1. Theraflu Effects on Microbial Flora
William Gatti
Central Catholic High School
Grade 9

2. Microbial Flora
Little known about association between humans and their flora.
Provide nutritional and digestive benefits, secrete vitamins, stimulate antibody production, and protect against pathogenic molecules.
Supplements and other consumables taken by humans might have unintended effects on the flora populations and their functions.

3. Gram (+) vs. Gram (-) Bacteria
-Most pathogenic bacteria in humans are Gram (+) organisms.
-Simple cell wall.
-Some antibiotics work against the formation of the cell wall.
-Cell wall contains an extra layer of lipopolysaccharides for extra protection.
-Outer membrane protects bacteria from several antibiotics.

4. Staphylococcus Epidermidis (Staph)
Common surface symbiotant in many mammals (humans).
Gram (+) bacteria
Most strains considered non-pathogenic.
Pathogenic strains can be life threatening.

5. Theraflu Over-the-counter flu medicine.
Designed to treat viruses, not bacteria.
Composed of antihistamines and decongestants.
These include Acetaminophen, Diphenhydramine, and Phenylephrine.
Normally comes in powder form, but is also sold as a liquid.

6. Purpose
To determine if Theraflu Daytime ExpressMax has an effect on Staphylococcus epidermidis survivorship.

7. Alternative Hypothesis
Null Hypothesis
Theraflu will not have a significant effect on the survivorship of the Staph.
Alternative Hypothesis
Theraflu will significantly alter the survivorship of the Staph.

8. Materials Sharpie Theraflu Daytime ExpressMax Sterile tips
Sterile spreader bars
SDF (Sterile Dilution Fluid)
Ethanol
Bunsen Burner
Vortex
LB Media
Theraflu Daytime ExpressMax
Staphylococcus epidermidis
4 test tubes
36 LB agar plates (1% Tryptone,
0.5% Yeast extract, 1% NaCl)
Macro-pipette
Micropipette
Klett Spectrophotometer
Sidearm Flask

9. Procedures 1. Staph e. was grown overnight in sterile LB media.
2. Samples of the overnight cultures were added to fresh media in sterile sidearm flasks.
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 108 cells/mL.
4. The cultures were diluted in sterile dilution fluid to a concentration of approximately 105 cells/mL.

10. Procedures Continued
5. Four test tubes were set up as follows: 10% 1% 0.1% 0% 6. Tube A was vortexed mL of the solution in tube A was pipetted into 6 agar plates. 8. Steps 6-7 were repeated using tubes B, C, and D. 9. The plates were placed in an incubator for 24 hours, and then the colonies were counted visually. Each colony was assumed to have arisen from one cell. A B C D
Sterile Water
8.9 mL
9.7 mL
9.89 mL
9.9 mL
Theraflu
1 mL
0.1 mL
0.01 mL
0 mL
Microbe
Total
10 mL

11. Procedures (Infused Plates)
0.2mL of diluted Theraflu concentrations (0%, 10%, 100%) were spread on LB agar plates and incubated for an hour to absorb the solutions.
100uL aliquots of bacterial suspensions from the control tube were spread onto the infused plates.
The plates were incubated at 37o Celsius for 24 hours.
The resulting colonies were counted visually. Each colony was assumed to have arisen from one cell.

12. Theraflu Effects on Staph Survivorship (Liquid Pulse)
P-Value: 1e-06

13. Dunnett’s Test T-crit Value: 3.10 T > T-crit: Significant Variation
P-value: 1e T < T-crit: No Significant Variation
Concentration
T Value
Result
0.1%
4.7
Significant 1%
6.9
10%
8.6

14. Theraflu Effects on Staph Survivorship (Infused Plates)
P-Value:

15. Dunnett’s Test (Infused Plates)
T-crit Value: 3.03
P-Value:
Concentration
T Value
Result
10%
3.5
Significant
100%
6.8

16. Liquid Pulse LD50

17. Infused Plates LD50
The LD50 Value is approximately 55%.

18. Conclusion
Theraflu in concentrations of 0.1%, 1%, and 10% had significant effects on Staph survivorship.
Theraflu infused exposure had significant effects at 10% and 100%.

19. Limitations Only one species of bacteria was tested
Slight variation in synchronization of plating
Only three different concentrations of Theraflu tested
Only 1 exposure time

20. Future Studies Vary exposure time More replicates
Increase concentration range to reach LD50
Synergistic effects of drugs

21. References https://en.m.wikipedia.org/wiki/Theraflu

22. Single Factor Anova

23. Single Factor Anova for Infused Plates