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

2. 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.

3. 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.

4. 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

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

6. 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)

7. 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.

8. Liquid Pulse Tube Makeup0% 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

9. Liquid Pulse Procedure Cont.
4. The test tubes were mixed through inversion for 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.

10. 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 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.

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

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

13. 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.

14. 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

15. 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

16. 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.

17. 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.

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

19. Thanks You
Any Questions?

20. ANOVA (LP)

21. ANOVA (AI)