Study Title Antimicrobial Activity and Efficacy of Seal Shield's Electroclave Test Method Custom Device Study Study Identification Number NG7233 Study Sponsor Christian Davis Seal Shield 3105 Riverside Avenue Jacksonville, FL 32205 (904) 428-9497 christian.davis@sealshield.com Test Facility Microchem Laboratory 1304 W. Industrial Blvd Round Rock, TX 78681 (512) 310-8378
Test Device Information The test device, pictured below, was received on 25MAY2016. Test Microorganism Information The test microorganism(s) selected for this test: Electroclave Staphylococcus aureus (MRSA) This bacteria is a Gram-positive, cocci shaped, aerobe which is resistant to the penicillin-derivative antibiotic methicillin. MRSA can cause troublesome infections, and their rapid reproduction and resistance to antibiotics makes them more difficult to treat. MRSA bacteria are resistant to drying and can therefore survive on surfaces and fabrics for an extended period of time and therefore makes this bacteria an excellent representative for antimicrobial efficacy testing on surfaces. Enterobacter cloacae This bacteria is a Gram-negative, rod shaped, facultative anaerobe commonly found in the gastrointestinal tract of humans. It is not usually a primary pathogen although it is sometimes associated with urinary and respiratory tract infections. Although relatively susceptible to disinfection and desiccation when dried on a surface, it can be a challenging microorganism to mitigate in solution. Page 2 of 8
Summary of the Procedure Test microorganisms are prepared in TSB and incubated overnight. The test culture is diluted as necessary in PBS targeting a concentration 1x10 6 CFU/carrier. Sterile glass slides (test carriers) are inoculated with 0.020 ml of test culture over approximately 2 square inches and incubated at 36 ± 1 C for approximately 20 minutes, or until visually dry. Test carriers are treated according to device manufacturer's instruction. Treated and control test carriers are harvested in 20 ml of neutralizing broth or PBS and enumerated using standard dilution and plating techniques. Microbial concentrations are determined at time zero by analysis of control materials immediately after inoculation with microorganisms. Additional microbial concentrations are determined at each contact time to account for potential desiccation due to drying. All plates are incubated at 36 ± 1 C for approximately 24-48 hours. After incubation, microbial concentrations are determined and the reduction of microorganisms is calculated relative to the control materials. Study Timeline Culture Initiated Culture Harvested Carrier Inoculated Carrier Harvested Carriers Evaluated Report Delivered 26MAY2016 27MAY2016 27MAY2016 27MAY2016 29MAY2016 15JUN2016 16JUN2016 16JUN2016 16JUN2016 17JUN2016 17JUN2016 Note: Dates in row 1 above detail progression of MRSA testing. Row 2 details progression of CRE testing. Page 3 of 8
Criteria for Scientific Defensibility of a Custom Device Study For Microchem Laboratory to consider a Device Study 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 10 6 cells/carrier or greater. 2. Positive/Growth controls must demonstrate growth of the appropriate test microorganism. 3. Negative/Purity controls must demonstrate no growth of test microorganism. Passing Criteria Because of the nature of the study, passing criteria may be determined by the Study Sponsor. Testing Parameters used in this Study Test Device Mode of Use: UV Light Carrier: Sterile 1 x 3 Glass Slides Replicates: 3 (test), 1 (control) Culture Growth Media: TSB Culture Growth Time: 24 ± 4 hours Culture Dilution Media: PBS, if applicable Culture Supplement: None Inoculum Concentration: 1.0 x 10 6 CFU/Carrier Inoculum Volume: 0.020ml/2in 2 : 1 contact times, see notes Contact Temperature: Ambient Neutralizer (Vol.): D/E Broth (20 ml) Enumeration Media: TSA Enumeration Plate Incubation Temperature: Enumeration Plate 36 C ± 1 C Incubation Time: 24-48 hours Page 4 of 8
Study Notes Antimicrobial efficacy was evaluated at 3 contact times: 13 minutes and 33 seconds Overnight test cultures were not modified (other than dilution in PBS for S. aureus) prior to testing. Study Photographs Left Image: Inoculated carrier placement prior to treatment. Right Image: Bays 2 and 3 during treatment. Page 5 of 8
Control Results Neutralization Method: N/A Growth Confirmation: Confirmed Media Sterility: Confirmed Calculations Where: B = Number of viable test microorganisms on the control carriers A = Number of viable test microorganisms on the test carriers after the contact time Where: B = Number of viable test microorganisms on the control carriers A = Number of viable test microorganisms on the test carriers after the contact time Page 6 of 8
Results of the Study Table 2. S. aureus (MRSA) results at 13 minutes and 33 seconds Microorganism Test Device (minutes:seconds) Carrier Identification Replicate CFU/Carrier Average CFU/Carrier Percent Reduction vs. Control at Log 10 Reduction vs. Control at S. aureus ATCC 33592 MRSA Electroclave Control at Time Zero 5.80E+06 5.80E+06 13 minutes 33 seconds Control 9.40E+06 9.40E+06 Left Middle 2.10E+03 1.84E+03 N/A 1.94E+03 99.98% 3.69 Right 1.88E+03 1.00E+08 1.00E+07 1.00E+06 Replicate CFU/Carrier 1.00E+05 1.00E+04 1.00E+03 1.00E+02 1.00E+01 1.00E+00 Control at Control at Replicate 1 Replicate 2 Replicate 3 Time Zero 13 minute and 33 second contact time Page 7 of 8
Results of the Study Table 5. E. cloacae (CRE) results at 13 minutes and 33 seconds Microorganism Test Device (minutes:seconds) Carrier Identification Replicate CFU/Carrier Average CFU/Carrier Percent Reduction vs. Control at Log 10 Reduction vs. Control at E. cloacae ATCC BAA-2468 CRE Electroclave Control at Time Zero 1.20E+07 1.20E+07 13 minutes 33 seconds Control 6.30E+06 6.30E+06 Left Middle 1.69E+03 9.30E+02 N/A 1.44E+03 99.98% 3.64 Right 1.71E+03 1.00E+08 1.00E+07 1.00E+06 Replicate CFU/Carrier 1.00E+05 1.00E+04 1.00E+03 1.00E+02 1.00E+01 1.00E+00 Control at Control at Replicate 1 Replicate 2 Replicate 3 Time Zero 13 minute and 33 second contact time Page 8 of 8