Investigating the Synergistic Antibacterial Properties of Different Honeys

1. Investigating the Synergistic Antibacterial Properties of Different Honeys Team Members Teo Yi Fan 4A3 Tan Jun Ming 4A3 Sia Ke Xun 4A3 Kenneth Phang 4A2 Group : 01-085

2. Outline • Background • Objective and Hypothesis • Materials and Apparatus • Methodology • Variables • Results • Timeline • References

3. Background • Honey has long been utilised by humans for its antimicrobial and wound healing properties. (Angie Knox et al, 2004) • Traditional remedies containing honey were used in the topical treatment of wounds by numerous ancient civilisations. (Molan Peter C. et al, 1992).

4. Background • Such properties are derived from the peroxide content of honeys, of which a higher content, results in a greater amount of anti-bacterial inhibine. (Jonathan W. White Jr. et al, 1973). • Some honeys (e.g. spruce honey and fir honey) have shown synergistic effects when combined together, resulting in a greater antibacterial effect. (J.H. Dustmann et al, 1979)

5. Rationale • Bacterial immunity to antibiotics is on the rise calling about a search for an alternative treatment for bacterial infections (Ricki Lewis et al,1995) • Honey being long been recognised for its antimicrobial and wound healing properties is now being rapidly researched as an alternative to bacterial infections. (Angie Knox et al, 2004)

6. Rationale • If honey from different floral sources is proven to exhibit greater antibacterial properties when combined together, this can benefit greatly to the research of honey as an alternative solution for bacterial infections.

7. Objectives and Hypothesis • Objectives • Determine the best combination of honey for a synergistic antibacterial property • Hypothesis • A combination of honeys from different floral sources will exhibit a greater antibacterial effect as compared to that of a single source

8. Materials • Honey Samples • Manuka Honey • Manuka honey is proven to be the most effective at curing strains of Staphylococcus aureus from infected wounds. (R.A.Cooper et al 1999) • Tualang Honey • Malaysian Tualang honey has displayed antibacterial properties against wound and enteric organisms including E. Coli (Hern T Tan et al 2009)

9. Materials • Honey Samples • Buckwheat Honey • Studies indicate buckwheat honey to be an effective antimicrobial wound-healing product. Buckwheat honey’s phenolic constituents, which are present in relatively large amounts, is responsible for its antibacterial activity.

10. Materials • The honey samples will then be combined together in a 1:1 ratio in the following combinations. • The combinations will be known as C1, C2 and C3.

11. Materials • Bacterial Suspension • Staphylococcus epidermis • S. epidermidis strains produce slime that interferes with immune defenses (phagocytosis). S. epidermis are often associated with implanted devices (for example, catheters, prosthetic devices) • Escherichia coli • E. Coli infections are once again on the rise and doctors quickly learned that using antibiotics against E. Coli actually increased tissue damage rather than preventing it, calling a search for an alternative solution to E. Coli Infections (Mead PS, Slutsker L, Dietz V, McCaig LF, Bresee JS, Shapiro C, Griffin PM, Tauxe RV 2011)

12. Materials • NA Plates • Cell Spreader • Syringe and Micro filter

13. Apparatus Spectrophotometer Centrifuge Incubator Autoclave

14. Variables • Independent Variables • Combination of Honey • Concentration of Honey • Dependent Variables • Number of CFUs

15. Variables • Controlled Variables • Volume of Honey Sample • Volume of Bacteria Suspension • Time of Incubation • Type of Agar

16. Methodology • S. epidermis and E. coli suspensions will be produced by overnight cultures grown at 37°C then adjusted to 0.5 McFarland in nutrient broth. • The suspensions will then be diluted to 10-6 of its initial concentration.

17. Methodology • A serial 1:1 dilution of all honeys with sterile water will then be conducted to produce honey solutions of concentrations 20%, 10% and 5%

18. Methodology • 25 μl of honey and 50 μl of bacteria suspension will be added to each agar plate and evenly spread. • Each plate will be incubated for 24 hours at 36.5 oC

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39. Further Works • Increase variety of bacteria • Increase variety of honey • Research for and carry out other methodologies

40. References • 10 February, 1982). Method for the assay of antibacterial activity of honey. New Zealand: Microbiology Standard Methods Manual. Retrieved from http://www.fewstersfarmhoney.com.au/userfiles/file/1_waikato%20antibacterial%20honey%20test.pdf • Honey could be effective at treating and preventing wound infections. (31 January, 2012). (Society for General Microbiology) Retrieved 20 March, 2012, from Science Daily: http://www.sciencedaily.com/releases/2012/01/120131205919.htm • Wound Infection - Care Guide. (2012). Retrieved 10 February, 2012, from Drug Information Online: http://www.drugs.com/cg/wound-infection.html • Dustmann, J. (1979). Antibacterial Effect of Honey. Portorož, Slovenia. From Apimondiaonline: http://www.apimondiafoundation.org/foundation/files/1979/J.H.%20DUSTMANN.pdf

41. References • Knox, A. (8 June, 2004). Harnessing honey's healing power. Retrieved February, 2012, from BBC | Health: http://news.bbc.co.uk/2/hi/3787867.stm • Sherlock, O., Dolan, A., Athman, R., Power, A., Gethin, G., Cowman, S., & Humphreys, H. (2010). Comparison of the antimicrobial activity of Ulmo honey from Chile and Manuka honey against methicillin-resistant Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Dublin, Ireland: BioMed Central Ltd. From: http://www.biomedcentral.com/1472-6882/10/47 • Tan, H. T., Rahman, R. A., Gan, S. H., Halim, A. S., Hassan, S. A., Sulaiman, S. A., & Kaur BS, K. (2009). The antibacterial properties of Malaysian tualang honey against wound and enteric microorganisms in comparison to manuka honey. Kelantan, Malaysia: BioMed Central Ltd. From: http://www.biomedcentral.com/1472-6882/10/31 • Valgas, C., Machado de Souza, S., Smânia, E. F., & Smânia Jr., A. (2007). Screening methods to determine antibacterial activity of natural products. Brazillian Journal of Microbiology, 369-380. From: www.scielo.br/pdf/bjm/v38n2/v38n2a34.pdf

42. Thank You! Q & A