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

2. 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

3. 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

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

5. 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.

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

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

8. 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.

9. 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

10. 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 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.

11. 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.

12. 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%

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

14. 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% yes Tea Tree: 0.1% 4.74 yes 1% 8.59 yes 10% yes

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

16. 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

17. References
supplements/ingredientmono tea%20tree%20oil.aspx?activeingredie ntid=113
ee_oil
benefits/
/19/sea_breeze_is_back
d/fda/fdaDrugXsl.cfm?id=41925&type =display

18. Tea tree brand Oil ANOVA

19. Sea Breeze Brand Astringent ANOVA