1. Vitamin E Remediation Of UV Stressed Staph
Jonathan Krystopolski-Czernics
Both bacteria were grown until a density of 50 klett spectrophotometer density was reached. This was approximately 108 cells/mL

2. Vitamin E - fat soluble nutrient. - Known as tocopherol
- Acts as an antioxidant
- Protects cells from free radicals.

3. Ultraviolet light
Ultraviolet light - form of radiation not visible to our eyes
- Invisible on the electromagnetic spectrum
- Can damage living tissues by creating free radicals
- UV wavelength used was 254nm, germicidal

4. free radicals Free radicals
- Unstable molecules created by chemical changes in the body
- Can harm living tissues and can be created with UV light.
- Can be fought with antioxidants in the immune system.
- Might be neutralized by antioxidants

5. Microbial Flora
• Little is known about the association between humans and their
flora
• Effects are mutualistic, parasitic, pathogenic and commensal
• Provide nutritional and digestive benefits, secrete vitamins,
stimulate antibody production, and protect against pathogenic
microbes

6. Staph Staphylococcus epidermidis - bacteria found on the skin.
What is Staph
Staph
Staphylococcus epidermidis
- bacteria found on the skin.
- Can cause multiple infections.
- gram positive
- easy to grow in lab setting
- can produce biofilms

7. Purpose
This experiment was conducted to test the remedial effects of vitamin E on UV stressed staph.
The question that was asked was:
“Can Vitamin E protect Staph cells from ultraviolet light?”

8. Hypothesis
Null Hypothesis: Vitamin E will not appear to have a remedial effect on UV stressed staph.
Alternative Hypothesis: Vitamin E will have a remedial effect on UV stressed Staph.

9. Similar Experiments
An experiment was conducted by Wen-Hsing Cheng, Beth A. Valentine , and Xin Gen Lei
They tested the ability of vitamin E to replace Cellular Glutathione Peroxidase(GPX1)
They observed that there was no evidence to say that vitamin E was able to replace GPX1

10. Materials Liquid vitamin E Staphylococcus epidermidis Micropipette
Sterile Dilution Fluid [SDF} (100mM KH2PO4, 100mM K2HPO4, 10mM MgSO4, 1mM NaCl)
UV culture hood
Incubator
15ml conical tubes
Vortex
Sterile water
Sharpie
Liquid vitamin E
Staphylococcus epidermidis
Micropipette
Sterile Spreader bars
Ethel alcohol
Bunsen burner
Sterile Micropipette tips
LB agar (0.5% yeast extract, 1% tryptone, 1% sodium chloride)
Plates

11. Procedure
1. The bacteria was grown until a density of 50 klett spectrophotometer density was reached. This was approximately 10^8 cells/mL.
2. All equipment was sterilized.
3. The desired amount of amount of liquid vitamin E was pipetted into each conical that contained sterile fluid.
ml of staph was pipetted into the each of the conical tubes.
5. The mixture was incubated for 10 minutes at room temperature while being rolled back and forth in order to mix the solution with constant inversion.

12. Table of Concentrations
Materials In Tubes
Control Group
Concentration 1
Concentration 2
Sterile Fluid
9.9ml
9.8 ml
8.9 ml
Vitamin E
0ml
.1ml
1ml
Staph
.1 ml
Total
10ml

13. Procedures Continued 6. .1ml of solutions was pipetted onto LB auger
7. the plates were exposed to uv light for selected intervals.
8. The plates were incubated for 1 day and the colonies were counted (the colonies were recounted after a 48 hour period).

14. Vitamin e effects on uv stressed staph
Vitamin E Interaction P Value: .616
UV light Interaction P value:3e-06
Two Factor Interaction P Value: 2.92
Number of Colonies
Effect of vitamin above first 3 bars
Blue p value to show value for uv
Two factor interAction
UV Exposure Times

15. Dunnett's test analysis
Dunnet's Test formula:
T=m1-mc/square root of 2mse/n
Variable
T-crit
T Value
Interpretation
0ml vit E
3 seconds UV
3.90
8.05
Significant
6 seconds UV
8.43

16. Conclusion Hypothesis : The null hypothesis could not be rejected
Vitamin E did not appear to have a remedial effect on UV stressed staph.

17. Limitations
-Spread plating was not performed in perfect synchronization, therefore some bacteria had a slightly higher exposure time -Only 3 different UV exposure times were tested -Only 3 concentrations of vitamin E were tested - Only one species

18. Extensions -Increase the number of replicates.
-Alter concentration of vitamin
-Test different vitamins and/or models.

19. Bibliography
vitamin%20e.aspx?activeingredientid=954&activeingredientname=vitamin%20e

20. Single factor anova results

21. two factor Anova Results.