1. Synergistic Effects of Cooking Products on Microbial Life
Nathan Lucchino
Grade 11
Central Catholic High School
2nd Year in Pjas

2. Extra Virgin Olive Oil
Olive oil is a fat obtained from the olive of the Olea Europaea tree.
Besides cooking it is used in cosmetics, pharmaceuticals and soaps, and was used to light traditional oil lamps.

3. Agave Sweetener
A nectar derived from the sap of the plant during the inflorescence period of its growth, and can be used in place of sugar.
The sap can also be used to make pulque, a Mexican alcoholic beverage.

4. Escherisca coli
Escherisca coli is a large and diverse group of gram (-) bacteria
It is found in the intestinal tract of most animals, including humans.

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

6. Purpose
How will the two products in synergy affect the E.coli survivorship?
The purpose of the experiment is to test whether Extra Virgin Olive Oil and Agave Sweetener will have a significant effect on E.coli survivorship.

7. Hypothesis
Null Hypothesis: The cooking products will not have a significant effect on the E.coli survivorship
Alternative Hypothesis: The cooking products will have a significant effect on the E.coli survivorship

8. Materials E.coli Latex Gloves Micropippetes Micro tubes
Extra Virgin Olive Oil
Agave Sweetener
Sterile Fluid (SF)
63 LB Agar plates
LB media
1 syringe sterile filter

9. Procedure 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 culture was placed in an incubator (37 degrees C) until a density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 10 to the 7th cell/ml
4. The culture was diluted in sterile dilution fluid to a concentration of approximately 10 to the 5 cell/ml.
5. The solutions were prepared for pipetting on the plates.

10. Procedure (Cont.)
6. The solutions were inverted to allow mixing of the variables and the sterile fluid with bacteria for 10 minutes. Then the plates were vortexed in preparation of spreading.
7. The bacteria was pippeted from the tubes and spread on LB plates at a concentration of .1%.
8. The plates were incubated for 24 hours.
9.The colonies were counted visually, and each colony was assumed to have arisen from one cell.
10. The appropriate stat analysis was used to analyze the data and assess the hypothesis.

11. Concentration (ml)
Test Tube 1
Test Tube 2
Test Tube 3
Test Tube 4
E.coli .1
Olive Oil
.01
Agave
SDF
9.9
9.89
9.8
9.88
Total 10

12. Concentration (ml)
Test Tube 5
Test Tube 6
Test Tube 7
E.coli .1
Olive Oil
Agave
.01
SDF
9.89
9.8
9.7
Total 10

13. Concentration
T value
(T crit 2.75)
Significance
Agave .1%
6.70
Sig
Agave 1%
9.48
Olive Oil .1%
1.86
Not Sig
Olive Oil 1%
5.68

14. Olive Oil Effects on E.coli
Number of colonies
Concentration

15. Agave effects on E.coli
Number of Colonies
Concentration

16. Synergistic effects of Agave and Olive oil on E.coli
Interaction P-value:
Numbers of colonies
Concentration

17. Dunnett's Test Results
The ANOVA showed that the data was significant and a Dunnett's test is needed to find out where the significance is. The Dunnett's test showed the Agave concentrations of 1% and .1% affected the bacteria survivorship significantly, along with the 1% concentration of the Olive Oil.

18. Conclusion
The null was rejected, for the .1% and 1% Agave along with the .1% concentration of Olive Oil significantly affected the bacteria survivorship.
The synergy did not affect the bacteria significantly, neither was there a significant effect between the two variables.

19. Limitations
The plating was not exactly synchronized, which could have resulted in extra replication in the tubes.
Only a few concentrations were tested.
Only one type of exposure was tested.

20. Future Changes
Use different concentrations of each variable, or different variables.
Test effects on growth rate of bacteria.
A different model could be used.

24. Sources http://www.allaboutagave.com