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Juul E-cigarette Effects on Human Flora

Juul E-cigarette Effects on Human Flora. Jack Catanzarite Pittsburgh Central Catholic HS Grade 12 1st Year in PJAS. Rationale. JUUL makes up about 68% of the e-cigarette’s $2 billion market

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Juul E-cigarette Effects on Human Flora

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  1. Juul E-cigarette Effects on Human Flora Jack Catanzarite Pittsburgh Central Catholic HS Grade 12 1st Year in PJAS

  2. Rationale • JUUL makes up about 68% of the e-cigarette’s $2 billion market • 20.8% of high school students and 4.9% of middle school students- over 3.6 million youth- were current users in 2018 • Medical professionals are very concerned because JUUL e-cigarettes deliver higher concentrations of nicotine than other e-cigarettes

  3. Juul E-cigarettes • Uses an internal, regulated heating mechanism that creates an easily inhaled aerosol • Requires a non-refillable juul pod cartridge, containing a nicotine e-liquid formula • 5% nicotine by volume (double the concentration in other e-cigs) • Nicotine in a single juul pod is equivalent to a pack of cigarettes • Pod cartridges come in multiple flavors, such as cool mint and mango

  4. Microbial Flora • Present in the respiratory, urinary, and digestive tracts as well as on skin. • Consists of mostly bacteria, few protists and fungi. • Most are harmless, some can cause diseases. • Influences human physiology, anatomy, and overall health. • Approximately 1014 bacteria are harbored on and within humans.

  5. Staphylococcus epidermidis • Bacteria found on healthy human skin • Gram (+) • Forms 1/2-2 mm colonies after incubation in agar • Widely studied microbial flora in or on humans • Forms biofilms • Can be a pathogen when inside the body

  6. Escherichia coli • Diverse and large group of bacteria • Found in the intestines of most mammals • Strains are mostly not pathogenic • Gram (-) • Widely used model of human microbial flora

  7. Question • Do smoke products effect human micro-flora of the body? • Purpose: To test the effects of mango JUUL e-cigarette juice on E-coli and Staph bacteria populations.

  8. Hypothesis • Null hypothesis: JUUL e-cigarette juice will not have a significant effect on E-coli and Staph survivorship • Alternate hypothesis: JUUL e-cigarette juice will significantly reduce E-coli and Staph survivorship

  9. Materials • Escherichia coli (E-coli) • Staphylococcus epidermidis (Staph) • Vortex • Incubator • Test tubes • Mango flavored JUUL e-juice pods • Sterile Dilution Fluid (100mM KH2PO4, 10mM MgSO4, 1mM NaCl) • Micro pipettes • Sterile pipette tips • Spreader bars • Ethanol • Bunsen burner • LB plates + media (1% tryptone, 0.5% yeast extract, 1% NaCl)

  10. Liquid Pulse Procedure • Bacteria (E.coli and Staph) were grown overnight in sterile LB media. • Samples of the overnight cultures were added to fresh media in a sterile sidearm flask. • 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. • The cultures were diluted in sterile dilution fluid to a concentration of approximately 10⁵ cells/mL. • A 10% stock solution of vapor juice and sterile water was made. • The following components were added to sterile test tubes.

  11. Table of Concentrations

  12. Liquid Pulse Procedure cont. • These solutions were then vortexed and allowed to sit at room temperature for 15 minutes. • After vortexing to allow even suspension, 100 µL aliquots were removed from the tubes and spread on LB-agar plates. • The plates were incubated at 37°C for 24 hours. • The resulting colonies were counted visually, each colony assumed to have arisen from one cell.

  13. E-coli p-value= 1.3E-10 Staph p-value= 6E-12

  14. Dunnett’s Test: Liquid Pulse

  15. Conclusions • The null hypothesis was REJECTED for all concentrations except 0.01% E.coli. • Experiment shows that mango JUUL e-juice (5% nicotine) does have a significant and harmful effect on E-coli • The experiment had a surprising positive effect on Staph growth

  16. Limitations • Only survivorship tested • Vitality of bacteria may have varied • One exposure period • Limited replicates

  17. Extensions • More assessments • Different flavors of JUUL e-juice • More exposure times • More levels of nicotine • Test the effects of the vapor

  18. Resources • www.center4research.org/the-dangers-of-juuling/ • www.tobaccofreekids.org • https://www.sciencedirect.com/topics/food-science/microbial-flora • http://web.uconn.edu/mcbstaff/graf/Student%20presentations/S%20epidermidis/sepidermidis.html • https://www.cdc.gov/ecoli/index.html

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