Transcript Average Amount of Copper in ppm

Colloidal Silver
Antibacterial Assessment
Alex Senchak
Grade 9
Central Catholic High School
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 Bacteria
that cause
infections are becoming
increasingly resistant to
antibiotics and other
antimicrobial agents.
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One of the most common forms of bacteria
found in many environments.
Symbiont in intestinal tracts of many
mammals.
Gram negative, rod shaped bacillus.
Most non-pathogenic.
Pathogenic strains can lead to life threatening
infections.
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Gram positive coccus.
 Common surface symbiont in many
mammals (human).
 Most forms considered nonpathogenic.
 Pathogenic forms can be life
threatening.
 Forms biofilms.

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Gram Positive (Staph)
Most pathogenic bacteria in
humans are gram-positive
organisms.
Simple cell wall.
Antibiotics such as penicillin
work against the formation of
the cell wall.
Gram Negative (E. Coli)
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Cell wall is thin extra layer of
lipopolysaccharide which adds
extra level of protection.
If the toxin enters the
circulatory system it causes a
toxic reaction.
This outer membrane protects
the bacteria from several
antibiotics.
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 Disables
enzymes that power
bacteria.
 Colloidal Silver as an antibiotic.
 Argyria: Caused by overuse of
Colloidal Silver.
Paul Karason “Blue Man”
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To
determine if Colloidal
Silver will affect the
survivorship of E. Coli and
Staphylococcus Epidermidis.
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Colloidal Silver exposure will not
significantly reduce the survivorship of E.
Coli or Staphylococcus Epidermidis.
Hypothesis
 Colloidal Silver exposure will significantly
reduce the survivorship of E. Coli and
Staphylococcus Epidermidis.
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Large glass jar
Beaker
Micro and macro pipettes
+ tips
Spreader bars
76 LB agar plates (1%
Tryptone, 0.5% Yeast
Extract, 1% NaCl)
Escherichia Coli bacteria
Staphylococcus
Epidermidis bacteria
Colloidal Silver making
kit:
 Generator
 Silver Electrodes
 Buffer
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TDS meter
Burner
Turn-table
Tube racks
Vortex
Incubator
Gloves\goggles
Klett
Spectrophotometer
SDF (sterile dilution
fluid).
20 sterile test tubes
Ethanol
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1.
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7.
8.
Sterile glass container, beaker and measuring cup were used to
transfer liquid.
Used measuring cup to pour approximately 470 ml of distilled
water into the glass container.
Attached silver electrodes to generator.
Electrodes were suspended in the solution and powered for 75
minutes.
This process was performed in the dark for efficiency.
The deposition of dark particles on the electrodes provided
evidence of colloidal silver.
After 75 minutes a TDS meter was used to determine the
concentration of silver in the solution.
The silver solution was stored in the dark until use.
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1. Bacteria (E. coli and Staph) 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°C) until a
density of 50 Klett spectrophotometer units was reached.
This represents a cell density of approximately 108
cells/mL.
4. The culture was diluted in sterile dilution fluid to a
concentration of approximately 105 cells/mL.
5. A colloidal silver suspension was mixed with the
appropriate amount of SDF to create colloidal silver
concentrations of 13%, 6.5%, 1.3%, and 0.13%.
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0%
Stock
Microbe
SDF
Colloidal
Silver
Total
0.1
ml
9.9
ml
0 ml
0.13%
Stock
1.3%
Stock
6.5%
Stock
0.1 ml 0.1 ml 0.1 ml
9.8 ml 8.9 ml 4.9 ml
0.1 ml
10 ml 10 ml
1 ml
10 ml
12.87%
Stock
0.1
ml
0 ml
5 ml
9.9
ml
10 ml 10 ml
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6. 100 µL of cell culture was then added to the silver solutions,
yielding a final volume of 10 mL and a cell density of
approximately 103 cells/mL.
7. The solutions were vortexed and allowed to sit at room
temperature for 15 minutes.
8. After vortexing to evenly suspend the cells, 100 µL aliquots
were removed from the tubes and spread on LB agar plates.
9. The plates were incubated at 37°C for 24 hours.
10. The resulting colonies were counted visually. Each colony
was assumed to have arisen from one cell.
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250.00
P= 7.53E-24
213.00
181.83
200.00
156.67
139.50
0
150.00
0.13
Colonies
1.3
6.5
100.00
12.87
15.17
50.00
0.00
0%
0.13%
1.3%
6.5%
12.87%
% of Silver in Solution
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Dunnett’s Test Analysis (E. coli)
T Critical = 3.26
(significant)
Alpha = .05
Silver
Concentration
T Value
Interpretation
0.13%
6.75
Significant
1.3%
12.92
Significant
6.5%
15.91
Significant
12.87%
42.82
Significant
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250.00
P= 3.75E-19
201.17
200.00
156.67
141.33
0
128.67
Colonies
150.00
110.67
0.13
1.3
6.5
100.00
12.87
50.00
0.00
0%
0.13% 1.3%
6.5%
12.87%
% of Silver in Solution
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Dunnett’s Test Analysis (Staph)
T Critical = 3.26
(significant)
Alpha = .05
Silver
Concentration
T Value
Interpretation
0.13%
13.69
Significant
1.3%
18.41
Significant
6.5%
22.31
Significant
12.87%
27.85
Significant
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Percentage of Surviving Colonies
90%
80%
70%
60%
50%
E. coli
40%
Staph
30%
20%
10%
0%
0.13%
1.30%
6.50%
12.87%
% of Silver in Solution
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
There appeared to be a dosage effect
of the silver on the bacteria.
Increasing amounts of silver led to
less bacteria surviving.
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Null Hypothesis
 Colloidal Silver exposure will not
significantly reduce the survivorship
of E. Coli or Staphylococcus
Epidermidis.
 REJECTED
Hypothesis
 Colloidal Silver exposure will
significantly reduce the survivorship
of E. Coli and Staphylococcus
Epidermidis.
 ACCEPTED
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
There was a lag
time when plating
the cells.
Exposure times to
the silver varied
slightly.
Test a higher
concentration of
colloidal silver on
bacteria.
 Test different
types of bacteria.
 Infuse the agar
with silver to
provide a
prolonged surface
exposure.
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Bukhari, Mohammad. "Student Presentation on Staphylococcus
Epidermis." Web. 2 Nov. 2009.
<http://web.uconn.edu/mcbstaff/graf/Student%20presentations/S%20ep
idermidis/sepidermidis.html>.
"Colloidal Silver General Information." Web. 25 Oct. 2009.
<http://www.lifedevice.com/General%20Info.htm>.
"Colloidal Silver Studies-Univiversity of North Texas and SilverKare." Web.
28 Oct. 2009.
<http://www.silvermedicine.org/colloidalsilverstudytexas.html>.
"Colloidal Slilver kills over disease causing bacteria." Kombacha Power
Products. Web. 28 Oct. 2009.
<http://www.kombuchapower.com/colloidal_silver.htm>.
"E. coli." Kids Health from Nemours. Web. 28 Oct. 2009.
<http://kidshealth.org/kid/stay_healthy/food/ecoli.html#>.
"Silver Poisoning & Argyria: FDA, EPA, WHO." Colloidal Silver Safety and
Toxicity Center. Web. 3 Nov. 2009.
<http://www.silvermedicine.org/safetyinformation.html>.
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Staph Results
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400 uL
200 uL
100 uL
100 uL
108 cells/mL
(Bacteria)
107 cells/mL
(Bacteria)
105 cells/mL
with silver
103 cells/mL
102 cells
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