1. James Brunner Pittsburgh Central Catholic High School Grade 11

2.  In this experiment, a culture of Escherichia Coli Bacteria was tested under the effects of two commercial metabolism booster supplements.  Model = E. coli  Variable = Metabolic Enhancer Products  Interaction = E. coli Survivorship

3.  Metabolism is the set of chemical processes in organisms that sustains life.  Most commercial diet pills (such as Lipodrene SR and X12 ) function by increasing the metabolic rate and burning fat.  Studies show that intestinal bacteria, such as E. coli, can play a major role in obesity.

4.  E.coli is a pathogen that is found in the lower intestines of warm blooded animals.  Found in the Gut Flora (intestinal bacteria)  Represents prokaryotic cell model of intestinal bacteria in this experiment.

5.  One of the most common ingredients in metabolism-boosting supplements is caffeine  Recent studies show that the amount of caffeine in a cup of coffee can kill the beneficial Gut Flora bacteria found in the colon. X12 and Lipodrene SR, the enhancers used in this experiment, contain close to the amount of caffeine found in a cup of coffee.

6.  Purpose = To determine the effects of metabolic enhancer products on the survivorship of E. coli.

7.  Hypothesis = The Metabolic Enhancers will inhibit E. coli growth; the plates containing metabolic enhancers will grow less cell colonies than those grown as a control.  Null Hypothesis = There will be no significant difference between the number of colonies grown as a control and the number of colonies grown under metabolic enhancers.

8.  48 LB agar plates( 1 % tryptone, 5 % yeast extract, 1% NaCl, 1.5 % agar)  LB media (1 % tryptone, 5 % yeast extract, 1% NaCl)  Klett spectrophotometer  Sterile pipette tips  Micropipettors  Vortex  Incubator  Sidearm flask  Spread plate  Spreader bar  Ethanol  20 mL Sterile capped test tubes  E.coli B  Sterile dilution fluid  Lipodrene SR  X12

9. 1. E. coli was grown overnight in sterile LB media. 2. A sample of the overnight culture was added to fresh media in a sterile sidearm flask. 3. The cultures were placed in incubators at 37°C until a density of 50 Klett spectrophotometer units were reached. 4. The cultures were diluted in sterile dilution fluid to a concentration of approximately 10 5 cells/mL. 5. Various concentrations (0.01%, 0.1%, and 1%) of X12 and Lipodrene SR were created in test tubes containing SDF, resulting in 9.9 mL per tube 6. 100 µL of cell culture was then added to the test tubes, yielding a final volume of 10 mL 7. The solutions were mixed by vortexing and allowed to sit at room temperature for 15 minutes. 8. After vortexing to evenly suspend cells, 100 µL were removed from the tubes and spread on LB plates. 9. The plates were incubated at 37 degrees for 24 hours. 10. The resulting colonies were counted. Each colony is assumed to have arisen from one cell.

10. Control0.01%0.1%1% SDF8.9 mL Enhancer1.0 mL0.01 mL0.1 mL0.0 mL SDF0.0 mL0.99 mL0.9 mL1.0 mL E. coli0.1 mL TOTAL10 mL

11. LipodreneControl AVERAGE#OFCOLONIESAVERAGE#OFCOLONIES Variable P value = 0.4923 P value = 0.0705 P value = 0.5319

12.  The data was analyzed using ANOVA Charts.  10 ANOVA charts were created-  1 encompassing all data sets.  1 comparing the effects of ethanol to the control.  1 comparing each concentration of metabolic enhancer back to the control.  1 for each Metabolic enhancer, comparing all concentrations to control

13. Anova: Single Factor SUMMARY GroupsCountSumAverageVariance Column 161425237.5967.9 Column 261491248.5764.3 Column 361398233647.2 Column 461342223.66671095.467 Column 561273212.1667914.9667 Column 661487247.8333918.9667 Column 761246207.66671997.867 Column 861205200.8333652.9667 ANOVA Source of VariationSSdfMSFP-valueF crit Between Groups13975.3171996.4732.0065980.0782122.249024 Within Groups39798.1740994.9542 Total53773.4847

14.  Following the results of the ANOVAS, the null hypothesis was accepted.  All ANOVAS performed produced P values >.05, indicating that there was no significant variation among the groups.  There is a clear trend that as [metabolic enhancer] increased, less cells survived; however, ANOVA tests conclude this trend was not significant and may have been due entirely to chance  ANOVA tests may have shown that null was accepted as decreases in E. coli survival not very large, but clearly exist.

15.  Ethanol did show to have a minor inhibitory effect on E. coli growth, could have resulted in inaccurate data  Difficult to dissolve metabolic enhancers, solutions may not have been entirely dissolved  Limited number of replicates  Limited incubation time

16.  Several of the p values obtained from the ANOVAS were close to the.05 cutoff, indicating that in future experiments the results could vary significantly.  More replicates could be added to the experiment  More metabolic enhancers could be added  Adding more concentrations could provide a more accurate view of the effects metabolic enhancers have on E. coli

17.  http://www.buzzle.com/articles/using-diet- pills-increase-metabolism-good-idea.html http://www.buzzle.com/articles/using-diet- pills-increase-metabolism-good-idea.html  http://www.nlm.nih.gov/medlineplus/ency/a rticle/002257.htm http://www.nlm.nih.gov/medlineplus/ency/a rticle/002257.htm  http://www.purenewyou.com/shop/index.ph p?main_page=index&cPath=162 http://www.purenewyou.com/shop/index.ph p?main_page=index&cPath=162  http://apnews.excite.com/article/20060614/ D8I7MBE00.html http://apnews.excite.com/article/20060614/ D8I7MBE00.html