2014, Mar 14). Dengue. Retrieved from

http://emedicine.medscape.com

The incidence of dengue has grown dramatically around the world in recent decades.

Over 2.5 billion people – over 40% of the world's population – are now at risk from dengue.

WHO currently estimates there may be 50–100 million dengue infections worldwide every year.

World Health Organization, (2014, Mar). Dengue and Severe Dengue. Retrieved from

http://www.who.int

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. Most of the

cases were from Metro Manila (10,775), Region III (Central Luzon) (7,024), and Region IV-A
 Surigao del Sur State University
Cantilan, Surigao del Sur
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(Cavite, Laguna, Batangas, Rizal and Quezon provinces, or Calabarzon) (6,219). Majority of the

deaths came from Regions IV-B (Mindoro island and the provinces of Marinduque, Romblon

and Palawan, or Mimaropa), VI (Western Visayas), XI (Davao region), XIII (Agusan del Norte,

Agusan del Sur, Surigao del Norte, Surigao del Sur and Dinagat Islands provinces, or Caraga),

and the Autonomous Region in Muslim Mindanao. Dela Cruz, J.M.N (2012, Jul 14). Dengue

cases continue to rise – DOH. Retrieved from http://twiwired.com

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. Merlone,S. (2012 Mar 30). Tagetes erecta.

Retrieved from http://flipper.diff.org

This study aims to determine the effectiveness of Marigold (Tagetes erecta) stem and

leaf extract as a potential larvicide against Mosquito Larvae. The researchers used standard

methods in extracting the leaves and stems.

 Surigao del Sur State University
Cantilan, Surigao del Sur
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Statement of the Problem:

This study aims to determine the larvicidal potency of Marigold (Tagetes erecta) leaf and

stem extract against Mosquito Larvae. Specifically it will look into:

1. What particular concentration of Marigold as a larvicide to Mosquito Larvae.

a. Synthetic/Commercial Larvicide

b. Pure Stem Extract

c. Pure Leaf Extract

d. Mixed (Stem and Leaf) Extract

e. No Extract

2. Mortality rate of Mosquito Larvae in every set-up.

Research Objectives:

1. Test the Larvicidal effectiveness or potency of Marigold (Tagetes erecta) leaf and

stem extract against Mosquito larvae.

2. Know if there is a significant difference between the concentrations of the Marigold

(Tagetes erecta) stem and leaf extract in killing Mosquito larvae.

Null Hypothesis:

There is no significant difference between the concentrations of Marigold (Tagetes

erecta) stem and leaf extract as a potential larvicide against Mosquito larvae.
 Surigao del Sur State University
Cantilan, Surigao del Sur
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Significance of the study:

The result of this study will benefit the following:

Community – the study provides information to the members of the community that

Marigold (Tagetes erecta) plant is a potential larvicide against Mosquito larvae.

Future Researchers – the study provides information to future researchers regarding

the use of Marigold (Tagetes erecta) as larvicide. It can be the key in creating a successful and

beneficial Health product with lesser expenses and with high quality result.

University Extension – the adequate dimension of the result of this study to the

accepted Barangay residents help them to have an alternative and convenient larvicide.

Medical Practitioners – the study can provide a new idea on how to prevent certain

mosquito transmitted disease particularly dengue.

Scope and Limitations

This research 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.

 Surigao del Sur State University
Cantilan, Surigao del Sur
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Definition of terms:

OPERATIONAL

1. Analysis- the separation of components into its constituents in order to find out what

it contains, to examine individual parts, or to study the structure of the whole.

2. Extraction- the process of extracting or being extracted.

3. Ovicidal/Larvicidal Trap- a device use to control Mosquito population.

CONCEPTUAL

4. Extract- to obtain a substance from a compound, in solid, liquid, or gas form by using

an industrial or chemical process.

5. Mosquito larvae- the wingless immature worm-shaped form of many mosquitoes that

develops into a pupa or chrysalis before becoming an adult mosquito.

6. Larvicide- a chemical used to kill larvae.

7. Phytochemicals- a naturally occurring plant substance. Some phytochemicals have

been shown in research to protect against disease.

8. Potency- the state or quality of being potent.

 Surigao del Sur State University
Cantilan, Surigao del Sur
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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
 Surigao del Sur State University
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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 drive

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.3 Marigold

Marigold or Tagetes is a genus of 56 species of annual and perennial mostly

herbaceous plant 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 common insect pests, as well

as nematoes. 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.

 Surigao del Sur State University
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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 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.

Ajaytao (2014 May 12). Eternal Beauty of Nature. Retrieved from

https://ajaytaobotanicalblog.wordpress.com

Since it has an insecticidal effect, it is hypothesized that the stem and leaf extracts can

be an effective larvicide against mosquitoes.

Below is the scientific classification of Marigold.

Kingdom: Plantae

Division: Magnoliophyta

Class: Magnoliopsida

Order: Asrerales

Family: Calenduleae

Genus: Calendula
 Surigao del Sur State University
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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 polycetylenes from Asteraceae indicated that α-

terthienyl, a phototoxic thiophene from Tagetes, had an insecticidal property against A. aegypti

when combined with near-UV radiation (Arnason et al. 1981 pp. 64-68). Morallo-Rejesus &

Decena (1982) isolated a-terthieny 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 and 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 (lyengar et al. 1987 pp.65-69). The topical LDso for Mancuda sexta (L.)

was 10 IJ-gIg, but for Heliothis virescens (F.) it was 470- IJ-gIgand for Ostrinia nubilalis (Hiibner)

the LDso was 700 IJ-g/g (lyengar et al. 1987 pp. 27-29.) This difference was likely cause by

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 (lyengar et al.

1987 pp. 28-30). This elimination was facilitated by higher levels of cytochrome P450 in those

species (lyengar et al. 1990 pp.28-30).

Generally, the efficacy of a α-terthienyl against various mosquito larvae may also be

related to limited evolutionary association between the compound and these insects, with the
 Surigao del Sur State University
Cantilan, Surigao del Sur
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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 naïve 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 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 been 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.

Essential oil from Tagetes erecta against Mosquito obtains compounds piperitone

(45.72%), D-limonene (9.67%), and piperitenone (5.89%) that are active against larvae of Ae.

aegypti, with LC50 of 79.78 microg/ml and LC90 of 100.84 microg/ml. The larvicidal thiophene

contents were higher in the roots and flowers as demonstrated by high-performance liquid

chromatography analysis. Thus, T. erecta constitutes a good source of varied compounds

showing larvicidal activity against Ae. aegypti. John Wiley & Sons (2008). Studies on the

antioxidant and analgesic activities of Aztec marigold (Tagetes erecta) flowers. Retrieved from

http://www.ncbi.nlm.nih.gov
 Surigao del Sur State University
Cantilan, Surigao del Sur
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CHAPTER III

Research Methodology

This study will primarily deal with the effectiveness of Marigold (Tagetes erecta) leaf and

stem extract as a potential larvicide against Mosquito larvae. It will be observed through the

researchers’ naked eye. The researchers are aiming to get the accurate and reliable result of

Marigold (Tagetes erecta) plant.

Preparation of Materials

Preparation and Collection of samples

Marigold (Tagetes erecta) leaves and stems will be gathered and will be confirmed by

the experts. The plants will be washed under running tap water. It will be pounded using mortar

and pestle. The excreted juice will be extracted through manual pressing using a piece of cloth.

The mosquito larvae will be cultured with the use of OL trap. In making an OL Trap, first,

plastic tumbler will be filled with tap water up to 250ml, and then OL pellets will be poured into

the plastic cup. Then, a lawanit stick will be placed in the solution with a rough surface facing

up. Plastic cup will be placed inside and outside of the house in a low-laying area. Lastly, let the

cup stand for six to seven days. Five mosquito OL trap will be exposed to same location within

24 hours. After, it will be covered with a mosquito net and let the eggs to became a mosquito

larvae for 5 days. It will be cultured for reproduction.

 Surigao del Sur State University
Cantilan, Surigao del Sur
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Application of Marigold leaf and stem extracts to the set-ups

Experiment 1

Beakers of the same sizes will be prepared. Each beaker will contain 100 ml water and

will be labeled set-up A for control set-up, set-up B Pure Stem Extract, set-up C Pure Leaf

Extract, set-up D Mixed (Stem and Leaf) Extract, set-up E Synthetic/Commercial Larvicide.

Then, approximately 50 mosquito larvae will be collected and 10 mosquito larvae will be

placed in each beaker. Marigold leaf and stem extracts will then be added to experimental set-

ups at the same time with 50ml, 75ml and 100ml levels of concentration except for Set-up E

since Synthetic/Commercial Larvicide will be used. No Marigold extract will be added in Set-up

A since it is the control set-up. Three trials in every set-up will be done.

The Marigold Stem and Leaf Extract as well as the Synthetic Larvicide will be added at

the same time on the beaker containing the Mosquito larvae. The behavior and health of the

mosquito larvae will be observed and will be recorded in tabulated form when the extract is

drained using the white cloth to clearly visualize the behavior and health of the Mosquito larvae.

Observations will be done within 2 hours in each trial.

Determination of effectiveness of the Marigold extract

The marigold stem and leaf extract will be added at exactly 9 o’clock in the morning.

Observations will be done within 2 hours and it will be observed every 1 hour starting from 9:00

a.m. until 10:00 a.m. in trial 1 and so forth. If the number of dead mosquitoes in the treated set-

ups will be greater than the number of dead mosquitoes in the untreated set-up, then the

experiment is a success.
 Surigao del Sur State University
Cantilan, Surigao del Sur
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Experimental Design

Experimental Set-up Control Group

Independent Dependent
Variable Variable
Level of Untreated
Concentration
Marigold Stem and Death of the Mosquito
Leaf Extract mosquito larvae Larvae

INPUT PROCESS OUTPUT

Marigold (Tagetes Application of Marigold Level of Concentration

erecta) Leaf and Stem (Tagetes erecta) Leaf of Marigold (Tagetes
Extract and Stem Extract erecta) Leaf and Stem
Extract
 Surigao del Sur State University
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Behavior
Distilled Time
Level of Concentration Total Volume and
Water Duration
Health
Synthetic K
50 75 100 100 150 175 200 1hr 2hrs
Larvicide

Pure Stem K
50 75 100 100 150 175 200 1hr 2hrs
Extract

Pure Leaf
50 75 100 100 150 175 200 1hr 2hrs K
Extract

Mixed
Stem and K
50 75 100 100 150 175 200 1hr 2hrs
Leaf
Extract

No Extract 0 0 100 100 100 100 100 1hr 2hrs

 Surigao del Sur State University
Cantilan, Surigao del Sur
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Schematic Diagram of the Research Methodology

Preparation and Collection of Materials

OL TRAP

EXTRACTION OF MARIGOLD LEAF AND

STEM

APPLICATION OF MARIGOLD
EXTRACT

OBSERVATION

DATA GATHERING: COUNTING THE

NUMBER OF DEAD MOSQUITO LARVAE

DATA ANALYSIS
 Surigao del Sur State University
Cantilan, Surigao del Sur
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Research Setting

The researchers will conduct study at Surigao del Sur State University Biology

Laboratory.

People Responsible in Gathering Data

The researchers were the one accountable in gathering the data of the experiment in

order to come up with accurate result. The researchers will be keen observers in order to note

every detail and changes that will occur in 3 trials in the experiment.

Statistical Tool

One way anova will be used as a statistical tool to find out the Larvicidal potency of

Marigold (Tagetes erecta) leaf and stem extract, and Scaling Method to get the mortality rate of

Mosquito larvae.
 Surigao del Sur State University
Cantilan, Surigao del Sur
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CHAPTER IV

Results and Discussion

This chapter presents the data gathered from the observations that were conducted. The

gathered data were analyzed and interpreted following the specific objectives enumerated in Chapter 1.

MORTALITY RATE OF MOSQUITO LARVAE USING SYNTHETIC PESTICIDE, 50ML, 70ML, 100ML MARIGOLD LEAF,
STEM, LEAF AND STEM EXTRACT IN 100ML DISTILLED WATER PER SET-UP WITHIN 2 HOURS.

Table 1: TRIAL 1
TIME
OF EXPERIMENTAL SET-UP CONTROL SET-UP
HOUR
SET-UP D
SET-UP B SET-UP C
SET-UP A MIXED MARIGOLD SET-UP E
PURE MARIGOLD PURE MARIGOLD
SYNTHETIC PESTICIDE LEAF AND STEM NO EXTRACT
STEM EXTRACT LEAF EXTRACT
EXTRACT
50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml
30
0 0 2 0 0 0 0 0 0 1 1 1 0 0 0
minutes
1 hour 1 2 2 0 0 0 0 0 1 1 2 3 0 0 0
1 hour
and 30 2 3 2 1 1 2 1 2 2 2 2 2 0 0 0
minutes
2 hours 4 3 4 2 4 4 4 3 4 3 3 4 0 0 0

Table 2: TRIAL 2
TIME
OF EXPERIMENTAL SET-UP CONTROL SET-UP
HOUR
SET-UP D
SET-UP B SET-UP C
SET-UP A MIXED MARIGOLD SET-UP E
PURE MARIGOLD PURE MARIGOLD
SYNTHETIC PESTICIDE LEAF AND STEM NO EXTRACT
STEM EXTRACT LEAF EXTRACT
EXTRACT
50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml
30
0 0 2 0 0 0 0 0 0 1 1 1 0 0 0
minutes
1 hour 1 1 3 0 0 0 0 0 0 1 1 2 0 0 0
1 hour
and 30 2 3 3 1 2 2 2 2 3 2 2 3 0 0 0
minutes
2 hours 5 5 2 3 2 4 3 4 3 3 3 4 0 0 0
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Table 3: TRIAL 3
TIME
OF EXPERIMENTAL SET-UP CONTROL SET-UP
HOUR
SET-UP D
SET-UP B SET-UP C
SET-UP A MIXED MARIGOLD SET-UP E
PURE MARIGOLD PURE MARIGOLD
SYNTHETIC PESTICIDE LEAF AND STEM NO EXTRACT
STEM EXTRACT LEAF EXTRACT
EXTRACT

50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml 50ml 75ml 100ml

30
0 0 3 0 0 0 0 0 0 1 1 2 0 0 0
minutes

1 hour 1 2 3 0 0 0 0 0 0 1 1 2 0 0 0

1 hour
and 30 3 3 3 1 1 3 2 3 4 3 3 3 0 0 0
minutes
2 hours 4 4 1 2 3 3 3 4 3 2 3 3 0 0 0

In set-up A (Synthetic Larvicide) approximately 80%-90% is the mortality rate of the

mosquito larvae at 50ml and 75ml level of concentration in each trial. In 100ml of Synthetic

Pesticide 100% mortality rate was observed in all trials.

In set-up B (Pure Marigold Stem Extract) and set-up C (Pure Leaf Extract) approximately

30%-60% mortality rate was observed in 50ml and 75ml level of concentration in each trial. Only

60%-70% mortality rate was noted in the 100ml level of concentration.

In set-up D (Mixed Marigold Stem and Leaf Extract), approximately 70%-80% is the

mortality rate of the mosquito larvae at 50ml-75ml level of concentration in each trial. With the

100ml Mixed Marigold Stem and Leaf Extract, it resulted to 100% mortality rate.

In control set-up, no mortality rate was observed since no extract was added.

The result shows that the Mixed Marigold Stem and Leaf Extract in 100ml water has

more larvicidal effect to the mosquito larvae.

 Surigao del Sur State University
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Table 4: EFFECTS OF SYNTHETIC PESTICIDE, 50ML, 70ML, 100ML MARIGOLD LEAF, STEM, LEAF AND STEM
EXTRACT IN 100ML DISTILLED WATER PER SET-UP TO THE HEALTH, BEHAVIOR, AND DEATH OF MOSQUITO
LARVAE AFTER 2 HOURS.

CONTROL
EXPERIMENTAL SET-UP
SET-UP

Set-up D
Set-up A Set-up B Set-up C
Mixed Marigold Set-up E
Synthetic Marigold Pure Marigold Pure
Stem and Leaf No Extract
Pesticide Stem Extract Leaf Extract
Extract
Most of them
All mosquito
Alive but weak died, some are All mosquito
HEALTH larvae are All are alive
and some died. alive but very, larvae died.
dead.
very weak.
Loss of Swam in a non-
equilibrium and Very slow wriggling
BEHAVIOR Restless. Very active.
finally led to movement. straight
death. pattern.

The mosquito larvae swim by means of wriggling movements, with insects flexing their

bodies from one side to the other to exhibit a zig-zag tracking pattern that is characteristically

detected in the untreated set-up. However, it did not exhibit the wriggling swimming pattern

when exposed to the Synthetic Pesticide and the Marigold Extracts. In response to exposure to

these set-ups, the mosquito larvae swam mainly in a non-wriggling straight pattern, which likely

reduced the swimming velocity.

In our study, swimming behavior was recorded after 2 hours of exposure, and the

inhibitory effects caused by the Synthetic Pesticide and Marigold Extracts tested prevailed.
 Surigao del Sur State University
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Table 5: SUMMARY OF DEATHS AFTER 2 HOURS

Total Standard
Mean Variance
Deviation

50 ml 75 ml 100 ml

Synthetic
23 26 30 59 8.78 1.09291 1.19444
Pesticide

Stem Extract 10 13 18 41 4.56 1.23603 1.52778

Leaf Extract 15 17 20 52 5.78 0.83333 0.69444

Mixed
21 23 30 74 8.22 1.39443 1.94444
(Stem and Leaf) Extract

No Extract 0 0 0 0 0 0 0

∑ Mean 6.835

Table 6: ONE WAY ANALYSIS OF VARIANCE TEST (Summary Table)

Sources of Degrees of
Sum of Squares Mean Square Computed F
Variation Freedom
MSB= 107.9091
Between Col. 107.9091 3 3
= 35.9697
F= 35.9697
8.57784
Within Col. 42.8892 5 MSW= 42.8892
5 = 4.1933
= 8.57784
Total 151.0001 8

Critical Fvalue= 3.78 and Fratio= 4.1933. Since obtained Fratio is greater than 3.78 at 5%

level of significance, there is a significant difference between the Marigold Extract and Synthetic

Pesticide.
 Surigao del Sur State University
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CHAPTER V

Findings, Conclusion and Recommendation

Findings:

Reject the Ho and accept the alternative hypothesis since 4.1933 is greater than the
critical F value which is 3.78 at 5% level of significance. Therefore, there is a significant
difference between the Marigold Stem and Leaf Extract and Synthetic Larvicide.

Conclusion:

The mixed Marigold leaf and stem extract has greater larvicidal potency and effect in
getting rid of mosquito larvae compared to using the pure leaf or pure stem extract. It was also
found out that the 100% extract poses greater potency in killing the mosquito larvae.

Recommendation:

In view of the aforementioned results the following are recommended:

 Further study using other plants as a potential larvicide against mosquito larvae.
 Analysis using dengue-carrying mosquito larvae.
 The usage of Synthetic Larvicide instead of Synthetic Pesticide may be implied.
 Surigao del Sur State University
Cantilan, Surigao del Sur
____________________________________________________________________________________

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