1. The Effects of Sports Drinks on E.coli Survivorship
Jonathan Farkos
Grade 9
Pittsburgh Central Catholic High School

2. Problem
Do sports drinks have adverse effects on microflora cells?

3. Human Symbionts, Pathogens, and Flora
More than 10% of your body mass is due to symbionts and pathogens
Mostly Prokaryotic cells
Some may cause diseases
Also protect the body from foreign invaders
Reinforce immune system, prevent allergies, and produce vitamins
Lie within skin, saliva, and gastrointestinal tracts
Break down abnormally large nutrients

4. E.Coli c600 Used in many cell/biochemical investigations
Easy to manipulate in labs
Normally found in the intestines of animals.
Some are pathogenic, but most are harmless
Large and diverse group of gram(-) bacteria
Utilized in this study as a human cell model and as a model of pathogens
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.

5. Gatorade
Very commonly used beverage amongst athletes and non- athletes
Provides electrolytes
Common uses include: During sports, Post-sport recovery, and as a common drink
Thought to be non-toxic substance with no effects on body
Ingredients:
Contains water, sucrose (table sugar), dextrose, citric acid, natural flavor, sodium chloride (table salt), sodium citrate, monopotassium phosphate, and flavoring/coloring ingredients, and sucrose acetate isobutyrate.

6. To assess the effects of Gatorade on the survivorship of E.coli
The human micro flora performs many functions vital  to human health.
Purpose

7. Hypotheses
Null Hypothesis: Gatorade will not affect the survivorship of E.coli Alternative Hypothesis: Gatorade will significantly reduce the survivorship of E.coli

8. Materials YEPD agar plates (1% Yeast Extract, 2% peptone, 2% dextrose)
Sterile pipette tips
Micropipettes
Vortex
Incubator
Spreading turntable
Spreader bar
Sports Drink (Gatorade)
Sterile capped test tubes with Sterile Dilution Fluid [SDF] (10mM KH2PO4, 10mM K2HPO4, 1mM MgSO4, .1mM CaCl2, 100mM NaCl)
E.Coli C600

9. Procedure
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.
5: LB agar plates(1% tryptone, .5% yeast extract, 1%NaCl) were prepared
6: Gatorade was pipetted into tubes containing SDF, to create the following concentrations: 0%, 1%, 10%, 25%, and 50%, yielding a final volume of 10 mL.

10. Procedure (Continued)
Concentration Chart 0% 1%
10%
25%
50%
Sterile Dilution Fluid
9.9 mL
9.8 mL
8.9 mL
7.4 mL
4.9 mL
Microbe
0.1 mL
Gatorade
0 mL
1 mL
2.5 mL
5 mL
Total Volume
10 mL
CHANGE
The cells were vortexed for even suspension.
8. After vortexing to evenly suspend cells, 0.1mL aliquots were removed from the tubes at time 10 minutes and 50 minutes and spread on YEPD agar plates (5 plates per concentration)

11. Procedure (Continued)
The plates were then incubated at 30 degrees for 48 hours.
The resulting colonies were counted. Each colony is assumed to have arisen from one cell.

12. P Value
P Value - Less than 1x10E-6

13. Variable Concentration Variable Concentration
Dunnett’s Test
T-Critical = 3.48
10 Min Exposure
Variable Concentration
T-Value
Interpretation
1% Gatorade
1.12
Not Significant
10% Gatorade
2.09
25% Gatorade
0.54
50% Gatorade
5.42
Significant
50 Min Exposure
Variable Concentration
T-Value
Interpretation
1% Gatorade
0.46
Not Significant
10% Gatorade
10.73
Significant
25% Gatorade
6.83
50% Gatorade
14.36
Significant change. Because anovas suggested that there was significant difference within varaition or groups. DON’T SAY PROVE significant variance from control CONFIDENCE

14. Analysis of Exposure Time
Did the duration of exposure have a significant effect?
Ratio of Survivorship of Variables to Control (10 Min Exposure)
1% Gatorade
10% Gatorade
25% Gatorade
50% Gatorade
Control (0% Gatorade)
96%
93%
102%
83%
Ratios of Survivorship of Variables to Control (50 Min Exposure)
1% Gatorade
10% Gatorade
25% Gatorade
50% Gatorade
Control (0% Gatorade)
101%
71%
81%
61%
Key Phrases: the less you say about stats the better it is. Increasing percentage survirorship was decreaed ANOVA anaylsis was performed to see im th emean within these groups varied significantly. The analysis revealed significance suggested that 2 of the means varied significantly. Because ANOVA showed significant variation exsts you want to find out of any variable group varied significantly from the control.
Dunnettts were performed to compare the variables back to the control to check for significance. Alpha is 95% confidence.
Don’t state too much.
Questions about stats.
T-test is a restricted test to two variables of an ANOVA.

15. Conclusions
Generally, increased concentrations of Gatorade lowered the amount of E.coli cells that grew while some concentrations provided aid to growth
Increased exposure did have a significant effect on the amount of surviving colonies.
Higher concentrations of Gatorade could also be effective in reducing the amount of E.coli colonies that could survive

16. Limitations Limited number of concentrations Only 1 species test
Only 2 exposure times
During the testing, plating could have possibly been unsynchronized.
Only one type of Sports Drink was used.

17. Further Studies Different types of Sports Drinks
More of a variety of concentrations
Experiment on other types of cells
Don’t read off of the screen. Become more confident.

18. Sources https://www.cdc.gov/ecoli/general/
you#1
Don’t use first person throughout the presentation. D

19. Amount of Colonies per plate (10 min Exposure)
Data Results
Amount of Colonies per plate (10 min Exposure)
Plate 1
Plate 2
Plate 3
Plate 4
Plate 5
Average 0%
Gatorade
300
284
297
355
326
312.4
1% Gatorade
293
298
306
311
301.2
10% Gatorade
302
286
280
291.6
25% Gatorade
348
310
307
303
321
317.8
50% Gatorade
235
238
279
261
258.6

20. Data Results (Continued)
Amount of Colonies per plate (50 min Exposure)
Plate 1
Plate 2
Plate 3
Plate 4
Plate 5
Average 0%
Gatorade
369
383
346
355
362
363
1% Gatorade
390
361
372
354
367.6
10% Gatorade
244
266
253
261
258
256.4
25% Gatorade
319
295
293
287
282
295.2
50% Gatorade
228
215
231
217
211
220.4

21. Survivorship of E.coli
Scatterplot arithmic graph Change colors.

22. Data Analysis (ANOVA)