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### Exploring Antifungal Properties in Termites: Insights on A. niger and T. kanei ###

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This research investigates the primary antifungal sources in termites, specifically analyzing their effectiveness against the fungi *Aspergillus niger* and *Trichoderma kanei*. Using body extracts mixed with phosphate-buffered saline (PBS) at varying pH levels, significant antifungal activities were observed, especially in extracts containing protozoa and bacteria. The study highlights the impact of pH on the antifungal properties, revealing that termites suspended in PBS at pH 7.3 show the most potent antifungal properties, particularly in the presence of protozoa. ###

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### Exploring Antifungal Properties in Termites: Insights on A. niger and T. kanei ###

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  1. Results Determining the main source of anti-fungal properties in termites

  2. AN OVERVIEW A. niger T. kanei Simulates conditions of the hemolymph Simulates conditions of the gut Contains bacteria, no protozoa. Presence of other small micro-organisms as well Contains nobacteria, no protozoa. Contains both protozoa and bacteria. Presence of other micro-organisms as well

  3. AN OVERVIEW A. niger T. kanei Simulates conditions of the hemolymph Simulates conditions of the gut Contains bacteria, no protozoa. Presence of other small micro-organisms as well Contains nobacteria, no protozoa. Contains both protozoa and bacteria. Presence of other micro-organisms as well

  4. Results for anti-fungal assay on A. niger, using body extracts of termites mixed in PBS (pH 10) Set B: Control (PBS) w/ extract (Not filtered) w/ extract (0.8um) w/ extract (0.45um)

  5. Results for anti-fungal assay on A. niger, using body extracts of termites mixed in PBS (pH 10) T-test p-value for Extract (NF): 0.0352 (Significant), Extract (0.8um): 0.148 (Insignificant), Extract (0.45um): 0.102 (Insignificant)

  6. Results for anti-fungal assay on T. kanei, using body extracts of termites mixed in PBS (pH 10) Set A: w/ extract (Not filtered) Control (PBS) w/ extract (0.45um) w/ extract (0.8um)

  7. Results for anti-fungal assay on T. kanei, using body extracts of termites mixed in PBS (pH 10) T-test p-value for Extract (NF): 0.0151 (Significant), Extract (0.8um): 0.0396 (Significant)

  8. AN OVERVIEW A. niger T. kanei Simulates conditions of the hemolymph Simulates conditions of the gut Contains bacteria, no protozoa. Presence of other small micro-organisms as well Contains nobacteria, no protozoa. Contains both protozoa and bacteria. Presence of other micro-organisms as well

  9. Results for anti-fungal assay on A.niger, using body extracts of termites mixed in PBS (pH 7.3) Set B: Control (PBS) w/ extract (Not filtered) w/ extract (0.8um) w/ extract (0.45um)

  10. Results for anti-fungal assay on A.niger, using body extracts of termites mixed in PBS (pH 7.3) T-test p-value for Extract (NF): 0.0452 (Significant), Extract (0.8um): 0.0780 (Insignificant)

  11. Results for anti-fungal assay on T. kanei, using body extracts of termites mixed in PBS (pH 7.3) Set B: Control (PBS) w/ extract (Not filtered) w/ extract (0.8um) w/ extract (0.45um)

  12. Results for anti-fungal assay on T. kanei, using body extracts of termites mixed in PBS (pH 7.3) T-test p-value for Extract (NF): 0.0303 (Significant), Extract (0.8um): 0.0295 (Significant)

  13. Comparison of anti-fungal properties of termites suspended in PBS of different pH values (against A. niger) • What we can conclude from this graph: • Protozoa has greater anti-fungal properties against A. niger as compared to bacteria. • Termites in PBS pH 7.3 has greater anti-fungal properties against A. niger as compared to termites in PBS pH 10. • Conclusion: Protozoa in the hemolymph has the greatest anti-fungal properties against A.niger. Protozoa, Bacteria, other small micro-organisms other small micro-organisms Bacteria, other small micro-organisms

  14. Comparison of anti-fungal properties of termites suspended in PBS of different pH values (against T. kanei) • What we can conclude from this graph: • Bacteria has greater anti-fungal properties against T. kanei as compared to protozoa. • Termites in PBS pH 7.3 has slightly greater anti-fungal properties against T. kanei as compared to termites in PBS pH 10. • Conclusion: Bacteria in the hemolymph has the greatest anti-fungal properties against T. kanei Bacteria, other small micro-organisms Protozoa, Bacteria, other small micro-organisms

  15. Methodology: Summary

  16. TEST ON AC SYSTEM Before (Control): Place a PDA plate in front of an air-conditioner for 15 min Switch on the air-conditioner to collect any spores Allow fungus to grow for three days.

  17. Methodology: Summary

  18. Methodology: Summary

  19. TEST ON AC SYSTEM • Pour the extracted body fluids into a spray bottle • Switch off the air-conditioner. • Open the vents. • Spray 8 times (3 sprays uses up 0.5 ml) at 8 different points, spread out evenly between each spray. • Leave the treated air-conditioner off for 1 day to allow for reaction.

  20. Methodology: Summary

  21. Methodology: Summary

  22. TEST ON AC SYSTEM • Place 8 PDA plates in front of the air-conditioner at respective locations for 15 minutes • Switch on the air-conditioner to collect the fungus • Allow fungus to grow for three days • Observe for reduction of mould growth at the end of the three days

  23. Results Determining the main source of anti-fungal properties in termites

  24. AC MOULD Frequently used air-con in SRC Before treatment After treatment

  25. AC MOULD Rarely used air-con in SRC Before treatment After treatment

  26. REFERENCES • Wong M.L.(2009) Air-conditioning can spread germs The Straits Times article, February 26. • Lankarge, V. (2003) What Every Home Owner Needs to Know about Mold and What to Do about It. Blacklick, OH, USA: McGraw-Hill Trade. p 14. • Yupaporn, S. (1996) Studies on Damage of Constructions Caused by Subterranean Termites and Its Control in Thailand. Bangkok: Forest Products Research Division, Royal Forest Department. • Bultman, D. and Southwell, C.R. (1976) Natural resistance of tropical American woods to terrestrial wood-destroying organism. Biotropica. V.8, p.71-95. • Scheffrahn, R. and Nan-Yao, S. (2000) Asian Subterranean Termite, Coptotermes gestroi. Document EENY-128, one of a series of Featured Creatures from the Entomology and Nematology Department, Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida. • US Patent 5866317 (1999) Method for collecting hemolymph of insects • Chapman, R.F. (1998) The insects: structure and function, University of Cambridge, p.101 & 106 • Pasanen, A.L. et al. (1991) Laboratory studies on the relationship between fungal growth and atmospheric temperature and humidity. Environ Int 1991;17: 225–228. • Araújo, R.L. (1958) Contribuição à biogeografia dos térmitas de São Paulo, Brasil. Insecta – Isoptera. Arquivos do InstitutoBiológico de São Paulo 25: 185-217.

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