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Alka Vita Research Report

Antibacterial Susceptibility Test Results for the Alka Vita Product
July 17, 2006


The purpose of this series of experiments was to test the susceptibility of both Gram-positive and Gram-negative bacteria to the modified liquid silicon product Alka Vita.

Six organisms were tested. The Gram-negative bacteria were Escherichia coli K–12, Escherichia coli O157:H7 (ATCC 35150), and Salmonella enterica serovar Typhimurium LT2. The Gram-positive bacteria were Enterococcus faecalis (ATCC 19433), Staphylococcus aureus (ATCC 12600), MRSA and Streptococcus pyogenes (ATCC 19615).

The method used was the broth microdilution method for susceptibility testing of antibacterial agents as recommended by the Clinical and Laboratory Standards Institute (CLSI). The detailed procedure is as follows.


1.               A culture of the test organism was grown at 35°C with aeration in sterile Cation-adjusted Mueller-Hinton broth (Becton-Dickinson, Sparks, MD) until slightly turbid, and the turbidity was then adjusted to a 0.5 McFarland standard [equivalent to approximately 108 colony-forming units (cfu) per ml]. To improve cell yield in the Enterococcus faecalis and Streptococcus pyogenes cultures, the Cation-adjusted Mueller-Hinton broth was supplemented with 2.5mg per mL of yeast extract (Becton-Dickinson)
2.               Appropriate dilutions of the Alka Vita product were prepared in sterile Cation-adjusted Mueller-Hinton broth such that the final concentrations of the product were 0.5%, 1.0%, 2.0%, 3.0%, and 5.0%. A 0% (no-product) control was also included.
3.               Appropriate volumes of the standardised cell suspension were added to the dilutions of the product to achieve a final cell density of 5 x 105 cfu per mL. The dilutions were then aliquoted into the wells of a 96-well microtiter plate (with a volume of 0.2mL per well).
4.               The first two wells in each column of the mocrotiter plate were reserved for a no-cell control, and the remaining six wells were used for the cell-inoculated dilutions of the product. Columns 1-6 represented the 0%, 0.5%, 1.0%, 2.0%, 3.0%, and 5.0% concentrations, respectively.
5.               The microtiter plates were incubated at 35°C for 16–20 hours. The culture turbidity was measured by visible-light spectroscopy (i.e., the absorbance of the culture at a wavelength of 590 nm) using a microtiter plate reader, and the results were recorded automatically.
6.               The baseline absorbance of the wells containing the uninoculated dilutions (no-cell controls) was subtracted from the absorbances of the wells containing the cell-inoculated dilutions of the product. For each set of six replicate wells, the average (mean) absorbance, % inhibition, and standard deviation were calculated from the collected data.

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The results of the experiments are summarised in the appended data tables. The minimum inhibitory concentration (MIC) of the product was the 1.0% concentration for all three of the Gram-negative bacteria (E. coli K-12, E. coli O157:H7, and S. enterica). However, the three Gram-positive bacteria (E. faecalis, S. aureus, and S. pyogenes) showed greater resistance. The MIC of the product was the 2.0% for S. aureus and 3.0% for E. faecalis and S. pyogenes, although partial inhibition was seen at lower concentrations for all three of these organisms.

The conclusions drawn from the experimental results are that the product completely inhibits the growth of all bacterial strains under the test conditions at a concentration of 3.0%. Depending on the specific organism, complete inhibition can occur at a concentration as low as 1.0%.

Click here to view Data Tables of Research Report







Randall M. Jeter, Associate Professor
Department of Biological Sciences
Texas Tech University
Lubbock, TX 79409-3131


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