testing of the bactericidal action of products based on the “hydronic” technology (“informed...

Testing of the Bactericidal Action of Products Basedon the ‘‘Hydronic’’ Technology (‘‘Informed Glass’’) on ATCCStrains of Pathogenic Gram-Positive and Gram-Negative

Bacteria and Yeasts (Staphylococcus aureus,Escherichia coli, and Candida albicans)

Aleksandar Racz, Ph.D., M.P.H., M.D.1 and Josip Cipris, M.A.2

Abstract

Background: Scientific experiments were conducted with drinking water kept in ‘‘ordinary, everyday-use’’glasses (drinking tumblers) and the so-called ‘‘informed’’ glasses (drinking tumblers), a patent-protected productsupposed to have an effect on the ‘‘structure, vitality and memory of water,’’ for which the manufacturer claimsto have a wide range of positive effects on the health of patients with chronic medical problems, especially a‘‘revitalizing’’ effect on water and the body (blue informed glass), additional metabolic effects such as facilitatingweight loss (green informed glass), and a stress-relieving action (red informed glass). According to the claims ofthe patent owner, a Slovenian inventor Vili Poznik, by the use of the ‘‘orgone methodology,’’ various tran-scendental, vitalizing information is purportedly coded and inscribed into the glass; this action is additionallyenforced by the addition of the ‘‘magic life’’ symbol—a specially designed energy condenser which, togetherwith the selected information, is permanently introduced into the liquid contained in the glass. The process ofselection and transfer of information is a production secret known only to the inventors of the original ‘‘hydronictechnology’’ for the discovery of which they have been awarded numerous prizes at innovation fairs, amongothers, a gold medal and a Crystal Globe at INPEX XVI exhibition in Pittsburgh, PA, in 2000.Research objectives: The aim of this research was to test the claims for informed glasses with respect to thebacteriocidal actions that have been claimed for these products.Materials and methods: Given that the manufacturer attributes to the products produced by the ‘‘hydronic’’technology—besides the effects on organoleptic properties of the drinking water consumed (which are subjectiveand liable to suggestion)—a broad bactericidal action against bacteria, yeasts, and molds but does not state asingle scientific proof, the efficacy and bactericidal action of the products based on the hydronic technology weretested using standardized microbiological tests in a certified laboratory. Respecting the principle of a single-blinded test for each of 5 samples of each type of informed glass, the growth reduction factor (RF; difference log:colony forming unit per mL [cfu=mL] of control glass and log cfu=mL of each informed glass) was determinedafter arbitrarily set time intervals of 0, 2, 4, 6, and 8 hours in spring water experimentally contaminated withstandardized ATCC strains.Results: While the blue informed glass produced statistically significant bacterocidal action on all strains of thebacteria tested under normal daylight conditions, this effect did not occur with the red and green informedglasses. In the other conditions (complete darkness and exposures to ultraviolet light), any effects noted werenegligible.Conclusions: The results indicate rather a possible photocatalytic action of a currently unknown and undeclaredcomponent, introduced accidentally or added intentionally during the manufacture of the glass, rather the actioncaused by the effects of applying ‘‘orgone technology.’’

1University of Applied Health Studies, Zagreb, Mlinarska, Croatia.2Croatian National Institute of Public Health, Zagreb, Rockfellerova, Croatia.This is a full-text article of a presentation for the 5th International Congress on Complementary Medicine Research, Tromsø, Norway,

May 19–21, 2010.

THE JOURNAL OF ALTERNATIVE AND COMPLEMENTARY MEDICINEVolume 16, Number 4, 2010, pp. 463–471ª Mary Ann Liebert, Inc.DOI: 10.1089=acm.2009.0600

463

Introduction

A vast range of products for ‘‘revitalization of water’’—which are primarily aimed at people with chronic med-

ical conditions seeking to improve their quality of life bymeans of alternative medicine—are sold through the internet.A special place on the Slovenian market (European Union;EU) is held by products based on the ‘‘hydronic technology,’’for which a Slovenian inventor Vili Poznik was awarded agold medal and a Crystal Globe at the INPEX XVI exhibitionin Pittsburgh, PA, in 2000. The subject matter of this researchis his leading product, the so-called ‘‘informed’’ glasses(drinking tumblers), which are advertised as the handiestproduct intended for revitalization of water. The product isa ‘‘vitreous,’’ manually blown and shaped glass that, it hasbeen claimed, ‘‘improves health, strengthens immunity, andprevents growth of pathogenic bacteria and all kinds ofyeasts.’’1

The technology of the manufacture of this glass has notbeen explained and is not available to expert members of thepublic, and what explanations of its efficacy that have beenprovided are not in conformance with generally acceptedscientific notions. Although supposedly protected by a pat-ent, the patent number cannot be found on the manufac-turer’s internet sites. Moreover, the patent number is neitherstated on the original packaging or on the product itself, noris it available on the official internet sites of the INPEX XVIexhibition. The inventor claims that these are unique see-through glasses (drinking tumblers), on the external side ofwhich a blue, green or red round seal of a few centimetres indiameter is placed, and a ‘‘magic life’’ symbol resembling theTibetan mandala is embedded on the bottom.2 Blue glass(Fig. 1) is his main, core product, the market success of whichwas followed by rapid production and launching on themarket of two other glasses (with red and green seals) and,after the completion of this research, a so-called gold in-formed glass was introduced. The glasses differ in color ofthe embedded seal, which does not only serve a differenti-ation purpose, but purportedly has specific powers, as well.The glasses are then exposed to bombarding with concertedtranscendental vitalizing information by means of an orgonetransmitter in the inventor’s laboratory. It should be pointedout that this information is not ‘‘written’’ (i.e., published), butthat these are purportedly transcendental instructions, ofcourse completely invisible to the eye or the microscope,which, once memorized in the glass, have a permanent effectof ‘‘vivification of tired or dead water.’’ The inventor claimsthat changes occur in the molecular structure of water (i.e.clusters or clouds of molecules are broken down and theangle between hydrogen bonds shifts by 108), which in-creases the quality of water.3 Depending on the ‘‘embeddedinformation,’’ in each type of glass (drinking tumbler), it hasa specific effect; thus, according to the manufacturer, the so-called blue informed glass (Blue-IG) neutralizes detrimentaleffects of substances dissolved in water and ‘‘increasesthe person’s vital abilities, improves health and enhancesthe vitality of the body, improves the taste of bread, softensfacial skin and alleviates acne‘‘4,5 the so-called green glass(Green-IG) facilitates healthy weight loss,6 and the so-calledred glass (Red-IG) has an antistress effect.7

The results of the Allium test allegedly prove ‘‘reducedgenotoxicity of municipal water’’ in major Slovenian cities; in

Ljubljana from 15.53% to 10.76%, and in Celje from 18.75% to11.76%.2 Successful application in agriculture is supportedby the ‘‘evidence’’ that, visually, the growth of wheat irri-gated with informed water was *10% better than that of acontrol group.8 In support of the effect on baker’s yeast, it isstated that bread made with revitalized water ‘‘leavens muchslower.’’5,9

Except for unsubstantiated allegations about widespreadbactericidal efficacy, no technical data about the product,composition, and material can be found in internet data-bases, and there is no reference to the efficacy of revitalizedwater. The only measurable effects of ‘‘informed water’’ onthe human body were recorded by the gas discharge visu-alization (GDV) method, which showed that the human aurais increased and its fragmentation reduced after drinkingwater from an informed glass.10

Research Objectives

Considering that a very broad bactericidal action is attrib-uted to informed glasses by the claim that they ‘‘efficientlyrevitalize water thus preventing growth of pathogenic bacteriaand all kinds of yeasts,’’ the objectives of this research were asfollows:

The main objective of the research was to test the bacteri-cidal efficacy of all three types of informed glasses with re-spect to ordinary glasses, by monitoring the reduction in thenumber of colonies of pathogenic strains of Gram-positive

FIG. 1. Blue ‘‘informed glass’’ shown in black and whitephoto.

464 RACZ AND CIPRIS

and Gram-negative bacteria and yeasts, in arbitrarily set timeintervals of 2, 4, 6, and 8 hours, including the effect on theinitial microbial load (0 interval) of samples of water.

An additional objective of the research was to investigatethe potential photocatalytic effect of possibly intentionallyused or accidentally introduced (nondeclared) compoundsduring the manufacture of one or more types of informedglasses.

Materials and Methods

The experimental part of the research was carried out inthe microbiological laboratory of the Section for Micro-biology of Foodstuffs and General Use Items of the CroatianInstitute of Public Health, which has been accredited by theCroatian Accreditation Agency. Five sets of glasses wereused in the experiment, with each set containing an ordi-nary (control) glass and a so-called blue, green, and redinformed glass. Before the experiment, the glasses weresterilized in a MELAG Euroklav 23 V-S autoclave to preventinitial contamination by microorganisms. Nonchlorinatedspring water in a glass bottle, tested bacteriologically by themembrane filtration method, was used in the experiment.

The testing of bactericidal efficacy was conducted in aTELSTAR Bio-II-A biologic cabinet, using standard strains ofmicroorganisms grown on appropriate selective media:Staphylococcus aureus ATCC 6538—Baird-Parker (with eggyolk tellurite agar), Biolife; Escherichia coli ATCC 29922—TBX(Tryptone Bile X-Gluc agar), Biolife; Candida albicans ATCC10231—Sabouraud Chloramphenicol agar, Biolife.

The culture preparation was carried out in the followingway: standard (ATCC) strains on ready-made carriers weregrown on the Triptone-Soy Broth liquid medium, in order torevitalize them. After 24 hours of incubation, standard bac-terial strains grown on the liquid medium were placed ontothe abovementioned solid differential media, which wasincubated for 24 hours at 378C, whereas the medium con-taining yeasts was incubated for 72 hours at 308C. Coloniesgrown on differential media were suspended in 10 mL ofsterile saline solution. Turbidity was determined by a densi-tometer, and corresponded to &1Eþ8 cfu=mL [colony form-ing units per milliliters] on the McFarland scale. Workingsuspensions were prepared from stock suspensions. One (1)mL of suspension was transferred using an automatic cali-brated pipette (Eppendorf ) from the first test tube where theconcentration of microorganisms was &1Eþ8 cfu=mL into9 mL of sterile saline solution (0.9% NaCl). The suspensionwas homogenized, using a vibration agitator. Further dilu-tions were made using the same procedure, until a suspensionconcentration of &1Eþ4 cfu=mL was obtained. In the exper-iment, to reduce potentization by vortexing on sucking liquid,only automatic calibrated pipettes (Eppendorf )—and notsyringes—were used for dilution.

Furthermore, the experimental protocol was designed sothat the prepared working suspensions should be inoculatedinto differential media. One (1) mL was taken from eachprepared working suspension of pure culture of Staphylo-coccus aureus (&1Eþ4 cfu=mL) and inoculated into eachglass containing water. The suspension was homogenizedon a horizontal agitator and 0.1 mL of suspension was takenfrom each glass by reusing an automatic calibrated pipette(Eppendorf ) and transferred onto the selective medium.

Sterile L-rods were used to distribute the suspension uni-formly on the medium. Media were placed in the incubatorat 378C for 24 hours in accordance with the valid HRN ISO6888-1 norm, and Candida albicans was incubated at 308C for72 hours.11

The first inoculation of contaminated water from glassesonto selective media was marked as zero hour and used forcontrol of the introduced microorganism count. Water ininformed glasses contained approximately the same micro-organism count=ml as that in the control glass. The aboveprocedure was repeated after having arbitrarily set time in-tervals of 2, 4, 6, and 8 hours, which are usually used instandardised methods in human microbiology, not takinginto account the mean-generation-times of the cells underexperimental conditions, which limits the generally stan-dardised method. After 24 hours of incubation the number ofgrown colonies was counted for each time interval and ex-pressed in cfu=ml (colony forming unit). During all timeintervals glasses were covered by a Petri dish, directly ex-posed under normal daylight conditions which allowed UVto penetrated into the water through the material (glass)from which the glasses was made and not through an openwater surface, and kept at the temperature of 248C. The sameprocedure was used with all microorganisms, with pertain-ing selective media, in the same time intervals, in accordancewith applicable norms HRN EN 1040, ISO 6579:2002, EN ISO6579:2002 and ISO=TS 16649-3:2005. To exclude a photo-catalytic effect, the experiment was repeated in the condi-tions of darkness and in the conditions of daylight exposurewith the change of UV filters (UV Filter film 2 Mm that block98,7% UV).*

Measurements were performed on all five sets of glasses,three times on each glass, and the results, for easy reference,were expressed only as an arithmetic mean based on 15measurements of the number of colonies in each informedglass as an average cfu=mL.

The statistical significance of the number of colonies in thenormal daylight conditions, and darkness and daylightconditions with the use of UV filters was determined byt-test using the SPSS 15.0 program.

Results

Surprisingly, there was statistically significant bacteri-cidal action of the blue informed glass on all strains—Staphylococcus aureus–ATCC 6538 (RF 1.02=1.27), Escherichiacoli–ATCC 29922 (RF 0.88=1.35) and Candida albicans–ATCC 10231 (RF 0.5=0.60)—especially after 6 and 8 hoursunder normal daylight conditions as opposed to whatoccurred when using the red and green informed glasses,where this effect was negligible (RF< 0.1). However, in thetests repeated in complete darkness, where the effect ofUV rays was excluded, the monitoring of reduction in timeintervals did not show a tendency of growth within blueglass as under the normal daylight, with a statisticallysignificant difference in RF (light=darkness) after 8 hoursconfirmed by a t-test with a risk of less than 1% (Staphy-lococcus aureus [t¼ 91.799; df¼ 8; p¼ 0000], Escherichia coli[t¼ 72.139; df¼ 8; p¼ 0000], and Candida albicans [t¼ 52.931;

*The UV Filter film specification is available at: www.hanitafolije.com=files=arh=uv=UV%20Filter%20Film,%202%20Mil.pdf

INFORMED GLASS AND BACTERIA AND YEASTS 465

df¼ 8; p¼ 0000]). The other two glasses produced, again, anegligible bactericidal action. The same result was obtainedalso after repeating the experiment in which glasses wereexposed under normal daylight conditions and isolatedusing an ultraviolet (UV) filter that blocked 98.7% of UVlight.

These results are presented in Tables 1–3 and in Histo-grams 1–9 (Figs. 2–10), containing basic parameters of de-scriptive statistics and the arithmetic mean of the reductionfactor value (RF¼difference log cfu=mL of the ordinaryglass and log cfu=mL of each informed glass [RF]).

The results of the research are very challenging. Whenwater was contaminated with a standard strain of Staphy-lococcus aureus at daily light levels, the greatest bactericidalefficacy with respect to the control glass was recorded withBlue-IG. The greatest value of RF was observed (RF¼ 1.02and RF¼ 1.27) in time intervals of 6 and 8 hours, with atendency to increase in time. Bactericidal efficacy related tocell apoptosis was confirmed by intracellular morphologicchanges observed by electronic microscope. Bactericidalefficacy of Red-IG and Green-IG in the same time intervalwas very low and did not differ significantly from the ef-ficacy of the control glass (Table 1 and Fig 2; Histogram 1).However, in the conditions of darkness, the number ofcolonies of Staphylococcus aureus with respect to the controlglass was not reduced both in Red-IG and Green-IG, andcontrary to the properties attributed to it, neither in Blue-IG, where reduction factor values were insignificant andranged from RF¼ 0 to RF¼ 0.05 without showing a ten-dency of increase (Fig. 3; Histogram 2). The difference inthe number of colonies determined by t-test after 6 hoursin Blue-IG at daily light compared to that in the condi-tions of darkness (normal daylight conditions=darkness)was statistically significant with a risk lower than 1% (t¼

53.463; df¼ 8; p¼ 0.000). The same results were also con-firmed after 8 hours, when the difference in bacterial countdetermined by t-test was also statistically significant with arisk lower than 1% (t¼ 91.799; df¼ 8; p¼ 0.000) and with agreater t-ratio that in the 6-hour interval. In the repeatedtest, when all four glasses were kept under UV filters, thenumber of colonies of Staphylococcus aureus with respect tothe control glass was not reduced both in Red-IG, Green-IG, and, again, not in Blue-IG, when RF values were in-significant without showing a tendency to increase (Fig 4;Histogram 3).

The reduction of the number of colonies in water con-tamination with a standard strain of Escherichia coli exposedto the action of informed glasses at daily light levels was thegreatest with Blue-IG, for which the following reductionfactors were recorded (RF¼ 0.88 and RF¼ 1.35) in the timeintervals of 6 and 8 hours (Table 2 and Fig. 5; Histogram 4).The bactericidal effect of Red-IG and Green-IG differed in-significantly from that of the control glass.2 When condi-tions were changed (no daily light), the number of coloniesof Escherichia coli with respect to the number of colonies inthe control glass (normal daylight condition=darkness) wasnot significantly reduced both in Red-IG and Green-IG, andeven in Blue-IG, which is displayed in Histogram 5 (Fig. 6)showing that RF values were insignificant in all threeglasses and range from RF¼ 0.02 to RF¼ 0.09, and that notendency to increase was recorded in different time intervalsas was also done in daily light. It was also established that,under changed conditions (normal daylight conditions=darkness), the difference in the reduced number of coloniesafter 6 hours in Blue-IG was statistically significant with arisk lower than 1% (t¼ 38.572; df¼ 8; p¼ 0.000), as it wasafter 8 hours (t¼ 72.139; df¼ 8; p¼ 0.000) and with a greatert-ratio than in the 6-hour interval. In the repeated test, when

Table 1. Reduction Factor (RF) for Staphylococcus aureus–ATCC 6538 in Normal Daylight Conditions

After 0, 2, 4, 6 and 8 Hours

Control Glass Green I-Glass Red I-Glass Blue I-Glass

Time cfu=mL Log cfu=mL Log cfu=mL RF Log cfu=mL RF Log cfu=mL RF

0 hours 3000 3.47 3.47 0.00 3.47 0.00 3.47 0.002 hours 2300 3.36 3.34 0.02 3.32 0.04 2.70 0.904 hours 2100 3.32 3.32 0.00 3.32 0.00 2.60 0.726 hours 2100 3.32 3.27 0.05 3.30 0.02 2.30 1.028 hours 1900 3.27 3.20 0.07 3.25 0.02 2.00 1.27

cfu, colony-forming units.

Table 2. Reduction Factor (RF) for Escherichia coli–ATCC 29922 in Normal Daylight Conditions

after 0, 2, 4, 6, and 8 Hours

Control Glass Green I-Glass Red I-Glass Blue I-Glass


0 hours 2200 3.34 3.32 0.02 3.32 0.02 3.34 0.002 hours 1300 3.11 3.04 0.07 3.02 0.09 3.06 0.054 hours 2300 3.36 3.25 0.11 3.28 0.08 2.90 0.466 hours 3800 3.58 3.30 0.28 3.32 0.26 2.70 0.888 hours 5600 3.75 3.52 0.23 3.44 0.31 2.40 1.35


466 RACZ AND CIPRIS

all four glasses were kept under UV filters, the numberof colonies of Escherichia coli with respect to the controlglass was not reduced both in Red-IG, Green-IG, andBlue-IG, in which RF values were insignificant (Fig 7; His-togram 6).

Given that fungicidal efficacy was also attributed to theproduct, the results obtained in experiments with pathogenicbacteria were checked on one standard strain of yeast. Inexperiments with Candida albicans that were designed in thesame manner and performed in daily light, the greatestfungicidal effect was expressed by the RF of the number ofcolonies with respect to the number of colonies in the controlglass obtained with Blue-IG, with a minimum increase in RFin time intervals, in which the greatest values were deter-mined in the time intervals of 6 and 8 hours (RF¼ 0.59 andRF¼ 0.60). Fungicidal efficacy of the other two glasses didnot differ significantly from that of the control glass (Table 3and Fig. 8; Histogram 7). In the changed conditions (nodaily light) the number of Candida albicans colonies withrespect to those in the control glass (normal daylightconditions=darkness) was not significantly reduced in Red-IG and Green-IG, and not in Blue-IG. It can be concludedfrom Histogram 8 that RF values for all three types of glassesare insignificant, range from RF¼ 0 to RF¼ 0.06, and do notshow a tendency to increase as what occurred in those ob-tained under daily light. A t-test on Blue-IG established astatistically significant difference in efficacy with a risk lowerthan 1% expressed as RF after 6 hours (normal daylightconditions=darkness) (t¼ 34.888; df¼ 8; p¼ 0.000) as well asafter 8 hours (t¼ 52.931; df¼ 8; p¼ 0.000), with a greatert-ratio then in the 6-hour interval. Also, in this test, when allfour glasses were kept again under UV filters, the number of

colonies of Candida albicans with respect to the controlglass was not reduced both in Red-IG, Green-IG, and notin Blue-IG, where RF values were insignificant (Fig. 10;Histogram 9).

Discussion

When interpreting the obtained results from the method-ological point of view, it should be emphasized that only theinitial suspension was prepared and determined turbidi-metrically. However, checks were made at greater dilutionsby counting on selective plates. During the experiment, col-ony count was determined only by the method of cultivationon solid selective media (depending on the type of micro-organism), and not turbidimetrically.

It should also be kept in mind that the variations in themicroorganism count observed in the controls are not con-sidered relevant in microbiology because microorganismscan never be entirely distributed homogeneously. Significantvariations cannot be observed when the number of coloniesis expressed in logarithmic values, as is conventionally donein microbiology (the variation and large numbers wouldotherwise give a false picture of the deviation if expressed inabsolute values).

Regretfully, we have nothing with which we could com-pare the obtained results. The inventor did not publishanywhere the results upon which he based his conclusionsregarding the bactericidal action of the product, and wecould not find a single published paper on any related re-search. The only results that were available to us were thoseregarding a nonscientific experiment in which, when indus-trially contaminated water was kept for a short period of

Table 3. Reduction Factor (RF) for Candida albicans –ATCC 10231 in Normal Daylight Conditions

After 0, 2, 4, 6, and 8 Hours

Control glass Green I-Glass Red I-Glass Blue I-Glass


0 hours 1230 3.08 3.08 0.00 3.08 0 3.07 0.012 hours 980 2.99 2.81 0.18 2.90 0.09 2.59 0.404 hours 800 2.90 2.77 0.13 2.90 0.00 2.47 0.436 hours 790 2.89 2.77 0.12 2.87 0.02 2.30 0.598 hours 600 2.77 2.76 0.01 2.77 0.00 2.17 0.60


0

0.5

1

1.5

2

2.5

3

3.5

log

cfu

/ m

l

0 hours 2 hours 4 hours 6 hours 8 hours

CONTROL GLASSGREEN INFORMED GLASSRED INFORMED GLASSBLUE INFORMED GLASS

FIG. 2. Histogram 1. Graphic display of reduction factor (RF) for Staphylococcus aureus-ATCC 6538 in normal daylightconditions after 0, 2, 4, 6, and 8 hours. See key for glass colors shown in black and white.


FIG. 4. Histogram 3. Graphic display of reduction factor (RF) for Staphylococcus aureus–ATCC 6538 under ultraviolet filterafter 0, 2, 4, 6, and 8 hours. See key for glass colors shown in black and white.

FIG. 3. Histogram 2. Graphic display of reduction factor (RF) for Staphylococcus aureus-ATCC–6538 in darkness after 0, 2, 4,6, and 8 hours. See key for glass colors shown in black and white.

FIG. 5. Histogram 4. Graphic display of reduction factor (RF) for Escherichia coli–ATCC 29922 in normal daylight conditionsafter 0, 2, 4, 6, and 8 hours. See key for glass colors shown in black and white.

FIG. 6. Histogram 5. Graphic display of reduction factor (RF) for Escherichia coli–ATCC 29922 in darkness after 0, 2, 4, 6, and8 hours. See key for glass colors shown in black and white.

468

time in the product that was a precedent of the informedglass (Blue-IG 2000), after 48 hours of incubation (at 378C in acupboard!) there was a 25% decrease in the initial bacterialcount and a 100% destruction of molds compared to asample that was not kept in the informed product.12 Theobtained results are, in any case, interesting. If experimentswere carried out only in daily light and only with the basicmodel of Blue-IG, we could speculate that the bactericidalaction of the informed glass and the efficiency of the hy-dronic technology were undoubtedly proven. The compar-ative results showing no bactericidal and fungicidal effects ofRed-IG and Green-IG glass, if this is not ‘‘disregarded in thename of a higher purpose,’’ might be explained by a possibleuncoordinated effect of subsequently embedded informa-tion, and by the absence of the foreseen synergistic effect.However, the fact that a reduction in the number of colonieswas recorded in daily light conditions in one type of glassand the complete absence of reduction in all three types ofglasses in the conditions of darkness and use of UV filtersprovided a bases for an assumption that the reduction ofbacteria and yeasts in Blue-IG could have been the result of aphotocatalytic effect of a component that was not statedanywhere by the producer.

The objective of this study was to test the justification ofthe claim of the bactericidal effect of the product. This wasdone only by comparing colony counts in ordinary glassesand the so-called informed glasses under the same condi-tions and during the same time intervals. Further exhaus-tive attempts to find an explanation for the absence of thereduction effect observed under normal daylight conditionswith the blue informed glass in the conditions of darknessand under an UV filter and for the complete absence of thereduction effect with the other two types of glasses go be-yond the context of this article.

However, for the sake of scientific correctness, and wish-ing to provide direction for future investigations, prelimi-nary initial measurements were made, the results of whichmay be an incentive to investigators in the future. Namely,one of the possible hypotheses to explain the proven effectspoints to the possible presence of subsequently applied na-nolayers of a compound with photocatalytic properties. Thepresence of nanoparticles could not be proven by using a setof different grade Millipore filters, neither could their pres-ence be detected by examining the water from the blue glassunder an electronic microscope.

The comparison of the results of measurements in theconditions of daylight, darkness, and under an UV filter in-dicates a possible photocatalytic process; therefore, it is log-ical to suspect the presence of an undeclared substance witha photocatalytic effect in the material from which the blueglass is made, and which is not present in the other two typesof informed glasses and in the control glass. This is the rea-son why all glasses were subjected to X-ray diffractionanalysis in a certified laboratory, where a clear differencewas obtained in one segment between the diffractogramsrecorded with the blue glass compared to those recordedwith the other two types of glasses, which showed no dif-ferences. Further investigations will have to be directed to-ward qualitative and quantitative determination of theunknown component in the glass itself. On the basis of ex-amination of literature references on the use of specific ma-terials with photocatalytic properties in medicine for wallcoatings of operating rooms, compounds such as TiO2 orSb2O3 ,13,14 could hypothetically be present, which should bedetermined in future investigations by qualitative andquantitative analysis, using a sophisticated method such asAAS. Namely, these are widely used compounds that causephotocatalysis as a result of the effect of UV light that excites

FIG. 7. Histogram 6. Graphic display of reduction factor (RF) for Escherichia coli–ATCC 29922 under an ultraviolet filter after0, 2, 4, 6, and 8 hours. See key for glass colors shown in black and white.

FIG. 8. Histogram 7. Graphic display of reduction factor (RF) for Candida albicans–ATCC 10231 in normal daylight con-ditions after 0, 2, 4, 6, and 8 hours. See key for glass colors shown in black and white.


electrons to shift to a higher energy state and to a conduc-tivity band, whereby free electrons and free electron holesappear. All of them react with oxygen and water, resulting insuperoxide ions and hydroxide radicals. The latter ones arehighly reactive and oxidize (i.e. destroy organic molecules),the results of which are carbon dioxide and water. Con-sidering that, in this way, microorganisms are also de-stroyed, all coatings of this kind are also called antibacterialor antimicrobial. This might explain the reduction effectobserved with Blue-IG. However, at this moment, this con-clusion is only hypothetical, as there is no clear explanationof the bactericidal efficacy of the Blue-IG in the conditions ofdaily light, which represents a strong limitation of this in-vestigation, but should also be an invitation and an incentivefor future extensive investigations.

In the evaluation of the efficacy of application of suchtheoretical constructions, it is not possible to avoid ques-tioning the principal thesis on revitalization of ‘‘tired’’ waterby hydronic technology by crushing large molecule clustersof up to 400 molecules of water to smaller groups which, insuch form, enter more easily into cells through the ‘‘hexag-onal door.’’ Namely, according to scientific notions, mutualbinding of neighboring molecules of water occurs throughhydrogen bonds that break up continuously and are formedagain where a typical bond has a life span that is measuredin picoseconds, which excludes the possibility of formationand maintenance of large clusters of molecules in the so-called tired waters. Water also enters cells by individualmolecules through aquaporin transcellular channels for wa-ter transport, which makes dubious, from the scientific point

of view, the thesis that, when we ‘‘pour water into an in-formed glass, clusters of molecules are de-grouped intosmaller groups of molecules which penetrate cells moreeasily’’2 as an explanation of the beneficial effect of revitali-zation of water.

Given that the manufacturer does not make any declara-tions of the composition of the product, and as there are noavailable data on the applied technologic procedures, andnot a single piece of scientific evidence of the efficacy of thisvery expensive product is provided, additional material isprovided for the critics of alternative medicine who generallyspeak of this as a simple fraud,15 or quackery.16 The value ofsuch investigations lies in the endeavor to establish verystrict and appropriate methodology that would be applicablealso in the evaluation of efficacy of similar products on themarket, aiming to distinguish valuable products from thosethat are not,17 because giving false hope to chronically andseriously ill patients is the most cruel form of quackery, as itmay stray the victims far away from other proven and effi-cient complementary or alternative treatments.

Conclusions

Antimicrobial efficacy of the hydronic technology wastested using pure strains of Gram-positive bacteria Staphy-lococcus aureus–ATCC 6538, Gram-negative Escherichia coli–ATCC 29922 and the yeast Candida albicans–ATCC 10231.The following conclusions can be made:

1. The research did not prove any antimicrobial efficacy oftwo out of three informed glasses (Red-IG and Green-

0

0.5

1

1.5

2

2.5

3

3.5

log

cfu

/ m

l


CONTROL GLASSGREEN INFORMED GLASSRED INFORMED GLASSBLUE INFORMED GLASS

FIG. 9. Histogram 8. Graphic display of reduction factor (RF) for Candida albicans–ATCC 10231 in darkness after 0, 2, 4, 6,and 8 hours. See key for glass colors shown in black and white.

0

0.5

1

1.5

2

2.5

3

3.5

log

cfu

/ m

l


CONTROL GLASS

GREEN INFORMED GLASSRED INFORMED GLASSBLUE INFORMED GLASS

FIG. 10. Histogram 9. Graphic display of reduction factor (RF) for Candida albicans–ATCC 10231 under an ultraviolet ilterafter 0, 2, 4, 6, and 8 hours. See key for glass colors shown in black and white.

470 RACZ AND CIPRIS

IG) with respect to an ordinary glass, under normaldaylight conditions, in time intervals of 2, 4, 6, and 8hours, including the initial microorganism load of waterin all glasses.

2. The research proved antimicrobial efficacy of Blue-IGwith respect to an ordinary glass, under daily lightconditions, in time intervals of 2, 4, 6, and 8 hours,including the initial microorganism load of water in allglasses. The reduction of all standard strains resultingfrom the apoptosis used in the testing of antimicrobialefficacy by Blue-IG at daily light was statistically sig-nificant.

3. The research did not prove antimicrobial efficacy of anyof the three informed glasses with respect to an ordi-nary glass, under the conditions of darkness and whenusing UV filters in time intervals of 2, 4, 6, and 8 hours,including the initial microorganism load of water, be-cause there was no significant reduction of microor-ganisms in the given time intervals.

4. Taking into consideration the statistically significantdifference in RFs between the normal daylight condi-tions on the one hand and darkness and under UV filteron the other hand with one tested informed glass, it canbe supposed that the bactericidal action exerted byBlue-IG is rather a result of the photocatalytic effect ofan unknown compound used in the manufacture of theglass, which is also supported by initial diffractometrictests, then based on modern science through an unex-plainable effect of hydronic technology of ‘‘informingglasses and revitalising tired water.’’ Future sophisti-cated investigations directed toward qualitative andquantitative identification of the possible presence of anundeclared compound with photocatalytic propertiesshould be performed via the AAS method, with theobjective to explain the observed antimicrobial efficacyof Blue-IG only in the conditions of normal daily light,for which this investigation has not provided a con-clusive explanation at this point.

Disclosure Statement

No competing financial interests exist for both authors.

References

1. Hydro Protect Water Technologies. Hidroprotect. Onlinedocument at: www.hydroprotec.com=informed-glasses.htmlAccessed August 1, 2009.

2. Informed Products Enhance Molecular Structure of theWater [in Croatian]. Online document at: www.terapeut-u-kuci.huped.hr=Accessed August 11, 2009.

3. Vetrovec J. Glass for better water [in Croatian]. Misteriji2006;153:19–21.

4. Informed Glass—Blue [in Croatian]. Online document at:Available from: www.zdrava.hr=art=0=plava_informirana_casa Accessed August 11, 2009.

5. Hydronic Products—Blue Glass [in Croatian]. Natura-Igm.Online document at: www.natura-igm.hr=informirani_proizvodi.php?url¼Plava%20Casa&ID¼ 1 Accessed August 15, 2009.

6. Hydronic Products—Green Glass [in Croatian]. Natura-IgmOnline document at: www.natura-igm.hr=informirani_proizvodi.php?url¼Zelena%20casa&ID¼ 2 Accessed August18, 2009.

7. Hydronic Rroducts—Red Glass [in Croatian]. Natura-Igm.Online document at: www.natura-igm.hr=informirani_proizvodi.php?url¼Crvena%20Casa&ID¼ 3 Accessed Au-gust 11, 2009.

8. Memory of Water [in Croatian]. Misteriji 2008;174:19–22.9. Vetrovec J. Informed glass 2000 [in Slovenian]. Misteriji

2001;98:29–30.10. Kononenko I, Zrimec T, Sadikov A, Skocaj D. GDV images:

Current research and results. In: Proceedings of New Scienceof Consciousness: 3rd International Conference on CognitiveScience. Ljubljana, Slovenia, October 2000:65–65. Onlinedocument at: http:==lkm.fri.uni-lj.si=xaigor=slo=clanki=KOGN2000.pdf Accessed August 11, 2009.

11. McDonnell G, Russell AD. Antiseptics and disinfectants:Activity, action, and resistance. Clin Microbiol Rev 1999;12:147–179.

12. Vetrovec J. Less bacteria and no moulds [in Slovenian].Misteriji 2001;97:21–22.

13. Kamat PV, Meisel D. Nanoscience opportunities in en-vironmental remediation. CR Chimie 2003;6:999–1007.

14. Todini and Co. s.p.a. Antimony Trioxide. Online documentat: www.todiniandco.com=products=antimony=antimony_trioxide.html Accessed August 10, 2009.

15. Marusic M. Complementary and glternative medicine—a measure of crisis in academic medicine. CMJ 2004;45:684–688.

16. Lower S. Water Pseudoscience and Quackery. Oneline doc-ument at: www.chem1.com=CQ Accessed August 11, 2009.

17. Sackett DL, Rosenberg WM, Muir Gray JA, et al. Evidencebased medicine: What it is and what it isn’t. BMJ 1996;312:71–72.

Address correspondence to:Aleksandar Racz, Ph.D., M.P.H., M.D.

Mlinarska 38Zdravstveno veleuciliste (University of Applied Health Studies)

Zagreb 10000Croatia

E-mail: [email protected]


testing of the bactericidal action of products based on the “hydronic” technology (“informed...

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