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The Antibacterial Properties of Honey. Megan Wasson Oakland Catholic High School. Problem. Can honey significantly inhibit the survival of bacteria colonies?. Motivation and Background.
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The Antibacterial Properties of Honey Megan Wasson Oakland Catholic High School
Problem • Can honey significantly inhibit the survival of bacteria colonies?
Motivation and Background • Observations of healing properties of honey have been noted by many ancient scientists and scholars including Hippocrates. • Using a natural substance such as honey for antimicrobial purposes offers a holistic solution for modern medicine. • Majority of raw honeys have the enzyme glucose oxidase which produces hydrogen peroxide that has definite antibacterial properties. • Manuka Honey is noted in the scientific literature for its antibacterial properties but does not have the enzyme glucose oxidase.
Purpose • To observe the effects of various honey solutions on E. coli colonies-selected bacteria medium. • Specifically, to investigate if the honey solutions will hinder the survival rate of the E. coli colonies.
Hypothesis • The various honey solutions will inhibit the survival rate of E. coli colonies. Null Hypothesis • The various honey solutions will not significantly impact the survival rate of E. coli colonies.
Materials • LB (luria broth) • LB agar plates • DH5- alpha E. coli • Spreader bar • Turn table • Ethanol • Sterile test tubes • Micropipettes and tips • Incubator • Sterile water • Organic Honeydew Honey • Bunsen burner • Hot plate • Stirrer Bar • Vortex • Microsoft Excel
Experimental Thrusts • Principal Experiment: Collect data to confirm or negate hypothesis of honey impacting E. coli colony survival rate. • Additional Experiment: Conduct test to prove that the honey was absorbed into the agar plate and to further confirm or negate the hypothesis.
Procedure for Principal Experiment • 250 mL of honey was heated to reduce the high viscosity. • Designated amount of water, honey, and E. coli was micropipetted into a tube which was vortexed.
Procedure for Principal Experiment (Continued) • 0.1 mL of each solution was micropipetted onto a designated agar plate • Total of eight plates for each solution • Solution was spread on plate with spreader bar and turn table • Plates were incubated at 37ºC overnight
Principal Experiment Results: Average Number of E. coli Colonies vs. Honey Solution P-value= 7.69x10^-3
Principal Experiment Results: Dunnett Test Results T-Critical=2.88
Procedure for Additional Experiment: Direct Exposure to Heat and Cold • Conducted to ensure honey was inside agar. • Specific steps included the following: • 1 mL of pure honey was pipetted on agar plate. • 12 plates were placed in incubator at 37ºC for 30 minutes and 12 plates were placed outside in 0ºC for 30 minutes. • 0.1 mL of 0%, 10%, or 50% solution was placed and spread on designated dish. • Total of 4 dishes for each solution for each temperature • Dishes were placed in an incubator at 37ºC overnight.
Additional Experiment Results: Direct Exposure to Heat: Average Amount of E. coli Colonies vs. Honey Solution P-Value= 1.74x10^-2
Additional Experiment Results: Direct Exposure to Cold: Average Amount of E. coli Colonies vs. Honey Solution P-Value=2.13x10^-2
Conclusions • Hypothesis was supported and null hypothesis was rejected as the various honey solutions significantly inhibited the survival rate of E. coli colonies.
Limitations • Honey was extremely viscous and pipettes were not designed to distribute precise and accurate amounts of liquid. • Some plates did not receive immediate transport to the incubator which provides the necessary environment to for the growth of E. coli.
Future Experimentation • Test the survival rate of other bacteria with honey solutions. • Compare the effects of honeys with glucose oxidase enzyme and the effects of Manuka honey on bacteria. • Create mixtures of honey and vitamins and apply the mixtures to bacteria and observe its survival rate.
Sources and Acknowledgements • Active Manuka Honey Association. AMHA. 2002. Web. 3 Oct 2010. • Cimolai, Nevio. Sweet Success? Honey as a Topical Wound Dressing. BC Medical Journal. 2 March 2007. Web. 28 Sept 2010. • Knox, Angie. Harnessing Honey's Healing Power. BBC. 4 June 2004. Web. 30 Sept 2010. • Sivasubramaniam, Lakshmi. Medicianl Properties of Liquid Gold: Honey. Pharmainfo.net. 16 Dec 2005. Web. 28 Sept 2010. • Slater, Michael. Does honey have healing benefits? MSN. 27 Nov 2006. Web. 30 Sept 2010. • A special thank you to Mr. Krotec and Central Catholic High School for help and use of facilities