Effect of PowerAde on E. Coli Survivorship By Nicholas Weising Grade 9 Central Catholic High School First Year in PJAS

PowerAde Sports drink manufactured by Coca-Cola company and the main competitor of Gatorade. Mainly composed of water Contains electrolytes (Magnesium, sodium, potassium) roughly 10% of daily intake Commonly consumed while playing sports, exercising, or doing tiring physical labor Contains small amounts of various Vitamin Bs, in total 30% of a daily intake Carbohydrates and sugars make up 42 grams in total.

Escherichia coli Gram-negative microflora Prokaryotic Widely-used in experiments Found in the intestines of warm-blooded animals as well as excrement Most varieties are harmless, though some can cause illnesses.

Rationale This experiment was conducted to discover the unknown effects of PowerAde on internal microflora. There were two methods of exposure: Liquid Pulse Agar Infusion

Hypotheses Null hypothesis- PowerAde will not significantly affect the survivorship of E. coli. Alternative Hypothesis- PowerAde will significantly affect the survivorship of E. coli.

Materials 1 bottle of grape Powerade(100% PowerAde) E.coli Sterile spreaders Sterile pipettes Test tubes Bunsen burner Ethanol Incubator YEPD Agar Petri ( 1% yeast extract, 2% peptone, 2% glucose Vortex Sterile Dilution Fluid (SDF) (per 1 liter) (100mm KH204, 10mm MgSO4, 1mm NaCl)

Liquid Pulse Procedure E. coli C600 was grown overnight The culture of cells was diluted to a concentration of 10^5 cells per ml. The next slide shows the ingredients were combined in the tubes to reach the desired exposures.

Liquid Pulse Tube Makeup 0% 5% 10% 25% 50% E. Coli 0.1 ml 0.1 ml Sterile Water 9.9 ml 9.4 ml 8.9 ml 7.4 ml 5 PowerAde 0 ml 0.5 ml 1 ml 2.5 ml Total Amount 10 ml

Liquid Pulse Procedure Cont. 4. The test tubes were mixed through inversion for 10 minutes to throughly emulsify the solution. 5. Samples of 0.1 ml were then taken from each tube and plated onto YEPD agar plates. 6. 8 plates were made for each concentration. 7. The plates were incubated at 37 Celsius for approximately 18 hours. 8. The resulting colonies were then counted by eyesight.

Agar Infusion Procedure 1) E. coli was used from the control group tube shown before The following volumes of of Powerade was added to the plates: 20 microliter PowerAde + 180 microliters sterile water, and 200 microliters Powerade. 2) 100 μL of cell culture from the control tube was then pipetted onto an infused plate. 3) The plates were incubated at 37°C for 24 hours. 4) The resulting colonies were counted visually.

Concentration of PowerAde P-value is 1.71 E-08 Concentration of PowerAde

Effect of PowerAde on E.coli, Agar Infusion P-value is 0.042 Concentration of PowerAde

Statistics ANOVA Dunnett’s Test Stands for "analysis of variance". This is a statistical test that compares variation within and between multiple groups.. This other statistical test allows you to determine what specific means varied from the control group. It is shown below.

Dunnett’s Test Results (LP) T crit = 3.02 Dunnett’s Test Results (LP) Group T- value Significant? 5% 0.6 No 10% 3 25% 5.3 Yes 50% 7.37

Dunnett’s Test Results (AI) T crit= 2.67 Dunnett’s Test Results (AI) Group T-value Significant? 180,20 0.77… No 200 1.9

Interpretation Two of the groups in Liquid Pulse showed a significant effect. Therefore, we can confidently reject the null hypothesis. It appears as though PowerAde negatively affects the survivorship of my microbial flora when in large concentrations.

Possible Improvements The method of just visually counting colonies isn't always accurate. There are various other types of microbial flora within humans. This experiment could use other species and get a better picture of PowerAde's Effect. Exposure time of the PowerAde should've been tested as well, because liquids can stay within the stomach for a long time.

Bibliography General Information." Centers for Disease Control and Prevention. Centers for Disease Control and Prevention, 06 Nov. 2015. Web. 15 Nov. 2016. "MicrobeWorld." Bacteria. Microbe World, 2014. Web. 15 Nov. 2016. Powerade Keeps You Hydrated. Sports Drink, Electrolyte Drink, Hydration Drink." POWERADE - The Official Website for Powerade. Coca-Cola Company, n.d. Web. 30 Nov. 2016. Important?" BuiltLean. N.p., 20 Feb. 2016. Web. 30 Nov. 2016 "What Are Vitamins and How Do They Work?" HowStuffWorks. N.p., 01 Apr. 2000. Web. 30 Nov. 2016.

Thanks You Any Questions?

ANOVA (LP)

ANOVA (AI)