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Household Cleaning Products Effects On Yeast Survivorship
Jack LeechPittsburgh Central Catholic
High SchoolGrade 10
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Ammonia• Common chemical used in cleaning
products and fertilizers.• In small amounts, ammonia can cause eye
and skin irritation. • In more concentrated amounts, ammonia
can cause severe skin burns. It can also lead to permanent lung damage if inhaled or blindness if it comes into direct contact with the eyes.
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Bleach
• Widely used household cleaning product.
• Bleach can also cause skin burns and eye irritation.
• One of the most harmful effect of bleach is its tendency to react with other cleaning products (ammonia or vinegar) to form lethal gases and byproducts.
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Organic Product: Seventh Generation
• Well-known for providing organic, environmentally safe products for over 18 years.
• Claims to be non-toxic, hypo-allergenic, emits no fumes, and contains no phosphates.
• Only listed harmful effect is possible eye irritant resulting from direct contact with the solution on the eye.
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Why were these variables used?
• Known to have some kind of negative effect on living cells.
• Now these results will be compared to an organic product and its effect.
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Why is Yeast the Test Subject?
• Cell used was a species of yeast known as Saccharomyces cerevisiae.
• Easy to manipulate in the laboratory.• Very similar in structure to human cells. • In addition, this cell is a eukaryotic cell but
buds in single colonies, so they can be easily counted.
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Purpose:
• Discover if any household cleaning products have a significant effect on the growth of yeast colonies.
• If the cleaning products have an effect on the yeast growth, which has the greatest effect; ammonia, bleach, or the organic product.
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Hypothesis
• Null-the cleaning products will have no significant effect on yeast survivorship.
• Alternative-based on the numerous negative effects that the products can cause, ammonia will have the greatest effect on the yeast growth, followed by bleach and finally Seventh Generation.
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Materials
• 42 YEPD agar plates (1% yeast extract, 2% glucose, 1.5% agar)
• YEPD media (1% yeast extract, 2% peptone, 2% glucose)• Sterile capped test tube sterile dilution fluid (SDF) (10 mM
KH2PO4, 10 mM K2HPO4, 1 mM MgSO4, 0.1 mM CaCl2, 100 mM NaCl)
• Ammonia• Chlorine bleach• Seventh Generation disinfectant spray• Micropipette• Permanent marker• Plate spreader• Ethanol
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Procedure (Indirect Exposure)• S.c. yeast was grown overnight in sterile YEPD media.• A sample of the overnight culture was added to fresh media in a
sterile sidearm flask.• The culture was placed in a shaking water bath (30 C) until a
density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 107 cells/mL.
• The plates were labeled and placed over the cleaning products fumes for intervals of 1 minute and 2 minutes.
• After vortexing to evenly suspend cells, 0.1mL aliquots were removed from the tubes and spread on 18 plates.
• The cell culture was diluted in sterile dilution fluid to a concentration of approximately 10 3 cells/mL.
• The plates were incubates at 30 C for 48 hours.• The resulting colonies were counted. Each colony is assumed to
have risen from one cell.
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Procedure (Direct Exposure)• S.c. yeast was grown overnight in sterile YEPD media.• A sample of the overnight culture was added to fresh media in a
sterile sidearm flask.• The culture was placed in a shaking water bath (30 C) until a
density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 107 cells/mL.
• The cell culture was diluted in sterile dilution fluid to a concentration of approximately 10 3 cells/mL.
• After vortexing to evenly suspend cells, 0.1mL aliquots were removed from the tubes and spread on 18 plates.
• The plates were labeled and placed over the cleaning products fumes for intervals of 1 minute and 2 minutes.
• The plates were incubates at 30 C for 48 hours.• The resulting colonies were counted. Each colony is assumed to
have risen from one cell.
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Yeast Survivorship of the Indirect Exposure
Time Exposed to Fumes
Resulting Colonies
p=0.03 p=0.27
p=
0.0
19 p
=0
.24
3
p=
0.7
07
p=
0.0
07
p=
0.9
36
p=
0.7
43
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Yeast Survivorship of the Direct Exposure
Resulting Colonies
Time Exposed to Fumes
p=0.002 p=0.006
p=
7.7
9 *
10
-6
p=
2.3
2 *
10-5
p=
0.2
29
p=
1.9
6 *
10
-6
p=
.00
02
p=
0.1
89
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Fumes of Cleaning Products
Ammonia
Seventh Generation
Bleach
Results
P-value( .019) < .05 (Significant effect on control)
P-value( .243) > .05 (Insufficient data to confirm an effect on control)
P-value( .707) > .05 (Significant effect on control)
Ammonia
Bleach
Seventh Generation
P-value( .007) < .05 (Significant effect on control)
P-value( .936) > .05 (Insufficient data to confirm an effect on control)
P-value( .743) > .05 (Insufficient data to confirm an effect on control)
Ind
ire
ct 1
min
. exp
osu
reIn
dir
ect
2 m
in. e
xpo
sure
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Fumes of Cleaning Products
Ammonia
Seventh Generation
Bleach
Results
P-value (7.79 * 10-6) < .05 (Significant effect on control)
P-value( 2.23 * 10 -5) < .05 (Significant effect on control)
P-value( .229) > .05 (Insufficient data to confirm an effect on control)
Ammonia
Bleach
Seventh Generation
P-value( 1.96 * 10-6) < .05 (Significant effect on control)
P-value( .0002) < .05 (Significant effect on control)
P-value( .189) > .05 (Significant effect on control)
Dir
ect
1 m
in. e
xpo
sure
Dir
ect
2 m
in. e
xpo
sure
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What Does This Mean?
• Direct exposure had a much more destructive effect on the yeast survivorship.
• Time exposed had little to do with the indirect exposure, but the time exposed did effect the direct exposure.
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Conclusions• Null hypothesis partially rejected. • Sufficient evidence to see that the direct
exposure had a negative effect on the yeast cells; indirect exposure was not as effective.
• Based on the p-values of the variables compared to the control, ammonia was the most destructive source followed by chlorine bleach and Seventh Generation.
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Conclusions (cont.)
• Indirect exposure did have some effect based on the cell count of the plates.
• Not enough to prove that it was the effect of the cleaning products fumes’ outside of chance.
• The most effective form of cell destruction resulted from the direct exposure.
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Limitations in the Experiment
• Not exposing the yeast completely to the fumes.
• Environmental contamination could have been a factor.
• The plates were not vortexed between each trial.
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Continuing the Experiment
• Use different kinds of fumes and expose to yeast.
• Test the separate ingredients of the product instead of the whole product.
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Works Cited
• www.epa.gov
• www.sciencedirect.com
• www.bbc.co.uk
• www.seventhgeneration.org