Transcript Metabolic Enhancer Effects on E Coli

James Brunner
Pittsburgh Central Catholic High School
Grade 11
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
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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.
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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.
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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.
 Purpose
= To determine the
effects of metabolic enhancer
products on the survivorship
of E. coli.
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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.
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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
1.
2.
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10.
E. coli was grown overnight in sterile LB media.
A sample of the overnight culture was added to fresh media
in a sterile sidearm flask.
The cultures were placed in incubators at 37°C until a
density of 50 Klett spectrophotometer units were reached.
The cultures were diluted in sterile5dilution fluid to a
concentration of approximately 10 cells/mL.
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
100 µL of cell culture was then added to the test tubes,
yielding a final volume of 10 mL
The solutions were mixed by vortexing and allowed to sit at
room temperature for 15 minutes.
After vortexing to evenly suspend cells, 100 µL were
removed from the tubes and spread on LB plates.
The plates were incubated at 37 degrees for 24 hours.
The resulting colonies were counted. Each colony is
assumed to have arisen from one cell.
Control
0.01%
0.1%
1%
SDF
8.9 mL
8.9 mL
8.9 mL
8.9 mL
Enhancer
1.0 mL
0.01 mL
0.1 mL
0.0 mL
SDF
0.0 mL
0.99 mL
0.9 mL
1.0 mL
E. coli
0.1 mL
0.1 mL
0.1 mL
0.1 mL
TOTAL
10 mL
10 mL
10 mL
10 mL
A
V
250
P value =
0.4923
P value =
0.0705
P value =
0.5319
E
R
A
G
200
E
#
150
0.01%
0.10%
O
F
100
1%
C
O
L
0%
50
O
N
I
E
S
0
X12
Lipodrene
Ethanol
Variable
Control
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The data was analyzed using ANOVA
Charts.
10 ANOVA charts were created1 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
Anova: Single Factor
SUMMARY
Groups
Count
Sum
Average
Variance
Column 1
6
1425
237.5
967.9
Column 2
6
1491
248.5
764.3
Column 3
6
1398
233
647.2
Column 4
6
1342
223.6667
1095.467
Column 5
6
1273
212.1667
914.9667
Column 6
6
1487
247.8333
918.9667
Column 7
6
1246
207.6667
1997.867
Column 8
6
1205
200.8333
652.9667
ANOVA
Source of Variation
SS
df
MS
Between Groups
13975.31
7
1996.473
Within Groups
39798.17
40
994.9542
Total
53773.48
47
F
2.006598
P-value
0.078212
F crit
2.249024
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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.
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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
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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
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http://www.buzzle.com/articles/using-dietpills-increase-metabolism-good-idea.html
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://apnews.excite.com/article/20060614/
D8I7MBE00.html