Study Title

Antibacterial Activity and Efficacy of Magic Textiles Fabric

Test Method
American Association of Textile Chemists and Colorists Method 100
Assessment of Antibacterial Finishes on Textile Materials

Study Identification Number

NG13210

Study Sponsor
Steve Kubec
MicroShield 360
4700 Rockside Rd Suite 608
Indepence, OH 44131
(216) 393-3945
steve.kubec@microshield360.com

Test Facility
Microchem Laboratory
1304 W. Industrial Blvd
Round Rock, TX 78681
(512) 310-8378
Testing performed by: Chris Craney
AATCC 100: General Information
The American Association of Textile Chemists and Colorists (AATCC) is a well established non-
profit organization that provides education, develops test methods, and sets standards for the
textile industry. The AATCC method 100 is a quantitative test method designed to assess the
performance of antimicrobial finishes on textiles. It can be conducted using contact times ranging
from ten minutes up to 24 hours. For an AATCC 100 test, non-antimicrobial control textiles are
used as the baseline for calculations of microbial reduction. The method is versatile and can be
used to determine the antimicrobial activity of a diverse array of porous materials in addition to
textiles. Because the method allows a great degree of latitude with regard to how the procedure is
carried out, some scientists consider it to be more similar to a testing guideline than a test method.

Laboratory Qualifications Specific to the AATCC 100

Microchem Laboratory began conducting the AATCC 100 test method in 2007. Since then, the
laboratory has performed thousands of AATCC 100 tests on a broad array of test substances,
against a myriad of bacterial, fungal, and viral species. The laboratory may also modify the
AATCC 100 test method as needed in order to accommodate customer needs. Every AATCC 100
test at Microchem Laboratory is performed in a manner appropriate to the test substances
submitted by the Study Sponsor, while maintaining the integrity of the method.

Study Timeline
Articles Articles Enumeration
Culture Report
Inoculated Harvested Plates
Initiated Delivered
& Incubated & Plated Evaluated

19JUN2019 20JUN2019 21JUN2019 24JUN2019 26JUN2019

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Test Substance Information
The test substance was received 14 JUN 2019. The following photographes were taken of the test
substance analyzed for this study.

Photo: On left (as sample was received),

On right (sample cut into appropriate size for study)

Test Substance Received: Leather Sample

Test Substance arrived in dimensions that were not optimal for the conduct of the Study. Test
substance was cut down to ideal sizes for the Study.

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Test Microorganism Information
The test microorganism(s) selected for this test:

Staphylococcus aureus 6538

This bacterium is a Gram-positive, spherical-shaped, facultative
anaerobe. Staphylococcus species are known to demonstrate resistance to
antibiotics such as methicillin. S. aureus pathogenicity can range from
commensal skin colonization to more severe diseases such as pneumonia
and toxic shock syndrome (TSS). S. aureus is commonly used in several
test methods as a model for gram positive bacteria. It can be difficult to
disinfect but does demonstrate susceptibility to low level disinfectants.

Klebsiella pneumoniae 4352

This bacteria is a Gram-negative, rod-shaped, facultative anaerobe. K.
pneumoniae is in the Enterobacteriaceae family which has developed
resistance to carbapenem class based antibiotics. Although K.
pneumoniae is considered normal flora of the human gastrointestinal
tract, this bacterium can also cause serious diseases such as
pneumonia. K. pneumoniae is relatively easy to disinfect and usually
serves as a good representation of an antimicrobial agent's efficacy
against Gram-negative bacteria.

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Summary of the Procedure
The following is general summary of the procedures used at Microchem, additional notes and
specific modifications made to the method or internal operating procedures are described later in
the report.

• Test articles were provided by the Study Sponsor to the performing laboratory. Upon receipt
and review of the test articles, the articles were considered to be ready for use in testing or
were processed per the method or per the Study Sponsor’s recommendations.
• For articles processed by the performing laboratory, articles were cut into 4.8 cm diameter
circles. Control fabric was steam sterilized and test fabrics were steam sterilized only upon
the request of the Study Sponsor.
• Once processed, fabric articles were stacked with 4-6 swatches per stack, or a sufficient
number of swatches to absorb ~1.0 ml of inoculum.
• The microorganisms were prepared by initiating a test culture in appropriate growth broth
and incubating for ~24 hours.
• The inoculum was prepared from the 24 hour incubated culture by diluting to the specified
concentration. The culture diluent that may be used in this method range from full strength
nutritive broth to reverse osmosis/deionized water or anything in between. For hydrophobic
articles, a surfactant such as Triton X-100 was added to the culture diluent.
• Once prepared, the inoculum was plated to confirm the starting concentration.
• A milliliter of inoculum was added to each stack of fabric articles or an appropriate volume
as designated by the nature of the article or specified by the Study Sponsor.
• A subset of control articles was harvested following inoculation to determine the starting
concentration on the fabric.
• Following inoculation, the test articles and parallel control articles were incubated in closed
sealed Petri dishes inside a sealed plastic bag/container under incubation conditions
optimal for the microorganism, typically 36°C.
• Upon completion of the contact time, dictated by the length of incubation, the test and
control articles were harvested.
• Carriers were harvested by aseptically folding stacks and placing them into neutralizer
media, then vortex mixing for ~1-2 minutes.
• The neutralizer was then plated using standard dilution and plating techniques.
Enumeration plates are incubated at the appropriate conditions for 24-48 hours.
• Sterility controls, including test and control articles, were performed on each day of testing.
The microorganisms used in the study were checked for purity.
• Neutralization verifications were performed if requested by the Study Sponsor prior to testing
initiation.

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Criteria for Scientific Defensibility of an AATCC 100 Study
For Microchem Laboratory to consider an AATCC 100 study to be scientifically defensible, the
following criteria must be met:

1. The average number of viable bacteria recovered from the time zero samples must be
approximately 1 x 105 cells/carrier or greater.
2. Ordinary consistency between replicates must be observed for the time zero samples.
3. The number of viable bacteria recovered from the control surface after the contact time
must not be significantly less (>2-Log10) than the original inoculum concentration.
4. Purity streaks must demonstrate growth of the appropriate test microorganism.
5. Negative/Purity controls must demonstrate no growth of test microorganism.

Passing Criteria
AATCC does not specify performance criteria, therefore it may be established by the Study
Sponsor. A similar test method, ISO 20743, recommends a 2-Log 10 or 99% reduction. The United
States Environmental Protection Agency (US EPA) often recommends a 3-Log 10 or 99.9% reduction.

Testing Parameters
Test Substance Swatch Size: 4.8 cm diameter Number of Swatches per Stack: See Notes
Replicates: Two

Culture Growth Media: Trypic Soy Broth Culture Growth Time: 24-48 hours
Culture Dilution Media: Trypic Soy Broth Culture Supplement: 0.1% Triton X-100
Inoculum Concentration: ~1.0 x 105 CFU/Carrier Inoculum Volume: 1.0 ml
Contact Time: 24 Hours Contact Temperature: Ambient
Neutralizer (Vol.): D/E Broth (20 ml) Enumeration Media: TSA
Enumeration Plate Enumeration Plate
Incubation Temperature: 36°C ± 1°C Incubation Time: 24-48 hours

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Study Modifications
There was no study modification.

Study Notes
The Microchem control swatches and test swatches were 4.8 cm in diameter. There were 4
swatches used per test replicate for the Microchem control. The leather test replicates each had 1
swatch.

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Control Results
Neutralization Method: Not performed Media Sterility: Sterility confirmed
Growth Confirmation: Target morphology, confirmed pure

Calculations
• CFU/carrier = x 10df x M
◦ df = The dilution factor used in the dilution and plating method used
◦ M = The media dilution factor for the volume of media used for harvesting the fabric

• Percent Reduction = [(B – A) / B] x 100

◦ B = Number of viable test microorganisms on the control carriers immediately after
inoculation, time zero
◦ A = Number of viable test microorganisms on the test carriers after the contact time

• Log10 Reduction = Log (B / A)

◦ B = Number of viable test microorganisms on the control carriers immediately after
inoculation, time zero
◦ A = Number of viable test microorganisms on the test carriers after the contact time

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Results of the Study of S. aureus 6538

Percent Reduction Log 10 Reduction

Test Average Compared to Compared to
Contact Time Carrier Type CFU/Carrier
Microorganism CFU/Carrier Control at Control at
Time Zero Time Zero

3.10E+05
Microchem
Time Zero 7.30E+04
Control
3.00E+05
N/A
>6.00E+08
S. aureus Microchem
>6.00E+08
ATCC 6538 Control
>6.00E+08
24 hours
1.00E+01
Leather 1.83E+03 99.84% 2.80
9.50E+02

Note: The upper limit of detection is 6.00E+08 CFU/Carrier. Values observed above this limit are denoted as
>6.00E+08 CFU/Carrier in the table and as 6.00E +08 in the graph.

1.00E+09

1.00E+08

1.00E+07

1.00E+06

1.00E+05
CFU/Carrier

1.00E+04

1.00E+03

1.00E+02

1.00E+01

1.00E+00
Microchem Control Microchem Leather
Control
Time Zero 24 hours

Carrier Type

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Results of the Study of K. pneumoniae 4352

Percent Reduction Log 10 Reduction

Test Average Compared to Compared to
Contact Time Carrier Type CFU/Carrier
Microorganism CFU/Carrier Control at Control at
Time Zero Time Zero

6.70E+04
Microchem
Time Zero 7.30E+04
Control
7.90E+04
N/A
>6.00E+08
K. pneumoniae Microchem
>6.00E+08
ATCC 4352 Control
>6.00E+08
24 hours
3.02E+03
Leather 1.83E+03 97.50% 1.60
6.30E+02

Note: The upper limit of detection is 6.00E+08 CFU/Carrier. Values observed above this limit are denoted as
>6.00E+08 CFU/Carrier in the table and as 6.00E +08 in the graph.

1.00E+09

1.00E+08

1.00E+07

1.00E+06

1.00E+05
CFU/Carrier

1.00E+04

1.00E+03

1.00E+02

1.00E+01

1.00E+00
Microchem Control Microchem Leather
Control
Time Zero 24 hours

Carrier Type

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The results of this study apply to the tested substances(s) only. Extrapolation of findings to related materials is the
responsibility of the Sponsor.

Copyright © Microchem Laboratory, 2019. Reproduction and ordinary use of this study report by the entity listed as
“Sponsor” is permitted. Other copying and reproduction of all or part of this document by other entities is expressly
prohibited, unless prior permission is granted in writing by Microchem Laboratory.

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