Pure H2o BioTechnologies

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PRHB - New Video Format August 2010


Pure H2O Bio Technologies, Inc.
New Video Format
August 2010

Market Opportunity – Treatment of Emerging Water-Borne Disease Pathogens

Over the past 20-years, increasing demands for pure drinking water by hospitals, clinics, nursing homes, residential consumers as well as recreational, educational, military, and commercial facilities have generated the need for water treatment technology that can disinfect organisms that are refractory to conventional water treatment and filtration. 286 million people in the US receive their water from public drinking water systems. In 2007, the EPA determined that over 51,252 or 32.9% of the drinking water systems in the US reported at least one health-based standard, monitoring and reporting, or other violation. Moreover, there is an additional need for the 5,747 registered hospitals in the US to provide safe drinking water in the heath care delivery setting. International opportunities are also another market sector particularly in developing countries. Pure Water Bio-Technologies intends to leverage its novel Silver TTO™ disinfection technology to produce water treatment systems that effectively treat chlorine-resistant water-borne pathogens such as Cryptosporidium and related enteric bacteria. The proprietary Silver TTO™ technology has the potential to disrupt the existing water treatment market in commercial, institutional as well as residential market sectors.

Silver-TTO™ Water Disinfection of Resistant Cryptosporidium



Crytosporidiosis – Disease Epidemiology

Infection by C. parvum has been reported in six continents and identified in patients aged 3 days to 95 years old ( Flanigan and Soave, 1993) and results in the gastro-intestinal disorder known as Crytosporidiosis. Transmission is usually fecal-oral, often through water contaminated by mammalian livestock. Persons most likely to be infected by Cryptosporidium are:

infants and younger children in day-care centers
those whose drinking water is unfiltered and untreated
those who have oral contact with feces of an infected individual
patients in a nosocomial (hospital patients) or healthcare setting with other infected patients or health-care employees
veterinarians who come in contact with farm animals
immunocomprised individuals such as those with AIDS, undergoing irradiation or chemotherapy
Consumers of water continue to express concerns over their water quality in response to several driving factors: (1) a growing population and industrial concentration in urban areas which place increased demand on existing water treatment facilities, (2) reoccurring outbreaks of chlorine-resistant water-borne pathogens, and (3) the potential impact of water-borne parasites that are refractory to conventional chlorine water-treatment particularly to immune-compromised patient cohorts. The 1993 outbreak of Cryptosporidiosis in Milwaukee, WI and the 1998 outbreak of Coliform (E. coli) bacteria in Atlanta, GA, (4) and in 2000 the Cryptosporidium outbreak in the Northwest UK are prime examples. In FY2004, the EPA reported that were 159,796 public drinking water systems in the United States (Factoids: Drinking Water and Ground Water Statistics for 2004) and 9,099 of these systems reported violations of TCR/T (Total Coliform Rule/Turbidity) that affected an estimated 11,382,966 people.




Images
Photomicrograph of Crpytosporidium.


Immunofluorescence image of Cryptosporidium parvum oocysts
Courtesy of H. D. A. Lindquist, U.S. EPA
Transmission Electron Micrograph of E. coli O157:H7 Bacterium
Courtesy of E. H. White – Centers for Disease Control & Prevention


Images:
Silver-TTO™ molecule, flowing rivers, ocean waves, glass filling with water.
Hospital and IV administration, high-tech lab fluid handling.

Cryptosporidium Cell Wall Structure and the Basis of Resistance
Cryptosporidium oocysts have also been shown to be resistant to all halogen compound treatments, including chlorine, iodine, bromine and fluorine, since they are designed to treat microorganisms in their vegetative state and not the oocyst stage of the parasite life cycle. Within the membrane-enclosed oocyst large amylopectin carbohydrate granules (0.1-0.3 micron) can be found besides as well as a large lipid bodies and a crystalline protein inclusion.

Specifically, classic disinfectants have been shown to have little effect on the carbohydrate component of the oocyst cell wall. Oocyst cell wall proteins can also contribute to resistance. In the past decade, the retail market has seen the introduction of iodine-based carbon resins for residential water treatment at the point-of-use. These systems merely entrap microorganisms and provide no disinfection capability to Cryptosporidium parvum oocysts..

Pure H20 Bio-Technologies (PRHB) has developed proprietary Silver-TTO™ point-of-entry and point of-use systems which can effectively disinfect chlorine-resistant organisms such as Cryptosprodium oocysts and water-borne bacteria.

Existing Water Treatment Limitations

Water treatment facilities, although operated correctly and remove the vast majority of microbial contaminants, can be breached by some recently emerging pathogens, as evidenced by the large number of waterborne disease outbreaks. This has created considerable concern for response to several driving factors:

¦ A dramatic increase in the number of cases of waterborne diseases caused by newly emerging pathogens
¦ The need for added protection against potential bio-terrorism

¦ Public awareness of waterborne outbreaks, e.g., the well-known outbreaks of cryptosporidiosis in Milwaukee, 1993, in Las Vegas, 1992-1994, and Lancashire, UK , 2002. There were 7,212 cases of cryptosporidiosis; 17,256 cases of giardiasis; 2,368 cases of E. coli 0175:H7; 9,877 cases of hepatitis; and 1,952 cases of legionellosis during 2005, proving that the threat of outbreaks still exists.
¦ Growing population and industrial concentration in urban areas

¦ Population demographics - cancer and other immune-compromised population cohorts.

¦ The Centers for Disease Control and Prevention have estimated that there are on the order of 940,000 cases of waterborne enteric (intestinal) disease in the USA annually (GFL & Associates, 1993).

Existing hyper-chlorine disinfection and carbon filter-based water treatment systems appear to have limits that suggest a need for improved potable water supplies in military installations or remote locations, residences, hospitals, nursing homes, day-care centers, commercial facilities, recreational facilities, cruise ships and hotels..

Presently, ozone and ultraviolet water treatment procedures are approved by the USEPA for disinfection of Cryptosporidium oocysts, however, there are practical problems associated with their use to deliver pure and safe drinking water. Ozone is very expensive to use as noted in the Southern California Water District study (G.Quist, I.C. Felkner and R. DeLeon. Evaluation of a Real-Time Online Monitoring Method for Cryptosporidium. J. AWWA, USA 2004, pp 1-103.) Ozone is readily absorbed by particulate organic matter, so that it's disinfectant capacity can be significantly reduced in turbid source water. In addition, water treated with ozone can become re-contaminated at or before the point of entry or point of use because ozone decays to oxygen leaving no residual disinfection capacity and requiring further augmentation by other disinfectants such as chlorine products, which are not effective against Cryptosporidium oocysts.

Ultraviolet light has a limited penetration capacity and does not work well except in very shallow water streams. It also does not leave a residual protection for water at the point of entry or point of use and penetration is blocked by turbidity, as is ozone. In addition, it is mutagenic so that resistant forms of microbes can be created when killing is not complete.

Although halogens such as iodine, chlorine, and fluorine can be very effective in killing pathogenic bacteria, our intended use of Silver TTO™ disinfectant is far superior for overall removal and/or killing of the newly emerging pathogenic strains

Unmet Need – Effective Water Disinfection
As a result of the current unmet market need to successfully disinfect Cryptosporidium parvum oocysts in drinking water, Pure H20 Bio-Technologies, Inc. intends to introduce its proprietary combination water purification systems, using its Silver TTO™ disinfection agent. It will be used in combination with absolute 1-micron porosity activated-carbon filter elements facilitating the removal of the Silver TTOTM along with other organic and metallic contaminants. This means that water pure enough to supply hospital needs as well as private homes is generated. These system components have been evaluated by independent U.S. environmental labs and demonstrate antimicrobial activity against E. coli and Cryptosporidium oocysts. These results were first produced in a bench-level disinfectant study and were reproduced successfully in the complete automated disinfectant delivery system. This means that the technology has been successfully converted to a product that has the potential for serving hospitals and private individual systems. It should be especially important when the municipal water gets contaminated at or before the point of entry or point of use, e.g., under a "boil water" directive from the city water management. The Silver TTOTM kills not only bacteria and viruses, but also Cryptosporidium oocysts (emerging pathogenic threat organism in water). The system can regulate the amount of Silver TTOTM present in the final water so that a low residual is present (use in private homes) or can remove it entirely when it is designated for hospital use where sensitive individuals could otherwise be at risk. It is not mutagenic, and can remain as a residual disinfectant at the point of entry or point of use. The PRHB Silver TTO™ technology can now be viewed as very competitive and a significant improvement over all current water treatment products from a business development perspective.

Laboratory System Studies


The disinfection of E. coli and Cryptosporidium parvum oocysts using the Pure H20 Water Purification System was performed by Clancy Environmental Consultants in St. Alban, VT in 2008 to demonstrate the effectiveness of the Silver TTO™ technology. It meets the challenge posed in a major AWWA publication (P. Rochelle and J. Clancy 'The Evolution of Microbiology in the Drinking Water Industry' J. AWWA, March 2006, pp 163-191) that enumerates newly emerging pathogen threats to safe drinking water. After successfully demonstrating the ability of Silver-TTOTM to kill E. coli and Cryptosporidium oocysts in a bench level study, an automated delivery system (patent pending) was designed and used to test the the efficacy of silverTTOTM for killing of high concentrations of E. coli and Cryptosporidium parvum, thus producing water suitable for drinking.

E. coli disinfection tests: The Silver-TTO disinfection of water inoculated with E. coli bacteria were measured by Colony Forming Units (CFU/mL) with mEndo media at 2 and 10 minute intervals using the PRHB water purification system.
Disinfection of E. coli bacteria by PRHB Silver-TTO™.


E. Coli Sample
Silver-TTO™
Contact Time


Average
E. coli CFU/mL

Log10 E. coli
Inactivation

0 min


1.1 x 105

0

2 min


1.1 x 104

1.1

10 min


< 1

> 5.0




*< 0.5 mg/L of dissolved silver Conclusions:

Low Silver-TTO concentrations will reduce the concentration of E. coli bacteria by greater than a factor of 5 log 10 in compliance with current US EPA's regulations.
Combination of AC-carbon filtration and Silver TTO™ disinfection is a functional system design for water purification.


In the next series of studies, samples of PRHB Silver-TTO™ purified water was incubated with human HCT-8 ileocecal adenocarcinoma or colon cells. They were subsequently stained for IgG-FITC labeled antibody for intracellular reproductive stages of Cryptosporidium parvum oocysts, and scored for infectivity.

Disinfection of Cryptosporidum Oocysts by PRHB Silver-TTO™ over 60 minutes.


Crytosporidium parvum Oocyts Sample ID


Cryptosporidium Oocysts per Innoculum


Average Percentage
Infectious Post-Treatment by Immunofluorescence


Oocyst Stock Culture


9.4 x 102

5.2 %

Heat-Shock Control


1.0 x 104

0.0 %

Process Control


8.6 x 102

4.0 %

Silver TTO™ – Test A*


1.2 x 103

< 0.08 %


Silver TTO™ – Test B*


1.3 x 103

< 0.08 %


Silver TTO™ – Test C*


1.2 x 103

< 0.08%




Silver-TTO Log Disinfection of Cryptosporidium parvum oocysts.



< 0.5 mg/L dissolved silver
Conclusions:

* Low Silver-TTO (less than 0.5 mg/L) can effectively reduce the infectivity of Cryptosporidium oocysts.
* Combination of AC-carbon filtration and Silver TTOTM disinfection is a functional system design for water purification.


PHBT Silver TTO™ Systems
The current point-of-use and point-of-entry systems are primarily based on carbon or fiber filters that merely entrap, but do not disinfect drinking water. The Pure H2O technology system combines use of absolute micron carbon pre- and post-filters with controlled release of oxygenated tetrasilver-tetraoxide (TTO) to treat waterborne pathogens and remove heavy metals and objectionable biomass. The use of TTO in the system is a great improvement over use of halogenated compounds such as chlorine, bromine and iodine because it is capable of killing the oocysts of Cryptosporidium and cysts of Giardia with an already proven ability to destroy pathogenic bacterial and viral species, including Escherichia coli and waterborne viral species specified by the US Environmental Protection Agency (US EPA). The US EPA, for treatment of swimming pools, has already approved TTO for its efficacy on microbes. Its efficacy for treatment of Cryptosporidium oocysts was successfully tested in 2008 for PRHB by a laboratory nationally recognized for its well-established expertise on Cryptosporidium as well as other pathogenic microbes. A non-provisional patent for use of TTO to disinfect Cryptosporidium oocysts has recently been submitted to the US Patent Office. The Company plans to certify the system through NSF using updated NSF/ANSI standards.

Intellectual Property



The Company’s decision to utilize new Silver TTO™ in combination with Activated-Carbon in its treatment of water is based upon known properties, including the novel Silver TTO™ capability to inhibit the proliferation of Cryptosporidium parasitic oocysts..

Pure H20 has 2 patents pending, i.e., tetrasilver tetraoxide application in the area of micro-biotechnology and mechanical engineering with the U.S. Patent & Trademark Office.
PRHB Products





The Pure H2O water purification systems include the latest Silver TTO™ disinfection technology designed to provide microbiologically pure and safe drinking water. Recent independent laboratory studies demonstrate the PRHB Silver TTO™ technology disinfection of resistant Crytosporidium oocysts. Pure H2O Bio-Technologies, Inc. has designed this product to purify water at point-of-entry (POE) and point-of-use (POU) to remove: chlorine, lead and organic chemicals, chloramines, and unpleasant tastes and odors. This reliable potable water system combines the proven unique benefits of absolute AC and absorption removal of offending contaminants with microbiological disease control and prevention.

Model 7000 Hospital Water Purification Unit

The Model 7000 Hospital Water Disinfection Unit incorporates the leading Pure H20 granular activated carbon water purification technology for applications in healthcare facilities. The Pure H20 Model 7000 SilverTTOTM storage and dispensing process, in conjunction with Activated-Carbon, generates a near 100% disinfection rate against various disease causing waterborne microorganisms. The final water generated is pure and will not contain organic or inorganic contaminants that will be harmful to sensitive patients, such as those undergoing kidney dialysis or those who are seriously immunocompromised (e.g., radiation or chemotherapy treated patients).
Independent Testing
Performance evaluations of the PRHB Silver TTO™ disinfection system were conducted under established protocols at Clancy Environmental Consultants, Inc. (CEC) which is an independent environmental testing laboratory based in the United States. CEC is a USEPA-approved laboratory for Cryptosporidium and E. coli analysis for the EPA Long Term 2 Enhanced Surface Water Treatment Rule and provides laboratory services to federal agencies and private sector clients through contracts with the U.S. General Services Administration (GSA) and corporate entities. The CEC laboratory is certified in Vermont and is NELAC (National Environmental Laboratory Accreditation Conference) accredited for microbiological analysis of drinking water. Independent validation of silverTTOTM and dissolved silver concentrations on these studies were conducted by Genapure, Inc. (formerly US Biosystems, Boca Raton, FL)

Product

The Pure H20 Bio-Technology potable water disinfection system utilizes a non-heat reliant, power independent method known to kill Cryptosporidium, which has been found in municipal water supplies and recreational facilities. As a result, the PHBT systems are designed with a technology combination of Silver TTO™ and absolute micron porosity GAC or similar final finish absorbent provide an additional level of protection for both residential and commercial / institutional applications.
Pure H2O Bio-Technologies has refined the traditional drinking water purification processes by utilizing Silver TTO™ disinfection systems, thereby achieving a purity unequalled in the water purification and bottled water markets. The Company offers four different structural point-of-entry (POE) & point-of-use (POU) disinfection / purification systems designed to meet various market and client preferences.
PHBT Silver TTO™ Models (images)
PHBT 75: Under-the-Counter System
PHBT 4000 & 3000: Residential and Commercial Systems
PHBT 7000: Hospital and Medical System
Competitive Landscape
Existing carbon-based iodine resin water treatment systems were initially certified without the use of live Cryptosporidum oocysts. These ion-exchange systems appear to have limited value or performance in disinfecting water. They temporarily entrap curtain organisms that may pass through the AC blocks during certain stages of the Cryptosporidium or bacteria life cycle. Ozone and UV treatment systems have shortfalls that were previously explained and involve cost, penetration, and lack of residual disinfectant protection. The SilverTTOTM system at present is not intended to replace existing municipal water treatment systems, but for use at the point-of-entry and point-of-use.
Pathway to Market Launch
The ability to launch a new drinking water purification system onto the market requires certification of both the system and manufacturing facility by NSF International in Ann Arbor, MI. NSF is a non-profit, non-governmental organization that is recognized as an authority in product certification and risk-management that is focused on food, water, indoor-air and other environmental standards and independent assessment services that serves manufacturers in over 80 countries. This independent NSF certification of the PRHB Silver TTO™ water purification system and manufacturing facilities will require additional funds for approximately “6” months.

Images:
Lab video elements from NSF or other stock video.

Financial Needs
· Silver-TTO engineering specifications list and Certification
· Manufacturing Facility Certification
· Beta Model Manufacturing.
· IP fillings.










Investment Opportunity

In the event voice mail box is full, please write to
Mr. Joseph P. Doxey, President at his E-mail address:
[e-mail]
or contact him directly at telephone number [phone]:


Contact Information
Mailing Address
Pure H20 Bio-Technologies, Inc.
370 W Camino Gardens Blvd Suite 332
Boca Raton, FL 33432

Telephone
Phone: [phone]
Fax: [phone]

Management Team


Joseph P. Doxey – Chairman, President & CEO

Dennis P. Boudreaux, P.E. - Director and COO

C. Eric Jones, Ph.D. - Director and Corporate Communication

Todd D. Stong, Ph.D, P.E - Technical Advisor

Cecil Felkner, Ph.D. - Consulting Microbiologist, Biochemist, & Toxicologist

EdwardC.West,P.E.-TechnicalConsultant

MichaelSember,V.P.-SalesandMarketing


MissionStatement

Pure H20 Bio-Technologies, Inc. (Pure H20) is an emerging, publicly traded (OTC-BB Pink Symbol PRHB), Florida based corporation that designs, develops, manufactures and markets potable water disinfection systems for residential and commercial applications. Potential uses include water treatment for whole house (under the counter), medical point-of-use, and well watersystems.