1. Luke Nedrow 9 th Grade Central Catholic High School

2.  Alcohol -the hydroxyl functional group is bound to a saturated carbon atom.  Ethanol –  1.Very common alcohol  2.Very toxic substance  3.Two main metabolites, acetaldehyde and acetic acid.

3.  Alcoholic beverages are one of America’s favorite drinks.  57% of American adults over the age 18 said they consumed alcohol in the past month.  88 thousand people die annually from drinking related issues.  223.5 billion dollars are spent in the USA alone on alcohol misuse problems.

4. Most DangerousSecond Most DangerousLeast Dangerous Takes a long time to digest and byproducts are very dangerous Most are ethanol based- Byproducts are dangerous if consumed in large quantities Tertiary and Secondary Alcohols Vodka, Moonshine, Methanol (Wood alcohol) Some beers and most wines Quickly digested- Most Beers

5.  The variables for this experiment were beer, liquor, and wine.  Beer-Coors Light  Alcohol Level- 4.2%  Wine- Sutter Home Sauvignon Blanc  Alcohol Level – 12.5%  Liquor- Smirnoff Vodka  Alcohol Level- 37.5%

6.  About 10% of the biomass on the body are symbionts  E.coli is one of the most prominent and well known models  E.coli was chosen for this experiment because of it’s diversity, it lives in the intestinal tract of humans and many other mammals, and it serves as a common prokaryotic cell model.

7.  Large and diverse group of gram(-) bacteria  One of the most common forms of bacteria: free living, symbiotic, or pathogens  Most strains are not dangerous, but the strains that are can produce fatal diseases.  Common prokaryotic cell model

8.  The human micro flora performs many functions vital to human health.  Close to 60% of the legal population drink alcohol on a monthly basis.  Does alcohol effect the micro flora in the digestive system?  Which type of alcohol effects the microflora the most?

9.  Null- The alcohol will have no significant effect on E.coli survivorship.  Alternate- The liquor (Smirnoff Vodka) will have the most significant effect on E.coli survivorship.

10.  Coors Light  Smirnoff Vodka  Sutter Home Sauvignon Blanc  LB agar plates (1% tryptone,.5% yeast extract, 1%NaCl)  E.coli  Spreader plates  Open flame and ethanol (for disinfecting spreader bars)  Various sized pipettes  Sterile Fluid  Test tubes  Test tube rack  Heat plate/stirrer  Vortex  Incubator  Bunsen Burner

11. 1.Bacteria (E.coli) were grown overnight in sterile LB media. 2. Samples of the overnight cultures were added to fresh media in a sterile sidearm flask. 3. The cultures were placed in an incubator (37˚C) until a density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 10^8 cells/mL. 4. The cultures were diluted in sterile fluid to a concentration of approximately 10^5 cells/mL.

12. 1.LB agar plates(1% tryptone,.5% yeast extract, 1%NaCl) were prepared 2. Alcohol was pipetted into tubes containing SDF, to create the following concentrations: 0%, 1%, 10%, and 50%, yielding a final volume of 10 mL. 3.The cells were vortexed for even suspension.

13. Control (0%)1%10%50% Variable (Type of Alcohol) 0 mL.1 mL1 mL5 mL Sterile Fluid9.9 mL9.8 mL8.9 mL4.9 mL E.coli.1 mL Final10 mL

14. 4. Removed test tubes from test tube rack and 0.1mL was pipetted into each plate. 5. Plates were incubated at 37°C overnight. The resulting colonies were counted.  Each colony is assumed to have risen from one cell.

15. Amount of Surviving Cells Smirnoff Vodka Concentrations

16. P- Value =2.45 ^ -08 Amount of Surviving Cells Coors Light Concentrations

17. P-Value= 1.06 ^-10 Amount of Surviving Cells Sauvignon Blanc Concentrations

18. Replicates Avg. of Group – Avg. of Control (2) (MS Within Group) A Dunnett’s quantifies significant change in data between the control and experimental groups.

19. BeveragesConcentrationsT-valueT-critConclusions Sauvignon Blanc 1% 10% 50% 13.7 18.1 24.5 3.49Sig Coors Light1% 10% 50% 2.2 12.7 15.1 3.58Not Sig Sig Smirnoff Vodka1% 10% 50% 3.2 6.1 7.5 3.56Not Sig Sig

20.  Reject Null Hypothesis for all the exposures except:  1% Coors Light  1% Smirnoff  Accept Alternative Hypothesis for all other trials except for the aforementioned  The alcohol appeared to have a negative effect on E.coli survivorship.

21.  Only tested one type of microbe  Only one exposure time  Possible contamination from spread-plating  Only three concentrations of alcohol  Only thee alcoholic drinks  Only survivorship quantified, not growth

22.  “E. Coli Food Poisoning.” About. About E. Coli, n.d Web. http://www.about- ecoli.com/  Melton, Lisa. “Alcohol.” Wikipedia, Web. http://en.m.wikipedia.org/wiki/Alcohol. http://en.m.wikipedia.org/wiki/Alcohol  Special Thanks to Mr. Krotec

23. Percent Survivorship Concentrations

24. 0% 566438486422 1% 370424378296 10% 224304286258 50% 192194202206 Surviving Colonies

25. 0%872828582 (Outlier)738 1%686804744676 10%394318346322 50%212204246238 Surviving Colonies

26. 0%606654618722 1%360371313362 10%264268258240 50%102112134126 Surviving Colonies