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

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

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

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

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

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

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?”

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.

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

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

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. 4. 0.1ml 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.

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

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).

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

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

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

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

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

Bibliography http://householdproducts.nlm.nih.gov/cgi-bin/household/brands?tbl=chem&id=2211 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2807625/ http://web.uconn.edu/mcbstaff/graf/Student%20presentations/S%20epidermidis/sepidermidis.html https://ods.od.nih.gov/factsheets/VitaminE-Consumer/ http://www.webmd.com/vitamins-supplements/ingredientmono-954- vitamin%20e.aspx?activeingredientid=954&activeingredientname=vitamin%20e http://www.ncbi.nlm.nih.gov/pubmed/10364708 http://www.livescience.com/50326-what-is-ultraviolet-light.html http://science.ksc.nasa.gov/mirrors/msfc/description/ultraviolet.html http://jn.nutrition.org/content/129/11/1951.short

Single factor anova results

two factor Anova Results.