1. The study tested the effectiveness of supernatant from the fungus Metharhizium anisopliae in eliminating the termite species Coptotermes curvignathus, which damages Para rubber crops in Thailand. 2. The results showed that the supernatant was just as effective at eliminating termites as the fungal spores themselves in a short period of time. 3. Using the supernatant could help control termite damage to Para rubber with less environmental pollution than chemical pesticides.

1. Efficiency of Supernatant from Metarhizium anisopliae for Eliminating Termite Coptotermes curvignathus in Para Rubber Mahidol wittayanusorn school, Thailand

2. Introduction

3. Introduction The Para rubber is the important economical goods in Thailand.

4. The table shows Thailand top 20 exported goods in 2006 Each cost: million dollar Market share: percents Introduction 3.5 4,500.7 5 . plastic grain 4.2 5,393.6 4 . Para rubber 5.4 7,028.7 3 . electrical circuit 7.4 9,540.8 2 . car and other components 11.5 14,876.3 1 . computer and electronic equipment market share price list

5. Introduction In general, Para rubber has often been injured by the group of herbivorous insects such as white grub, parasitic mite and termite.

6. Introduction The chemical insecticides are effective to solve problems in Para rubber. However, they remains high pollutions to surroundings.

7. Introduction Nowadays, the biological controls using insecticidal fungi are accepted. The fungi which have implicit qualities are Metharhizium spp.

8. Materials and methods

9. Methods Search for the information Dissolve supernatant from fungus Prepare termites Analyze the proteins in supernatant Inject supernatant to termites

10. Study the morphology of Metharhizium anisopliae Study the shape of Coptotermes curvignathus Method :search for information

11. Potato Dextrose Broth (PDB) Supplemented with Chitin and N-acetyl glucosamine Potato Dextrose Agar (PDA) Media Methods : Preparing medium

12. Culture in liquid medium 1x10 8 5x10 7 2.5x10 7 2 3 1 Culture on solid medium Methods : Culturing fungus spore/ml

13. On solid medium In liquid medium 5x10 7 spores/ml Prepare for injecting to the termites Methods : Culturing fungus

14. Filter spores and hyphae out Inject remains to termites Methods : Taking samples for analysis

15. Methods : Preparing termites Pick out worker termites 30 30 30 30 30 30 Experimental set 30 termites per group

16. Methods : Eject supernatant to termites 5x10 7 spores/ml 1x10 8 spores/ml 5x10 7 spores/ml 2.5x10 7 spores/ml PDB medium Sterile water Supernatant Spore control set experimental set 6 5 4 3 2 1

17. Methods : Eject supernatant to termites Inject the termites by micropipette Record the number of dead termites for three day. Repeat the experiment six times

18. Results

19. Results Mortality of termites on first day The number of termites Experimental set

20. Mortality of termites on second day Experimental groups Results The number of termites

21. Mortality of termites on third day Experimental groups Results The number of termites

22. The result of Bradford’s Assay Results 4.208x10 -5 2.5x10 7 5.936x10 -5 5x10 7 5.116x10 -5 1x10 8 Concentration of protein ( mg/l ) Initial concentration of spores ( conidia/ml)

23. Conclusions and Discussions

24. Supernatant from Metarhizium anisopliae is efficacious for eliminating Coptotermes curvignathus. The efficiency of supernatant and of spores are not significantly different to eliminate termites. 1. 2. Conclusions

25. The efficiencies of supernatant from varied initial concentrations of spores are not significantly different. 3. Conclusions

26. Discussions Percentage of mortality of termites Percent of mortality (%) Spore 1x10 8 5x10 7 2.5x10 7 Experimental groups

27. Application Basic knowledge Industrial development Effective termite elimination + Decrease the damage of Para Rubber

28. References Caroline Krieger de Moraes, Augusto Schrank, Marilene Henning Vainstein. (2003). Regulation of Extracellular Chitinases and Proteases in the Entomopathogen and Acaricide Metarhizium anisopliae. Current Microbiology 1 (46):205-210. Patcharin Krutmuanga,b and Supamit Mekchayb. (October 2005). Pathogenicity of Entomopathogenic Fungi Metarhizium anisopliae Against Termites. Conference on International Agricultural Research for Development. Richard J. Milner ( 2000 ) . Current status of Metarhizium as a mycoinsecticide in Australia. Biocontrol News and Information 2 (21):47-50 MAGNUS WA" and BRADLEY S. DAVIDSON~ ( 1993). NEW DESTRUXINS FROM THE ENTOMOPATHOGENIC FUNGUS metarhizium anisopliae. Journal of Natural Pro duct 4 (56):643-647. Fran~oise Odier j, Alain Vey , jp Bureau. (1992). In vitro effect of fungal cyclodepsipeptides on leukemic cells: study of destruxins A, B and E. Biol Cell 1 (74):267-271.

29. Sincerely thanks to: Dr. Panadda Bunseum The Institute of Molecular Biology and Genetics, Thailand Miss Chanikarn bunchuay The Institute of Science and Technology for Research and Development, Thailand Mr. Umnart Chanama The Institute of Science and Technology for Research and Development, Thailand Miss Orawan Piyabun Mahidol wittayanusorn school, Thailand Acknowledgement

30. Thanks for your attention