1. The Effects of Olive Oil on E. coli Survivorship
Galen Lovejoy
9th Grade
Central Catholic High School
1st Year participating in PJAS

2. Problem
How does Olive Oil effect the survivorship of the bacteria Escherichia coli?

3. Olive Oil Scientific Name: Olea europaea
Regarded as one of the world’s healthiest fats
Used for cooking
One of the world’s oldest cooking ingredients

4. Chemical Makeup of Olive Oil
Olive Oil fat is a triacylglycerol.
Triacylglycerols are formed by three fatty acids joining to a glycerol molecule.
Olive Oil also has free fatty acids that are mostly monounsaturated.

5. Microbial Flora
Refers to microorganisms living in the human body, including the gastrointestinal tract. 
Little is known about the association between humans and their microbial flora.
The four effects are mutualistic, parasitic, pathogenic, and commensal.
Perform functions beneficial to the host, including the manufacturing of essential vitamins, and the prevention of colonization by undesirable microbes.

6. Previous Studies
Previous studies show that olive oil has negative effects on bacteria. Though studies also show that gram (-) bacteria are more resistant to it.

7. Escherichia coli Largest group of known bacteria
Resides in the lower human tract system
Gram (-) Bacteria
Common prokaryotic cell model used in scientific studies

8. Purpose
To determine if olive oil has a significant effect on on E. coli survivorship.

9. Hypotheses
Null Hypothesis (only used for statistical analysis)- The Olive Oil will not have a significant effect on E. coli survivorship.
Alternate Hypothesis (formal hypothesis)- The Olive Oil will significantly alter survivorship of E. coli.

10. Materials E. coli C600 100% Virgin Olive Oil
YEPD agar plates (1% yeast extract, 2% peptone, 2% glucose)
E. coli C600
Sterile dilution fluid (100mM KH2PO4, 100mM K2HPO4, 10mM MgSO4, 1mM NaCl)
Sterile Test Tubes
Sterile Spreader bars
Sterile Micropipets
Sterile Macropipets
Incubator
100% Virgin Olive Oil
Ethanol

11. Procedure 1. A culture of E. coli C600 was grown overnight.
2.The culture of cells was diluted to a concentration of 10^8 cells/ml.
3. The following ingredients were used to create the desired exposures.

12. Concentration Table 0% (Control) 0.1% Olive Oil 1% Olive Oil
Microbe
0.1 ml
SDF
9.9 ml
9.89 ml
9.8 ml
8.9 ml
Olive Oil
0 ml
0.01ml
1 ml
Total Volume (ml)
10 ml

13. Procedure
4. The test tubes were then mixed through inversion for 10mins to emulsify the solution.
5. Samples of 0.1ml were taken from each tube and plated onto YEPD agar plates.
6. 8 replicates were plated for each tube.
7. The plates were incubated at 37˚C for approximately 18 hours.
8. The resulting colonies were counted visually. Each colony was assumed to have arisen from one cell.

14. Effects of Olive Oil on E. coli Survivorship
P-Value 3.309E-08

15. Dunnett’s Test Results
Concentration Groups
T-Value
T-Crit Value
Results
0.1%
3.11
2.95
Significant 1%
4.8
10%
8.62

16. Conclusion
The null hypothesis was rejected for all concentrations of olive oil.
The alternative hypothesis was accepted for all concentrations of olive oil.
The results of the statistical analysis indicates that all concentrations of olive oil significantly increased E. coli survivorship. (Growth stimulant?)

17. Only one type of olive oil was tested.
Limitations
Extensions
Only one type of olive oil was tested.
The cells could have had longer or shorter exposure times.
The plating was not perfectly synchronized.
Only one type of cell was tested.
Other types of olive oil could be use as a variable.
Prolong the amount of time the cells were exposed to the olive oil.
Increase effort to plate all concentrations as close as possible.
Use another cell model, such as yeast.

18. Works Cited
epidermidis/sepidermidis.html coli/basics/definition/con eating/expert-answers/food-and-nutrition/faq right/1103-whats-wrong-with-olive-oil.html#.VKiIqUc8KrU

19. ANOVA Anova: Single Factor SUMMARY Groups Count Sum Average Variance
Row 1 8
573
71.625
Row 2
705
88.125
Row 3
776 97
Row 4
939
ANOVA
Source of Variation SS df MS F
P-value
F crit
Between Groups 3
3.309E-08
Within Groups 28
Total 31