Time Kill Test

Test Objective:
To determine the effectiveness of SyDERMA® to destroy of bacteria within 30 seconds of contact time.

Test Method:
The samples were screened for sterility prior to testing by streak plating 0.10 gram of material onto Microbial Content Test Agar plates. The plates were incubated at 37°C for 48 hours. The plates were then observed for bacterial growth after 48 hours incubation. All samples were found to be in good microbial condition. Ten grams of sample were transferred to sterile 25 x 150 mm test tubes, inoculated with 0.05 milliliter of pure inoculum suspension and mixed thoroughly. A one-milliliter aliquot was withdrawn using a sterile 3 cc syringe. At 30 seconds, post-inoculation one milliliter was transferred to 9ml D/E Neutralizing Broth and dilution plate counted to recover number of viable organisms. D/E Neutralizing Agar was used to plate the dilutions. Plates were incubated at 37°C for 48 hours and then counted to recover number of viable organisms. Inoculum suspension concentrations of the test microorganisms were determined prior to testing by dilution plate count. The initial inoculum count in the samples was determined mathematically. Percentage reduction is calculated as the difference between the initial count in the sample and the number of surviving Colony Forming Units per milliliter at thirty seconds divided by the initial inoculum count and multiplied by 100.

Interpretation of Assay:
At the end of 30 seconds of contact time, the number of Colony Forming Units (CFU) of the test bacterial were counted and compared to the Initial Concentration for each of those bacterial. The results are expressed as a percentage reduction of CFU's.

Test Results:
SyDERMA®, reduced 99.99% of bacteria tested at 30 seconds contact time.

Observations:

Bacteria	Test	Initial Concenteration in Sample	*CFU's/ML of Test Microorganism Recovered at 30 Seconds	Percent Reduction at 30 Seconds
Pseudomonas aeruginosa	ATCC 15442	1.9 x 10 7	<10	>99.99%
Salmonella typhimurium	ATCC 6994	3.4 x 10 7	<10	>99.99%
Staphylococcus aureus	ATCC 6538	3.3 x 10 7	<10	>99.99%