THE TERMITICIDAL POTENTIAL OF Chromolaena odorata (L.)
R.M. KING & H. ROBINSON (HAGONOY)
RALPH STEPHEN E. CACAPIT
IZAVELO RAPHAEL J. LLOREN
CALEB MARK D. QUIAMBAO
DENNIS BRYAN A. VALDEZ
Don Mariano Marcos Memorial State University
South La Union Campus
College of Education
Laboratory High School
Agoo, La Union
Elective II (Research)
March 2009
THE TERMITICIDAL POTENTIAL OF Chromolaena odorata (L.)
R.M. KING & H. ROBINSON (HAGONOY)
RALPH STEPHEN E. CACAPIT
IZAVELO RAPHAEL J. LLOREN
CALEB MARK D. QUIAMBAO
DENNIS BRYAN A. VALDEZ
Submitted to
Tessie Q. Peralta, Ph. D.
Don Mariano Marcos Memorial State University
South La Union Campus
College of Education
Laboratory High School
Agoo, La Union
In Partial Fulfillment of the Requirements
in Elective II (Research)
March 2009
APPROVAL SHEET
This is to certify that this study, entitled: “THE TERMITICIDAL POTENTIAL OF
Chromolaena odorata (L.) R.M. KING & H. ROBINSON (HAGONOY)” presented and
submitted by Ralph Stephen E. Cacapit, Izavelo Raphael J. Lloren, Caleb Mark D. Quiambao,
and Dennis Bryan A. Valdez, to fulfill part of the requirements in Elective II (Research), was
examined and passed on March 4, 2009 by the Research Committee composed of:
TESSIE Q. PERALTA(Sgd) FLORDILIZA B. DALUMAY(Sgd)
Adviser Critic
LILIA L. MADRIAGA(Sgd)
Chairman
MARICON C. VIDUYA(Sgd)
Member
Accepted and approved in partial fulfillment of the requirements in Elective II
(Research) on March 4, 2009.
MERCEDITA A. MABUTAS(Sgd) PURIFICACION B. VERCELES(Sgd)
OIC – Principal Chairman
Laboratory High School Bachelor of Secondary Education Department
MANUEL T. LIBAO(Sgd)
Dean
College of Education
ii
ACKNOWLEDGMENT
The researchers would like to extend their thanks to the people who helped in the
realization and success of this study:
Dr. Tessie Q. Peralta, their subject teacher for her encouragement and motivation
throughout the development of this manuscript;
Prof. Mercedita A. Mabutas, the principal of the DMMMSU – SLUC, College of
Education, Laboratory High School, Agoo, La Union, for her encouragement toward a
deeper pursuit of knowledge;
Ms. Flordiliza B. Dalumay, their critic, and Prof. Lilia L. Madriaga, the Chairman of
the panel, for sharing their ideas, professional expertise, constructive criticisms, and
invaluable assistance towards the refinement of this study;
Ms. Maricon C. Viduya, the member of the panel, for sharing her expertise in
English grammars, and checking and editing this manuscript;
Dr. Raquel D. Quiambao, for sharing her expertise in the statistical part of the study;
Mr. Tristan B. Villanueva, the Laboratory Custodian of the DMMMSU – SLUC,
College of Sciences, for making the laboratory equipment and materials available to the
researchers;
Dr. Wilfredo F. Vendivil, the Senior Researcher of the National Museum, Botany
Division, for sharing his precious moments in determining and verifying the plant material
used in the study;
Ms. Adoracion A. Ceniza, the Chief, Baguio Pesticide Analytical Laboratory Satellite,
Bureau of Plant Industry, Baguio National Crop Research and Development Center, for the
bulk extraction of the hagonoy methanol leaf extract;
Their parents, for their love, guidance, encouragement, cooperation, inspiration,
and financial support;
Their brothers, sisters, friends, and relatives, for their unstinted support and
cooperation;
Their classmates, for the encouragement, moral support, help, and assistance; and
Above all, the Almighty God, for the spiritual enlightenment, guidance and support;
and for the strength and wisdom that enabled the researchers to achieve this undertaking.
The Researchers
iv
TABLE OF CONTENTS
Page
Title Page . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i
Approval Sheet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ii
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii
Table of Contents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
List of Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
List of Figures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . viii
List of Experimental Plates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
List of Appendices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x
Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
Chapter I Introduction
Background of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Objectives of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Null Hypotheses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Significance of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Scope and Limitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Time and Location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Definition of Terms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Conceptual Framework . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Chapter II Review of Literature
Termites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Chromolaena odorata . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Solignum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Chapter III Methodology
Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Research Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Collection and Preparation of Plant Material . . . . . . . . . . . . . . . . . . . . . . . 26
Preparation of Decoction from C. odorata Leaves . . . . . . . . . . . . . . . . . . . 26
Preparation of Expressed Juice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Preparation of the Methanol Leaf Extract . . . . . . . . . . . . . . . . . . . . . . . . . 27
Test Organism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Application of the Test Substances . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Data Gathered . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Data Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Chapter IV Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Chapter V Summary, Conclusion, and Recommendation
Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Recommendation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Experimental Plates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
Appendices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
vi
LIST OF TABLES
Table Title Page
1 Length of time in minutes for the termites to die due to various 30
treatments
2 One‐Way Analysis of Variance (ANOVA) Summary Table 32
3 Pairwise Comparison of the Mean Time using Scheffé's Test 33
vii
LIST OF FIGURES
Figure Title Page
1 Problem Tree (Termites) 4
2 Problem Tree (Dependence on Insecticides) 5
3 The Conceptual Framework 9
4 Research Flowchart 29
5 Length of time in minutes for the termites to die due to various 31
treatments
viii
LIST OF EXPERIMENTAL PLATES
Plates Title Page
1 Chromolaena odorata in its natural environment 42
2 Fresh leaves of Chromolaena odorata 42
3 The experimental set‐up 43
4 The separation of test organism from the soil 43
5 The test organisms placed on the dish 44
6 Decoction of Chromolaena odorata leaves 44
7 Pounding of fresh Chromolaena odorata leaves 45
8 Air dried Chromolaena odorata leaves 45
9 Preparation of Extract 46
10 Laboratory personnel of the Bureau of Plant Industry – 46
National Pesticide Analytical Laboratory performing the Rotavation
11 The test organisms administered by positive control (Solignum) 47
12 The test organisms administered by leaf decoction from hagonoy 47
13 The test organism administered by expressed juice from hagonoy 48
14 The test organism administered by 30% leaf extract solution 48
15 The test organisms administered by 60% leaf extract solution 49
16 The test organisms as affected by the various treatments 49
17 The researchers introducing the various test substances to the termites 50
18 Researcher of the National Museum, Botany Division determining the 50
plant material used in the study
ix
LIST OF APPENDICES
Appendix Title Page
A Sample Computation for Methanol Leaf Extract Solutions 51
B Statistical Formula 52
C Sample Computation of ANOVA and Scheffé’s Test 54
D Tables 58
E Figures 60
F Letter to the Senior Museum Researcher of the National Museum 65
G Certification of Plant Material 66
H Letter to the Chief, Baguio PAL Satellite‐BNCRDC 67
I Certificate of Appearance 68
(issued by the Chief, Baguio PAL Satellite‐BNCRDC)
x
THE TERMITICIDAL POTENTIAL OF Chromolaena odorata (L.)
R.M. KING & H. ROBINSON (HAGONOY)
Researchers: Ralph Stephen E. Cacapit
Izavelo Raphael J. Lloren
Caleb Mark D. Quiambao
Dennis Bryan A. Valdez
Adviser: Tessie Q. Peralta, Ph. D.
A B S T R A C T
This study aimed to analyze and determine the termiticidal potential of
Chromolaena odorata (L.) RM King & H Robinson (Hagonoy).
This study utilized the experimental method of research which focused on the use
standard laboratory diagnostic procedures implicated in termiticidal potential of hagonoy in
terms of the length of time for the termites to die upon application of the various
treatments.
The methods used were collection and preparation of plant material, the
preparations on decoction from hagonoy leaves, expressed juice, methanol extract,
Solignum, and the application of the various substances to the termites.
Specifically, this study tested if there were significant differences in the length of
time the termites were killed between pair of treatments and after the application of the
various treatments which were T1 – positive control, commercial pesticide (Solignum), T2 ‐
leaf decoction from C. odorata, T3 ‐ expressed leaf juice from C. odorata, T4 ‐ 30% C.
odorata leaf extract solution, and T5 ‐ 60% C. odorata leaf extract solution.
The data gathered were tabulated and computed using One‐Way Analysis of
Variance (ANOVA) to determine whether significant differences exist in the length of time
the termites were killed after the exposure to the five treatments. Scheffé's Test was used
to find out where the difference lies among the five treatments and which among the
treatments had the greatest termiticidal potential.
The study revealed that there were significant differences in the length of time the
termites were killed among the various treatments and between the pair of treatments. The
data gathered showed that sample termites in 60% C. odorata leaf extract solution has the
least length of time with a mean of 1.29 minutes. This was followed by T1 (positive control,
Solignum) with a mean 1.45 minutes, 30% C. odorata leaf extract solution with a mean of
1.47, and leaf decoction from C. odorata with a mean of 1.77. The treatment with the
longest time for the sample to die was the expressed leaf juice from C. odorata with a mean
of 2.15 minutes.
The F‐test showed that there were significant differences among the means of the
five treatments at 0.05 significance level. The Scheffé's Test implies that the positive control
and 30% C. odorata methanol leaf extract solution were equally efficient. Furthermore,
since all comparisons with 60% C. odorata leaf extract solution which has the shortest mean
time are significantly different, then it could be concluded that 60% hagonoy methanol leaf
extract solution is the best among the five treatments.
xii
Chapter I
INTRODUCTION
Background of the Study
Plants are considered as one of the most significant endeavors to study. All life on
Earth depends directly or indirectly on the plants for food and for other purposes. Some of
these contain thousands of medicinal compounds (Quisumbing, 1978) while the others have
potential as insecticide (http://encarta.msn.com).
Chromolaena odorata, locally known as Hagonoy or Hagonoi, is a shrub that is
widely distributed to many parts of the tropics. However, many farmers do not seem to
mind its benefits because these are problems in agricultural land and commercial
plantations. Chromolaena odorata (hagonoy) is a pest abundant in our country (Padilla, et.
Al., 2006)
It is proven that hagonoy causes diarrhea when ingested or even fetal deaths among
animals. It also reduces the carrying capacity of the field to grow more beneficial weeds.
(CPD Technoguide No. 8, 2003).
People even regard this as unusable. Because of this perception, the researchers
decided to make something out of Chromolaena odorata (Hagonoy). The plant could be
used as a termiticide.
Termites are some of the loathed and hated pests in the Philippines because they
feed on woods and weaken its structure and they are considered pest by local people. For
this reason, the researchers developed termiticide out of Chromolaena odorata (hagonoy)
plant.
2
According to Padilla, et al., the use of pesticide is really prevalent to industrial
countries like the Philippines. They are used in comforts of their business establishments
and even at home to eliminate the huge number of pests. And one of the pests, considered
as problem of Filipinos today are the destructive termites.
With these concerns, the researchers were motivated to investigate the termiticidal
potential of Chromolaena odorata (hagonoy).
Objectives of the Study
The main purpose of this study is to formulate a potential solution to the focal
problems presented in Figures 1 and 2, which are the destructive termites and the
dependence on insecticides by developing a termiticide out of C. odorata.
The study purported to evaluate the termiticidal potential of Chromolaena odorata
(L.) R.M. King & H. Robinson (Hagonoy).
In the study reported herein, it sought to:
1. Test termiticidal potential of Chromolaena odorata in terms of the length of time
the termites are killed after the application of the five treatments.
2. Determine the significant differences in the length of time the termites are killed
when exposed to the five treatments.
3. Analyze the significant differences in the length of time the termites are killed
between pairs of treatments.
3
In this regard, the researchers were encouraged to investigate the termiticidal
potential of Chromolaena odorata (hagonoy) and to develop ways of putting C. odorata to
beneficial uses as an effective alternative termiticide.
Null Hypotheses
The following null hypotheses were tested in the study:
1. There are no significant differences in the length of time the termites are killed
after application of the five treatments.
2. There are no significant differences in the length of time the termites are killed
between pairs of treatments.
4
PROBLEM TREE
Structural Damage and
Additional Expenses to
Repair Damaged
Properties Use of Expensive
Chemical Termiticide
TERMITES
Lack of Knowledge of
Destructive
Plant Alternative as
Termiticide
Figure 1. Problem Tree
5
PROBLEM TREE
Risk to Human
and Animal Health
Expensive
Pesticide Toxicity to Natural Enemies
Resistance and Other Non-target
Organisms
DEPENDENCE
ON
INSECTICIDES
Highly Easy to Use
Effective
Readily Lack of Knowledge on
Available its Impacts to Health
and Environment
Figure 2. Problem Tree
6
Significance of the Study
Results of this study may be beneficial to the following:
a. People and industry. This study may serve as a benchmark on termite
eradication. This study may also serve as a springboard towards the discovery of potentially
new natural foundations of pesticides from plants. After several years of research efforts,
the problems on C. odorata remained unsolved. In this unfolding scenario, it is necessary to
develop ways of putting C. odorata to beneficial uses (Bamikole, 2004). It can be used as a
source alternative foundations as termiticide which is health and environment friendly.
People living in far‐flung areas and places that cannot be reached by modern products may
also benefited by this study.
b. Future researchers. The results of this study would provide baseline information
that may serve as a frame of reference to the future researchers who become ardently
absorbed in this study; and
c. The Researchers. It is hoped that this study will contribute to the pool of
knowledge about the termiticidal properties of plants like Chromolaena odorata.
Ultimately, the results of this study will serve as the potential solution to the focal
problems on the destructive termites and the people’s dependence on insecticides.
Scope and Limitation
The concern and focus of this study was limited to the termiticidal potential of C.
odorata making use of 5 (five) treatments which can be found in Figure 3. Effects of the
7
positive control, hagonoy leaf decoction, expressed juice, and methanol extracts were
determined based on the length of time the termites are killed upon the application.
The extracting solvent used was a technical grade methanol. Thus, the component
that can only be extracted using methanol was considered. Distilled water was used as the
solvent for the treatments that used expressed juice and decoction (Biagtan, De Guzman,
Mangay‐ayam, 2007).
Time and Location
This study was conducted at Don Mariano Marcos Memorial State University‐South
La Union Campus, Laboratory High School, Agoo, La Union from December 2008 to February
2009. Crude extraction of the termiticidal components was done at the Bureau of Plant
Industry (BPI) – Baguio Pesticide Analytical Laboratory (BPAL), Baguio City. Verification of
the plant material was done at the Botany Division of the National Museum, Manila.
Definition of Terms
For better understanding of the study, the following terms were operationally
defined and used .
Commercial Pesticide. e.g. Solignum; This refers to the positive control used in the
study, which is a synthetic chemical mostly used as termite killer and wood preservative.
Decoction. This refers to the preparation made by boiling the leaves in water.
Eradication. This refers to the elimination or destruction of a population of
termites as part of pest control.
8
Expressed Juice. The preparation made by pounding C. odorata leaves using mortar
and pestle afterwich, fresh leaf juice was obtained.
Hagonoy. A local term of Chromolaena odorata; plant used in the study.
Insecticide. A type of pesticide used to eliminate insects.
Leaf Extract. A concentrated preparation of C. odorata (Hagonoy) leaf obtained by
separating the active constituent of the plant through extraction using methanol.
Methanol. A colorless organic substance used as a solvent also known as methyl
alcohol, carbinol, wood alcohol, wood naphtha or wood spirits. It is a chemical compound
with chemical formula CH3OH (often abbreviated MeOH) and boils at 64.7 °C (337.8 K).
Methanol Extract . It is a concentrated preparation of C. odorata obtained by using
methanol as the extracting solvent.
Termites. Social insects that are destructive to properties, and used as test organism
in this study.
Termiticide. It is a specific type of insecticide used to kill termites.
Pesticide. The agent known to eliminate different types pests like termites, fungi,
mollusks.
Conceptual Framework
This research study focused on the termiticidal potential of C. odorata.
Proper and careful laboratory method and techniques such as collection and
shredding of leaves, extraction, preparation of solutions and test for the efficacy of the
termiticide were done.
9
There were five treatments namely: T1 ‐ positive control, commercial pesticide
(Solignum), T2 ‐ leaf decoction from C. odorata, T3 ‐ expressed leaf juice from C. odorata, T4
‐ 30% C. odorata leaf extract solution, and T5 ‐ 60% C. odorata leaf extract solution.
The paradigm, which explains the conceptual framework of this study, is presented
in Figure 3.
INDEPENDENT VARIBLE DEPENDENT VARIABLE
Treatment: Length of time the termites are
killed after the exposure to:
a. T1 ‐ positive control, (Solignum)
a. leaf decoction from hagonoy
T2‐ leaf decoction from
b.
C. odorata b. expressed leaf juice from
hagonoy
c. T3‐ expressed leaf juice from
C. odorata c. methanol leaf extract
solutions
T4‐ 30% C. odorata leaf extract
d.
solution
e. T5‐ 60% C. odorata leaf extract
solution
Figure 3. Conceptual Paradigm showing the Independent and Dependent Variables
of the study.
Chapter II
REVIEW OF LITERATURE
Plants are used as insecticides since before the time of Ancient Romans. People
applied the crude mixture from natural products in insecticidal activities nowadays
(Franzios, 1997).
Many plants such as vetiver grass oil (Zhu, 2001); seeds (Vanucci, 1992); wood
leaves, such as leaves of tarbush (Flourensia cernua) (Tellez, 2001); mature leaves of 7
species of Eucalyptus (Lin, 1998); leaves of Azadirachta excelsa (Jack) Jacobs (Sajap, 2000);
and heartwood from trees have been investigated for termiticidal preservatives. Many
species of wood have been extracted, and the efficacy for termite resistance has been
tested. These woods include Taxodium distichum (L.), (Scheffrahn, 1988); Sawara wood
(Chamaecyparis pisifera D. Don) (Saeki, 1973); Port‐Orford‐cedar [Chamaecyparis
lawsoniana (A. Murr.) Parl.] (McDniel, 1989), bogwood of yakusugi (Yatagai, 1991);
Diospyros virginiana L. (common persimmon) (Carter, 1978), Mansonia altissima and
Piptadeniastrum africanum (Bultman, 1979); Juniperus procera (African pencil cedar)
(Kinyanjui, 2000); Taiwania (T. cryptomerioides) (Chang, 2001); Melaleuca (Melaleuca
quinquenervia) heartwood blocks (Carter, 1982); Lophopetalum wightianum,
Microcerotermes beesoni Snyder, Neotermes bosei (Snyder) and Heterotermes indicola
(Wasm.) (Sen‐Sarma, 1979); Brazil wood (Prosopis juliflora), Anadenanthera macrocarpa;
Astronium urundeuva; Schinopsis brasiliensis, Senna siamea, Tabebuia aurea, Amburana
cearensis, Tabebuia impetiginosa and Aspidosperma pyrifolium (Paes, 2001). Wang (1989)
compared the effects of eight species which included (a) Chamaecyparis obtusa,
11
(b) Cunninghamia lanceolata, (c) Taiwania cryptomerioides, (d) Cryptomeria japonica, (e)
Acacia confusa, (f) Castanopsis carlesii, (g) red oak (Quercus sp.) and (h) Gonystylus
bancanus.
These materials were extracted with organic solvents or hot water to get crude
mixture to test against termites. Most of them were tested in a mixture, and effective
compositions were isolated and identified by some of the researchers. The extracted
chemicals may act as feeding deterrent, ovipositor deterrents, repellent and toxicant, and
trail‐following compounds. Many kinds of chemicals are found to be termiticidal due to
different resources used. These chemicals include ferruginol and manool (Scheffrahn,
1988), nootkatone (Zhu, 2001; Maistrello, 2001), chamaecynone and isochamaecynone
(Saeki, 1973), 2‐phenoxyethanol (Laine, 1998), eugenol (Cornerlius, 1997), Terpene
(Sharma, 1994), α‐terpineol and three sesquiterpene alcohols T‐cadinol, torreyol (δ‐
cadinol), and α‐cadinol (McDaniel, 1989), 7‐methyljuglone, isodiospyrin (Carter, 1978),
cedrol and alpha‐cadinol (Chang, 2001), alpha–pinene, p‐cymene and 1,8‐ cineole (Lin,
1998), and aphthalene (Henderson, 1998).
These compositions may have different effects on different species of termites.
Most of the experiments were carried out with the FST (Chen, 1996; Valles, 2002). Other
species such as Reticulitermes flavipes (Green, 2000), viz. Microcerotermes beesoni,
Neotermes [Kalotermes] bosei and Heterotermes indicola (Sen‐Sarma, 1979; Gupta, 1978;
Jain, 1990), and white termite (O. obesus) (Singh, 2001) collected from sugarcane fields in
India were also employed. (http://etd.lsu.edu/docs/available/etd‐01172004‐
25708/unrestricted/Liu_thesis.pdf)
12
Termites
Termites are light‐colored social insects that form large colonies. Many species live
in warm or tropical regions, feed on wood, and are highly destructive to trees and wooden
structures (www.thefreedictionary.com).
The termites are a group of social insects usually classified at the taxonomic rank of
order Isoptera. As truly social animals, they are termed eusocial along with the ants and
some bees and wasps which are all placed in the separate order Hymenoptera. Termites
mostly feed on dead plant material, generally in the form of wood, leaf litter, soil, or
animal dung, and about 10% of the estimated 4,000 species (about 2,600 taxonomically
known) are economically significant as pests that can cause serious structural damage to
buildings, crops or plantation forests. Termites are major detrivores, particularly in the
subtropical and tropical regions, and their recycling of wood and other plant matter is of
considerable ecological importance.
As eusocial insects, termites live in colonies that, at maturity, number from several
hundred to several million individuals. They are a prime example of decentralized, self‐
organized systems using swarm intelligence and use this cooperation to exploit food
sources and environments that could not be available to any single insect acting alone. A
typical colony contains nymphs (semi‐mature young), workers, soldiers, and reproductive
individuals of both genders, sometimes containing several egg‐laying queens
(http://en.wikipedia.org/wiki/Termite).
13
A colony may number from 100 to more than 1 million termites. Excluding the
immature forms, called nymphs, a colony consists of several structurally differentiated
forms living together as castes with different functions in community life. In socially
advanced species, three principal castes exist: the reproductives, the soldiers, and the
workers. Both the reproductives and the soldiers occur in two or three distinguishable
forms, each specialized for a role in the division of labor in the colony. All forms comprise
individuals of both sexes, but only in the reproductives do the sexual organs undergo
complete development.
Among the reproductives are dark‐colored males and females with fully developed
wings and compound eyes. At maturity, they leave the parental nest in swarms. After the
flight, they shed their wings and mate. A new colony is then established by a male and
female who become primary reproductives, that is, the king and queen, whose sole
occupation is the production of eggs. Termite kings and queens are longer‐lived than other
termites, and the queens are larger than other members of their colony. In certain tropical
species, the king and queen live for ten years, and the queen grows to an enormous size,
sometimes as much as 20,000 times the size of the worker. The abdomen becomes so
distended with eggs that the queen is unable to move about. The eggs are laid at a
prodigious rate that totals about 30,000 a day in some species. Most termite colonies have
only one royal pair.
Apart from the reproductives, all castes are sterile and wingless and have whitish
bodies. Typically, the workers constitute the most numerous castes and are the smallest of
14
the adult forms. Workers build and provision the nest, tend the eggs, and feed and groom
all the other inmates of the community. In some species, no true worker caste exists;
undifferentiated nymphs do the work in the colonies of such species. All species have
soldiers with greatly enlarged heads. The soldiers of certain species are equipped with
huge jaws for defense of the colony; in some species, the soldiers have long snouts, from
which they can eject a sticky, poisonous substance to envelop an enemy and render it
helpless.
Termites feed mainly on wood or other materials containing cellulose. The
cellulose is partially digested by a protozoan ciliate living symbiotically in the intestines of
the worker. Enzymes produced by the protozoan break down the cellulose into
components that can be assimilated by the termites. Digested cellulose is distributed by
the workers to members lacking the protozoan. Some species feed on vegetable molds
that they cultivate. Other species obtain a special fluid secreted by beetles, known as
termitophiles, which live as guests within the termite community.
Termite nests, called termitaries, vary widely. The nests of certain tropical species
are huge mound‐like structures, often 6 m (20 ft) in height. These mounds have extremely
hard walls, constructed from bits of soil cemented with saliva and baked by the sun. Inside
the walls are numerous chambers and galleries, interconnected by a complex network of
passageways. Ventilation and drainage are provided, and heat required for hatching the
eggs is obtained from the fermentation of organic matter, which is stored in the chambers
serving as nurseries (www.encarta.msn.com).
15
Of the more than 55 species common in the United States, majority build their
nests underground. The subterranean termites are extremely destructive, because they
tunnel their way to wooden structures, into which they burrow to obtain food. Given
enough time, they will feed on the wood until nothing is left but a shell.
Termites are classified under Phylum: Arthropoda, Class: Insecta, Subclass:
Pterygota, Infraclass: Neoptera, Superorder: Dictyoptera, Order: Isoptera. Subterranean
termites came from family Termitidae. In the Philippines: Nasutitermes luzonicus Oshima,
Macrotermes gilvus Hagen, and Microcerotermes losbanosensis Oshima. Are the most
common (http://insectscience.org/7.26).
"Higher termites" (Family Termitidae) comprise three quarters of all described
termite species. They differ from "lower termites" (all families except Termitidae) by
having bacteria rather than protozoa in their guts. Many in the deserts of the American
Southwest are important foragers of plant litter, grass and dung. Most are harmless, not
known to cause serious structural damage, even though species of Amitermes feed mostly
on wood ( http://www.utoronto.ca).
Termitidae rely on symbiotic bacteria rather than protozoa (as in all other termite
families, the ‘lower’ termites) for the digestion of cellulose. Most Termitidae inhabit soil
nests. The complex architecture of the large mounds of the Macrotermitinae ensures
ventilation and regulation of the microclimate of the nest. There are five families, with
about 1500 species, occurring world‐wide but mainly in the tropics.(A Dictionary of
Zoology, 1999).
16
Chromolaena odorata
Chromolaena odorata (L.) R.M. King & H. Robinson, is a weed belonging to the
Asteraceae family which has now become so common in the Philippines since its
introduction from South America through the country's southern backdoor in the 1960's.
(CPD Technoguide No. 8, 2003)
C. odorata is a native of South and Central America but thoroughly naturalized in
parts of Africa, India, Ceylon, Indochina, Malaysia and Indonesia. It migrated into the
Philippines some 20 years ago. It might have found its way into the country either through
some air packing materials during the World War 11 or through traders from South
Borneo where it is growing abundantly in borders of abaca plantations
(http://www.ehs.cdu.edu.au).
Hagonoy is classified under Kingdom: Plantae; Division: Magnoliophyta;
Class : Magnoliopsida; Order: Asterales; Family: Asteraceae; Tribe: Eupatorieae; Genus:
Chromolaena.
Locally in the Philippines, the plant is known as Agonoi (Tagalog, Iloko, Bisaya);
Hagonoi (Tagalog, Monobo); Haluhagonoi (Tagalog); Haluka (Tagalog); Salonai (Iloko);
Malasili (Iloko); Larogon (Bagobo); Anoioi (Ivatan); Palunag (Pampangan); Palunai
(Pampangan); Lahunai (Sulu); and Hagonoy or Hagonio in many parts of the country. The
common names in English are devil weed, bitter bush, Chromolaena, jack in the bush, siam
weed, and triffid weed. The other names are verba de Maluco (Spanish); herbe de Laos
17
(French); kasengisil, masigsig (Guam); ngengesil (Palau); Utuot (Chuuk); rumput pirtih
(Indonesian); and siam kraut (German).
It is an economically significant weed because it grows rapidly under any
agricultural situations, in cattle ranches, and under plantations; it is avoided by all
livestock; it causes diarrhea when ingested or even fetal deaths among cows; and it
reduces the carrying capacity of the field to grow more beneficial weeds. (CPD
Technoguide No. 8, 2003).
C. odorata was first noted in Zamboanga and then shortly thereafter in Palawan
and Mindoro. From there, it spread very rapidly northward to Luzon, covering thousands
of acres of range and crop lands per year. Recent survey of the weed showed that it has
spread extensively in Mindanao island (except the northeastern region), in most islands in
the Visayas, Palawan and in areas surrounding Manila. In Mindanao, the spread of the
weed appears to be originating from important port areas of Davao and Zamboanga, and
progressing toward the northeastern region (http://www.ehs.cdu.edu.au).
C. odorata is an herbaceous perennial that forms dense tangled bushes 1.5‐2.0m in
height. It occasionally reaches its maximum height of 6m (as a climber on other plants). Its
stems branch freely, with lateral branches developing in pairs from the axillary buds. The
older stems are brown and woody near the base; tips and young shoots are green and
succulent. The root system is fibrous and does not penetrate beyond 20‐30 cm in most
soils. The flowerheads are borne in terminal corymbs of 20 to 60 heads on all stems and
branches. The flowers are white or pale bluish‐lilac, and form masses covering the whole
18
surface of the bush (Cruttwell and McFadyen, 1989). C. odorata is a big bushy herb with
long rambling (but not twining) branches; stems terete, pubescent; leaves opposite,
flaccid‐membranous, velvety‐pubescent, deltoid‐ovate, acute, 3‐nerved, very coarsely
toothed, each margin with 1‐5 teeth, or entire in youngest leaves; base obtuse or
subtruncate but shortly decurrent; petiole slender, 1‐1.5cm long; blade mostly 5‐12cm
long, 3‐6cm wide, capitula in sub‐corymbose axillary and terminal clusters; peduncles 1‐
3cm long, bracteate; bracts slender, 10‐12mm long; involucre of about 4‐5 series of bracts,
pale with green nerves, acute, the lowest ones about 2mm long, upper ones 8‐9mm long,
all acute, distally ciliate, flat, appressed except the extreme divergent tip; florets all alike
(disc‐florets), pale purple to dull off‐white, the styles extending about 4mm beyond the
apex of the involucre, spreading radiately; receptacle very narrow; florets about 20‐30 or a
few more, 10‐12mm long; ovarian portion 4mm long; corolla slender trumpet form;
pappus of dull white hairs 5mm long; achenes glabrous or nearly so (Stone 1970). The
seeds of Siam weed are small (3‐5mm long, ~1mm wide, and weigh about 2.5mg seed‐1
(Vanderwoude et al., 2005).
Moreover, the proximate, amino acid and phytochemical composition of
Chromolaena odorata was investigated. A high total carbohydrate (20.58% WW and
50.82% DW), crude fibre (10.76% WW and 26.57% DW) and protein (6.56% WW and
16.20% DW) content was observed. The protein is rich in the essential amino acids (with
histidine and phenylalanine being very high) and has a protein score of 88.24% with
methionine as the limiting amino acid. The phytochemical screening revealed the
presence of alkaloids, cyanogenic glycosides, flavonoids (aurone, chalcone, flavone and
19
flavonol), phytates saponins and tannins. The anti‐nutrients composition includes
cyanogenic glycosides (0.05% WW and 0.13% DW), phytates (0.22% WW and 0.54% DW),
saponins (0.80% WW and 1.98% DW) and tannins (0.15% WW and 0.37% DW). Our result
suggests that C. odorata is a source of high quality protein which could serve as a potential
source of protein supplement (Ngozi, et Al., 2009).
The PCA‐Davao Research Center, after having been granted a permit, imported the
stem‐galling fly, Procecidochares connexa Macquart (Diptera : Tephritidae) from Indonesia
ACIAR project CS2 96/91 "Biological Control of Chromolaena odorata in Indonesia, Papua
New Guinea and the Philippines". A series of host specificity tests on ‘hagonoy’ and several
other weed and tree species were conducted under the supervision of the National
Biosafety Committee of the Philippines (NCBP) of the Department of Science and
Technology (DOST). A year of testing showed that the gall fly was highly host specific to
‘hagonoy’. In a parallel project in Indonesia which imported the flies in 1994 and released
in 1995 the flies are now widely spread among Chromolaena in several islands.
The gall fly lays its eggs on practically all of the growing tips of ‘hagonoy’. As much
as 50 eggs are laid by a single female throughout its short lifetime of 1 to 2 weeks. In
about 12 to 15 days the tips begin to swell as manifestation of the activity of the maggots
inside. The gall attains a maximum of 9 by 13 mm in size and each contains from 2 to 10
pupae. Two months after egg laying the flies start to emerge. Egg –laying starts almost
immediately after mating. Death to ‘hagonoy’ is often as a result of extreme pressure
exerted by the fly where almost all shoots develop galls. The final dried‐up ‘hagonoy’ show
galls in every axil and in chains at tips (CPD Technoguide No. 8 Series of 2003).
20
The shrub is reported to be highly allelopathic to nearby vegetation (Muniappan
1994), a fact that has been demonstrated in controlled studies (Sahid and Sugau 1993). It
reduces the diameter growth of teak in infested plantations (Daryono and Hamzah 1979).
It was thought to be useful in the control of Imperata grass and for this reason was
deliberately introduced into the Ivory Coast, but the results were disappointing (Binggeli
1999). Because of the abundance of dead leaves and dry shoots, C. odorata stands are a
fire hazard (Muniappan 1994). Cattle do not eat C. odorata; however, it is browsed by
white‐tailed deer (Meyer and others 1984). In herbal medicine, leaf extracts with salt are
used as a gargle for sore throats and colds. It is also used to scent aromatic baths (Liogier
1990). Extracts of C. odorata have been shown to inhibit or kill Neisseria gonorrhea (the
organism that causes gonorrhoea) in vitro (Caceres and others 1995) and to accelerate
blood clotting (Triratana and others 1991). A satisfactory medium‐density particleboard
was prepared from Christmas bush stems (Kaleta and others 1999). During fallows
between cultivation, Christmas bush adds copious amounts of organic matter to the soil
and may reduce the populations of nematodes (M’Boob, 1991). It is also useful as mulch
for row crops (Swennen and Wilson 1984).
In addition, the allelopathic effects of Chromolaena odorata L. aqueous leaf extract
and residues incorporated in the soil on the growth and water status of Lycopersicon
esculentum Mill were studied. Significant growth reductions in L. esculentum were
observed from additions of C. odorata aqueous leaf extract at concentrations as low as 1g
fresh weight in 40ml of water. Reduction in growth was accompanied by decreases in leaf
water potential. Incorporation of C. odorata leaf material into the soil in which
21
Lesculentum Mill seedlings were germinated and grown caused significant depression in
growth over the 2‐week test period with addition of 2g residue to 80g soil. Allelochemicals
released from C. odorata plants and residues are suggested as a possible explanation for
yield reductions in crops in fields where C. odorata plants are present. One mechanism of
toxic action on seedlings involved interference with water balance.
(http://bioline.utsc.utoronto.ca/archive/00001932/)
Furthermore, essential oil extracts of Chromolaena odorata, and Lantana camara
from their leaves were tested for efficacy on the morality of the maize grain weevil,
Sitophilus zeamais (Coleoptera, Curculionidae). Concentrations of the essential oils relative
to the maize grains of 0.013, 0.025, 0.05 and 0.1% (v/w) were used for A. conyzoides and
0.063, 0.125, 0.25 and 0.50% (v/w) for C. odorata and L. camara. Twenty 7‐day old adult
weevils were fed on maize grains treated with the above concentrations of the essential
oils in Petri dishes. Control dishes contained insects and maize grains without essential
oils. The experiment was repeated three times. Dishes were incubated in the laboratory
for 7 days at 26°C and 75‐‐85% relative humidity. Insect mortality was recorded every 24
h. Graphs of percentage mortality versus the duration of exposure were constructed and
the LD50 was computed for each oil. Significant insect mortality was obtained with all the
essential oils used. The mortality of S. zeamais increased with the concentration of the
essential oils of the three plants and the duration of exposure of the weevils on the
treated substrates. The essential oil extract of Ageratum conyzoides was the most
effective insecticide (LD50 = 0.09% in 24 h), followed by that of L. camara (LD50 = 0.16%)
and C. odorata (LD50 = 6.78%). These results show that the essential oils of the leaves of
22
some of these weed species may be exploited for insect control in stored products
(http://cat.inist.fr/?aModele=afficheN&cpsidt=855900).
Furthermore, studies on Chromolaena odorata oils from the flowers and leaves
showed toxicity to mosquitoes, although the extent of toxicity varied from each oil. Lt50
determined for the C. odorata flower and leaf oils used on the larvae gave 10.6 minutes
and 12.0 minutes respectively. Adult treatments also gave the LD50 of 2.5 and 4.0 minutes
for the flower and leaf oils respectively. The C. odorata oils were effective against both the
larvae and adults of the mosquitoes and their effects found to be persistent. The oil
extracts therefore have good prospects of being developed as insecticides for mosquito
control (http://esa.confex.com/esa/2004/techprogram /paper_14489.htm).
Solignum
Solignum, the No. 1 wood preservative that provides sure protection against
termites (anay), woodborers (bukbok) and fungi (amag). Solignum has been tried, tested
and trusted in the Philippines for over 50 years now and is registered under Fertilizer and
Pesticide Authority (FPA). Solignum is available in two variants: Solignum Colorless and
Solignum Brown (http://www.jardinedistribution.com/construction.php ?cid=10).
According to Fertilizer and Pesticide Authority (FPA), the major termiticidal
component of Solignum is permethrin. (http://fpa.da.gov.ph/List%20of%20Registered%2
0Agricultural%20Pesticide%20Products.pdf) Permethrin is a common synthetic chemical,
widely used as an insecticide and acaricide and as an insect repellent. It belongs to the
family of synthetic chemicals called pyrethroids and functions as a neurotoxin, affecting
neuron membranes by prolonging sodium channel activation. It is not known to harm
23
most mammals or birds. It generally has a low mammalian toxicity and is poorly absorbed
by skin. In agriculture, permethrin is mainly used on cotton, wheat, maize, and alfalfa
crops, and is also used to kill parasites on chickens and other poultry. It is also extensively
used in Europe as a timber treatment against wood boring beetle (woodworm). Its use is
controversial since, as a broad‐spectrum chemical, it kills indiscriminately; as well as the
intended pests, it can harm beneficial insects including honey bees, aquatic life, and small
mammals such as mice. Permethrin is toxic to cats and many cats die each year after being
given flea treatments intended for dogs, or by contact with dogs that have recently been
treated with permethrin.
Permethrin is also used in healthcare, to eradicate parasites such as head lice and
mites responsible for scabies, and in industrial and domestic settings to control pests such
as ants and termites.
Permethrin kills ticks on contact with treated clothing. According to the
Connecticut Department of Public Health, it "has low mammalian toxicity, is poorly
absorbed through the skin and is rapidly inactivated by the body. Skin reactions have been
uncommon.”
Permethrin is used in tropical areas to prevent mosquito‐borne disease such as
dengue fever and malaria. Mosquito nets used to cover beds may be treated with a
solution of permethrin. This increases the effectiveness of the bed net by killing parasitic
insects before they are able to find gaps or holes in the net. Malaria kills 1‐3 million people
per year, and permethrin is believed to have very low if any toxicity in humans. Military
24
personnel training in malaria‐endemic areas may be instructed to treat their uniforms with
permethrin as well. An application should last several washes.
Recently, in South Africa, residues of permethrin were found in breast milk,
together with DDT, in an area that experienced DDT treatment for malaria control, as well
as the use of pyrethroids in small‐scale agriculture.
Permethrin is extremely toxic to fish. Extreme care must be taken when using
products containing permethrin near water sources. Permethrin is also highly toxic to cats.
Flea and tick repellent formulas intended (and labeled) for dogs may contain permethrin
(like k‐9 Advantix and Advantage Multi for Dogs) and cause feline permethrin toxicosis in
cats: specific flea and tick control formulas intended for feline use, such as those
containing fipronil, should therefore be used for cats instead.
Permethrin is classified by the US EPA a likely human carcinogen, based on
reproducible studies in which mice fed permethrin developed liver and lung tumors.
Carcinogenic action in nasal mucosal cells for inhalation exposure is suspected due to
observed genotoxicity in human tissue samples, and in rat livers the evidence of increased
preneoplastic lesions lends concern over oral exposure (http://en.wikipedia.org/
wiki/Permethrin).
Chapter III
METHODOLOGY
This chapter deals with the materials, methods, and procedures used in the
preparation of the different test substances and its application to termites. It also includes
the data gathered and the statistical analysis of the data. Research flowchart is also
presented in Figure 3.
Materials
Plant material – Chromolaena odorata leaves collected from Aringay, La Union
Test organism – termites
Laboratory apparatus – triple beam balance, spatula, stirring rod, beaker,
Erlenmeyer flask, graduated cylindeccr, petri dish, scissor, timer, blender, rotary evaporator,
desiccators, sprayer, clean containers
Chemicals ‐ chemical termiticide (Solignum), technical grade methanol
Others – distilled water
Research Design
The study did not use any design for the reason that studies such as this one,
concerned treatment of the experimental animals done by batches or done one after the
other and not at the same time.
In line with this, this study utilized the experimental method of research using
standard laboratory diagnostic procedures involved in termiticidal potential of hagonoy.
26
The study used five treatments, which are as follows:
a. T1 – positive control, commercial pesticide (Solignum)
b. T2 – leaf decoction from C. odorata
c. T3 – expressed leaf juice from C. odorata
d. T4 – 30% C. odorata leaf extract solution
e. T5 – 60% C. odorata leaf extract solution
Collection and Preparation of Plant Material
Fresh leaves of C. odorata were removed from the plant. Only leaves without
adhering dirt particles or associated with insect or insect bites were used. The leaves were
washed by running water.
Preparation of Decoction from C. odorata Leaves
About 100 grams of dried C. odorata leaves were boiled in 250 mL water. Boiling was
done for about 15 minutes. Afterwards, the mixture is allowed to cool then placed in a clean
container. Eventually, the mixture was filtered using filter paper. The filtrate collected was
the decoction used in the treatment.
Preparation of Expressed Juice
About 100 grams of fresh C. odorata leaves were pounded using mortar and pestle
and squeezed using a clean cheese cloth. The obtained juice was filtered using filter paper.
Finally, the expressed leaf juice was placed in a clean container.
27
Preparation of the Methanol Leaf Extract
Fresh C. odorata leaves were dried, cut into small pieces, and homogenized using a
blender. The ground leaves were soaked in technical grade methanol for two days with
occasional stirring. The volume of solvent was just enough to immerse the material. The
methanol extract was filtered through a filter paper. After filtration, the methanol was
removed by concentrating at 400C in vacuo using a rotary evaporator, and kept in a
dessicator at room temperature to remove the moisture of the specimen.
Test Organism
A total of 225 subterranean termites in more or less uniform in sizes were collected
from Agoo, La Union. The researchers dug up termitaries and collected the termites
including the soil where they thrived on and placed in a box. After the collection, the test
organisms were separated from the soil and placed them in a petri dish. Each petri dish
contained 15 termites.
Application of the Test Substances
Five (5) treatments namely T1 – positive control, commercial pesticide (Solignum); T2
‐ leaf decoction from C. odorata, T3 ‐ expressed leaf juice from C. odorata, T4 ‐ 30% C.
odorata leaf extract solution, and T5 ‐ 60% C. odorata leaf extract solution were utilized in
the study. The test substances were sprayed once to the sample termites by using sprayer.
28
These doses were administered once for each treatment. The termiticidal effects
were observed as to the length of time the termites are killed. The treatment was
replicated three (3) times.
Data Gathered
The following data was collected during the course of the study:
The length of time the termites were killed. The data on the length of time the
termites were killed after the application of the test substances was determined using timer
in minutes. This was done for every replication.
Data Analysis
The data gathered were tabulated and computed using the following statistical
tools:
1. Analysis of Variance (ANOVA) to determine whether significant differences exist
in the length of time the termites are killed among the five treatments; and
2. Scheffé's Test to find out where the difference lies and determine which among
the five treatments has the greatest termiticidal potential.
29
Preparation and collection of Preparation of various test
resources used in the study: substances:
a. plant material a. Positive control
b. test organisms b. leaf decoction from
c. laboratory apparatuses hagonoy
and equipment c. expressed leaf juice from
d. chemicals, etc. hagonoy
d. methanol extracts
Gathering of data:
a. length of time for the test Application of the various test
organisms to die as substances to the test
affected by various test organisms (termites)
substances
Analyzing the data gathered
using the following statistical
tools:
a. Analysis of Variance
b. Scheffé's Test
Figure 4. The Research Flowchart depicting chronologically the methods and procedures
used in the study.
Chapter IV
RESULTS AND DISCUSSION
This chapter presents the analysis, discussion, and interpretation of the data
gathered from the test organisms that were treated with the Chromolaena odorata leaf
decoction, expressed juice, and various concentration levels. It also discusses the findings in
relation to the problems in Chapter I.
Table 1. Length of time in minutes for the termites to die due to various treatments
Replication Treatment
Treatment 1 2 3 Mean
T1 1.43 1.48 1.45 1.45
(positive control, Solignum)
T2 1.73 1.83 1.75 1.77
(leaf decoction from C. odorata)
T3 2.05 2.25 2.15 2.15
(expressed leaf juice from C. odorata)
T4 1.33 1.58 1.50 1.47
(30% C. odorata leaf extract solution)
T5 1.18 1.42 1.28 1.29
(60% C. odorata leaf extract solution)
GRAND MEAN 1.626
Table 1 presents the data on the length of time in minutes for the sample termites
to die upon application of the various treatments. The results reveal that the sample
termites in T5 (60% Chromolaena odorata leaf extract solution) has the shortest length of
time with a mean of 1.29 minutes. This is followed by T1 (positive control, Solignum) with a
31
mean 1.45 minutes, T4 (30% Chromolaena odorata leaf extract solution) with a mean of
1.47, and T2 (leaf decoction from Chromolaena odorata) with a mean of 1.77. The treatment
with the longest time for the termites to die is T3 (expressed leaf juice from Chromolaena
odorata) with a mean of 2.15 minutes. The shorter the number of mean time, the higher its
termiticidal potential, so this implies that the T5 (60% Chromolaena odorata leaf extract
solution) which has the lowest mean is the most effective and it has the greatest
termiticidal potential among the various treatments.
2.5
2
1.5
Length of time in R1
minutes
1 R2
R3
0.5
0
1 2 3 4 5
Treatment
Figure 5. Length of time in minutes for the termites to die due to various treatments
32
Table 2. One‐Way Analysis of Variance (ANOVA) Summary Table
Source of Degrees of Sum of Squares Mean Square F‐ratio
Variation Freedom
Within 4 1.39 0.35 38.89*
Treatments
Between 10 0.09 0.009
Treatments
TOTAL 14 1.48
Legend: *significant at ∞ = .05 CV = 5.83 %
The One‐Way Analysis of Variance is used to compare the means of the five
treatments is presented in Table 2. The F‐test shows that there are significant differences
among the means of the five treatments at 0.05 significance level. This implies that there
are treatments that are more efficient than the others in terms of the length of time for the
sample termites to die upon application of such treatment. Since the Coefficient of
Variation (CV) is 5.83 %, the study is highly reliable. Hence, to determine where the
difference lies, was further tested using Scheffé's Test.
Furthermore, Table 3, on the next page, depicts that all pairwise comparisons exhibit
significant differences with the exception of T1, the positive control and T4, 30% C. odorata
leaf extract solution. This implies that T1 and T4 have comparable effect in terms of the
length of time it takes the sample termites to die. Thus, the application of the positive
control, Solignum (T1) and the application of 30% C. odorata leaf extract solution (T4) are
equally efficient in killing termites. Furthermore, since all comparisons with T5 which has the
33
lowest mean time, are significantly different, then it can be derived that T5 (60% C. odorata
leaf extract solution) is the best among the five treatments.
Table 3. Pairwise Comparison of the Mean Time using Scheffé's Test
Between Treatments F’ F(.05)(k‐1) ∞ = .05
T1 vs. T2 1 896.30 13.44 significant
T1 vs. T3 9 074.10 13.44 significant
T1 vs. T4 7.41 13.44 not significant
T1 vs. T5 474 13.44 significant
T2 vs. T3 2 674 13.44 significant
T2 vs. T4 1 666.7 13.44 significant
T2 vs. T5 4 266.7 13.44 significant
T3 vs. T4 8 563 13.44 significant
T3 vs. T5 13 696.3 13.44 significant
T4 vs. T5 600 13.44 significant
Conclusively, the null hypotheses, indicating that there are no significant differences
on the length of time the termites are killed upon the application of the five treatments,
and between the pairs of treatments, are rejected. This is because each treatment is
significantly different from one another. This strongly indicates that 60% C. odorata
methanol leaf extract solution is the best among the five treatments however; the positive
control and 30% C. odorata methanol leaf extract solution are equally efficient.
Chapter V
SUMMARY, CONCLUSIONS, AND RECOMENDATIONS
This chapter presents a summary of the experimental study, the drawn conclusions,
and the offered recommendations.
Summary
This study aimed to analyze and determine the termiticidal potential of
Chromolaena odorata (L.) King & Robinson (Hagonoy).
It study utilized the experimental method of research which focused on the use
standard laboratory diagnostic procedures implicated in termiticidal potential of Hagonoy in
terms of the length of time for the termites to die upon application of the various
treatments.
The study was conducted at DMMMSU ‐ South La Union Campus, Agoo, La Union
from December 2008 to February 2009. Crude extraction of the termiticidal components
was done at the Bureau of Plant Industry (BPI) – Baguio Pesticide Analytical Laboratory
(BPAL), Baguio City. Verification of the plant material was done at the Botany Division of the
National Museum, Manila.
The methods used were collection and preparation of plant material, the
preparations on decoction from Chromolaena odorata leaves, expressed juice, methanol
extract, and the application of the various substances to the termites.
Specifically, this study tested if there were significant differences in the length of
time the termites were killed among and between the five treatments which were T1 –
positive control, commercial pesticide (Solignum), T2 ‐ leaf decoction from C. odorata, T3 ‐
35
expressed leaf juice from C. odorata, T4 ‐ 30% C. odorata leaf extract solution, and T5 ‐ 60%
C. odorata leaf extract solution. The extracting solvent used was technical grade methanol.
Thus, the components that could only be extracted with methanol were considered. For the
treatments that used expressed juice and decoction, distilled water was used as the solvent.
The data gathered were tabulated and compute using One‐Way Analysis of Variance
to determine whether significant differences exist in the length of time the termites are
killed after the exposure to the five treatments. Scheffé's Test was used to find out where
the difference lies among the five treatments had the greatest termiticidal potential.
The study revealed that there were significant differences in the length of time the
termites were killed after exposure to the various treatments and between the pair of
treatments. The data gathered showed that sample termites in T5 (60% Chromolaena
odorata leaf extract solution) had the least length of time with a mean of 1.29 minutes.
This was followed by T1 (positive control, Solignum) with a mean 1.45 minutes, T4 (30%
Chromolaena odorata leaf extract solution) with a mean of 1.47, and T2 (leaf decoction from
Chromolaena odorata) with a mean of 1.77. The treatment with the longest time for the
sample to die was T3 (expressed leaf juice from Chromolaena odorata) with a mean of 2.15
minutes.
The result of the One‐Way Analysis of Variance was used to compare the means of
the five treatments. The F‐test showed that there are significant differences among the
means of the five treatments at 0.05 significance level. This implied that there were
treatments that are more efficient than the others in terms of the length of time for the
sample termites to die upon application of such treatment. The Scheffé's Test implies that
36
T1 and T4 had comparable effects in terms of the length of time it took the sample termites
to die. Furthermore, since all comparisons with T5 (60% Chromolaena odorata leaf extract
solution) which had the lowest mean time are significantly different, it was concluded that
T5 was the best among and the most effective the five treatments. Besides, T5 (60% C.
odorata leaf extract solution was the best substitute for Solignum, the positive control, for
the reason that C. odorata is environment and health friendly in contrast with Solignum.
Conclusions
Within the limits of the study, the following conclusions were drawn:
1. Based on the results of the study in terms of the length of time for the termites to
die, 60% Chromolaena odorata leaf extract solution is effective in killing the sample
termites with the shortest length of time. The shorter the number of mean time, the
higher its termiticidal potential. However, the positive control (Solignum) and 30%
Chromolaena odorata leaf extract solution have comparable effect in terms of the
length of time it takes for the sample termites to die. This indicates that the T1 and
T4 are not significantly different from each other which implies that Solignum and
30% C. odorata leaf extract solution have the same effect (same length of time) for
the termites to die.
2. In terms to the length of time for the termites to die, there are significant
differences between the pairwise comparisons of T1 (positive control, Solignum) vs.
T2 (leaf decoction from C. odorata) , T1 (positive control, Solignum) vs. T3 (expressed
leaf juice from C. odorata), T1 (positive control, Solignum) vs. T5 (60% C. odorata leaf
extract solution), T2 (leaf decoction from C. odorata) vs. T3 (expressed leaf juice from
37
C. odorata), T2 (leaf decoction from C. odorata) vs. T4 (30% C. odorata leaf extract
solution) , T2 (leaf decoction from C. odorata) vs. T5 (60% C. odorata leaf extract
solution), T3 (expressed leaf juice from C. odorata) vs. T4 (30% C. odorata leaf extract
solution), T3 (expressed leaf juice from C. odorata) vs. T5 (60% C. odorata leaf extract
solution), and T4 (30% C. odorata leaf extract solution) vs. T5 (60% C. odorata leaf
extract solution). However, the positive control (Solignum) and 30% C. odorata leaf
extract solution are comparable with each other. Based from the results, leaf
decoction, expressed leaf juice, and the different extract solutions from C. odorata
are efficient in terms of their termiticidal potential.
3. The positive control and the 30% C. odorata methanol leaf extract solution have
comparable effect in terms of the length of time it takes for the sample termites to
die. Thus, the application of the positive control, Solignum and the application of
30% C. odorata leaf extract solution are equally efficient in killing termites.
Furthermore, since all comparisons with T5 which has the lowest mean time are
significantly different, then it can be derived that T5 is the best and the most
effective in eradicating termites among the five treatments.
Recommendations
According to results of the study, the following recommendations were offered:
1. In view of the fact that there was a great termiticidal potential of Chromolaena
odorata, which has a comparable effects to the commercial chemical termiticides, it
is recommended that this plant be used as the best alternative termiticide for it is
health and environment friendly.
38
2. While leaf decoction, expressed leaf juice, and the different concentration levels of
Chromolaena odorata have almost the same efficacy, decoction is better to use for
practical reasons. And it is more economical to use and is easy to be obtained by the
people especially those living in far‐flung areas and places that cannot be reached by
modern products.
3. Additional studies should utilize the same preparations of Chromolaena odorata to
investigate its effects on other type of pest as well as plants.
4. Further studies should be conducted by oil extraction and fractionation of the
various chemical constituents of Chromolaena odorata to analyze which of these
substances have the potential to cause such termiticidal effect.
5. Other researchers may employ the similar arrangement and set‐up of treatments of
the study using variety of plants in analyzing and determining their potential not
only as termiticide but also another type of pesticide.
6. Ultimately, a great termiticidal potential of Chromolaena odorata was found out. For
this reason, the researchers recommend the patenting of termiticidal product out of
Chromolaena odorata which is efficient, cheap, economical, organic, and
environment and health friendly.
39
REFERENCES
A Dictionary of Zoology 1999, Oxford University Press 1999. ISBN: 0192800760
http://www.encyclopedia.com/doc/1O8‐Termitidae.html
Acda, M. 2007 Toxicity of Thiamethoxam Against Philippine Subterranean Termites. 6pp.
Journal of Insect Science 7:26, available online: insectscience.org/7.26
Ahiati, JT. 2004. A Study of the Insecticidal Effects of Chromolaena odorata Oil Extracts
on the Larvae and Adults of Mosquitoes (Family Culicidae)
http://esa.confex.com/esa/2004/techprogram /paper_14489.htm
Aterrado E.D.,Sanico R.L, Status of Chromolaena Odorata Research in the Philippines.
Davao Research Center, Davao City, Philippines & Visayas State College of
Agriculture, Baybay, Leyte 6521‐A, Philippines. http://www.ehs.cdu.edu.au/
chromolaena/proceedings/first/Status%20of%20Chromolaena%
20odorata%20research%20in%20the%20Philippines.htm
Bamikole, M.A et. Al. 2004. Converting Bush to Meat: A Case of Chromolaena odorata
Feeding to Rabbits. Department of Animal Science, University of Benin, Nigeria
Bouda, H. (et al). Effect of Essential Oils from Leaves of Ageratum conyzoides, Lantana
camara and Chromolaena odorata on the Mortality of Sitophilus zeamais
(Coleoptera, curculionidae).
http://cat.inist.fr/?aModele=afficheN&cpsidt=855900
Broto, Antonio S. 2006. Statistics Made Simple. Second Edition. National Bookstore,
Quezon City, Philippines.
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Chromolaena odorata. http://www.ehs.cdu.edu.au/ chromolaena/proceedings/first/
Status%20of%20Chromolena%20odorata%20research%20in%20the%20Philippin
es.htm
Chromolaena odorata.www.fs.fed.us/global/iitf/pdf/shrubs/
Chromolaena%20odoratum.pdf
Chromolaena odorata. http://www.issg.org/database/species/ecology.asp?fr=1&si =47
Eleyinmi, Afolabi. 2007. Chemical composition and antibacterial activity of Gongronema
latifolium .Department of Agricultural, Food and Nutritional Sciences, University
of Alberta, Edmonton T6G 2P5, Canada
Enyi, J., 2001. Allelopathic Effects of Chromolaena Odorata L. (R. M. King and Robinson –
(Awolowo Plant’ Toxin on Tomatoes (Lycopersicum esculentum Mill)
http://biblioteca.universia.net/html_bura/ficha/params/id/3920514.html
Guidebook to Grassland Plants (A Resource Material for Biology Teachers). 2003
Foundation for the Advancement of Science Education,Inc., Science
Education Center, University of the Philippines Press, Diliman, Quezon City.
List of Registered Agricultural Pesticide Products as of 31 Dec 2007. 2008 PRSD.
Fertilizer and Pesticide Authority, FPA Bldg. B.A.I. Compound Visayas Ave. Diliman,
QuezonCity.http://fpa.da.gov.ph/List%20of%20Registered%20Agricultural%
20Pesticide%20Products.pdf
Ngozi, Igboh, Jude, Ikewuchi and Catherine, Ikewuchi. 2009. Chemical Profile of
Chromolaena odorata L. (King and Robinson) Leaves. Pakistan Journal of
Nutrition, University of Port Harcourt, Department of Biochemistry, Port
Harcourt, Nigeria
41
Plants. http://encarta.msn.com
Padilla, et. Al., Chromolaena odorata as Termite Eradicator. 2006, Agoo Kiddie Special
School, Agoo, La Union
Primer on Biological Control of Hagonoy (Chromolaena odorata).CPD Technoguide No.8,
2003. Crop Protection Division, Davao Research Center,Agricultural Research &
Development Branch, Philippine Coconut Authority,Bago‐Oshiro, Davao City.
Quisumbing, E. 1978. Medicinal Plants of the Philippines, Katha Publishing Co., Inc.
Quezon City, Philippines.
Termite. http://en.wikipedia.org/wiki/Termite
Termites. www.encarta.msn.com
Termites. www.thefreedictionary.com
Permethrin. http://en.wikipedia.org/wiki/Permethrin.
Protect Your Wood, Preserve Your Home. http: /www. jardinedistribution.com/
construction.php ?cid=10.
Yaojian, L. 2004. Study on the Termiticidal Components of Juniperus virginiana,
Chamaecyparis nootkatensis and Chamaecyparis lawsoniana.
http://etd.lsu.edu/docs/available/etd‐01172004‐
225708/unrestricted/Liu_thesis.pdf
42
EXPERIMENTAL PLATES
Plate 1. Chromolaena odorata in its natural environment
(from http://en.wikipedia.org/wiki/File:Chromolaena_odorata.jpg)
Plate 2. Fresh leaves of Chromolaena odorata
43
Plate 3. The experimental set‐up
Plate 4. The separation of test organism from the soil
44
Plate 5. The test organisms placed on the dish
Plate 6. Decoction of Chromolaena odorata leaves
45
Plate 7. Pounding of fresh Chromolaena odorata leaves
Plate 8. Air dried Chromolaena odorata leaves
46
Plate 9. Preparation of Extract
Plate 10. Laboratory personnel of the Bureau of Plant Industry – National Pesticide
Analytical Laboratory performing the Rotavation
47
Plate 11. The test organisms administered by positive control (Solignum)
Plate 12. The test organisms administered by leaf decoction from hagonoy
48
Plate 13. The test organism administered by expressed juice from hagonoy
Plate 14. The test organism administered by 30% leaf extract solution
49
Plate 15. The test organisms administered by 60% leaf extract solution
Plate 16. The test organisms as affected by the various treatments
50
Plate 17. The researchers introducing the various test substances to the termites
Plate 18. Researcher of the National Museum, Botany Division determining the plant
material used in the study
APPENDICES
Appendix A. Sample Computation for Methanol Leaf Extract Solutions
I. 30% Methanol Leaf Extract Solution
100 mL hagonoy methanol leaf extract solution
= 100 mL × 30%
= 100 mL × .30
= 30 mL pure hagonoy methanol leaf extract
= 100 mL – 30 mL
= 70 mL distilled water
= 70 mL distilled water + 30 mL pure hagonoy methanol leaf extract
II. 60% Methanol Leaf Extract Solution
100 mL hagonoy methanol leaf extract solution
= 100 mL × 60%
= 100 mL × .60
= 60 mL pure hagonoy methanol leaf extract
= 100 mL – 60 mL
= 40 mL distilled water
= 40mL distilled water + 60mL pure hagonoy methanol leaf extract
52
Appendix B. Statistical Formula
I. Analysis of Variance
1. Degrees of freedom (df)
dfwithin = total number of treatments -1
dfbetween = total df – dfwithin – replication df
Total df = total number of observations – 1
2. Sum of Squares (SS)
Correlation Factor (CF)
( grand total ) 2
CF =
total number of observatio ns
Total Sum of Squares = ∑X 2
−CF
Block Sum of Squares = ∑ (treatment total ) 2
− CF
replication
Sum of Squares between = Total SS – Block SS – SSwithin
3. Mean Square (MS)
SS within
MSwithin =
Treatment − 1
SS between
MSbetween =
t ( r − 1)
53
4. Observed F – Value
MSwithin
Observed F (within) =
MSbetween
5. Coefficient of Variation (CV)
MS between
CV = × 100
grand mean
II. Scheffé's Test
(T1 − T2 ) 2
F'= SW 2 ( n1 +n2 )
n1n2
where:
T1 = Mean of 1st sample
T2 = Mean of 2nd sample
SW= MSbetween
n1= Total of 1st sample
n2= Total of 2nd sample
54
Appendix C. Sample Computation of ANOVA and Scheffé’s Test
I. Analysis of Variance (ANOVA)
A. Degrees of Freedom (df)
df between = 5-1 = 4
df within = 10
Total df= 15 – 1 = 14
B. Sum of Squares (SS)
( grand total ) 2 ( 24 .41) 2 595 .85
CF = = =
total number of observatio ns 15 15
CF = 39.72333333
Total SS = ∑x 2
− CF
Total SS = 1.48
SSbetween= ∑ (treatment total) 2
− CF
SSbetween= 0.09
SSwithin= total SS – SSbetween
SSwithin= 41.11 – 37.72 = 1.39
55
C. Mean Square (MS)
SS within 1.39
MSwithin= =
t −1 (5 − 1)
MSwithin= 0.35
SS between .09
MSbetween= =
t ( r − 1) 5(3 − 1)
MSbetween= 0.009
MS within 0.35
F – value = =
MSbetween 0.009
F – value = 38.89
MS between
CV= × 100
Grand Mean
0.009
CV= × 100
1.626
CV= 5.83%
56
II. Scheffé's Test
(T1 − T2 ) 2
F'= SW 2 ( n1 +n2 )
n1n2
1. T1 vs. T2
(1.45 − 1.77) 2 0.1024
F '= 2
= = 1,896 .30
(.009) (6) 0.000054
9
2. T1 vs. T3
(1.45 − 2.15) 2 0.49
F '= 2
= = 9,074 .10
(.009 ) (6) 0.000054
9
3. T1 vs. T4
(1.45 − 1.47 ) 2 0.0004
F'= 2
= = 7.41
(.009) (6) 0.000054
9
4. T1 vs. T5
(1.45 − 1.29) 2 0.0256
F'= 2
= = 474 .00
(.009) (6) 0.000054
9
5. T2 vs. T3
(1.77 − 2.15) 2 0.1444
F'= 2
= = 2,674
(.009 ) (6) 0.000054
9
57
6. T2 vs. T4
(1.77 − 1.47 ) 2 0.09
F'= 2
= = 1,666 .70
(.009 ) (6) 0.000054
9
7. T2 vs. T5
(1.77 − 1.29) 2 0.2304
F '= 2 = = 4,266.70
(.009) (6) 0.000054
9
8. T3 vs. T4
(2.15 − 1.47) 2 0.4624
F '= 2 = = 8,563 .00
(.009) (6) 0.000054
9
9. T3 vs. T5
(2.15 − 1.29) 2 0.7396
F '= 2 = = 13,696 .30
(.009) (6) 0.000054
9
10. T4 vs. T5
(1.47 − 1.29) 2 0.0324
F '= 2
= = 600 .00
(.009) (6) 0.000054
9
58
Appendix D. Tables
Table 1. Length of time in minutes for the termites to die due to various treatments
Replication Treatment
Treatment 1 2 3 Mean
T1 1.43 1.48 1.45 1.45
(positive control, Solignum)
T2 1.73 1.83 1.75 1.77
(leaf decoction from C. odorata)
T3 2.05 2.25 2.15 2.15
(expressed leaf juice from C. odorata)
T4 1.33 1.58 1.50 1.47
(30% C. odorata leaf extract solution)
T5 1.18 1.42 1.28 1.29
(60% C. odorata leaf extract solution)
GRAND MEAN 1.626
Table 2. One‐Way Analysis of Variance (ANOVA) Summary Table
Source of Degrees of Sum of Squares Mean Square F‐ratio
Variation Freedom
Within 4 1.39 0.35 38.89*
Treatments
Between 10 0.09 0.009
Treatments
TOTAL 14 1.48
Legend: *significant at ∞ = .05 CV= 5.83%
59
Table 3. Pairwise Comparison of the Mean Time using Scheffé's Test
Between Treatments F’ F(.05)(k‐1) ∞ = .05
T1 vs. T2 1 896.3 13.44 significant
T1 vs. T3 9 074.1 13.44 significant
T1 vs. T4 7.41 13.44 not significant
T1 vs. T5 474 13.44 significant
T2 vs. T3 2 674 13.44 significant
T2 vs. T4 1 666.7 13.44 significant
T2 vs. T5 4 266.7 13.44 significant
T3 vs. T4 8 563 13.44 significant
T3 vs. T5 13 696.3 13.44 significant
T4 vs. T5 600 13.44 significant
60
Appendix E. Figures
PROBLEM TREE
Structural Damage and
Additional Expenses to
Repair Damaged
Properties Use of Expensive
Chemical Termiticide
TERMITES
Lack of Knowledge of
Destructive
Plant Alternative as
Termiticide
Figure 1. Problem Tree
61
PROBLEM TREE
Risk to Human
and Animal Health
Expensive
Pesticide Toxicity to Natural Enemies
Resistance and Other Non-target
Organisms
DEPENDENCE
ON
INSECTICIDES
Highly Easy to Use
Effective
Readily Lack of Knowledge on
Available its Impacts to Health
and Environment
Figure 2. Problem Tree
62
INDEPENDENT VARIBLE DEPENDENT VARIABLE
Treatment: Length of time the termites are
killed after the exposure to:
a. T1 ‐ positive control,
(Solignum) a. leaf decoction from
hagonoy
b. T2‐ leaf decoction from
C. odorata b. expressed leaf juice
from hagonoy
c. T3‐ expressed leaf juice
from C. odorata c. methanol leaf extract
solutions
d. T4‐ 30% C. odorata leaf
extract solution
e. T5‐ 60% C. odorata leaf
extract solution
Figure 3. Conceptual Paradigm
63
Preparation and collection of Preparation of various test
resources used in the study: substances:
a. plant material a. Positive control
b. test organisms b. leaf decoction from
c. laboratory apparatuses hagonoy
and equipment c. expressed leaf juice from
d. chemicals, etc. hagonoy
d. methanol extract
Gathering of data:
a. length of time for the test Application of the various test
organisms to die as substances to the test
affected by various test organisms (termites)
substances
Analyzing the data gathered
using the following statistical
tools:
a. Analysis of Variance
b. Scheffé's Test
Figure 4. The Research Flowchart
64
2.5
2
1.5
Length of time in R1
minutes
1 R2
R3
0.5
0
1 2 3 4 5
Treatment
Figure 5. Length of time in minutes for the termites to die due to various treatments
65
Appendix F. Letter to the Senior Museum Researcher of the National Museum
66
Appendix G. Certification of Plant Material
67
Appendix F. Letter to the Chief, Baguio PAL Satellite‐BNCRDC
68
Appendix I. Certificate of Appearance (issued by the Chief, Baguio PAL Satellite‐BNCRDC)