GMO Effects on Microbial Flora
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Transcript GMO Effects on Microbial Flora
Blair Bean
Grade 9
Pittsburgh Central Catholic Highschool
Variety of organisms that live in and on the
body
◦
◦
◦
◦
◦
◦
Skin
Gastrointestinal tracts
Nose
Pharynx
Mouth
Conjunctiva
Provide moisture, nourishment, and a stable
pH for body to properly function
Fungi
Yeast
Malassezia fungi
Bacterium
Enterobacteriae
Streptococcus
salivarius
Staphylococcus
epidermidis
Neisseria
meningitides
Enterobacteriae
One of the most used models in history
Found in intestines
Gram negative
◦ Cell wall
◦ Mostly nonpathogenic
Rod-shaped
Important part of healthy intestines
Vitamin B12 and K
Aerobic
Common model
Lives on skin
Infects cuts, pimples, and acne
Gram positive
◦ Cell wall
◦ More pathogenic
Round and clumped together
Aerobic
Cucumis sativius
97% water
Vitamin K2
◦ Helps blood coagulate
Molybdenum
◦ Needed for sulfite enzymes
Detoxes body
Source of energy through complex
carbohydrates
Reduces risk of estrogen cancers
Not a GMO
Contains none or few seeds capable of
reproduction
Triploid (three sets of chromosomes)
Altered by using growth hormones and gene
transfer on the triploid DNA strain
Contains seeds capable of reproduction
Diploid or tetraploid
Horticulturists grow the perfect strain
Unaltered by hormones and gene transfer
Do cucumber extracts affect the survivorship
of microbial flora?
Determine if there is a difference in the
effects of seedless and seeded cucumber on
microbial flora
Microbial flora is important to the overall
health of the human body
Humans eat both seedless and seeded
cucumber everyday
How do cucumbers affect the flora?
Null: Cucumber extract will not significantly
affect microbial flora survivorship.
Alternative: Cucumber extract will
significantly affect microbial survivorship
Alternative: There will be a significant
difference in the effects of seedless and
seeded cucumber on microbial flora.
Micropipettes
Sterile pipette tips
Escherichia coli
(nonpathogenic)
Staphylococcus epidermidis
(nonpathogenic)
Seedless cucumber
Seeded cucumber
Sterile dilution fluid (100mM
KH2PO4, 100mM K2HPO4,
10mM MgSO4, 1mM NaCl)
LB Agar Plates (1% Tryptone,
0.5% Yeast Extract, 1% NaCl)
LB Media (0.5% yeast extract,
1% tryptone, 1% sodium
chloride)
Spreader bars
Vortex
Test tube rack
Sidearm flask
Klett Spectrophotometer
Ethanol
Bunsen burners
Latex gloves
Permanent markers
Filter paper
Cheese cloth
Glass jars with lids
Both cucumbers used in the experiment were
ground in separate food processors
The resulting slurry was separately filtered
through cheese cloth into two different glass jars
with lids
The jars were refrigerated overnight
Escherichia coli and Staphylococcus epidermidis
were also grown overnight in sterile LB media.
A sample of each culture was added to separate
LB media in a separate sidearm flask.
The cultures were incubated until a density of 50
Klett spectrophotometer units was reached. This
represents a density of 10^8 cells/ml
The cultures were then diluted in a sterile
dilution fluid to a density of 10^5 cells/mL
The individual cucumber juices were drained
through filter paper and sterile filtered through
0.2 micron filters into separate test tubes until a
consistency that could easily be micro pipetted
was met
The following ingredients were mixed to create
the desired cucumber extract exposures
Contents
0x
Concentration
1x
Concentration
10x
Concentration
Sterile Dilution
Fluid
8.9ml
8.9ml
8.9ml
E. Coli or
Staph
0.1ml
0.1ml
0.1ml
Additional
Sterile Fluid
1ml
0.9ml
0ml
Seedless or
Seeded
Cucumber Juice
0ml
0.1ml
1ml
10ml
10ml
10ml
Total
Volume
Tubes were allowed to sit for 10 minutes. Then
after vortexing each of the 8 tubes to evenly
suspend cells, 0.1 mL aliquots from each tube
was spread evenly on 50 LB agar plates
Tubes were allowed to sit for another 20 minutes
and the plating process was then repeated
The plates were incubated overnight
The resulting colonies were counted and
recorded. Each colony was assumed to have risen
from one cell.
Number of surviving colonies
P-value
0.000789973
E. coli 10 Minute
700
0.4626
600
0.9870
500
400
300
Seeded
200
Seedless
100
0
Control
1x
Concentrations of Extract
10x
Number of surviving colonies
E. coli 30 Minute
0.2496
700
P-value
0.001199133
0.8641
600
500
400
300
Seeded
200
Seedless
100
0
Control
1x
Concentration of Extract
10x
Number of surviving colonies
P-value
2.50902E-07
Staph 10 Minute
700
600
0.9349
0.8461
500
400
300
Seeded
200
Seedless
100
0
Control
1x
Concentration of Extract
10x
P-value
0.02884179
Number of surviving colonies
Staph 30 Minute
700
0.5817
600
500
0.3608
400
300
Seeded
200
Seedless
100
0
Control
1x
Concentration of Extract
10x
Cucumber did significantly affect microbial
survivorship across all concentrations
Seeded and seedless cucumber did not show
significant variance in their effects on
microbial survivorship. The different
cucumbers did not affect the microbial flora
differently
The E. coli graphs suggest a trend of growth
while the Staph graphs suggest a trend of
decreasing colonies
Limitations
Plating was not completely synchronized
Only two exposure times
Only two concentrations
Only one type of exposure
Extensions
More replicates
Infuse agar instead of just liquid pulse
More concentrations
Test GMO plants and compare them to
cucumbers
"California Watermelon Statistics, Facts." California Watermelon Statistics, Facts.
See California, n.d. Web. 29 Dec. 2014.
"Cucumbers." Cucumbers. The George Mateljan Foundation, n.d. Web. 01 Jan.
2015.
"E. Coli – the Biotech Bacterium | Biotech Learning Hub." Biotechnology Learning
Hub RSS. The University of Waikato, 25 Mar. 2014. Web. 29 Dec. 2014.
"GMO Facts." The NonGMO Project RSS. Non-GMO Project, n.d. Web. 29 Dec. 2014.
"How Products Are Made." How Seedless Fruits and Vegetables Is Made. Advameg,
Inc., n.d. Web. 29 Dec. 2014.
Moder, Justine. "Escherichia Coli." Escherichia Coli. University of Wisconsin, La
Crosse, Apr. 2008. Web. 01 Jan. 2015.
"What Are Bacteria? » Staphylococcus Aureus." What Are Bacteria. N.p., n.d. Web.
01 Jan. 2015.