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The Effect of Crumb-Turf Microbial Survivorship

This study investigates the potential health problems related to crumb-turf in artificial grass fields. It examines the impact of crumb-turf pellets on the survivorship of microbes such as E. Coli and Staph. The results show that crumb-turf has an effect on microbial survivorship, with different effects observed for E. Coli and Staph. The limitations of the study include variations in turf pellet size and unknown interactions between bacteria and turf pellets.

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The Effect of Crumb-Turf Microbial Survivorship

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  1. The Effect of Crumb-Turf Microbial Survivorship Nate Matthews Central Catholic Grade 9

  2. Modern Artificial Turf Fields The purpose is to simulate grass rooted in soil The second layer of pellets, or crumb-turf, tend to be displaced during play Players can come in contact with the turf during play Can be transferred to other locations, like homes Artificial grass crumb-turf

  3. Crumb-Turf • Made from granulated virgin rubber or recycled tires • Virgin rubber may contain hazardous chemicals • Rubber from tires has a different chemical makeup, including carbon-black and other potentially dangerous substances • The granular pellets, being very small, are thought to expose the chemicals inside

  4. Potential Health Problems Related to Crumb-Turf • Soccer players sick, some cancers • Linked to crumb-turf fields (theory) • EPA now involved, collecting samples and reaching out to players • Other agencies and lawsuits involved

  5. Problem • Carcinogenicity hard to test • Can test for microbial effect • Information may aid athletes and parents • 3 different fields were assessed (CCHS, CMU, and Schenley Oval)

  6. Hypotheses • Null: the crumb-turf pellets will noteffect the microbial survivorship • Alt: the crumb-turf pellets will alter microbial survivorship

  7. Microbes E. Coli (gram negative) Staph (gram positive) Staphylococcus epidermidis(staph) is a normal surface symbiont found on humans and other species Patients with compromised immune systems at risk of developing infection, can be lethal • Escherichia coli (E. coli) present in many environments, normally found in the intestines of mammals • Some are pathogenic, and cause diarrhea or illness outside of the intestinal tract • Can be transmitted through contaminated water or food, or through contact with animals or persons

  8. Materials • 1.5 mL micro tubes • Ethanol • Spreader bars • Bunsen burner • Weigh boats • Electronic scale (10e-4) • Bags • Gloves • Autoclave • Crumb-turf • Escherichia coli (C-600) • Staphylococcus epidermidis • Sterile Dilution Fluid (100mM KH2PO4, 100mM K2HPO4, 10mM MgSO4, 1mM NaCl) • LB Agar plates x 112 • LBMedia f(0.5% yeast extract, 1% tryptone, 1% sodium chloride) • Pipettes

  9. Procedure • Turf pellets were collected from fields (kept in bags for sterility) • Turf pellets were sterilized through autoclave • E. coli and Staph 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.

  10. Procedure (cont) • Pellets were weighed into individual masses of 0.01g, 0.05g, and 0.1g • The following ingredients including the turf pellets were added to 1.5 mL micro tubes • 0.1 mL samples were taken from each tube and plated onto LB media plates • Plates incubated at 37°C for 1 day • Colonies were quantified

  11. Procedure Conc. Table

  12. P-value<10e-6 2e-6 <10e-6 0.045

  13. Dunnett's tests E. coli

  14. P-value = 0.0003

  15. Dunnett's tests Staph

  16. Conclusion • All p-values (for e. coli and staph) were well below 0.05, so the null was rejected, and the turf had an effect on the microbes • For E. coli, the crumb-turf reduced survival at the lowest mass, but increased it at the higher masses • For staph, all masses of crumb-turf reduced survivorship

  17. Limitations • The turf pellets varied in size, so it could have affected surface exposure • Plating was not exactly synchronized • Unknown interaction between bacteria and turf pellets (adherence)?

  18. Extensions • Liquefying the turf pellets so that the microbes would not stick • Attaching microbes to the turf and seeing how long they survive • It would be interesting to do this again, as the CMU data showed strange results at highest mass of turf

  19. Bibliography • https://www.epa.gov/chemical-research/federal-research-recycled-tire-crumb-used-playing-fields • https://en.wikipedia.org/wiki/Crumb_rubber • http://www.nbcnews.com/storyline/artificial-turf-debate/rubber-mulch-safe-surface-your-childs-playground-n258586 • https://www.cdc.gov/ecoli/general/index.html

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