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Researchers:




Research Adviser:
     TESSIE Q. PERALTA, Ph. D.

 Don Mariano Marcos Memorial State University
                         South La Union Campus
                            College of Education
                     LABORATORY HIGH SCHOOL
                                 Agoo, La Union

 2009 CHEMREZ Green Chemistry Award [SIBOL Category]
THE PROBLEM &
ITS BACKGROUND


                 4
5
Chromolaena odorata (HAGONOY)

> a weed belonging to the Asteraceae family

> occurs in agricultural areas, natural
forests, planted forests, range /grasslands,
riparian zones, ruderal /disturbed, scrub/
shrublands

> considered invasive weed of field crops in
its introduced range, and has been reported
to be the most problematic invasive species

> reduces the carrying capacity of the field
to grow more beneficial weeds (allelophatic)

> causes diarrhea when ingested or even
fetal deaths among cows

> contains carcinogenic pyrrolizidine
alkaloids (hepatotoxic-potentially damaging
to the liver).
                                                The bright green regions indicate where
                                               Chromolaena has invaded forest gaps and
                                                                                   6
                                                         replaced the forest.
TERMITES
> group of social insects usually classified
at the taxonomic rank of order Isoptera

> prefer to feed on dead plant material,
wood, leaf litter, or soil


> live in colonies; prime example of
decentralized, 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

> typical termite colony contains nymphs
(semi-mature young), workers, soldiers, and
reproductive individuals of both sexes,
sometimes containing several egg-laying
queens [insect castes]
4 Major Termite Species in the Philippines
       Nasutitermes luzonicus
       (Luzon Point Headed termite)


       Coptotermes vastator
       (Philippine Milk termite)


       Microcerotermes losbanosensis
       (Los Banos termite)


       Macrotermes gilvus
       (Mound Building termite)
                                             8
Additional Expenses
                                         to Repair Damaged
Structural Damage                            Properties




                    TERMITES




   Destructive                         Use of Expensive
                                     Chemical Termiticide


                 Lack of Knowledge of
                  Plant Alternative as
                      Termiticide
                                                               9
10
Risk to Public Health
           and Environment                                     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
                                                        its Impacts to Health
                    Available
                                                          and Environment
                                                                           11
Objectives of the Study

1. To test termiticidal potential of C. odorata
   in terms of the length of time the termites
   are killed after the application of the five
   treatments.
2. To determine the significant differences in
   the length of time the termites are killed
   when exposed to the five treatments.
3. To analyze the significant differences in
   the length of time the termites are killed
   between pairs of treatments.
                                                12
Null Hypotheses
The following statements were tested experimentally
   at .05 level of significance:
1. There are no significant differences in the
    length of time the termites are killed after
    the application of the five treatments.

2. There are no significant differences in the
    length of time the termites are killed
    between pairs of treatments.
                                                  13
Conceptual Paradigm
INDEPENDENT VARIABLE   DEPENDENT VARIABLE




                                            14
Significance of the Study
Results of the study may be significant to the following:

   a. People and Industry

   b. Other Researchers

   c. The Researchers




                                                            15
RESEARCH METHODS
17
a. plant material


       b. Test organisms




                           c. Laboratory apparatuses and
d. chemicals, etc                   equipments           18
19
Rotary Evaporator




                                                         20
http://chem-ilp.net/labTechniques/RotaryEvaporatorApparatus.htm
Rotavation




             21
Rotary Evaporation


                   C. odorata MeOH
                      leaf extract



Methanol




                                     22
Hagonoy Methanol Leaf Extract Solutions
                                 (sample computation)
    30% MeOH Leaf Extract Solution           60% MeOH Leaf Extract Solution
             (100 mL)                                 (100 mL)




                100 mL 30% MeOH Leaf                      100 mL 60% MeOH Leaf
                    Extract Solution                          Extract Solution




                +                =                            +               =
30 mL pure hagonoy   70 mL H20               60 mL pure hagonoy   40 mL H20
 MeOH leaf extract                            MeOH leaf extract

                                                                                  23
24
The test organisms as affected by the various treatments




T1 - positive control   T2 – leaf decoction   T3 – expressed leaf juice




    T4 – 30% hagonoy methanol     T5 – 60% hagonoy methanol
                                                                  25
        leaf extract solution         leaf extract solution
26
27
DATA GATHERED:
Length of time in minutes for the termites to die due to various treatments
One-Way ANOVA Summary Table
 Source of        Degrees of     Sum of        Mean          F-ratio
 Variation         Freedom       Squares      Square


 Between                 4        1.39          0.35         38.98*
Treatments

  Within                10        0.09         0.009
Treatments

    TOTAL               14        1.48

Legend: *significant at ∞ =.05                    CV = 5.83 %
                                                                       29
                                  Click here to see sample computation
Pairwise Comparison of the Mean Time using Scheffé's Test

     Between          F’        F(.05)(k-1)     ∞ = .05
     Treatments
     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
                                                                30
Null Hypotheses
1. There are no significant differences in
    the length of time the termites are killed
    after the application of the five
    treatments.
2. There are no significant differences in the
    length of time the termites are killed
    between pairs of treatments.

Decision: Reject Ho
                                             31
Conclusions:
• 60% C. odorata leaf extract solution is
  effective in killing the sample termites with
  the shortest length of time

• The positive control and 30% C. odorata
  leaf extract solution are not significantly
  different from each other


                                                32
Conclusions:

• There are significant differences between the
  pairwise comparisons of:
   • T1 vs. T2           • T2 vs. T5
   • T1 vs. T3           • T3 vs. T4
   • T1 vs. T5           • T3 vs. T5
   • T2 vs. T3           • T4 vs. T5
   • T2 vs. T4

• T1 vs. T4 = not significant between .05 level
                                                  33
Recommendations:
•   C. odorata be the best alternative
    termiticide for it is health and environment
    friendly.

•   Leaf decoction and expressed leaf juice
    are better to use for practical reasons. It
    is more economical to use and easy to
    obtain.

                                                  34
Recommendations:
• Further studies should be conducted

   > utilize the same preparations of C.odorata to
     investigate its effects on other type of pest (ants,
     bedbugs, cockroaches, mosquitoes, flies, mice, rats,
     etc.) as well as on plants (weeds, herbs, etc.)

   >    oil extraction and fractionation of the various
       chemical constituents of C. odorata to analyze
       which of these substances have the potential to
       cause such termiticidal effect


                                                       35
Recommendations:
• 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
• Develop a patented termiticidal product
  out of C. odorata which is efficient, cheap,
  economical, organic, and environmental
  and health friendly
                                             36

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The termiticidal potential of c. odorata

  • 1. Researchers: Research Adviser: TESSIE Q. PERALTA, Ph. D. Don Mariano Marcos Memorial State University South La Union Campus College of Education LABORATORY HIGH SCHOOL Agoo, La Union 2009 CHEMREZ Green Chemistry Award [SIBOL Category]
  • 2. THE PROBLEM & ITS BACKGROUND 4
  • 3. 5
  • 4. Chromolaena odorata (HAGONOY) > a weed belonging to the Asteraceae family > occurs in agricultural areas, natural forests, planted forests, range /grasslands, riparian zones, ruderal /disturbed, scrub/ shrublands > considered invasive weed of field crops in its introduced range, and has been reported to be the most problematic invasive species > reduces the carrying capacity of the field to grow more beneficial weeds (allelophatic) > causes diarrhea when ingested or even fetal deaths among cows > contains carcinogenic pyrrolizidine alkaloids (hepatotoxic-potentially damaging to the liver). The bright green regions indicate where Chromolaena has invaded forest gaps and 6 replaced the forest.
  • 5. TERMITES > group of social insects usually classified at the taxonomic rank of order Isoptera > prefer to feed on dead plant material, wood, leaf litter, or soil > live in colonies; prime example of decentralized, 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 > typical termite colony contains nymphs (semi-mature young), workers, soldiers, and reproductive individuals of both sexes, sometimes containing several egg-laying queens [insect castes]
  • 6. 4 Major Termite Species in the Philippines Nasutitermes luzonicus (Luzon Point Headed termite) Coptotermes vastator (Philippine Milk termite) Microcerotermes losbanosensis (Los Banos termite) Macrotermes gilvus (Mound Building termite) 8
  • 7. Additional Expenses to Repair Damaged Structural Damage Properties TERMITES Destructive Use of Expensive Chemical Termiticide Lack of Knowledge of Plant Alternative as Termiticide 9
  • 8. 10
  • 9. Risk to Public Health and Environment 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 its Impacts to Health Available and Environment 11
  • 10. Objectives of the Study 1. To test termiticidal potential of C. odorata in terms of the length of time the termites are killed after the application of the five treatments. 2. To determine the significant differences in the length of time the termites are killed when exposed to the five treatments. 3. To analyze the significant differences in the length of time the termites are killed between pairs of treatments. 12
  • 11. Null Hypotheses The following statements were tested experimentally at .05 level of significance: 1. There are no significant differences in the length of time the termites are killed after the application of the five treatments. 2. There are no significant differences in the length of time the termites are killed between pairs of treatments. 13
  • 13. Significance of the Study Results of the study may be significant to the following: a. People and Industry b. Other Researchers c. The Researchers 15
  • 15. 17
  • 16. a. plant material b. Test organisms c. Laboratory apparatuses and d. chemicals, etc equipments 18
  • 17. 19
  • 18. Rotary Evaporator 20 http://chem-ilp.net/labTechniques/RotaryEvaporatorApparatus.htm
  • 20. Rotary Evaporation C. odorata MeOH leaf extract Methanol 22
  • 21. Hagonoy Methanol Leaf Extract Solutions (sample computation) 30% MeOH Leaf Extract Solution 60% MeOH Leaf Extract Solution (100 mL) (100 mL) 100 mL 30% MeOH Leaf 100 mL 60% MeOH Leaf Extract Solution Extract Solution + = + = 30 mL pure hagonoy 70 mL H20 60 mL pure hagonoy 40 mL H20 MeOH leaf extract MeOH leaf extract 23
  • 22. 24
  • 23. The test organisms as affected by the various treatments T1 - positive control T2 – leaf decoction T3 – expressed leaf juice T4 – 30% hagonoy methanol T5 – 60% hagonoy methanol 25 leaf extract solution leaf extract solution
  • 24. 26
  • 25. 27
  • 26. DATA GATHERED: Length of time in minutes for the termites to die due to various treatments
  • 27. One-Way ANOVA Summary Table Source of Degrees of Sum of Mean F-ratio Variation Freedom Squares Square Between 4 1.39 0.35 38.98* Treatments Within 10 0.09 0.009 Treatments TOTAL 14 1.48 Legend: *significant at ∞ =.05 CV = 5.83 % 29 Click here to see sample computation
  • 28. Pairwise Comparison of the Mean Time using Scheffé's Test Between F’ F(.05)(k-1) ∞ = .05 Treatments 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 30
  • 29. Null Hypotheses 1. There are no significant differences in the length of time the termites are killed after the application of the five treatments. 2. There are no significant differences in the length of time the termites are killed between pairs of treatments. Decision: Reject Ho 31
  • 30. Conclusions: • 60% C. odorata leaf extract solution is effective in killing the sample termites with the shortest length of time • The positive control and 30% C. odorata leaf extract solution are not significantly different from each other 32
  • 31. Conclusions: • There are significant differences between the pairwise comparisons of: • T1 vs. T2 • T2 vs. T5 • T1 vs. T3 • T3 vs. T4 • T1 vs. T5 • T3 vs. T5 • T2 vs. T3 • T4 vs. T5 • T2 vs. T4 • T1 vs. T4 = not significant between .05 level 33
  • 32. Recommendations: • C. odorata be the best alternative termiticide for it is health and environment friendly. • Leaf decoction and expressed leaf juice are better to use for practical reasons. It is more economical to use and easy to obtain. 34
  • 33. Recommendations: • Further studies should be conducted > utilize the same preparations of C.odorata to investigate its effects on other type of pest (ants, bedbugs, cockroaches, mosquitoes, flies, mice, rats, etc.) as well as on plants (weeds, herbs, etc.) > oil extraction and fractionation of the various chemical constituents of C. odorata to analyze which of these substances have the potential to cause such termiticidal effect 35
  • 34. Recommendations: • 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 • Develop a patented termiticidal product out of C. odorata which is efficient, cheap, economical, organic, and environmental and health friendly 36