Decon7 Crushes Glutex in Study (Salmonella on Wood and Concrete Carriers)

Study TitleAntibacterial Activity and Sanitizing Efficacy of Decon7 Systems, LLC Test Substances

Test MethodASTM International Method E1153

Test Method for Efficacy of Sanitizers Recommended for Inanimate Non-Food Contact Surfaces

Study Identification NumberNG6240

Study SponsorJoe Drake

Decon7 Systems, LLC7575 E Redfield Rd., Suite 235

Scottsdale, AZ 85260(480)-339-2858

[email protected]

Test FacilityAntimicrobial Test Laboratories

1304 W. Industrial BlvdRound Rock, TX 78681

(512) 310-8378

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History of the Laboratory

Antimicrobial Test Laboratories was launched in 2006 to provide testing services to theantimicrobial industry. The company has grown considerably since then but its focus remains thesame: Test antimicrobial agents, test them well, and test them fast! Antimicrobial Test Laboratoriesoperates a 15,000+ square foot facility near Austin, Texas, where hundreds of studies areconducted annually by a staff of friendly, knowledgeable, and experienced microbiologists andvirologists.

Laboratory Qualification Statement

Antimicrobial Test Laboratories was founded by microbiologist Dr. Benjamin Tanner. Thelaboratory ensures consistent, reproducible results by utilizing a well-trained and educated scientificstaff who work from a comprehensive system of Standard Operating Procedures, official standardmethods from ASTM, AOAC, and other organizations, and custom study protocols. The laboratoryprovides testing services to dozens of Fortune 500 companies and has been inspected for GLPcompliance by the US government.

Scientist Qualifications

Your Study was designed, conducted and reported by: Kelli Jo Kuntzman, B.S.

If you have any questions about your study, please don't hesitate to contact Kelli Jo at:

[email protected](512) 310-8378

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Kelli Jo graduated from Texas State University with a Bachelors of Science degree in Microbiology.

Kelli Jo is a hardworking, detail oriented, and dedicated microbiologist. As a Microbiology Associate at Antimicrobial Test Laboratories, she participates in a broad array of studies and test methods. Her strong work ethic and interest in comprehensive and accurate testing make her an asset to the laboratory and clients alike.

ASTM E1153: General Information

ASTM International, formerly the American Society for Testing and Materials (ASTM), is aninternationally recognized organization that develops and publishes product and testing standards.ASTM E1153 is a quantitative test method designed to evaluate the antimicrobial efficacy ofsanitizers on pre-cleaned inanimate, nonporous, non-food contact surfaces. The method istypically used with a maximum contact time of 5 minutes, during which the sanitizer reduces theconcentration of viable test microorganisms. ASTM E1153 utilizes non-antimicrobial agents ascontrols to establish baselines for microbial reductions. The ASTM E1153 method is a benchmarkmethod for non-food contact surface sanitizers and is recognized by several regulatory agencies asan approved method for claim substantiation.

Laboratory Qualifications Specific to ASTM E1153

Antimicrobial Test Laboratories began conducting the ASTM E1153 test method in 2007. Sincethen, the laboratory has performed hundreds of ASTM E1153 tests on a broad array of testsubstances, against a myriad of bacterial and fungal species. The laboratory is also experiencedwith regard to modifying the test method as needed in order to accommodate customer needs.Every ASTM E1153 test at Antimicrobial Test Laboratories is performed in a manner appropriate forthe test substances submitted by the Study Sponsor, while maintaining the integrity of the method.

Study Timeline

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SurfacesInoculated

SurfacesTreated

SurfacesEvaluated

ReportDelivered

CultureInitiated

Wood26JUL2015 27JUL2015 27JUL2015 28JUL2015 17AUG2015Concrete

09AUG2015 10AUG2015 10AUG2015 11AUG2015 17AUG2015

Test Substance Information

The test substances were received on 23JUL2015 and the following pictures were taken.(note: photos depict the test substances that were analyzed in this study)

Test Substances Received: Decon 7 Auto-Marine-RV Part 1, Decon 7 Auto-Marine-RV Part 2, EasyDecon DF200 Part 3 (Booster), and Glutex GQ1 Sanitizer and DIsinfectant

Test Substances arrived and required dilution. Test substances were diluted prior to use in thestudy.

Test Microorganism Information

The test microorganism(s) selected for this test:

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Salmonella enterica ATCC 10708This bacteria is Gram-negative, rod-shaped, facultative anaerobe. Like the closely related Escherichia genus, Salmonella are common to all parts of the world and share habitats in the digestive systems of cold and warm-blooded animals. S. enterica is one of the most common bacteria associated with zoonotic and foodbourne illness. Because of it's regular occurrence and pathogenicity, S. enterica is a common bacteria for measuring disinfectant efficacy.

Diagram of the Procedure

Summary of the Procedure

• The test microorganism is prepared, usually by growth in liquid culture medium.• The test culture may be supplemented with an artificial soil load, such as horse or fetal bovine

serum, for one-step cleaner/sanitizer claims.• Sterilized carriers are inoculated with a volume of the test culture. Inoculated slides are dried in

an incubator. Only completely dried carriers are used in the test.• Test carriers are treated with the test substance and incubated for the predetermined contact

time.• Control carriers are treated with a buffered saline solution and are allowed to sit for the

predetermined contact time.• At the conclusion of the contact time, test and control carriers are chemically neutralized.• Dilutions of the neutralized test substance are evaluated using appropriate growth media to

determine the surviving microorganisms at the respective contact time.• The effect of the test substance is compared to the effect of the control substance in order to

determine microbial reductions.

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Criteria for Scientific Defensibility of an ASTM E1153 Study

For Antimicrobial Test Laboratories to consider an ASTM E1153 study to be scientifically defensible,the following criteria must be met:

1. The average number of viable microorganisms recovered from the control carriers must beapproximately 7.5 x 105 cells/carrier or greater.

2. Ordinary consistency between replicates must be observed for the control carriers.3. Positive/Growth controls must demonstrate growth of appropriate test microorganism.4. Negative/Purity controls must demonstrate no growth of test microorganism.

Passing Criteria

ASTM International defines passing criteria to be a 3 Log10 or 99.9% reduction in the treated testcarriers when compared to the control carriers.

Testing Parameters used in this Study

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Test Carrier Size: 1 inch x 1 inch Replicates: TripleTest Substance Volume: ~2.5 ml (Three pumps)

Culture Growth Media: Tryptic Soy Broth Culture Growth Time: 18 hoursCulture Supplement: N/A Carrier Inoculum Volume: 0.050 mlInoculum Concentration: Carrier Inoculum Area: 1 inch x 1 inchCarrier Dry Temp: Ambient (25°C ± 2°C) Carrier Dry Time: 10 minutesContact Temp.: Ambient (25°C ± 2°C) Contact Humidity.: N/AContact Time: 10 minutes & 8 hours Neutralizer: See Notes

36°C ± 1°C 24-48 hours

Incubation Conditions: Aerobic

1 x 107 CFU/Carrier

Enumeration Plate Incubation Temperature:

Enumeration Plate Incubation Time:

Study Modifications

This study was modified for use on concrete and wooden carriers.

Study Notes

The neutralizer chosen for this assay was D/E broth supplemented with 0.1% Calalase.

Per the Sponsor's request, the test substance was prepared in the following way:The 1:1 solution was prepared by adding 80 ml of Part A to 80 ml of Part B. 3.2 ml was removed from this solution and 3.2 ml of Part 3 (Booster) replaced what was removed. The 1:2.5 solution was prepared by adding 20 ml of the prepared 1:1 solution to 30 ml of 200 PPM Hard Water. The 1:5 solution was prepared by adding 10 ml of the prepared 1:1 solution to 40 ml of 200 PPM Hard Water. The 1:10 solution was prepared by adding 5 ml of the prepared 1:1 solution to 45 ml of 200 PPM Hard Water. When tested against wooden carriers, the Glutex test substance was prepared by adding 0.25 ml of test substance to 63.75 ml of prepared 200 PPM Hard Water. When tested against concrete carriers, the Glutex test substance was prepared by adding 0.512 mlof test substance to 64.0 ml of prepared 200 PPM Hard Water.

Study Photographs

Photos above show the scientist inoculating a wooden carrier and a concrete carrier.

Photo depicts the foaming action of of the test substance. of 15

Control Results

Calculations

Where:B = Number of viable test microorganisms on the control carriers after the contact timeA = Number of viable test microorganisms on the test carriers after the contact time

Where:B = Number of viable test microorganisms on the control carriers after the contact timeA = Number of viable test microorganisms on the test carriers after the contact time

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Neutralization Method: Validated Media Sterility: SterileGrowth Confirmation: Morphology on appropriate growth agar

Results of the Study For Wooden Carriers

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Contact Time Carrier Type Replicate CFU/Carr ier

Time Zero1 3.30E+06

4.40E+06 N/A2 4.20E+063 5.70E+06

10 Minutes

1 4.05E+067.85E+06 N/A2 5.40E+06

3 1.41E+07

1 2.90E+016.70E+01 99.9991% 5.072 1.27E+02

3 4.50E+01

Decon7 1:2.51 3.00E+01

2.87E+01 99.9996% 5.442 3.00E+013 2.60E+011 1.92E+02

2.81E+03 99.96% 3.452 1.50E+033 6.75E+031 9.00E+02

3.45E+03 99.96% 3.362 6.30E+033 3.15E+031 3.72E+06

3.92E+06 50.13% 0.302 5.28E+063 2.75E+06

TestMicroorganism

AverageCFU/Carr ier

Average Percent

Reduction Compared to

TreatedControl

Average Log1 0


TreatedControl

S. enterica ATCC 10708

Numbers Control

TreatedControl (Wood)

Decon7 1:1

Decon7 1:5

Decon71:10

Glutex1:256

Results of the Study For Wooden Carriers

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1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3Numbers Control

TreatedControl (Wood)

Decon7 1:1

Decon7 1:2.5

Decon7 1:5

Decon71:10

Glutex1:256

Time Zero 10 Minutes

1.00E+00

1.00E+01

1.00E+02

1.00E+03

1.00E+04

1.00E+05

1.00E+06

1.00E+07

1.00E+08

Carrier Type

CFU

/Car

rier

Results of the Study For Concrete Carriers

The limit of detection for this assay was 1 CFU/ml. Observations below this limit are representedas <1.00E+00 in the table above.

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Time Zero1 1.95E+07

2.55E+07 N/A2 2.85E+073 2.85E+07

10 Minutes

1 1.65E+076.55E+07 N/A2 1.13E+08

3 6.75E+071 <1.00E+00

<1.00E+00 >99.999998% >7.822 <1.00E+003 <1.00E+00

Decon7 1:2.51 <1.00E+001

<1.33E+00 >99.999998% >7.692 <1.00E+003 2.00E+001 3.15E+03

2.90E+03 99.996% 4.352 5.55E+033 2.00E+001 7.50E+02

4.20E+03 99.994% 4.192 1.17E+043 1.57E+021 <1.00E+00

<1.00E+00 >99.999998% >7.822 <1.00E+003 <1.00E+00

TestMicroorganism

AverageCFU/Carr ier

Average Percent


TreatedControl

Average Log1 0


TreatedControl


Numbers Control

TreatedControl

(Concrete)

Decon7 1:1

Decon7 1:5

Decon71:10

Glutex1:256


The limit of detection for this assay was 1 CFU/ml. Observations below this limit are representedas zero in the graph above.

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1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3Numbers Control

TreatedControl (Concrete)

Decon7 1:1

Decon7 1:2.5

Decon7 1:5

Decon71:10

Glutex1:256

Time Zero 10 Minutes

1.00E+00

1.00E+01

1.00E+02

1.00E+03

1.00E+04

1.00E+05

1.00E+06

1.00E+07

1.00E+08

1.00E+09

Carrier Type

CFU

/Car

rier


The limit of detection for this assay was 1 CFU/ml. Observations below this limit are representedas <1.00E+00 in the table above.

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Time Zero1 3.90E+07

4.45E+07 N/A2 6.45E+073 3.00E+07

8 Hours

1 <1.00E+00<1.00E+00 >99.999998% >7.652 <1.00E+00

3 <1.00E+001 <1.00E+00

<1.00E+00 >99.999998% >7.652 <1.00E+003 <1.00E+00

Decon7 1:2.51 <1.00E+00

<1.00E+00 >99.999998% >7.652 <1.00E+003 <1.00E+001 <1.00E+00

<1.00E+00 >99.999998% >7.652 <1.00E+003 <1.00E+001 <1.00E+00

<1.00E+00 >99.999998% >7.652 <1.00E+003 <1.00E+001 <1.00E+00

<1.00E+00 >99.999998% >7.652 <1.00E+003 <1.00E+00

TestMicroorganism

AverageCFU/Carr ier

Average Percent


NumbersControl

Average Log1 0


NumbersControl


Numbers Control

TreatedControl

(Concrete)

Decon7 1:1

Decon7 1:5

Decon71:10

Glutex1:256


The limit of detection for this assay was 1 CFU/ml. Observations below this limit are representedas zero in the graph above.

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

Copyright © Antimicrobial Test Laboratories, 2015. Reproduction and ordinary use of this study report by the entitylisted as “Sponsor” is permitted. Other copying and reproduction of all or part of this document by other entities isexpressly prohibited, unless prior permission is granted in writing by Antimicrobial Test Laboratories.

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1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3Numbers Control

TreatedControl (Concrete)

Decon7 1:1

Decon7 1:2.5

Decon7 1:5

Decon71:10

Glutex1:256

Time Zero 8 Hours

1.00E+00

1.00E+01

1.00E+02

1.00E+03

1.00E+04

1.00E+05

1.00E+06

1.00E+07

1.00E+08

Carrier Type

CFU

/Car

rier

Decon7 Crushes Glutex in Study (Salmonella on Wood and Concrete Carriers)

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