SUMMARY OF TESTS BEING EXECUTED WITH OXILITE OR NEUTRAL OXILITE PRODUCED ON WPT WATER-MASTER EQUIPMENT.

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1 SUMMARY OF TESTS BEING EXECUTED WITH OXILITE OR NEUTRAL OXILITE PRODUCED ON WATER-MASTER EQUIPMENT. 2 LABORATORY TEST EXECUTED WITH OXILITE. Bactericidal effect of (ph 2-3, ORP>11mV, 3mg/l) inocolum 1.7 x 1 4 cfu/ml is shown in table 1. Bacteria Disinfectant 1 s 6 s 18 s MSSA methicillin sensitive Staphylococcus aureus MRSA methicillin resistant Staphylococcus aureus Staphylococcus epermidis Pseudomonas aeruginosa Escheria coli Serratia marcencens Burkholderia cepacia >5 >5 >5 >5 >5 >5 41 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 27 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 We tested the microbial activity of STEL water against methicillin-sensitive Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Staphylococcus epermidis, Serratia marcencens, Escheria coli, Pseudomonas aeruginosa and Burkholderia cepacia. The number of bacteria was reduced below detection limit following incubation in STEL water for 1s. The bactericidal activity of STEL water was similar to that of 8% ethanol, but superior to that of,1 chlorhexidine and,2% povidine iodone. We conclude that STEL water is a low cost but powerful disinfectant.

2 Bactericidal effect of (ph 2-3, ORP>11mV, 3mg/l) inocolum 1.7 x 1 6 cfu/ml is shown in table 2. Bacteria Disinfectant 1 s 6 s 18 s MSSA methicillin sensitive Staphylococcus aureus MRSA methicillin resistant Staphylococcus aureus Staphylococcus epermidis Pseudomonas aeruginosa Escheria coli Serratia marcencens Burkholderia cepacia 8 >5 >5 15 >5 >5 >5 >5 >5 >5 71 >5 >5 >5 >5 3 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 1 >5 >5 >5 237 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 The tuberculocidal activity of (ph2-3.5, ORP>115mV, 5mg/l) under clean and dirty conditions is shown in table 3. Suspension tests with Mycobacterium tuberculosis H37 Rv NCTC 7416 Contact time Mean log1 reductions obtained Clean conditions Dirty conditions Pre disinfection count (challenge) 1 min 2 mins 5 mins 1 mins 2 mins 3 mins >6.24 >6.24 >6.24 >

3 A > 5log1 reduction in Mycobacterium tuberculosis is indicative of tuberculocidal activity i.e. high level disinfection. This was achieved under clean conditions (absence of organic matter) in 5. Little or no reduction in test mycobacteria was detected in 3 mins when Superoxidised water was tested in the presence of 1% serum (dirty conditions). The sporicidal activity of (ph2-3.5, ORP>115mV, 5mg/l) under clean and dirty conditions is shown in table 4. Log 1 spores (Bacillus subtill var Niger NCTC 173) remaining after exposure to Anolyte or 2% glutaraldehyde. (suspension tests) Contact time 2% glutaraldehyde Clean conditions Dirty conditions Clean conditions Dirty conditions Pre disinfecting challenge min 2 min 5 min 1 min 2 min 3 min 1 hour 2 hours A 6 log1 reduction in test spores was achieved with freshly generated solution in 5. This is far more rapid than the widely used 2% glutaraldehyde. However, when 1% horse serum was added as an organic load no appreciable sporicidal activity was noted in 2 hours. Items must therefore be scrupulously clean before they are disinfected using this agent. Initial cleansing using validated automated system is advised.

4 LABORATORY TEST EXECUTED WITH NEUTRAL ANOLYTE(AN,AND or ANK) The bactericidal activity of Neutral Oxilate (ph 6-7, ORP>1mV, active chlorine 3-5mg/l) under clean and dirty conditions is shown in table 5. Suspension test with Pseudomonas aeruginosa NCTC 6749 Solution clean conditions fresh 24 hours dirty (1% serum) fresh 24 hours dirty (1% serum) fresh 2% glutaraldehyde clean 1%serum 1%serum Log 1 remaining after The bactericidal activity of Neutral (ph 6-7, ORP>1mV, active chlorine 3-5mg/l) under clean and dirty conditions is shown in table 6. Log 1 remaining after The sporicidal activity of Neutral (ph 6-7, ORP>1mV, active chlorine 3-5mg/l) under clean and dirty conditions is shown in table 7. Suspension tests with Bacillus subtills var Niger NCTC 173 spores Solution clean conditions fresh dirty (1% serum) fresh dirty (1% serum) fresh 2% glutaraldehyde clean 1%serum 1%serum Log 1 remaining after Precount 3 secs 1 min 2 mins 5 mins 1 mins 2 mins Suspension test with Enterococcus faecalis NCTC 775 Solution precount 3 secs 1 min 2 mins 5 mins 1 mins 2 mins clean conditions fresh 24 hours dirty (1% serum) fresh 24 hours dirty (1% serum) fresh 2% glutaraldehyde clean 1%serum 1%serum precount hr 2 hrs secs min mins mins mins mins mins

5 The sporicidal activity of Neutral (ph 6-7, ORP>1mV, active chlorine 3-5mg/l) under clean conditions is shown in table 8. Surface tests with Bacillus subtills var Niger NCTC 173 spores Solution precount 5 mins 1 mins 2 mins 3 mins 1 hr 2 hrs % glutaraldehyde (5/5) 4.38 (5/5) 4.29 (5/5) 3.86 (5/5) 1.3 (5/5) (/5) Log 1 remaining after The sporicidal activity of Neutral (ph 6-7, ORP>1mV, active chlorine 3-5mg/l) under clean and dirty conditions is shown in table 9. Endoscope tests with Bacillus subtills var Niger NCTC 173 spores Solution Channel Predisinfection count 2% glutaraldehyde Biopsy Suction Air Water Biopsy Suction Air Water Mean post disinfection count.49 (-2.3).23 (-3.8).24 (-2.48).15 (-2.28) Mean Log 1 RF No. of cycles The bactericidal activity of Neutral (ph 7-8, ORP>8mV, active chlorine 2-5mg/l) is shown in table 1. Virulent Cholera vibrio Avirulent Cholera vibrio 6365, 1444, ) Concentration of Concentration of 5ml testsample [mg/l] cholera vibrio /ml NEGATIVE NEGATIVE NEGATIVE 18/ml NEGATIVE NEGATIVE NEGATIVE 17/ml NEGATIVE NEGATIVE NEGATIVE Measured after 24hrs at 37 degrees C. The bactericidal activity of Neutral (ph 7-8, ORP>8mV, active chlorine 5mg/l) is shown in table 11. Entero-bacteria Cloacae and Staphilococcus aureus. Bacterial suspensions were in contact with Neutral (1:9 dilution) during 1, 2, 3. Active chlorine concentrati on, mg/l Type of bacteria Initial bacterial concentrati on, CFU/ml Percent of viable cell number reduction during contact period 5 5 E. Cloacae St. aureus 3.8x x min. 2 min. 3 min

6 E. Clocae St. aureus E. Clocae 1.9x x x

7 PRACTICAL TESTS WITH NEUTRAL (ANK) The bactericidal activity of Neutral (ph 7-8, ORP>8mV, active chlorine 2-5mg/l) under clean and dirty conditions is shown in table 12 to 15. The efficiency of ANK bactericidal action on three-day old Enterobacter cloaceae culture Active Material Number of viable cells E. cloaceae chlorine in ANK, mg/l Applied, CFU/ml of prospective washing sample Washed without ANK treatment, CFU/ml Washed after ANK treatment, CFU/ml 25 Sandstone plate 3.5x1log6 2.6x1log4 (no protein) 2.5x1log5 (protein) 4.2x1log2 1.6x1log5 Age of culture = 72 hours at 37 degrees C. Results after 4 exposure. The efficiency of ANK bactericidal action on tree-day old culture Pseudomonas Fluorencens in the presence of albuminous impurities. Active Material Number of viable cells Ps. Fluorencens chlorine in ANK, mg/l Applied, CFU/ml of prospective washing sample Washed without ANK treatment, CFU/ml Washed after ANK treatment, CFU/ml 35 Sandstone plate 3.2x1log8 2.4x1log6 (protein) < 5 (protein) Age of culture = 72 hours at 3 degrees C. Results after 4 exposure. The effect of ANK bactericidal action against Staphylococcus aureus, strain 96, on different surfaces Active Material Number of viable cells St. aureus chlorine in ANK, mg/l Applied, CFU/ml of prospective washing sample Washed without ANK treatment, CFU/ml Washed after ANK treatment, CFU/ml Sandstone plate Plastic Stainless steel Stainless steel 3.3x1log7 2.4x1log7 3.6x1log7 2.2xlog7 Age of culture = 72 hours at 3 degrees C. Results after 4 exposure. 1.4x1log7 (no protein) 1.8x1log7 (protein) 3.1x1log5 (protein) 1.6x1log6 (protein) 1.4x1log4 (protein) 4.2x1log2 7.5x1log6 3.5x1log1 <1 3.6x1log2 The efficiency of ANK bactericidal action in relation to Bacillus Cereus BKM 687 on different Active Material Number of viable cells B. cereus BKM 687 chlorine in ANK, mg/l Applied, CFU/ml of prospective washing sample Washed without ANK treatment, CFU/ml Washed after ANK treatment, CFU/ml Sandstone plate Dutch tile Stainless steel Stainless steel Age of culture = 72 hours at 37 degrees C. Results after 4 exposure. 2.8x1log6 1.4x1log6 8.4x1log5 2.2x1log6 3.x1log5 (no protein) 4.2x1log5 (protein) 2.6x1log5 (protein) 2.5x1log3 (protein) 1.8x1log3 (protein) 1.x1log2 1.x1log4 1.7x1log2 2.2x1log3 1.3x1log3

8 Neutral and was applied for washing and disinfecting of a poultry meat processing shop equipment. The treatment guarantees practically complete eradication of sanitary-indicative and specific microflora. The results are shown in table 16. For washing and disinfecting of equipment surfaces (1 minute each, respectively) we applied a water jet sprayer and a compression pump providing the pressure of 3-4 Atm. Presence of microflora, thousands of microbial bodies/cm Groups Technological tube Before treatment E. coli 1.1 Salmonella.74 After treatment Group 1 (Water 9 o C., septodor,,5% solution) E. coli Salmonella.5 Group 2 (Water 9 o C., acid E. coli ph 2-3, C.ac 2 Salmonella mg/l) Group 3 (Redulite 18 o C., E. coli.6 ph 1-11, acid, ph Salmonella.1 2-3, C.ac 2mg/l) Group 4 (Neutral, E. coli.3 18 o C., ph 6-7, C.ac Salmonella 2mg/l) Note: Disinfectant agent consumption as per 3ml/m2 Ground meat mincer Technological table Cutter

9 DRINKING WATER TESTS WITH (A) The bactericidal activity of (ph 2-3, ORP>11m, 5mg/l) on Escherichia coli in relation to concentration of in the treated water and concentration of amount of bacteria is shown in table 17. Concentration of Escherichia Coli [CFU/1mL] 78. up to 14. Concentration of in drinking water [mg/l],15,3,5 1,2 5 Minutes 1 Minute 1 Minute 1 Minute Minute 1 Minute 1 Minute 1 Minute Reaction time: no bacteria in 1 ml. The bactericidal activity of (ph 2-3, ORP>11m, 5mg/l) on Pseudomonas aeruginosa in relation to concentration of in the treated water and concentration of amount of bacteria is shown in table 18. Concentration of Escherichia Coli [CFU/1mL] 12. up to up to 5.6 Concentration of in drinking water [mg/l],15,3,5 1,2 5 Minutes 5 Minutes 5 Minutes 5 Minutes 5 Minutes 5 Minutes 1 Minute 1 Minute 3 5 Minutes Reaction time: no bacteria in 1 ml. The bactericidal activity of (ph 2-3, ORP>11m, 5mg/l) on Legionella pneumonas in relation to concentration of in the treated water and concentration of amount of bacteria is shown in table 19. Microbilogical contamination in Sample (CFU/ml*) Amount of dosed to sample, ml Active Chlorine in dosed to sample, mg/l > > > <2. CFU =TMC = total microbiological contamination. (1. TMC = 1log4 /ml) Microbilogical contamination in Sample after 6mins. (CFU/ml*)

10 The bactericidal activity of (ph 2-3, ORP>11m, 25mg/l) on Lactobacillus frigidus and Pediscoccus damnosus in relation to concentration of active chlorine in the treated water and concentration of amount of bacteria is shown in table 2. Bacteria With treated tapwater up to a measurement of,5mg/l active chlorine (1:5 dosing of ) Without the presence of organic load/ clean conditions After 1 L. frigidus 3.6 x 1 4 ~1 ~ P. damnosus 4.5 x Bacteria After 2 With treated tapwater up to a measurement of 2,5mg/L active chlorine (1:1 dosing of ) With the presence of organic load/ dirty conditions After 1 L. frigidus 1.1 x 1 5 >1 P. damnosus 2 x After 2 The sporicidal activity of (ph 2-3, ORP>11m, 25mg/l) on Sachh. diastaticus and Pichia anomaia in relation to concentration of in the treated water and concentration of amount of bacteria is shown in table 21. Spores Sacch. Diastaticus With treated tapwater up to a measurement of,5mg/l active chlorine (1:5 dosing of ) Without the presence of organic load/ clean conditions After x 1 4 >1 >1 ~1 ~7 Pichia anomaia 2.1 x 1 4 >1 >1 >1 ~6 Spores Sacch. Diastaticus After 2 With treated tapwater up to a measurement of 2,5mg/L active chlorine (1:1 dosing of ) With the presence of organic load/ dirty conditions After x 1 4 ~ Pichia anomaia 1.5 x 1 4 >1 >1 ~1 5 After 2

11 The bactericidal activity of (ph 2-3, ORP>11m, 25mg/l) on Pediscoccus damnosus in relation to concentration of in the treated water and concentration of amount of bacteria is shown in table 22. Bacteria With treated tapwater up to a measurement of 2,5mg/L active chlorine (1:1 dosing of ) Without the presence of organic load/ clean conditions After 1 P. damnosus 2.2 x 1 3 Bacteria After 2 With treated tapwater up to a measurement of 5mg/L active chlorine (1:5 dosing of ) With the presence of organic load/ dirty conditions After 1 P. damnosus 1.5 x 1 4 ~1 After 2 The sporicidal activity of (ph 2-3, ORP>11m, 25mg/l) on Sachh. diastaticus and Pichia anomaia in relation to concentration of in the treated water and concentration of amount of bacteria is shown in table 23. Spores With treated tapwater up to a measurement of 2,5mg/L active chlorine (1:1 dosing of ) Without the presence of organic load/ clean conditions After 1 Pichia anomaia 2.7 x Spores After 2 With treated tapwater up to a measurement of 5mg/L active chlorine (1:5 dosing of Oxililte) With the presence of organic load/ dirty conditions After 1 Pichia anomaia 2.3 x 1 4 >1 1 After 2

12 The bactericidal activity of Neutral (ph 7-8, ORP>8m, 25mg/l) on Lactobacillus frigidus and Pediscoccus damnosus in relation to concentration of active chlorine in the treated water and concentration of amount of bacteria is shown in table 24. Bacteria With treated tapwater up to a measurement of 2,5mg/L active chlorine (1:1 dosing of ) Without the presence of organic load/ clean conditions After 1 L. frigidus 3.2 x P. damnosus 3.2 x 1 3 Bacteria After 2 With treated tapwater up to a measurement of 5mg/L active chlorine (1:5 dosing of ) With the presence of organic load/ dirty conditions After 1 L. frigidus 4.5 x 1 4 >1 >1 ~8 ~8 P. damnosus 6 x 1 4 ~1 After 2 The sporicidal activity of Neutral (ph 7-8, ORP>8m, 25mg/l) on Sachh. diastaticus and Pichia anomaia in relation to concentration of in the treated water and concentration of amount of bacteria is shown in table 25. Spores With treated tapwater up to a measurement of 2,5mg/L active chlorine (1:1 dosing of ) Without the presence of organic load/ clean conditions After 1 After 2 Sach. diastaticus 5 x 1 4 ~ Pichia anomaia 1.8 x 1 4 ~4 ~ Spores With treated tapwater up to a measurement of 5mg/L active chlorine (1:5 dosing of ) With the presence of organic load/ dirty conditions After 1 Sach. diastaticus 5 x 1 4 >1 ~ After 2 Pichia anomaia 15.1 x 1 4 >1 >1 ~1 ~8

13 All the above given tables are extracted from scientific reports owned by the Aquastel group, by the authors or from articles made public. The Aquastel group has permission to use this documentation for marketing purposes. If you appreciate this summary of reports and want to make some copies of it, please take in mind that: These researches are the results of some intensive work: the authors and the Aquastel Group have invested quite a lot of time and money in it; Making copies without written permission is unlawful and punishable by law

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