THE INSECTICIDAL PROPERTY OF PANSIT-PANSITAN (Peperomia pellucida (Linn.) (HBK)) LEAF CRUDE EXTRACT AGAINST GREEN LEAFHOPPER (Nephotettix malayanus (Ishihara et Kawase)) A Thesis Presented to the College of Arts and Sciences University of Perpetual Help System DALTA Pamplona, Las Piñas City In Partial Fulfillment of the Requirements for the Degree in Bachelor of Science in Biology Gerard Majello B. Dalisay
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The Insecticidal Property of Pansit-Pansitan Leaf Crude Extract Against Green Leaf Hoppers
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THE INSECTICIDAL PROPERTY OF PANSIT-PANSITAN (Peperomia pellucida (Linn.) (HBK)) LEAF CRUDE EXTRACT AGAINST GREEN
LEAFHOPPER (Nephotettix malayanus (Ishihara et Kawase))
A Thesis Presented to the College of Arts and SciencesUniversity of Perpetual Help System DALTA
Pamplona, Las Piñas City
In Partial Fulfillment of the Requirements for the Degree inBachelor of Science in Biology
Gerard Majello B. Dalisay
March 2010
2
APPROVAL SHEET
The thesis entitled “The Insecticidal Property of Pansit-pansitan
(Peperomia pellucida (Linn.) (HBK)) Leaf Crude Extract Against Green
Leafhopper (Nephotetix malayanus (Ishihara et Kawase))”, prepared by Mr.
Gerard Majello B. Dalisay in partial fulfillment of the requirements for the
Degree in Bachelor of Science in Biology, is hereby recommended for oral
examination.
MARIA LOURDES C. AGUIRRE, MSc. Adviser
Approved in completion of the requirements for the Degree of Bachelor of Science in Biology by the Oral Examination Committee.
MIRIAM B. RAÑOLA, M.A. Member
RAQUEL MARI-ANN O. PAJE, M.A. Member
MARY GRACE B. MARIBAO, M. Ed.Chairman
Accepted in partial fulfillment of the requirements for the Degree in Bachelor of Science in Biology.
FLORENCIA C. MARFIL,Ph.D.Dean, College of Arts and Sciences
March 2010
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ACKNOWLEDGEMENT
First of all I would like to express my gratitude to the One God for the many
blessings that He has given. He is always there taking good care of me and always
comforts me whenever I was in trouble, pain or lonely. He guided me in all the
activities of my life including the construction of this study and He never lets me
go both in good times and in the bad.
I would also like to thank the following persons who helped me in making
this study.
To my parents, Mr. Honorio and Victoria Dalisay who greatly supported me
in physical, mental, emotional, spiritual and especially financial terms. I thank
them for the guidance, encouragement and love that they gave to me. To my
uncle, Mr. Teodoro Dalisay for the information about my test organism. Mrs.
Irenea Dalisay for assisting the construction of the insect cages. Mr. Gregorio
Gomez Jr., Mrs. Editha Roldan and the rest of the employees of Dalisay Poultry
Supply and Dalisay Piggery Farm for the construction of the insect cages and
assisting me whenever I go to the site of the study. To all the rice farmers of the
town of Lian, Batangas for looking after my experimental set ups whenever I am
unable to do so.
To the thesis adviser, Mrs. Maria Lourdes Aguirre for her extreme patience,
assistance and support. Truly the unmatched and ultimate “Mother of Perpetual
Help” for always listening to our complaints and problems and comforting us
when we are in need.
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To Ms. Miriam Rañola, for the patience and endurance in dealing with ones
papers and giving suggestions that can contribute to the betterment of the study.
To Mrs. Mary Grace Maribao, for the assistance and life-inspiring words
which has proven to be a valuable asset in achieving ones goal.
To Ms. Raquel Paje, for the wise and practical examples which enables one
to efficiently tackle the problems of life.
To Mr. Eduardo Jimmy Arma, for the humorous statements and antics that
never fails to enlighten anyone. Truly a walking piece of pure joy that makes you
feel happy and refreshed anytime, anywhere.
To Mr. Allan Alvarez, Dr. Vendivil and the rest of the staff of the Botany
Division of the National Museum for the plant identification. To Dr. Sinohin,
Mrs. Fe Alzona and the rest of the staff of the Crop Protection Cluster of the
University of the Philippines Los Baños, Laguna for the identification of test
organism. Dr. Timbang and the rest of the staff of the IHS Laboratory of the
University of Perpetual Help System DALTA for lending us laboratory
apparatuses required for the study. Also to Mr. Arturo Tobias Calizon for the
statistical analysis.
To Mr. Paolo Louis Almario for allowing me to be patient, kind and
enduring, I thank you for teaching me all of these. To Mrs. Kristina Ezra Salceda-
Dumdum for the tremendous amount of humor that she has given me. I thank you
for the huge amount of food that you are giving to us whenever we are at your
house. Ms. Monique Moldez for teaching me how to be strong, to be able to fight
back from other people’s insults whether in a casual or formal manner. I am very
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grateful for changing me from my doormat and submissive self to the strong
willed and firm individual that I am today. To Ms. Joanna Marie Valloso for
always reminding me of assignments, quizzes and other academic tasks that are
needed to accomplish. You truly are a genuine secretary in training. Thank you
also for sharing your food with me whenever you are unable to finish it.
To Mr. Gene LemuelAbanto, Olivier Thoenen and the rest of the B.S.
Biology students of the University of Perpetual Help System DALTA. I humbly
thank you all for the comedy, drama, horror, romance and all the emotions that
you let me experience. Truly I never have encountered such people and the
experiences that they gave to me are simply life changing and priceless.
Lastly to the people whom I failed to mention I thank you for your
generosity and effort in aiding me in this study. Once more I gratefully thank you
and God bless us all.
Gerard Majello Bulao Dalisay
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ABSTRACT
This study entittled “The Insecticidal Property of Pansit-pansitan
(Peperomia pellucida (Linn.)(HBK)) Leaf Crude Extract Against Green
Leafhopper (Nephotettix malayanus (Ishihara et Kawase))” aims to confirm the
active components present in Pansit-pansitan leaf crude extract using
phytochemical analysis. The study also aimed to determine the insecticidal
property of Pansit-pansitan based on the mortality rate of Green Leafhopper and
compare the different concentrations of Pansit-pansitan leaf crude extract with
Sevin ® 85 WP (positive control).
The leaf crude extract of the plant material was obtained through decoction
method. Phytochemical analysis was done to confirm the different constituents
present in Pansit-pansitan leaf crude extract.
In each treatment three trials have been carried out. These were Sevin ® 85
WP (positive control) and the varying concentrations of the leaf crude extract of
Pansit-pansitan (50%, 75% and 100%) have been prepared by dissolving 5g, 7.5g
and 10g of leaf crude extract in 10ml of distilled water, respectively. Insecticidal
property was determined based on the mortality rate of Green Leafhopper by
administering 10ml of each treatment and the positive control through spray
method.
Results showed that the highest mortality rate of Green Leafhoppers was
obtained for Sevin ® 85 WP (positive control) followed by 100% concentration of
Pansit-pansitan leaf crude extract and the least mortality rate has been observed at
50% concentration. Statistical analysis showed that the computed F value of
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3.303303 is greater than the critical value of F of 3.24 at 0.05 level of significance
thus, the null hypothesis was rejected. There is a significant difference in the
insecticidal property between Pansit-pansitan leaf crude extract and the positive
control, Sevin ® 85 WP.
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TABLE OF CONTENTS
TITLE iAPPROVAL SHEET iiACKNOWLEDGEMENT iiiABSTRACT viTABLE OF CONTENTS viiiLIST OF TABLES xLIST OF FIGURES xiLIST OF PLATES xiiLIST OF APPENDICES xiii
CHAPTERS
I THE PROBLEM AND ITS BACKGROUND
Introduction 1Theoretical Framework 2Conceptual Framework 3Statement of the Problem 4Hypothesis 4Significance of the Study 5Scope and Delimitation 5Definition of Terms 7
II REVIEW OF RELATED LITERATURE
Conceptual Literature 8Scientific Classification of Pansit-pansitan 8
Botanical Description 9Chemical Content of Botanical Extract 10Brief Background about Green Leafhopper 12
Classification 12Taxonomy 13Life Cycle 14
Sevin 85 ® WP 15Research Literature 15
III RESEARCH METHODOLOGY AND PROCEDURE
Research Design 18Research Setting 18Research Procedure 19
Collection and Preparation of Plant Specimen 19
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Identification of Plant Material 19Identification of Test Organism 20
Extraction Procedure 20Decoction Method 20Preparation of Different Concentrations
of Leaf Crude Extract 20Preparation of Sevin ® 85 WP (control) 20
Preparation of the Test Organism 21Preparation of Green Leafhopper Cage 21Collection of Green Leafhopper 21Preparation of Green Leafhopper for Mating 21Application of Leaf Extract to Green Leafhopper 22
Statistical Treatment 22
IV PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA 23
V SUMMARY, CONCLUSION AND RECOMMENDATION
Summary 28Conclusion 29Recommendation 30
BIBLIOGRAPHY
PLATES
APPENDICES
CURRICULUM VITAE
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CHAPTER 1
THE PROBLEM AND ITS BACKGROUND
Introduction
Rice is the staple food of the Filipino people. This particular crop has been
in the Philippine table for thousands of years from its early inhabitants up to the
modern age. It is an annual plant that is cultivated in warm and wet areas in which
the seeds are cooked and used for food.
The said crop is widely cultivated throughout the country, carefully planted
in well irrigated areas, tended by the most skilled farmers and efficiently
processed before going out to the market. However, rice continues to decrease in
yield due to pest infestations thus plant cultivation and harvesting is severely
affected.
The most effective method to get rid of pest infestations nowadays is to use
commercial insecticides however they may cause unwanted effects such as land
poisoning and may endanger human health if not properly handled. Today’s
farmers are running low on funds to buy insecticides which are too expensive.
Thus the use of botanical extracts have been encouraged as possible substitute for
commercially available insecticides.
It is interesting to note that certain plants have insecticidal properties.
Among these plants that have shown such properties is Pansit-pansitan
(Peperomia pellucida (Linn.) (HBK)) . The said herbaceous annual plant contains
chemical substances such as alkaloids that is proven to cause irritation and death
to most types of insects.
2
Theoretical Framework
Pansit-pansitan has long been used by people from all parts of the world in a
variety of ways. In the old days infusion of the leaves and stems are used for gout
and arthritis. The plant has a history of ethnomedicinal use. Anti-inflammatory,
chemotherapeutic, and analgesic properties have been found in crude extracts of
Pansit-pansitan. Data in Bolivia documents the whole plant being crushed, mixed
with water, heated, and then administered to stop hemorrhage (www.drugs.com).
Externally it is used as a facial rinse for complexion problems. Pounded
whole plant used as warm poultice for boils, pustules and pimples. It is also used
for headaches and rheumatic pains (http://www.stuartxchange.org/Pansit.html).In
Southeast Asia, Pansit-pansitan is reported to have analgesic, antibacterial and
antifungal activity. According to a recent study, the plant contains alkaloid which
is a plant-derived compound that is toxic or physiologically active. It has many
uses depending on its type such as quinine which has anti-malarial compound or
tropane alkaloids which have been used throughout history as poisons (Murray G,
Frank M, 1973). Phytochemical studies have also revealed the presence of
alkaloids and tannin in abundant amounts (De Padua LS, Pancho JV. 1983).
3
Conceptual Framework
Green Leafhoppers are a common pest problem throughout the Philippines.
Millions of pesos are spent to control them every year. Pansit-pansitan has been
used as an alternative to commercial insecticides that can be used to exterminate
such rice pests.
Independent variable Dependent variable
Figure 1. A schematic diagram showing the insecticidal property of Pansit-
pansitan against Green Leafhopper (N. malayanus (Ishihara et
Kawase))
Varying concentrations
of Pansit-pansitan leaf
crude extract (50%,
75%, 100%) and Sevin
® 85 WP (positive
control)
Insecticidal property of
the leaf crude extract
Mortality rate of Green
Leafhoppers
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Statement of the Problem
The study was conducted in order to determine the insecticidal property of
Pansit-pansitan (Peperomia pellucida (Linn.)(HBK)) leaf crude extract against
Green Leafhopper (Nephotettix malayanus (Ishihara et Kawase)).
Specifically it aimed to:
1. confirm the active components present in Pansit-pansitan (Peperomia
pellucida (Linn.)(HBK)) leaf crude extract using phytochemical analysis;
2. determine the insecticidal property of Pansit-pansitan (Peperomia
pellucida (Linn.)(HBK)) against Green Leafhoppers (Nephotettix
malayanus (Ishihara et Kawase)); and
3. compare the different concentrations of Pansit-pansitan (Peperomia
of 8-16 eggs and hatch within about 4-8 days. Upon hatching the nymph measures
0.9 mm long with whitish and shiny appearance and as it matures, it turns
yellowish. The shape of the nymph is similar to that of the adult except that the
nymph is smaller and is wingless (Estoy A, 2000). After about 10 to 14 days after
hatching the leafhopper turns to adult which has green appearance with black
markings on the head, thorax, and abdomen. A pair of black spots is either present
or absent on the forewings. The adult insect is 4.2-4.3mm long and the total life
Figure 4. Life Cycle of Green Leafhopper
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cycle from egg to adult is between 22-26 days (Joshi R, 2001). Green
Leafhoppers are most destructive at adult stage due to increased nutrient
consumption from rice plant in preparation for mating and reproduction (Shepard
BM, et al. 1995)
Sevin ® 85 WP
Sevin ® 85 WP is a wide-spectrum insecticide which controls over 100
species of insects on fruit, cotton and other crops as well as on poultry and pets.
Its active ingredient is carbaryl (C12H11NO2) from the chemical group
carbamate (1-naphthyl-N-methyl) (Instant EPA's Pesticide Facts.1996). It kills
insects by disrupting the normal function of their nervous system. Nerve impulses
are transmitted from one nerve to another by a chemical called acetylcholine.
Under normal conditions an enzyme called acetylcholinesterase destroys
acetylcholine so that another nerve impulse may be transmitted. Carbaryl stops
the functioning of the enzyme so acetylcholine builds up resulting in restlessness,
paralysis and death. Carbaryl affects human nerves in the same way as in insect
nerves. However in insects it affects the central nervous system while in humans
it affects the nerve and muscle interaction. Thus carbaryl poisoning symptoms are
different in humans and in insects (Ware G, 2000).
Research Literature
In a study conducted by Dasmariñas, Jover and Pasig (2001) Pansit-
pansitan was used as insecticide for termites. According to the study the plant
contains considerable amounts of monoterpenes, flavonoids, amides and
alkaloids. The chemicals mentioned are known to have insecticidal property in
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which it disrupts the nerve and motor functioning of insects thus causing death. In
the experiment mentioned, the researchers prepared four treatments: Treatment A
consisting of 100% Pansit-pansitan extract, Treatment B with 75% Pansit-
pansitan extract, Treatment C with 50% Pansit-pansitan extract, Treatment D with
25% Pansit-pansitan extract and Treatment E as positive control consisting of
unknown commercial pesticide. The treatments were tested on termites randomly
collected and placed in containers with 20 termites in each and three replicates are
made per treatment. Number of mortality was counted every 3min until 15min
was achieved. Results showed that Treatment A (100% Pansit-pansitan extract)
showed the highest mortality rate of 100% compared with the other plant extracts
and is comparable with Treatment E which is the positive control which also
showed 100% mortality rate.
Pansit-pansitan however has lethal activity according to Lim (2006). The
researcher assessed the plant’s lethal activity using the Brine Shrimp (Artemia
salina) Lethality Bioassay. The testing involved the exposure of the organism to
different aqueous extract concentrations of Pansit-pansitan (10%, 30% and 50%)
in three serial dilutions. Mortality among organisms was observed for 48 hours in
experimental and control groups in three replications and LC50 value was
computed using the Spearman-Karber Method. Significant differences among
sample observations were determined by employing One-Way Analysis of
Variance. Lethal activity of the test plant on the invertebrate model was proven
with 9.18 LC50 value and 50 percent extract concentration as the most lethal
treatment.
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Another study by Cambe, Delfin and Sales, (2003) is aimed to find out if
oleander (Thevetia peruviana) fruit extract can be used as insecticide against rice
bugs. The experimental design was Completely Randomized Block Design
(CRBD) with the following treatments: Treatment A - (25 ml extract + 25 ml
water), Treatment B - (50 ml extract + 25 ml water), Treatment C - (75 ml extract
+ 25 ml water), Treatment D - (100 ml extract), Controlled set-up (100% water)
and replicated three times. The crude extract was obtained by pounding,
squeezing, and filtering. Phytochemical analysis revealed that extract of the young
fruit contains alkaloid, thevetin, peruvoside and cyanogenic glycosides. The
chemicals have influenced the effectiveness of extract and using greater
concentrations can hasten and increase mortality to 100%. Using the One-way
Analysis of Variance and Duncan’s Test, results led to the conclusion that
oleander can be used as an insecticide against rice bugs.
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CHAPTER 3
RESEARCH METHODOLOGY AND PROCEDURE
This chapter presents the research design, methods and adapted procedures in the
study.
Research Design
An experimental method was used in the study to determine the insecticidal
property of different concentrations (50%, 75% and 100%) of Pansit-pansitan leaf
crude extract against Green Leafhopper.
This experiment used three treatments. Different concentrations (50%,
75% and 100%) of the plant’s leaf material have been prepared by dissolving 5g,
7.5g and 10g of Pansit-pansitan leaf crude extract in 10ml of distilled water,
respectively. Sevin ® 85 WP was used as positive control. The positive control
was prepared by dissolving 10g of the insecticide to 10ml of distilled water.
Research Setting
This study along with the collection of plant material and the culturing of the
test organism was conducted at the Dalisay Piggery Farm Compound located in
Lian, Batangas. Phytochemical analysis was done at the IHS Laboratory,
University of Perpetual Help System DALTA located in Las Piñas City. The leaf
crude extract was tested for the following:
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Test for Tannin
A few drops of Ferric Chloride T.S. was added to 2ml plant extract.
Test for Alkaloids
The addition of 1% HCL was used to modify the aqueous extract when one
or two drops of Mayer’s reagent was added.
Grease Spot Test
The plant material was pressed on a bond paper. A temporary presence of
grease spot indicates that the plant material contains volatile oil while
permanent spot indicates the presence of fixed oil.
The study was conducted from December, 2009 to January, 2010.
Research Procedure
Collection and Preparation of Plant Specimen
The plant material was collected at the Dalisay Piggery Farm Compound
in Lian, Batangas. The collected plant material was washed with distilled water
to remove dust and insects and air-dried for three days. Afterwards, it was pre-
weighed prior to submission for phytochemical analysis.
Identification of Plant Material
The plant material was initially identified by the researcher and verified
by Mr. Wilfredo F. Vendivil Ph.D, Senior Researcher of Botany Division,
National Museum located at P. Burgos St. Manila.
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Identification of Test Organism
The test organism was identified by Mrs. Fe D. Alzona, Researcher of
the College of Agriculture, Crop Protection Cluster, University of the
Philippines Los Baños located at Los Baños, Laguna.
Extraction Procedure
Decoction Method
The researcher used decoction method in order to obtain the leaf crude
extract. Five hundred ml of water was boiled first before adding the leaves. The
chopped leaves which weighed 500g was added. The heat was lowered to 50-
60 °C in order to maintain a slow simmer for 10 to 15 minutes. It was removed
from heat and strained before using.
Preparation of Different Concentrations of Leaf Crude Extract
The preparation of varying concentrations (50%, 75% and 100%) of
Pansit-pansitan leaf crude extract were as follows: for 50%, 5g of leaf extract
was dissolved in 10ml of distilled water, for 75%, 7.5g of leaf extract was
dissolved in 10ml of distilled water and for 100%, 10g of leaf extract was
dissolved in 10ml of distilled water.
Preparation of Sevin® 85 WP (control)
Ten grams (10g) of the powdered form of the control (Sevin 85 WP) was
dissolved in 10ml of distilled water.
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Preparation of the Test Organism
Preparation of Green Leafhopper Cage
Sixteen cages were prepared on the natural habitat of the test organism
using a fine net firmly paced on the site by supporting wooden pegs. The first
two cages were used to house the insects in the wild which measured 1m x 1m
x 0.46m. The second two cages were used to house the insect eggs laid by the
previous generation which have the same dimension as the cages that were
used previously. The remaining twelve cages which measured 0.30m x 0.30m
x 0.15m were then used to house the cultured adult forms of Green Leafhopper
in accordance to the number of treatments, trials and control that were used
(Hou, 1987 modified).
Collection of Green Leafhopper
Random wild Green Leafhoppers were collected at Dalisay Piggery
Farm Compound in Lian, Batangas. Insects were collected by constructing
cages on the natural habitat and were cultured to ensure uniformity.
Preparation of Green Leafhopper for Mating
Random Green Leafhopper adults were collected from the wild by
constructing 2 cages which measured 1m x 1m x 0.46m on the natural
environment of the insects. Green Leafhopper eggs were collected after 6 days
by gently brushing the rice plants located on the site with a soft brush to
remove the adult insect thus the insect eggs remained in the rice plant. The rice
plants along with the Green Leafhopper eggs were carefully transferred to
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another 2 cages in the natural habitat which measured the same as the other 2
cages that were used previously and was allowed to reach adulthood. After 11
to 13 days mature insects were collected and distributed to the remaining 12
cages which measured 0.30m x 0.30m x 0.15m in accordance with the number
of treatments, trials and control that were used (Hou, 1987 modified).
Application of Leaf Extract to Green Leafhopper
Spray method was done to apply the different treatments using atomizer.
About 10ml of each treatment was sprayed to the respective cages containing
10 cultured Green Leafhoppers. Each treatment had three replications, thus the
total number of cultured Green Leafhoppers used was 120 including the
control. Higher mortality rate of the insect indicates the effectiveness of the
plant extract. The mortality rate of Green Leafhopper was determined after the
extract was applied through spray method at intervals of every 6 minutes for a
total of 30 minutes.
Statistical Treatment
One way analysis of variance (ANOVA) was used to determine the
significant difference in the insecticidal property between Pansit-pansitan
(Peperomia pellucida (Linn.HBK)) leaf crude extract and the positive control
Sevin ®85 WP.
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CHAPTER 4
PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA
This chapter presents the data gathered, the analysis of data and the different
findings together with their respective analysis and interpretation of results.
In this study the insecticidal property of Pansi-pansitan leaf crude extract
was tested against Green Leafhopper through spray method. The presentation of
data includes the results of the phytochemical analysis, mortality rate of Green
Leafhopper using different concentrations of Pansit-pansitan leaf crude extract
and Sevin ®85 WP (control) and the result of the ANOVA test.
Table 1. Results of Phytochemical Analysis of Pansit-pansitan (Peperomia pellucida (Linn.HBK)) Leaf Crude ExtractTests Theoretical Result Actual Result
Tannins + +
Alkaloids + +
Grease Spot + (volatile oil) + (volatile oil)
Table 1 shows the different constituents present in Pansit-pansitan leaf
extract based on phytochemical analysis. Active components present were tannins
which were indicated by blue to black precipitate, alkaloids which was indicated
by the presence of precipitate and volatile oil which was indicated by a temporary
grease spot. The result justifies the research by De Padua LS and Pancho JV
(1983) on the chemical components of Pansit-pansitan in which the results also
showed the same active components.
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Table 2. Mortality Rate of Green Leafhopper Using Different Concentrations of Pansit-pansitan Leaf Crude Extract and Sevin ® 85 WP (Control)
Treatments Time in Minutes
6 12 18 24 30
Sevin 85 WP (control)
10% 26.67% 33.33% 16.67% 6.67%
100% 6.67% 13.33% 20% 6.67% 0%
75% 3.33% 10% 16.67% 3.33% 0%
50% 0% 3.33% 10% 6.67% 0%
Table 2 shows the mortality rate of Green Leafhoppers using varying
concentrations of Pansit-pansitan leaf crude extract and Sevin ® 85 WP (control).
Based on the results, the positive control Sevin ® 85 WP showed a mortality
rate of 10% after 6 minutes of application, 26.67% after 12 minutes, 33.33% after
18 minutes, 16.67% after 24 minutes and 6.67% after 30 minutes of application.
The 100% concentration of the leaf crude extract showed a mortality rate of
6.67% after 6 minutes of application, 13.33% after 12 minutes, 20% after 18
minutes, 6.67% after 24 minutes and 0% after 30 minutes of application. The 75%
concentration of the leaf crude extract showed a mortality rate of 3.33% after 6
minutes of application, 10% after 12 minutes, 16.67% after 18 minutes, 3.33%
after 24 minutes and 0% after 30 minutes of application. The 50% concentration
of the leaf crude extract showed a mortality rate of 0% after 6 minutes of
application, 3.33% after 12 minutes, 10% after 18 minutes, 6.67% after 24
minutes and 0% after 30 minutes of application.
25
It was observed that there is an increasing trend of mortality rate of Green
Leafhopper up to 18 minutes, a decrease in mortality rate after 24 minutes of
application and at 30 minutes, all the different concentrations of Pansit-pansitan
leaf crude extract showed 0% mortality rate. The positive control Sevin ® 85 WP,
showed 6.67% mortality rate from 6minutes onwards to 30 minutes after
application.
It can be inferred from the data that both the positive control, Sevin ® 85
WP and the different concentrations of Pansit-pansitan leaf crude extract provide
optimum mortality rate at 18 minutes after application of Pansit-pansitan leaf
crude extract on Green Leafhoppers.
26
Figure 5 shows the mortality rate of Green Leafhoppers using varying
concentrations of Pansit-pansitan leaf crude extract and Sevin ® 85 WP (positive
control). The red column represents Sevin 85 ® WP (positive control), the yellow
column represents 100% concentration of Pansit-pansitan leaf crude extract, the
green column represents 75% concentration of Pansit-pansitan leaf crude extract
and the blue column represents 50% concentration of Pansit-pansitan leaf crude
extract.
Figure 5. Mortality rate of Green Leafhopper using varying concentrations of Pansit-pansitan leaf crude extract and
Sevin 85 ® WP (control)
27
Based on the figure, the positive control Sevin 85 ® WP and the varying
concentrations (100%, 75% and 50%) of Pansit-pansitan leaf crude extract showed the
highest mortality rate of 33.33%, 20%, 16.67% and 10% respectively after 18 minutes of
application. The lowest mortality rate of 6.67%, 0%, 0% and 0% was attained for the
positive control Sevin 85 WP and the varying concentrations (100%, 75% and 50%) of
Pansit-pansitan leaf crude extract respectively which occurred after 30 minutes of
application.
Source df SS MS F Critical value of F
Decision Conclusion
Treatments 3 55 18.333
3.303303 3.24 Reject Ho
SignificantError 16 88.8 5.55
Total 19 143.8
Table 3. Summary of Computation of ANOVA Test to Determine the Significant Difference of Pansit-pansitan Leaf Extract and Sevin 85 WP (Control)
Table 3 shows the computed value of Pansit-pansitan leaf crude extract and
Sevin 85 ® WP (control). Since the computed F value of 3.303303 is greater than
the critical F value which is 3.24 at 0.05 level of significance, the null hypothesis
is rejected. This means that there is a significant difference in the insecticidal
property between Pansit-pansitan leaf crude extract and the positive control, Sevin
® 85 WP.
28
CHAPTER 5
SUMMARY, CONCLUSION AND RECOMMENDATION
This chapter discusses the summary, conclusions and recommendations of the
study.
Summary of Findings
This study entitled “The Insecticidal Property of Pansit-pansitan (Peperomia
pellucida Linn. HBK) Leaf Crude Extract against Green Leafhopper (Nephotettix
malayanus Ishihara et Kawase)” was conducted to confirm the active components
present in Pansit-pansitan leaf crude extract using phytochemical analysis,
determine the insecticidal property of Pansit-pansitan based on the mortality rate
of Green Leafhopper, determine the insecticidal property of Pansit-pansitan based
on the mortality rate of Green Leafhopper and lastly to compare the different
concentration of Pansit-pansitan leaf crude extract with Sevin ® 85 WP (positive
control).
Pansit-pansitan leaves were collected at the Dalisay Piggery Farm
Compound in Lian, Batangas. The collected plant material was washed with
distilled water to remove dust and insects and air-dried for three days.
Phytochemical analysis was conducted at the IHS Laboratory, University of
Perpetual Help System DALTA located in Las Piñas City. This experiment has
used three treatments. Different concentrations (50%, 75% and 100%) of the
plant’s leaf material have been prepared by dissolving 5g, 7.5g and 10g of Pansit-
pansitan leaf crude extract in 10ml of distilled water, respectively. Sevin ® 85 WP
29
had been used as positive control by dissolving 10g of the powdered form of the
insecticide to 10ml of distilled water.
Based on the results of the phytochemical analysis, Pansit-pansitan leaf
crude extract contains alkaloids, tannins and volatile oils.
The effects of the varying concentrations of Pansit-pansitan leaf crude
extract on mortality rate of Green Leafhopper showed that the highest mortality
rate obtained was that of the control (Sevin ® 85 WP) which has 28 deaths or
93.33% followed by the 100% concentration of Pansit-pansitan leaf crude extract
which has 14 deaths or 46.67%. The least mortality rate of Green Leafhoppers has
been observed at 50% concentration of Pansit-pansitan leaf crude extract which
only showed 6 deaths or 20%. The ANOVA test reveals that the null hypothesis
was rejected. There is a significant difference in the insecticidal property between
Pansit-pansitan leaf crude extract and the control, Sevin 85 ® WP.
Conclusions
Based on the above findings, the following conclusions were drawn:
1. active components present in Pansit-pansitan leaf crude extract are
alkaloids, tannins and volatile oil;
2. varying concentrations of Pansit-pansitan leaf crude extract on mortality
rate of Green Leafhoppers showed that the highest mortality rate
obtained was that of the control, Sevin ® 85 WP which showed 28
deaths or 93.33% followed by the 100% concentration of Pansit-pansitan
leaf crude extract which has 14 deaths or 46.67%. The least mortality
30
rate of Green Leafhoppers has been observed at 50% concentration of
Pansit-pansitan leaf extract which only showed 6 deaths or 20%; and
3. there is a significant difference in the insecticidal property between
Pansit-pansitan leaf crude extract and the positive control, Sevin 85 ®
WP.
Recommendations
Based on the experiment results, the researcher would like to put forward
the following recommendations:
1. a study regarding the insecticidal property of Pansit-pansitan on the
different stages in the life cycle of Green Leafhoppers must be
conducted;
2. test other parts of Pansit-pansitan plant for insecticidal property;
3. test other local plants that may have insecticidal property;
4. try other extensive methods of plant extraction; and
5. a thorough study on the physiological responses of Green
Leafhopper at different time of application.
1
BIBLIOGRAPHY
Cambe R, Delfin NC,Sales LO. 2003. Insecticidal Effect of Yellow Oleander Fruit Extract on Rice Bugs. Capiz National High School
Dales MJ. 1996. A Review of Plant Materials Used for Controlling Insect Pests. U.J Natural Resources Institute, Kent U.K p.84.
Dasmariñas JT, Jover EM, Pasig KC. 2001. Peperomia pellucida for Termite Control. University of Southern Mindanao Kabacan, Cotabato
De Padua LS, Pancho JV. 1983. Handbook on Philippine medicinal plants, Volume IV. Los Baños: UPLB Documentation and Information Section, Office of the Director of Research.
Estoy AB., LB Flor, EM Valdez et al. 2000. Occurance of Major Rice Insect Pest in PhilRice Stations. Philippine Rice Research Institute (PhilRice).
Harborne Jeffery, H. Baxter. 1983. Phytochemical Dictionary: A Handbook of Bioactive Compounds from Plants. Taylor & Frost, London. 791 pp.
Hou R. F. 1987. Mass Rearing of Green Rice Hopper (Nephotettix cincticeps). International Rice Research Institute
Joshi R, Estoy A, Arida G et al. 2001. Rice Technology Bulletin No: 37. Philippine Rice Research Institute (PhilRice).
Latorre P-Jay D. 2005. Insecticidal Property of Acacia Seeds and Bark Against Termites. Bureau of Soils Lipa City
Manalo JB, Byung HH, Yong NH, et al. 1983. Studies on Ether-Soluble Neutral Compounds of Peperomia pellucida. Annual Report of Natural Products Research Institute, Seoul National University; 22: 16-9.
1
Manalo JB, Recaña MP, Coronel VQ et al. 1984. Potassium Nitrate from Peperomia pellucida (L) HBK. NIST J. 1:25-31
Murray G. and Frank Morehouse. 1973. Organic Molecules in Action. Gordon and Breach.
Pathak MD, Khan ZR. 1994. Insect Pests of Rice. Manila (Philippines): International Rice Research Institute. 89 p.
Pathak MD, Vea E, John VT. 1967. Control of Insect Vectors to Prevent Virus Infection of Rice Plants. J. Econ. Entomol. 60:218-225.
Reissig WH, Heinrichs EA, Litsinger JA, Moody K, Fiedler L, Mew TW, Barrion AT. 1986. Illustrated Guide to Integrated Pest Management in Rice in Tropical Asia. Manila (Philippines): International Rice Research Institute. 411 p.
Shepard BM, Barrion AT, Litsinger JA. 1995. Rice-Feeding Insects of Tropical Asia. Manila (Philippines): International Rice Research Institute. 228 p
Ware, G.W. 2000. The Pesticide Book. Fresno, CA: Thomson Publications. p. 57, 83.
Unpublished sources
Manalo JB. 1983. A Phytochemical Investigation of Pepercmia pellucida (L) HBK.UP Manila
Juniper and Sage Essential Oils(http://www.juniperandsage.com/oils/toxicity.php)
Lans. 2007. Journal of Ethnobiology and Ethnomedicine 2007 3:3 doi: 10.1186/1746-4269-3-3(http://www.ethnobiomed.com/content/3/1/3/table/T4)
Pesticide Management Education Program (PMEP), Cornel University Ithaca, NY(http://pmep.cce.cornell.edu/profiles/extoxnet/carbaryl-dicrotophos/carbaryl-ext.html#top)
Sri Lanka Department of Agriculture(http://www.agridept.gov.lk/institutes_more_sub_links.php?id=184&sMenu=National+Botanic+Gardens+(NBG+)
United States Department of Agriculture Natural Resource Conservation Service( http://plants.usda.gov/java/profile?symbol=PEPE5 )
United States National Library of Medicine(http://chem.sis.nlm.nih.gov/chemidplus/ProxyServlet?objectHandle=DBMaint&actionHandle=default&nextPage=jsp/chemidheavy/ResultScreen.jsp&ROW_NUM=0&TXTSUPERLISTID=000523808)
Virginia Tech's Entomology Department. Species of leafhoppers. (http://everest.ento.vt.edu/~idlab/vegpests/vegi/LH.jpeg)
Plate 5. Application of Varying Concentrations of Pansit-pansitan Leaf Crude Extract to Cultured Insects
1
APPENDICES
1
Appendix A
Data showing the number of Green Leafhoppers died in varying concentrations of Pansit-pansitan leaf crude extract and the control (Sevin ® 85
WP) at different time intervals.
Treatments Number of Test
Organisms
Number of Dead Leafhoppers Based on Time
(minutes)
Total Number
of Deaths
Total Number of Deaths
in Percent6 12 18 24 30
Sevin ® 85 WP
(control) 30 3 8 10 5 2 28 93.33%
50% 30 0 1 3 2 0 6 20%
75% 30 1 3 5 1 0 10 33.33%
100% 30 2 4 6 2 0 14 46.67%
Appendix B
2
Outline of Research Procedure
Research Procedure Overview
C ollec tion of P lant Material
Phytoc hemic alAnalys is
tes t for tannin
tes t for alkaloid
g reas e s pot tes t
Preparation of Different C onc entrations of L eaf C rude E xtrac t
Preparation of T es t Org anis ms
preparation of g reen leafhopper c ag es
preparation of g reen leafhopper for mating
Applic ation of L eaf E xtrac t to G reen L eafh opper
S tatis tic al T reatment
Appendix C
3
Certification of Borrowing of Laboratory Equipment
University of Perpetual Help – Las Piñas Pamplona, Las Piñas City
TO: DR. VALERIANO V. TIMBANG JR. Director, IHS Laboratories This University
THRU: MRS. CLARISSE ROMULO Laboratory Coordinator
SUBJECT: As stated
DATE: November 21, 2009
Greetings!
We are fourth year BS Biology students from this University and we are currently enrolled in Thesis Writing. It is a requirement in this subject to conduct an experiment/analysis about biological problems. With this regard, may we ask your good office to allow us to use/borrow some materials and equipments needed for the conduct of our methodology.
Attached herewith is the list of materials to be used for the different experiments with the planned schedule.
Hoping for your kind consideration. Thank you so much.
Truly yours
DALISAY, GERARD MAJELLMOLDEZ, MONIQUE
VALLOSO, JOANNA MARIE
Noted:
MRS. MA.LOURDES C. AGUIRRE Adviser
4
DATE LIST OF MATERIALS
QUANTITY CHEMICALS
December 9, 20097:30-10:30(Class of Mr. Retardo-
Morphoanatomy)
Centrifuge 1 Trisodium citrate (anhydrous)
Micropipettor 1Test tube 1
Mortar and pestle 1 Ferric chlorideWater bath 1 Lead acetateBunsen burner 1 HClErlenmeyer flask 1 Mayer’s reagentFilter paper Sodium CarbonateBeaker(50 ml and 100ml)
This is to certify that the study entitled “The Insecticidal Property of Pansit-pansitan (Peperomia pellucida (Linn.) (HBK)) Leaf Crude Extract Against Green Leafhopper (Nephotettix malayanus (Ishihara et Kawase))” conducted by Mr. Gerard Majello B. Dalisay was statistically analyzed by the undersigned:
MR. ARTURO TOBIAS CALIZON
Faculty, Mathematics Department
Noted:
MRS. AIDA REGACHO
Chairman, Mathematics Department
6
Appendix E
Certification of Identification of Plant Material
7
Appendix F
Certification of Identification of Test Organism
8
CURRICULUM VITAE
Personal Background
Name: Gerard Majello B. Dalisay
Age: 19 years old
Birthday: April 26, 1990
Address: Herrera Subdivision Lian, Batangas
Name of Father: Honorio L. Dalisay
Name of Mother: Victoria B. Dalisay
Educational Background
School Year
Elementary Nasugbu East Central School 1996-2002
High School Blessed Christ Child Montessori Foundation 2002- 2006
College University of Perpetual Help System DALTA 2006- present