Effects of Acne Medication on Staphylococcus Epidermidis Grant Cardamone Pittsburgh central catholic Grade 9

Acne medications -Can protect the skin from bacterial infections -Helps kill bacteria pockets that form on the skin -Can protect the skin from bacterial infections -Made of a mixture of Benzoyl Peroxide and other antiseptic ingredients

Variable ingredients Water Starch Glycol Hydrogenated Castor Oil Tea Tree Brand Sea Breeze Brand Water Starch Glycol Hydrogenated Castor Oil Citral Lithium Hydroxide Glycerin Alcohol Leaf Oil Salicylic Acid Benzoic Acid Clove (eugenia caryophyllus) Oil Eucalyptus Globulus Oil Fragrance Glycerin Water

Rationale Approximately 80% of people between the ages 11-30 are affected by acne. Does a new acne medication have more positive effects on skin bacteria than the more traditional Tea Tree Oil?

Microbial flora Present in the respiratory, urinary, and digestive tracts, as well as the skin. Consists of mostly bacteria, and few protists and fungi. Most are harmless, but some can cause diseases. 1,014 bacteria are harbored within and on humans.

Staphyloccocus epidermidis Gram positive Common surface symbiont in many mammals. Most forms considered non- pathogenic. Potentially pathogenic upon systemic entry. Forms biofilms

Purpose To find the effects of acne medications on a skin bacteria linked to the creation of acne.

Hypotheses Null- Sea Breeze astringent and Tea Tree Oil will have no significant effects on the survivorship of Staph E. Alternative- Sea Breeze astringent and Tea Tree Oil will have a significant effect on the survivorship of Staph E.

Materials Vortex Incubator Bunsen Burner Sterile Test Tubes Ethanol Sterile Dilution Fluid (SDF) (per 1 liter)(100mM KH2PO4, 10mM MgSO4, 1mM NaCl) Sidearm Flask 32 YEPD Agar plates(1% yeast extract, 2% peptome, 2% glucose) Klett Spectrotometer Tea Tree Oil Sea Breeze Astringent LB media(0.5% yeast extract, 1% tryptone, 1% sodium chloride Micropipette(Sterile tips) Sterile Water Spreader Bars Staphylococcus Epidermidis

Procedure Staph was grown overnight in sterile LB media. Samples of the culture were added to fresh media in a sterile sidearm flask. The cultures were incubated until a density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 108-109 cells/ml. The acne medications were diluted with sterile dilution fluid in concentrations of 0%, 0.1%, 1%, and 10% to total 9.9 ml. 0.1 ml. of cell culture was then added to the test tubes, yielding a final volume of 10 ml. and a cell density of approximately 103 cells/ml.

Procedure(continued) 6. The tubes incubated for 15 minutes at room temperature. After vortexing to evenly suspend the cells, 100 µL aliquots were removed from the tubes and spread on LB-agar plates. The plates incubated in a 37°C incubator overnight. The resulting colonies were counted visually. Each colony was assumed to have arisen from one cell.

Procedure(concentration chart) Tube 1 Tube 2 Tube 3 Tube 4 Sterile Water 9.9 ml 9.89 ml 9.8 ml 8.9 ml Tea Tree Oil/ Sea Breeze 0 ml 0.01 ml 0.1 ml 1 ml Staph Total Volume 10 ml Concentration 0% 0.1% 1% 10%

Sea Breeze P value:<10^-6 Tea Tree P value:10^-6

Dunnett's test Comparison: T-crit=2.57 t value T value>T-crit?Significant? Sea Breeze: 0.1% 4.04 yes 1% 6.08 yes 10% 12.12 yes Tea Tree: 0.1% 4.74 yes 1% 8.59 yes 10% 18.59 yes

Conclusion The null hypothesis was rejected. The Sea Breeze had a significant effect on the survivorship, but less effective than Tea Tree Oil.

Limitations and extensions Use additional bacteria species, particularly other gram positive species Use additional concentrations Create more plates to ensure consistency in results Change exposure times (use Agar infusion) Platings could not be perfectly synchronized Potential slight errors in counting Limited possible number of plates Limited number of concentrations Limited exposure times

References http://www.webmd.com/vitamins- supplements/ingredientmono-113- tea%20tree%20oil.aspx?activeingredie ntid=113 https://en.m.wikipedia.org/wiki/Tea_tr ee_oil https://draxe.com/tea-tree-oil-uses- benefits/ http://www.happi.com/news/2010/02 /19/sea_breeze_is_back https://dailymed.nlm.nih.gov/dailyme d/fda/fdaDrugXsl.cfm?id=41925&type =display

Tea tree brand Oil ANOVA

Sea Breeze Brand Astringent ANOVA