Transcript CCHS Nate Lucchino synergistic effects of cooking products on

Synergistic Effects of Cooking
Products on Microbial Life
Nathan Lucchino
Grade 11
Central Catholic High School
2nd Year in Pjas
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.
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.
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.
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.
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.
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
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
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.
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.
Concentrat Test Tube
ion (ml)
1
E.coli
.1
Test Tube
2
.1
Test Tube
3
.1
Test Tube
4
.1
Olive Oil
0
.01
.1
.01
Agave
0
0
0
.01
SDF
9.9
9.89
9.8
9.88
Total
10
10
10
10
Concentratio Test Tube 5
n (ml)
E.coli
.1
Test Tube 6
Test Tube 7
.1
.1
Olive Oil
0
0
.1
Agave
.01
.1
.1
SDF
9.89
9.8
9.7
Total
10
10
10
Concentration
T value
(T crit 2.75)
Significance
Agave .1%
6.70
Sig
Agave 1%
9.48
Sig
Olive Oil .1%
1.86
Not Sig
Olive Oil 1%
5.68
Sig
Olive Oil Effects on E.coli
350
Number
of
colonies
P-value:
>0e-13
300
250
200
150
100
50
0
0.10%
1%
Concentration
Control
Agave effects on E.coli
350
P-value:
.0006
300
Number
of
Colonies
250
200
150
100
50
0
0.10%
1%
Concentration
control
Synergistic effects of Agave and Olive oil on E.coli
350
Interaction
P-value:
.303712
300
250
Numbers of
colonies
200
150
100
50
0
Both variables .1%
Both variables 1%
Olive oil 1% Agave .1% Agave 1% Olive oil .1%
Concentration
control
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.
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.
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.
Future Changes
• Use different concentrations of each variable, or different variables.
• Test effects on growth rate of bacteria.
• A different model could be used.
Sources
• http://www.allaboutagave.com
• http://www.oliveoiltimes.com/olive-oil-health-benefits
• http://www.cdc.gov/ecoli/
• http://www.medicalnewstoday.com/articles/266259.php