Please read and share this important technical report (below the action item and Dr. Rima’s comments) showing that nano silver CANNOT become a threat to aquatic life or soil life. The EPA’s assault is a transparent attempt to shore up the ability of Big Pharma to destroy smaller competitors using government regulatory agencies and “Poison Press” as their weapons.
Please click here (http://salsa.democracyinaction.org/o/568/t/1128/campaign.jsp?campaign_KEY=26405) to tell the EPA in no uncertain terms that you are not buying their junk science and you are continuing to buy nano silver.
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Go to Products and then to Silver Solutions (bottom of left side menu).
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Yours in health and freedom,
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Rima E. Laibow, MD
Medical Director
Natural Solutions Foundation
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© 2008 Colloidal Science Lab., Inc.
Silver Nanoparticles: No Threat to the Environment
George J. Maass, Ph.D. (Colloidal Science Laboratories)
Abstract:
Silver is an effective germ fighter and silver nanoparticles are widely recognized
as being especially effective because of their enormously high surface area. Due to the
large number of manufacturers using silver nanoparticles in their products, some concern
has arisen about the effects on the environment when these products are disposed of or
washed. This report will demonstrate that silver nanoparticles do not remain “nanosizeâ€
when they come in contact with normal environmental samples, such as soil and water,
but they agglomerate to form much larger, much less biologically effective, silver
particles which are non-toxic, non-ionic and have no history of being harmful to the
environment or aquatic life. Furthermore, there is no possibility that silver nanoparticles
can ever form silver ions, except in the presence of strong oxidizing substances.
Introduction:
Recently, the Environmental Protection Agency (EPA) issued a statement that
they were planning to regulate companies that produce nanoparticles for use as anti-
microbials. This gives rise to the question: why are the dietary supplement and
nanoparticle industries being targeted at this time and what is the rational for new
regulation of an industry which has previously had no reported harmful effects to
humanity or the environment?
The EPA knows that silver nanoparticles are effective as antimicrobials. The
reason given by the EPA for the current interest is that silver nanoparticles, or products
that claim to be silver nanoparticles, are now being produced by a number of
manufacturers, and the EPA is concerned that, when these particles are disposed of, there
might be an appreciable amount of silver nanoparticles suddenly appearing in the
environment. The proposed concern is due to the fact that the silver nanoparticles are so
small that their surface area per unit weight is very large, therefore, for a given weight of
product, the biological effectiveness, which is proportional to surface area, is far beyond
that which would be expected. This much is true and it is part of the reason that silver
nanoparticles are so attractive in biological applications. The EPA is not questioning the
fact that silver nanoparticles are effective in killing harmful bacteria, but that, because of
the high surface area/weight effectiveness parameter, by disposing these particles into
sewers or waterways, might there be harmful effects to the environment by eliminating
the bacteria which are useful in normal waste degradation?
The last statement shows a misunderstanding of what silver nanoparticles are and
what they do. Nanoparticle technology is relatively new to the scientific community for
good reasons: Nanoparticles are difficult to produce; they are difficult to stabilize once
they have been produced; they are not stable enough to exist in nature for very long. The
purpose of this work is to prove that normal interaction of nanoparticles with various
© 2008 Colloidal Science Lab., Inc.
soils and different water sources is sufficient to change the size and dramatically decrease
the biological activity. Specifically, the areas of examination will include experiments
which will establish that silver colloids, which start out as nanoparticles, upon contact
with the environment “grow†to much larger clusters, as indicated by their average
particle size distribution, (a nanoparticle size measurement), and zeta potential
measurements, which will establish that the zeta potential is outside of the range required
for nanoparticle stability.
There are several other areas which, as recent articles have indicated, show
misunderstandings about silver and its nanoparticles. At Arizona State University,
Westerhoff and Benn (1) have reported “findings†which have never been observed
during the last 10 years at Colloidal Science Laboratories (CSL). For example, they claim
that nanosilver particles produce ionic silver when exposed to moisture. This is NOT
true! This is tantamount to saying that silver metal is water soluble. At CSL, various
forms of silver, ranging from solid silver metal to fine silver powder to colloidal
nanoparticles, have been exposed to water for long periods of time with agitation. No
increase in conductivity or silver ion concentration has ever been observed when silver
metal in any form is treated with water. Silver metal requires chemical treatment with
nitric acid or Aqua Regia to make silver ions. Westerhoff and Benn were working with
commercially prepared socks which are prepared by treating the socks with solutions of
silver in questionable form. There may have been some actual colloidal silver in the
treatment solutions, but there most certainly were soluble silver compounds present, and,
once the socks are rinsed, these are the substances which put silver ions back into the
wash water.
Some people also claim that only silver ions have antimicrobial properties. This
is another misconception. Colloidal silver is a wonderful antimicrobial by itself, which is
a good thing, because silver ions are very reactive with chloride ion to form insoluble,
and biologically inert, silver chloride. This happens in the bloodstream and in nature
wherever halogen anions are present.
As this report will show, the high biological effectiveness of colloidal silver does
not persist in nature because the nanoparticles agglomerate as soon as they come in
contact with the environment, specifically soil and water. Westerhoff and Benn admit that
silver particles “clump†together in the fabric and in the wash water. That is precisely the
point to be considered for environmental safety. How much “clumping†does it take so
that the particles are no longer considered to be “nanoâ€, but much larger, therefore
eliminating the continuing effect of high biological activity? These researchers, and
others, are very quick to make the jump from colloidal nanoparticles, in socks for
example, to ionic silver and its toxic effect, especially in zebra fish, and they speak as if
the ions came from the colloidal nanoparticles. It is necessary to be very clear about this.
If the researchers are finding silver ions in the wash water or anywhere else, then the
silver ions were present in the original material. This cannot be stated too strongly.
Unless people have taken to washing their socks in nitric acid, the conversion of
colloidal silver nanoparticles to silver ions is not possible.
© 2008 Colloidal Science Lab., Inc.
On the other hand, it is not surprising that certain research institutions are being
encouraged, and perhaps even funded, to conduct research that implies toxicity for
colloidal silver. According to well known internet health forum host, Tony Isaacs, the
reason for this is that “The only thing colloidal silver is toxic for is the profits of big drug
companies.†Mr. Isaacs also states “Silver has been used effectively by mankind to fight
germs and ailments for thousand of years.†“It (colloidal silver) is far safer, more
effective and less expensive than the marginally effective and side effect laden
mainstream antibiotics.†(2)
The work at hand will examine four different environmental conditions which
change the morphology and stability of silver colloids:
1. The effect of drainage of silver colloids through several soil samples.
2. The effect of interaction of silver colloids with different water
samples.
3. The effect of exposure of silver colloids to sunlight.
4. The change in level of silver colloids with regard to biological activity.
Experimental:
Sample Selection
At the outset, the first two questions to be addressed were what environmental
samples should be used and to what concentration of colloidal silver should these
samples be exposed. Since this is a first attempt at this kind of information, it was
decided to limit the environmental samples to the following:
1. Sand, taken from the New Jersey shore
2. Dirt, taken from central New Jersey,
3. Dirt, taken from Northern Pennsylvania
4. Water, local tap water from Westampton, NJ
5. Water, sea water, taken from the New Jersey shore
6. Water, taken from a northern Pennsylvania well
The soil samples represent some of the most common types found on the Eastern
Coast of the United States. The sand is essentially an Entisol, which is a type of soil that
is not subject to a great deal of chemical change and is common to areas where deposition
and removal occur at regular intervals. The New Jersey soil is primarily an Ultisol,
which contains clay, quartz, kaolinite and various iron oxides. The Pennsylvania soil is
most likely a mixture of Alfisols and Inceptisols which are clays that are productive for
growing most crops and are common to many areas. (3)
The water samples are Sea water, rich in many salts, NJ tap water, which has been
through routine purification, and Pennsylvania well water, which most likely contains
carbonates and nitrates. Therefore, the selection of samples should be sufficient to
establish the effect of the environment on nanoparticles for this initial study.
© 2008 Colloidal Science Lab., Inc.
Approximately 8 to 10 lbs of each environmental sample were collected. From
these, 18 to 20 samples of 20.0 g each were selected, and these were randomized for the
testing.
Next, in looking at the quantitative amount of colloidal silver to be used, it was
decided that the initial test case should provide information with regard to an
overabundance of nanoparticles being released to the environment, rather than just a trace
amount. If the environment is not substantially altered by the overabundance, surely it
will not be influenced by smaller amounts.
Preliminary work indicated that, at concentrations of up to 6 ppm, and probably
higher, based on the weight of soil samples, no nanoparticles would survive. Therefore, a
more reasonable amount, but still an enormously high concentration for a natural
occurrence, was selected.
Colloidal silver samples at our disposal were of the dietary supplement type and
average at least 20 ppm of silver. Most soil samples require 0.5 to 0.75 their weight in
water to start draining. It was decided that the colloidal silver would be diluted 10 to 1
and then applied to each soil sample. This would make each sample contain a minimum
of 2 ppm of silver nanoparticles, based on the weight of the soil. This would correspond
roughly to dumping 27 liters of 20 ppm colloidal silver onto one ton of dirt. Since most
colloidal silver customers are concerned with teaspoon and tablespoon quantities, it
should be safe to say that this experiment covers something well above the worst case
scenario.
Measurements:
In each experimental case, the selected sample of colloidal silver was mixed with
the environmental sample and the change in particle size and zeta potential recorded after
a specified time. The instrument used for this work was the Malvern Zetasizer, Model
Nano ZS. Since the samples which were in contact with soil contained very large
macroparticles and rocks, the samples all required vacuum filtration through grade 601
Ahlstrom filter paper to eliminate the particles which are 3 to 4 orders of magnitude
greater than the ones of interest in this study. This filtration has no effect on
nanoparticles.
For the trials in which the environmental samples were water, the colloidal silver
was diluted 10 to 1 in the water in question.
Results:
The initial data in this section shows the properties of the colloidal silver used in
these trials. This sample, selected at random, had 81% of its particles at 1.74 nm, and a
Zeta potential of -31.7 mV. The data in Tables 1 through 6 show the results of the
particles found in the fluid after the specified time of contact with the environmental
© 2008 Colloidal Science Lab., Inc.
samples in question. For example, it is shown in Table 1 that when DI water was filtered
through the soil samples, no nanoparticles could be found, but only large particles on the
order of 300 nm or more.
Table 2 shows that, after only 15 minutes of contact with the soil samples, a
decrease in zeta potential, and the smallest particles have increased to the 3 to 8 nm
range, and they still represent 80 to 90% of the total.
Table 3 indicates that, after a full 7 days of contact with the soil, but kept away
from sunlight, the nanoparticles have increased 3 to 8 times in size.
In Table 4, these results are more dramatic, since the samples were all exposed to
the sunlight for the 7 days, with the increases in size being 7 to 20 fold, and the smallest
particles now representing only 30 to 40 % of the total.
For the data in Table 5, the colloidal silver was left in contact with the
environmental water sample for 21 days in sunlight. As can be seen from the table, the
particle sizes have significantly increased (3 orders of magnitude), with a corresponding
drop in the zeta potential.
In Table 6, the samples were left in contact with the water samples instead of the
soil samples for 7 days in the sunlight. The results of these tests show that each water
sample also decreased the zeta potential and increased the particle size.
PROPERTIES OF COLLOIDAL SILVER USED IN TESTING
Smallest Particles,
nm Zeta Pot., mV Total Ag, ppm Ionic Ag, ppm
1.74 -31.7 21.4 9.60
TABLE 1 – DEIONIZED WATER
Filtering
Medium Smallest
Particles, nm Zeta Pot., mV Total Ag, ppm Ionic Ag, ppm
Sand None found -20.2 0.00 0.00
NJ Soil None found -1.5 0.00 0.00
PA Soil None found -31.3 0.00 0.00
TABLE 2 – COLLOIDAL SILVER – 15 MIN. CONTACT – 7 DAYS LATER
Filtering
Medium Smallest
Particles, nm Zeta Pot., mV Total Ag, ppm Ionic Ag, ppm
Sand 3.53 -20.6 1.14 0.00
NJ Soil 4.35 -22.2 1.57 0.20
PA Soil 8.30 -21.7 1.05 0.20
© 2008 Colloidal Science Lab., Inc.
TABLE 3 – COLLOIDAL SILVER – 7 DAYS CONTACT – NO SUNLIGHT
Filtering
Medium Smallest
Particles, nm Zeta Pot., mV Total Ag, ppm Ionic Ag, ppm
Sand 5.4 -15.7 1.27 0.00
NJ Soil 9.7 -20.8 0.56 0.00
PA Soil 14.7 -2.8 0.17 0.00
TABLE 4 – COLLOIDAL SILVER – 7 DAYS CONTACT – SUNLIGHT
Filtering
Medium Smallest
Particles, nm Zeta Pot., mV Total Ag, ppm Ionic Ag, ppm
Sand 11.3 -22.8 0.94 0.00
NJ Soil 26.9 -22.2 0.41 0.00
PA Soil 34.2 -21.2 0.35 0.00
TABLE 5 – COLLOIDAL SILVER – 21 DAYS CONTACT – SUNLIGHT
Filtering
Medium Smallest
Particles, nm Zeta Pot., mV Total Ag, ppm Ionic Ag, ppm
Sand >2000 -11.3 0.03 0.00
NJ Soil >1900 -4.6 0.24 0.00
PA Soil >1700 -7.6 0.39 0.00
TABLE 6 – COLLOIDAL SILVER – 7 DAYS CONTACT
Filtering
Medium Smallest
Particles, nm Zeta Pot., mV Total Ag, ppm Ionic Ag, ppm
Tap Water 113 -11.3 0.03 0.00
Sea Water 631 -4.6 1.14 0.00
Well Water 32.1 -15.7 1.47 0.20
While some of the changes in particle size seem small, one must realize that they
represent large changes in loss of surface area and, since biological activity is
proportional to surface area, this would correspond to large losses in biological
effectiveness. In Figure 1, it can be seen that a change in particle size from 2 to 10 nm
represents about an 80% loss in surface area for the same weight of particles. This is an
approximation, since the exact morphology of the particles is not known. To make these
calculations possible, an assumption has to be made that the particles are spherical and
the spheres are close packed.
© 2008 Colloidal Science Lab., Inc.
Fig. 1 – Loss of Area
0
1
2
3
4
5
6
7
8
9
10
0 5 10 15 20 25 30 35 40
diameter, nm
su
r
f
.
a
re
a
/m
l
In a previous paper by F. Key and G. Maass (4), the nature of a colloid was
described as being a suspension of very small particles which are stabilized by having a
diffuse double layer of solution ions around them. The charge acquired by these particles
gives rise to a potential difference (i.e., mutual repulsion) between them that keeps them
separate and stabilizes the colloid. This potential difference is called the Zeta Potential,
and has been described in countless books on electrolytic effects in solutions. When the
colloid is composed of nanoparticles, the task of preventing the agglomeration is not an
easy one.
As the previous paper pointed out, if the zeta potential is more negative than
-30 mV, then the mutual repulsion between particles is sufficient to keep them separate
and stabilize the colloid. When the zeta potential is between -15 mV and 0 mV, however,
the particles agglomerate and flocculation or precipitation occurs.
In a 1996 report by the Department of the Interior by M. Elimelech and A. E.
Childress (5), it was pointed out that for world average fresh water rivers, the
concentration of common anions and cations across all normal pH ranges is sufficient to
change the zeta potential range from about -10 mV to +5 mV, making agglomeration of
nanoparticles occur. In sea water, the agglomeration would be even more pronounced.
Conclusions:
Theoretically, if a very large amount of silver nanoparticles from many sources
were to be dispersed into the same part of the environment at the same time, it might be
© 2008 Colloidal Science Lab., Inc.
possible that the concentration of some good bacteria, as well as the bad bacteria, would
be diminished, but this is not at this time considered a serious threat for the dietary
supplement nanoparticles. The points to be remembered are as follows:
1. This report has demonstrated that silver nanoparticles will grow to
biologically far less active “clumps†even if one dumps 27 liters of 20
ppm colloidal silver on each ton of soil. In practice, this is an
enormously high number which could not be expected to be reached
realistically.
2. In spite of the number of manufacturers producing silver nanoparticles
or claiming to be silver nanoparticles, because of the low concentrations
in which these products are sold, the total amount which could be
released in any part of the environment would still be expected to be
very low.
3. As shown by all the experiments above, nanoparticles do not last as
nanoparticles in nature for very long, but grow to harmless clumps of
silver metal.
4. Silver nanoparticles are not water soluble, and therefore, silver colloids
will not release silver ions into the environment.
Once agglomeration of the silver nanoparticles occurs, the result is simply
silver metal; a harmless metal which has existed in nature from the beginning of our
planet. Most people would not object to finding silver metal on their property.
References:
1. Environ. Sci. & Technol. 2008, 42, p 7025-7026
2. Tony M. Isaacs, www.americanchronicle.com/articles49272 , May 14, 2008
3. Dept. of Agriculture, Handbook 296, 2006
4. F. Key and G. Maass, “Ions, Atoms and Charged Particlesâ€, available at
www.silver-colloids.com publications
5. Zeta Potential of RO Membranes by M. Elimelech and A. E. Childress, contract
No. 1425-4-CR-81-19290
ABOUT THE AUTHOR
Dr. George Maass is the chief chemist for Colloidal Science Laboratories
and serves as senior scientific advisor to Purest Colloids, Inc. He holds a
BS in chemistry from Fordham University and a Ph.D. in physical
chemistry from Iowa State University.
For the last 12 years, Dr. Maass has been an adjunct professor of
chemistry at Camden County College, while operating his own consulting
business. He has authored papers and presented seminars on his work in
the all across the US, as well as in England and in Mexico.
© 2008 Colloidal Science Lab., Inc.
Dr. Maass, a recognized problem solver, has the ability to determine the
facts which cause phenomena, and to determine the methods by which
they can be controlled.
On November 18, 2008 the NY Times published the report of a health scorecard entitled “The Wrong Place to Be Chronically Ill” which follows. It once again documents, as does every survey of international health and health cost, that the US reliance on prescription medication rather than on healthy food, healthy life style, exercise and supplements, is a national disaster of the first water.
Only clean food, high potency natural supplements and nutrients and a huge pile of sceptical salt about what the FDA and the pharmaceutical industry, through its gullable spokespeople, uninformed doctors, will change that along with one more ingredient: common sense!
Eating GMO food is disastrous for your health. Organic food and supplements are essential. Go to www.Organics4U.org to stop using nutrients from GMO sources! Get hold of some high quality nano silver and stop using antibiotics. Check out http://www.Nutronix.com/naturalsolutions and go to the Products tab, then the Silver solutions button. Your health will thank you! Growing whatever you can and making contact with local growers to make sure you know what is in your food.
It’s your choice. Act now to make your life a healthy one! And act now to donate (https://staging.drrimatruthreports.com/index.php?page_id=189) to the Health Freedom organization that is there for you, the Natural Solutions Foundation, www.HealthFreedomUSA.org.
Yours in health and freedom,
Dr. Rima
Rima E. Laibow, MD
Medical Director
Natural Solutions Foundation
www.HealthFreedomUSA.org
www.GlobalHealthFreedom.org
www.NaturalSolutionsFoundation.org
www.Organics4U.org
www.NaturalSolutionsMarketPlace.org
www.NaturalSolutionsMedia.tv
The Wrong Place to Be Chronically Ill
Tuesday 18 November 2008
The New York Times | Editorial
Chronically ill Americans suffer far worse care than their counterparts in seven other industrial nations, according to a new study by the Commonwealth Fund, a New York-based foundation that has pioneered in international comparisons. It is the latest telling evidence that the dysfunctional American health care system badly needs reform.
The results of the study, published by the respected journal Health Affairs, belie the notion held by many American politicians that health care in this country is the best in the world. That may be true at a handful of pre-eminent medical centers, but it is hardly true for the care provided to a huge portion of the population.
The Commonwealth Fund’s survey of 7,500 patients in Australia, Canada, France, Germany, the Netherlands, New Zealand, Britain and the United States focused on patients who suffered from at least one of seven chronic conditions: hypertension, heart disease, diabetes, arthritis, lung problems, cancer or depression.
The care they received in this country – or more often did not receive – ought to be a cause for shame. More than half of the American patients went without care because of high out-of-pocket costs. They did not visit a doctor when sick, skipped a recommended test or treatment or failed to fill a prescription. The uninsured suffered most, but even 43 percent of those who had insurance all year skipped care because of costs.
Americans also were most likely to report wasting time because their care was so poorly organized. About a third reported that medical records and test results were not available when needed or that tests were duplicated unnecessarily. A third experienced a medical error, such as being given the wrong medication or test results. Some 40 percent found it very difficult to get after-hours care without going to an emergency room.
The United States did comparatively well in some areas, such as providing relatively prompt access to specialists and clear instructions to patients leaving the hospital. But the nation’s overall performance was abysmal.
By contrast, Dutch patients reported far more favorable experiences with their health care system, largely because the Netherlands provides universal coverage (through individual mandates and private health insurance), a strong primary care system and widespread use of electronic medical records. It should be possible to achieve the same level of performance here.
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There is a great deal of misinformation about silver solutions. While there are several kinds (ionic, colloidal, nano technology, protein silver), the safety of silver is extraordinary, despite the fear of turning blue (argyria) which has been promulgated over the years.
The truth is that virtually no one has ever turned blue because of a deposition of silver particles in the skin. In the documented case where this has happened, the person drank huge and irrational amounts of silver daily for decades. In the recent well-publicized case of a politician who was supposed to have turned blue from drinking silver, it turned out to be a hoax perpetrated by his political opponents in the campaign for office he was participating in.
The smaller the particle size, the more effective the silver is at disrupting the biological function of disease causing bacteria, mycoplasma and viruses. The smaller the particle size, the more efficiently the body can get rid of the silver.
The silver we recommend, ASAP silver sol solution is exceedingly small in size, has never been shown to provide any medical or health problems and has been used for people from infancy to people in their 90s.
Here are the facts as I see them, after extensive experience using silver for infections of all types:
1. Silver in nano sized particles does not cause argyria because the particles are small enough to pass easily out of the urine, rather than being deposited in tissues. No case of argyria has ever been reported with nano particle silver
2. As you said, the amounts you would have to consume border on the insane.
3. Nano silver is effective, without any known side effects, against every pathogen it has been tested against
4. Through its applications in Africa, we know that pregnant women, tiny babies, fragile elderly, people with diabetes, cancer, auto immune disease, etc., do not develop side effects when they take silver as directed.
5. No ulcers have ever been reported with nano silver
6. The silver we recommend has been shown NOT to kill beneficial bacteria, thus protecting the integrity of the gut and the immune system.
ASAP silver sol solution has been tested against more than 650 pathogenic organisms and has been effective in vitro (in the lab) against every one. It has also been tested against the normal bacteria (which we refer to collectively as “pro biotics”) and found, rather amazingly, to spare these beneficial bacteria.
I personally never travel without ASAP silver sol solution. I have used it for common ailments like a cold and uncommon ones like malaria. It can be taken via sub lingual administration directly under the tongue to speed absorption and prevent it from reaching the GI tract or it can be mixed in a small amount of water and taken by mouth. It has no flavor or taste so even babies can take it without distress.
We have concluded a special arrangement with Nutronix.com to allow us to offer you this silver sol solution. Please do yourself a favor and lay in a good supply of this outstanding health aid.
Click here, http://www.Nutronix.com/naturalsolutions, to get your supply. Click on the “Products” tab, then go to the left hand column and click on “Silver Solutions”.
It is not clear how long the FDA will allow this exceptional product to remain on the market. I suggest you stockpile it for pandemics, general disease use and immune system boosting.
Yours in health and freedom,
Dr. Rima
Medical Director
Natural Solutions Foundation
www.HealthFreedomUSA.org
www.GlobalHealthFreedom.org
www.NaturalSolutionsFoundation.org
www.Organics4U.org
www.NaturalSolutionsMarketPlace.org
www.NaturalSolutionsMedia.tv
Effect of Prophylactic Treatment with ASAP-AGX-32 and ASAP Solutions on an Avian Influenza A (H5N1) Virus Infection in Mice
Gordon Pedersen,American Biotech Labs
Sidwell, Robert W., The Institute for Antiviral Research of Utah State University Animal, Dairy and Veterinary Sciences Dept
Alan Moloff, D.O., M.P.H., Colonel, MC, U.S. Army (RET)
Robert W. Saum Ph.D., IDHA, Medical Science and Technology, American Biotech Labs
Introduction
Avian Influenza (H5N1, or Bird Flu) can be a fatal disease in humans and a serious threat to become a pandemic event. Since there is no pharmaceutical remedy for the Bird Flu it is essential that preventive treatments be tested and developed in order to enhance survivor rates in the human population.
Since 1973, Silver has been shown to have topical activity against 22 bacterial species (643 isolates) including gram positive and gram negative bacteria1. As an antimicrobial agent, Silver has been shown to be beneficial in the treatment and prevention of burn infections, post surgical wound infections, and gynecological infections2, 3. In addition, Silver has been shown to be active against black mold4, Anthrax5, Bubonic plague6, Malaria7, and numerous viruses such as Hepatitis 8.
Recently it was reported that the American Biotech Labs product, Silver Sol, demonstrated additive and synergistic effects when combined in individual trials with 19 different antibiotics9. The Silver
from ABL was shown to improve the effectiveness of the antibiotics even against
antibiotic resistant infections 9.
The Merck Index identifies the following medicinal uses of silver: Antiseptic particularly for mucous membranes and infectious sinusitis10. The Merck Manual and Centers for Disease Control, recommend that Silver nitrate drops should be placed in each infant eye as soon as possible or at least in the first half hour of life to prevent gonorrheal ophthalmia 11.
The safe use of Silver as an orally consumed preventive agent has been demonstrated and supported by reports from the EPA and the United States Department of Health and Human Services in a 76 week long study12,13. Dogs that inhaled Silver showed activity in the lung in one hour with 90% of the silver carried to the liver by the blood within 6 hours14.
Due to the increased risk from methicillin resistant bacteria, black mold, plasmodium and especially bird flu, the need for orally consumed, safe, daily prophylactic prevention exists. In this study, Silver Sol from American Biotech Labs demonstrates safe beneficial and preventive activity against H5N1 Bird Flu, when taken orally in mice.
The American Biotech Labs product, ASAP- AGX-32, as well as their product designated ASAP, to be virucidal against the avian influenza A/Vietnam/1203/2004 (H5N1) x A/Ann Arbor/6/60 hybrid virus, with an up to 2 log10 virus titer reduction occurring after a 6 h incubation of the product and the virus
(USU report dated March 28, 2006). In that same report, similar incubation with the avian influenza A/Duck/MN/1525/81 (H5N1) virus reduced the virus titer by approximately one-half log10 in the same
time period. This material is reportedly very well tolerated in human subjects when ingested orally, Dr Gordon Pedersen of American Biotech Labs designed a study with the Centers for Antiviral Research to
evaluate the potential for ASAP-AGX-32 and ASAP, to inhibit an avian influenza A (H5N1) virus infection of mice when administered orally to the animals beginning 1 week prior to virus exposure. This report describes the results of this experiment.
Materials and Methods
Animals: Female specific pathogen-free 18- 21 g BALB/c mice were obtained from Charles River Laboratories (Wilmington, MA). They were quarantined 5 days prior to use. They were housed in polycarbonate cages with stainless steel tops and provided tap water and mouse chow ad libitum.
Virus: Influenza A/Duck/MN/1525/81 (H5N1) virus was originally provided by Dr Robert Webster of the St. Jude Hospital (Memphis, TN). The virus was adapted to mice by passage twice through weanling animals and a large pool prepared in MDCK cells for use in this study. The virus was titrated in young
adult mice prior to use in the present experiment.
Compounds: ASAP and ASAP-AGX-32 were provided by Dr Pedersen. They were in blue bottles, so all studies run with each were performed using the materials in injection bottles covered with aluminum foil to avoid light exposure. All were stored at room temperature until used. It is understood that the ASAP solution contained a colloidal silver at a concentration of 10 ppm, and the ASAP-AGX-32 contained the same colloidal silver at a concentration of 32 ppm. Ribavirin, included as a known positive
control, was provided by ICN Pharmaceuticals, Inc. (Costa Mesa, CA); it was dissolved in sterile saline and stored at 4o C until used.
Arterial Oxygen Saturation (SaO2) Determinations: SaO2 was determined using the Ohmeda Biox 3800 pulse oximeter (Ohmeda, Louisville, OH). The ear probe attachment was used, the probe paced on the thigh of the animal. Readings were made after a 30 sec stabilization time on each animal. Use of an earlier Ohmeda Model (3740) for measuring effects of influenza virus on SaO2 in mice has been previously described by Dr.s Sidwell and Pedersen15.
Lung Score Determinations: Each mouse lung removed and placed in a petri dish which, using a permanent black marker, had been divided into sections which were pre- numbered from 1 through 3 or, for placebo controls, 1 through 5. Each lung was assigned a score ranging from 0 (normal appearing lung) to 4 (maximal plum coloration in 100% of lung). These scores were assigned blindly, with the individual doing the scoring not being aware of what group was being examined. An arithmetic mean was determined for each group.
Lung Virus Titer Determinations: Each mouse lung was homogenized and varying dilutions assayed in triplicate for infectious virus in MDCK cells as described previously16. Each lung homogenate was
centrifuged at 2000 g for 5 min and the supernatents used in these assays.
Experimental Design: Groups of 19 mice were treated by oral gavage (p.o.) with either ASAP-AGX-32 or ASAP twice daily (every 12 h) for 7 days, then infected intranasally (i.n.) with an LD70 dose of
influenza virus, then treated an additional 10 days. A similar group of mice were treated p.o. with ribavirin at a dosage of 75 mg/kg/day twice daily for 5 days beginning 4h pre-virus exposure. The infection was achieved by anesthetizing the mice with an intraperitoneal injection of Ketamine at a dosage of 100 mg/kg and instilling 90 µl of suspended virus in minimum essential medium on the nares of the animals. As controls, 35 mice were treated with water using the identical schedule as used for the
ASAP materials and infected as above. Ten infected, test substance-treated mice and 20 water-treated controls were observed daily for deaths for 21 days after virus exposure, and SaO2 levels ascertained on days 3-11, which were the times when this parameter usually declines. From the remaining infected, treated animals, 3 test substance-treated and 5 water-treated control mice were killed on days 1, 3 and 6, and their lungs removed, assigned a consolidation score, weighed, and assayed for virus titer.
As toxicity controls, 3 uninfected mice were treated in parallel with each test material and observed for signs of adverse effects for 21 days. The weights of these mice as well as 5 normal controls were determined prior to initial treatment and again 18 h after final treatment to determine if the treatments
affected host weight gain. Three normal controls were also sacrificed on days 3 and 6 to provide background lung data.
Statistical Analysis
Increases in total survivors were evaluated by chi square analysis with Yates’ correction. Increases in
mean day to death, differences in mean SaO2 values, mean lung weight, and mean virus titers were analyzed by t-test. Only animals dying up to day 21 were considered for mean day to death calculations. The Wilcoxon ranked sum analysis was used for mean lung score comparisons. Each
statistical test was run using Excel software on a MacIntosh computer.
Results and Discussion
The results of this experiment are summarized in Table 1 and in Figures 1-4. As seen in Table 1, the virus challenge in this experiment was lethal to 14 of the 20 placebo-treated mice, with the mean day to
death being 8.4 days. Such a pattern of death is considered ideal for evaluation of potential antiviral agents. This optimal condition was verified by the observation that Ribavirin was fully protective to the
mice, preventing any deaths from occurring (Table 1), significantly lessening SaO2 declines (Figure 1), inhibiting lung score development (Figure 2), lung weight increase (Figure 3), and lung virus titer
increases (Figure 4).
Treatment with ASAP-AGX-32 appeared to not affect the numbers of animals dying of influenza, although a half-day delay in mean day to death was seen (Table 1). SaO2 declines in this group of treated mice were almost at the same rate as those in the placebo controls, although it was interesting
that on the first day this parameter was assayed, a highly significant (P<0.001) difference was seen (Figure 1). SaO2 declines are a manifestation of declining lung function, suggesting that the lung consolidation in the lungs did not progress as rapidly as seen in the placebo controls. The treatment appeared to moderately lessen lung consolidation as seen by lower lung scores on each time evaluated, the day 6 mean lung score being significantly (P<0.05) less than the placebo treated controls (Figure 2). Lung weights, another indication of fluid developing in the lungs to cause pneumonia in the animal, were also less at each time point than seen in the placebos (Figure 3). The mean lung virus titers in the
mice treated with ASAP-AGX-32 were lower than the placebo controls on days 3 and 6 of the infection (Figure 4).
Treatment with ASAP, which we understand is a less-concentrated version of ASAP-AGV- 32, also provided some intriguing results. Especially of interest was the observation that 60% of the infected mice treated with this compound survived compared to the 30% in the placebo-treated controls. Although not statistically significant because of the number which survived in the latter controls, this effect is strongly suggestive a disease-inhibitory effect may have occurred. At two time points during the SaO2 assays, days 3 and 6, the declines normally seen were significantly lessened (P<0.01), and
there was a general lessening of decline throughout the times of assay (Figure 1). Modest inhibition in lung scores were seen in this treated group as well, especially on day 6 (Figure 2); the lung weight data did not correlate too well with the lung scores, however (Figure 3). Again, slight inhibition of lung virus titers were seen in the ASAP- treated, infected mice (Figure 4).
Both the ASAP formulations were well tolerated by the toxicity control mice as seen by no deaths occurring in them and host weight increases observed during time of therapy. Ribavirin, while not lethal to the mice, did result in a 0.4 g host weight loss (Table 1); this was an expected effect for the latter material, since the maximum tolerated dose is approximately 100 mg/kg/day.
It is difficult to attribute the effects seen in this experiment wholly to viral inactivation, since both test materials were administered orally to animals infected by direct nasal inhalation, although the treatments began one week before virus exposure, so it is possible that a portion of the Silver Sol may
have been able to be in the vicinity of the virus-exposed lung tissue. It is also possible that this material is exerting a mild immunomodulatory effect in the animals, which would provide modest protection
against the infection. If such a mechanism is indeed associated with the potential activity seen, then a different treatment schedule, perhaps limiting the number of treatments to one per day or once every other day, may enhance any immune modulatory effects, since it is recognized that too-frequent dosing may overtax the immune system. The greater protection seen by the lower- dosed ASAP material could be explained by immunomodulation, since the greatest immunologic effect is not necessarily at the
highest dose used.
Another mechanism whereby the ASAP materials may have inhibited the influenza virus infection in these studies may simply be one of coating the virion with Silver Sol to prevent attachment and penetration. Again, the material would need to be in the vicinity of the exposed lung tissues at the time
infection was initiated. The Silver material could also play a role in limiting apoptosis of the epithelial lining of the lung induced during acute lung inflammation. Apoptosis plays a causative role in acute lung injury in part due to epithelial cell loss.
Further studies would have to be conducted to more fully delineate the actions of this material.
It is acknowledged that the effects seen in the present study, while of considerable interest, would need to be repeated to confirm that the observations were not due to mere chance. Consideration of combined use of oral administration of the ASAP materials and intranasal instillation at near
the time of virus exposure would determine
whether the effects seen were indeed
associated with virucidal effects of these
materials.
Summary
Mice infected with avian influenza A/Duck/MN/1525/81 (H5N1) virus were treated with the Silver Sol-containing formulations ASAP-AGX-32 and ASAP provided by American Biotech Labs. Oral gavage treatment began 7 days prior to virus exposure and continued twice daily for a total of 17 days. Treatments with both formulations provided a suggested inhibitory and preventive effect on this virus infection as seen by either less animals dying in the treated groups than in the placebo-treated
controls, delay in mean day to death, lessened SaO2 decline, modest inhibition of lung consolidation, and/or lessened virus titers in the lungs. Ribavirin was included as a positive control drug, used orally at a dose of 75 mg/kg/day twice daily for 5 days beginning 4 h pre-virus exposure, and this treatment was markedly inhibitory to the infection as expected.
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Another theoretical “impossibility†becomes real by experiment. Water burning? Yes, it can!
Materials Research Innovations, March 2008, Vol 12, 3-5.
Table 1. Expt. ABLA-1. Effect of Oral Gavage Prophylactic Treatment with ASAP-AGX-32 and ASAP on an Influenza A (H5N1) Virus Infection in Mice.
Animals: Female 18-21 g BALB/c mice Virus: Influenza A/Duck/MN/1525/81 (H5N1) Drug diluent: Company diluent Treatment schedule: bid x 17 beg -7 days (Ribavirin: bid x 5 beg -4 h) Treatment route: p.o. Experiment. duration: 28 days Tax Controls Infected, Treated Mice Tax Controls Dosage
Surv/Total Mean Host Weight Change (g) Surv/Total Mean Day to Death ± SD Mean Day 11
SaO2 (% =± SD) ASAP- AGX- 32 32ppm 3/3 1.8 2/10 8.9 ± 1.4 75.4 ± 1.0
ASAP 10ppm 3/3 1.4 6/10 7.3 ± 1.0 76.7 ± 2.1 Ribavirin 75 mg/kg /day 3/3 -0.4 10/10*** >21.0 ± 0.0*** 86.6 ± 2.5** * H2O — — — 6/20 8.4 ± 1.8 76.0 ± 1.9 Norma l Contro ls — 5/5 2.3 — — 88.8 ± 3.0
Difference between initial weight and weight 18 h after final treatment.
Difference between initial weight and weight 18 h after final treatment.
*P<0.05; **P<0.01; ***P<0.001 compared to H2O -treated controls.