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

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.

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.

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

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.

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

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.

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

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.

Procedures Continued 5. Four test tubes were set up as follows: 10% 1% 0.1% 0% 6. Tube A was vortexed. 7. 0.1 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

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.

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

Dunnett’s Test T-crit Value: 3.10 T > T-crit: Significant Variation P-value: 1e-06 T < T-crit: No Significant Variation Concentration T Value Result 0.1% 4.7 Significant 1% 6.9 10% 8.6

Theraflu Effects on Staph Survivorship (Infused Plates) P-Value: 0.000286

Dunnett’s Test (Infused Plates) T-crit Value: 3.03 P-Value: 0.000286 Concentration T Value Result 10% 3.5 Significant 100% 6.8

Liquid Pulse LD50

Infused Plates LD50 The LD50 Value is approximately 55%.

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%.

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

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

References https://en.m.wikipedia.org/wiki/Theraflu https://theraflu.com/product/expressmax-daytime-severe-cold-and-cough https://en.m.wikipedia.org/wiki/Staphylococcus

Single Factor Anova

Single Factor Anova for Infused Plates