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The Effects of Gatorade on Microbial Survivorship

This study examines the impact of Gatorade on the survivorship of Staphylococcus epidermidis and Escherichia coli. Different concentrations of Gatorade were tested to determine its effects on the growth of these bacteria. The results showed that Gatorade had no effect on E. coli, but promoted growth in small quantities and killed the bacteria in larger quantities for Staphylococcus epidermidis.

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The Effects of Gatorade on Microbial Survivorship

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  1. The Effects of Gatorade on Microbial Survivorship Christopher Samek Grade 9 Pittsburgh Central Catholic High School 1st Year in PJAS

  2. Problem • Gatorade is constantly consumed to rehydrate the body. • At what concentrations does Gatorade affect the survivorship of human microflora?

  3. Sports Drinks • First sports drink created by a chemist, William Owen, in the U.K. in 1927. • In 1968, sports drinks began to revolutionize sports. • Used for rapid replacement of electrolytes and fluid lost during exercise. • Restores depleted muscle carbohydrate storages that cause muscle failure and fatigue.

  4. Gatorade Ingredients • Glycerol ester of rosin • Protein 0g • Sodium 270 mg • Potassium 75mg • Total Fat 0g • Water • Glucose • Sucrose • Dextrose • Total Carbohydrates 34g • Citric acid • Natural flavor • Sodium citrate • Monopotassium phosphate • Gum arabic • Red40

  5. Microbial Flora • Microorganisms that reside in or on the surface of the body • Respiratory, urinary, digestive tracts, and skin • Consists of mostly bacteria, fungus, protists, and archea • Most are harmless, but some can cause disease • Most studies based on ingested materials study human cells, but human are a collection of cells and symbiotants • Estimated 10% body mass is symbiotants

  6. Staphylococcus epidermidis • Human skin flora • Gram (+) bacterium • Surrounded by a simple cell wall • Most forms are non-pathogenic • Forms biofilms on plastic devices

  7. Escherichia coli • Large and diverse group of gram (-) bacteria • Surrounded by an extra cell wall composed of lipopolysaccharides • Free living, symbiotes, or pathogens • Most strains are non-pathogenic • Serves as a common prokaryotic cell model

  8. Purpose To determine what effect Gatorade has on Staph e. and E. coli survivorship

  9. Hypotheses Null : Gatorade will not have a significant effect on the survivorship of E. coli or Staph. e. Alternate : Gatorade willhave a significant effect on the survivorship of the E. coli and/or Staph. e.

  10. Materials • Matches • Incubator • Ethanol • Bunsen burner • Vortex • Gatorade • Micropipettes • Turn table • Labelling tape • 0.22 Micron sterile filter • 64 LB Agar Plates (0.5% yeast extract, 1% tryptone, 1% sodium chloride) • Escherichia Coli (DH5-alpha) • Staphylococcus Epidermidis • Sterile Dilution Fluid [SDF](100mM KH2PO4, 100mM K2HPO4, 10mM MgSO4, 1mM NaCl) • Sterile test tubes • Racks • Sterile spreader bars

  11. Procedure • E. coli and Staph. e. were grown overnight in sterile LB media. • Samples of the overnight culture were added to fresh media in a sterile sidearm flask. • The cultures were incubated until a density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 10^8 - 10^9 cells/mL. • The cultures were diluted in sterile dilution fluid to a concentration of approximately 10^5 cells/mL. • The Gatorade was diluted with sterile dilution fluid to concentrations of 0%, 5%, 10%, and 25% to total 9.9 mL.

  12. Table of Concentrations

  13. Procedure • 0.1 mL of cell culture was then added to the test tubes, yielding a final volume of 10 mL and a cell density of approximately 10^3 cells/mL. • The tubes were allowed to incubate at room temperature for 10 minutes. • After vortexing to evenly suspend cells, 0.1 mL.aliquots were removed from the tubes and spread on LB agar plates. • The plates incubated at 37°C overnight. • The resulting colonies were counted. Each colony is assumed to have arisen from one cell.

  14. P-Value = 0.49 Not Significant Results Surviving Colonies 0% Concentrations of Gatorade

  15. P-Value = 1.10E-34 Significant Results Survivorship of Staph. e. when Exposed to Various Concentrations of Gatorade 0% 5% 10% 25%

  16. Dunnett’s Test (Staph. e.) T-Critical – 2.88 Alpha – .05

  17. Conclusion • E. coli • The null hypothesis fails to be rejected. • The alternate hypothesis can be rejected. • Gatorade had no effect on the E. Coli in any amount. Staph. e. • The null hypothesis can be rejected. • The alternate hypothesis can be accepted. • Gatorade promoted bacterial growth in small quantities. • Gatorade killed the bacteria in larger quantities.

  18. Limitations / Extensions Extensions • More trials to create a better basis for evaluating the results • Vary exposure time in a two-factor experiment • Use agar infused plates Limitations • Plating not perfectly synchronized • Slightly shorter or longer exposure time • Only two species tested

  19. References https://en.wikipedia.org/wiki/Gram-positive_bacteria https://en.wikipedia.org/wiki/Gram-positive_bacteria https://www.gatorade.com https://www.cdc.gov/ecoli/general/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2807625/ http://www.livestrong.com/article/43161-list-ingredients-gatorade/

  20. Liquid Exposure Raw Data (E. coli)

  21. Liquid Exposure Raw Data (Staph. e.)

  22. E. coli Anova

  23. Staph. e. Anova

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