Independent Researcher who tested Aqualiv. He kept running the tests and Aqualiv kept working.
E.coli Experiment
Testing the ability of our ceramic water treatment media to program water with antibacterial properties. Status: Complete
The following experiment was performed by Dr. Sean Sleight, Ph.D., Department of Bioengineering, University of Washington (September 2010)
Dr. Sleight was provided with a ceramic ball containing our antibacterial programming. Dr. Sleight used the ceramic ball to program common purified water with antibacterial properties. Using E.coli bacteria, Dr. Sleight was able to demonstrate the antibacterial performance of programmed water vs. unprogrammed water. To verifiy that the ceramic did not add any chemicals to the water, a deprogrammed ceramic ball was tested. Results with the deprogrammed ceramic are identical to the control water.
The experiment was performed over a 24-hour period at 37C (three replicates each). The photo below shows the results. Control is on the left (non-programmed water), programmed water on the right.
Dr. Sleight's Official Report
PURPOSE
Validate the ability of AquaLiv's ceramic water treatment media to program ordinary water with antibacterial properties.
BACKGROUND
Bacterial antibiotic resistance is a major problem worldwide and our current treatments of antibiotics are causing multi-drug resistant strains to evolve at a rapid pace. Most antibiotics only require one or a few mutations to confer resistance and bacteria continuously exchange antibiotic resistance genes encoded on plasmids, making it very easy for pathogenic bacteria to evolve. Using multiple antibiotics for treatment may sometimes work on a short timescale, but this overuse of antibiotics will continue to make the problem worse long-term. One promising treatment option is to use water programmed with antibacterial properties, a non-chemical treatment. Like temperature, this treatment does not target just one DNA sequence, but the entire organism, making it difficult for bacteria to confer resistance. As a proof-of-principle experiment, we have measured the antibacterial properties with a common, non-pathogenic strain of E. coli.
METHODS
E. coli strain MG1655 engineered with a GFP-expressing plasmid was inoculated from a freezer stock in a test tube containing 5 mL of LB + 100 µg/mL ampicillin media and incubated at 37°C for 24 hours. Three replicates of the overnight culture were then diluted 1:1000 in treatment water (a 45mL glass vial containing one ceramic ball with the antibacterial programming) and control water (same water without the ceramic ball) in 1.5 mL Eppendorf tubes. The tubes were vortexed and incubated at 37°C for 24 hours. To measure cell density after the incubation, the tubes were again vortexed and diluted 1:1000 in LB media, then 0.1 mL was spread on LB + 100 µg/mL ampicillin agar plates. Colony Forming Units (CFU) were counted with the ImageJ program for each of the three treatment and control replicate plates.
RESULTS
The mean cell density after the 1:1000 dilution and 24 hour incubation was plotted (see Figure above) and the error bars represent one standard deviation of the three replicates. On average, there is a 158.64-fold decrease in cell density in the treatment water relative to the control water.
DISCUSSION
Although these results are preliminary, this same experiment has now been successfully replicated three times (using three replicates each on three different weeks), using both in the engineered strain and in the normal MG1655 strain not containing a plasmid, and the results are similar. The cell density was only measured for the experiment shown in the graph because the correct dilution was not made in the other experiments to accurately count colonies on the control plates. It should be noted that the initial cell density was not measured for this experiment to measure survival (final cell density / initial cell density), but this has been performed in subsequent experiments and survival is close to 100% for the control water. The experiment shown here indicates that survival is likely <1% for the treatment water.
It is possible that the antibacterial properties imparted by the ceramic ball do not cause cell death, but instead have a growth inhibition effect, such that the cells cannot grow after being spread on LB agar plates. The programmed water may even have both survival and growth inhibition effects, but this is currently unknown. We tested whether substances in the ceramic ball are responsible for the antibacterial effect, but the experiments containing deprogrammed ceramic balls show the same results as the control water, so this is highly unlikely. Therefore, at this time, it appears that the antibacterial properties come from the water itself, having been successfully programmed by the ceramic ball.
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