1. Effects of Road Salt Deicers on Yeast Survivorship James Brunner Pittsburgh Central Catholic High School Grade 9

2. Introduction  In this experiment, a yeast culture was tested under the effects of various chloride salts  Model- yeast, Variable- chloride salts, Interaction- yeast survivorship  Idea arose from wondering what happens to ecosystems in winter with road salt

3. Yeast  Eukaryotic organism used in foods and drinks  Most commonly studied organism in labs  Easy to use- grows quickly, easy to count colonies, and nonpathogenic (safe to use)  Less tolerable to extreme conditions- achieves better data

4. Sodium Chloride  Standard salt used to de-ice roads  Known damages- harmful to water, plants, metals, and even the roads it’s used on  One of the cheapest salts to melt ice  Past studies- as salt content increased, yeast survivorship decreased  Chemical Symbol: NaCl

5. Magnesium Chloride  Also used as road salt de-icer  Less harmful then Sodium Chloride  Known to damage metal and concrete, but still known as best for metal and concrete  Contains 17-56% more chloride ion than other salts  Melts ice at slower rate than other chloride salts  Chemical Symbol: MgCl 2

6. Potassium Chloride  Another salt used as de-icer  Thought to be one of the “safer” salts to melt ice with  Used as sodium-free substance for table salt  Known as best for use with vegetation  Chemical Symbol: KCl

7. Purpose & Hypotheses  Purpose- to determine the effects of various chloride salts on a yeast culture  Null Hypothesis- The survivorship of the yeast cells will not be affected by any of the three chloride salts  Alternative Hypothesis- all three chloride salts will be more harmful to yeast then if the yeast cells were left alone; Sodium Chloride will be the most harmful and Potassium Chloride will be the least harmful

8. Methods and Materials Materials:  60 agar plates  Yeast Culture  NaCl  KCl  MgCl 2  Pipette  10 test tubes  Water  Ethanol  Ethanol Spreader  Burner  Weigh boat  Scale  Incubator  Vortex Genie  3 mixers

9. Methods and Materials (continued) 1..1%,.5%, and 1% concentrations of NaCl, KCl, and MgCl 2 were created in test tubes 2. 1 test tube without any salt for control 3. Yeast was added to test tubes 4. 1 ml from each test tube was pipetted onto 6 agar plates, resulting in a total of 60 plates 5. Colonies counted after 2 days

10. Data Analysis  The data was analyzed using ANOVA Charts  9 ANOVA charts were created-  1 for all data  1 for each concentration of salt with the control (no.5% KCl)  The Dunnett Test was used on all ANOVA charts for stronger data

11. ANOVA Chart

12. Results  All salts affected the yeast; yeast survivorship declined as salt concentration rose  NaCl was the least harmful, while MgCl 2 was the most harmful  All plates grew yeast well; except for the plates containing.5% KCl, which was contaminated and did not grow any yeast  The results in this experiment pertained to those of past studies of this nature

13. Effects of Salt on Yeast Survivorship #ofcolonies#ofcolonies Type of Salt Salt Concentration

14. Pictures NaCl 1%KCl.1%

15. Pictures (continued) MgCl 2.5%No Salt Added

16. Final Interpretations  After the data was collected, the alternative hypothesis was overall correct  All colonies affected negatively by the salts  Some of alternative hypothesis incorrect- Sodium chloride least instead of most harmful, MgCl 2 most harmful, KCl in middle  According to data, road salt de-icers inhibit the growth of yeast colonies

17. Final Interpretations (continued)  After ANOVA and Dunnett tests, null hypothesis rejected  Most ANOVA tests showed null hypothesis accepted (most p values over.05), but Dunnett test showed that it was rejected (most t values less than t critical)  Dunnett test more powerful than ANOVA- compares better over two groups of data  ANOVA tests may have shown that null was accepted as decreases in yeast survival not very large, but clearly exist

18. Limitations and Extensions Possible flaws of experiment:  Only complete set of data-NaCl  All 5% KCl contaminated-data could not be obtained; 1 plate in 5% MgCl 2 contaminated  After removal from incubator, lids from 3 plates broken when placed in stacks  May have caused colonies to be smudged around, resulting in incorrect data

19. Limitations and Extensions (continued) Extensions:  More concentrations and salts added  More plates used for each concentration of salt  Another model, such as bacteria, added to achieve better data

20. References  “Rock Salt (NaCl) Sodium Chloride.” Peterschemical.com Dec. 21, 2007. <http://www.peterschemical.com/sodium-chloride/  “Potassium Chloride and Urea.” Peterschemical.com Dec. 21, 2007. < http://www.peterschemical.com/potassium-chloride/  “Magnesium Chloride.” Peterschemical.com Dec 21, 2007. <http://www.peterschemical.com/magnesium-chloride/  “Yeast.” Wikipedia.org Dec. 21, 2007. <http://en.wikipedia.org/wiki/Yeast  “Sodium Chloride.” Wikipedia.org Dec. 21, 2007. <http://en.wikipedia.org/wiki/Sodium_chloride  “Magnesium Chloride.” Wikipedia.org Dec. 21, 2007 <http://en.wikipedia.org/wiki/Magnesium_chloride

21. References (continued)  “Potassium Chloride.” Wikipedia.org Dec. 21, 2007 <http://en.wikipedia.org/wiki/Potassium_chloride  “Road Salt and Water Quality.” Des.state.nh.us Dec. 21, 2007 <http://www.des.state.nh.us/factsheets/wmb/wmb-4.htm  “Field and laboratory investigations on the effects of road salt (NaCl) on stream macroinvertebrate communities.” Sciencedirect.com Dec. 21, 2007 <http://www.sciencedirect.com/science?_ob=ArticleURL&_u di=B6VB5- 455VM763&_user=10&_rdoc=1&_fmt=&_orig=search&_sort =d&view=c&_acct=C000050221&_version=1&_urlVersion= 0&_userid=10&md5=c9d9b919f172d211bbad1839fd5d0afe  “Effect of Sodium Chloride on Bakers' Yeast Growing in Gelatin.” Aem.asm.org Dec. 21, 2007 http://aem.asm.org/cgi/content/abstract/43/4/757