• Bacteriophage: When E. coli has a Virus.
• Problem, Solution, Phages, Difficulties.
• Germ Warfare: How Viruses May Help.
• Functional and Genomic Gold from Dirt.
• Novel Way to Kill Streptococci Bacteria.
• Water treatment: Phages, H202, Silver.
• Emergence of new Salmonella Enteritidis phage types in Europe?
• Exploding Anthrax. Lysins to Kill.

1

http://www.cellsalive.com/phage.htm
Bacteriophage: When E. coli has a Virus.
Author: anonymous
2000, or later.

T4 bacteriophage is a virus that looks like an alien landing pod.
With its six legs, the bacteriophage attaches to the surface of the much larger bacteria Escherichia coli (E. coli). Once attached, the bacteriophage injects DNA into the bacterium. The DNA instructs the bacterium to produce masses of new viruses. So many are produced, that the E. coli bursts.

2

http://www.phage-biotech.com/links.html
Problem, Solution, Phages, Difficulties .
by Anonymous
2000, 0r later.

The Clinical Problem
Today's escalating antibiotics crisis is the direct result of the large scale and indiscriminant over-use of antibiotics and disinfectants, which has triggered an increase in the quantity, variety, and proliferation of multidrug-resistant and, consequently, particularly virulent bacterial pathogens. These pathogens have developed resistance to virtually all extant antibiotics, including the antibiotic of last resort, vancomycin.

Pathogens, once considered clinically under control or obliterated, are re-emerging as prolific killers (e.g., Mycobacterium tuberculosis).

Highly virulent and resistant nosocomial infections are rampant in hospitals everywhere. In US hospitals alone, more than 2,000,000 patients succumb to infectious diseases every year, and over 90,000 die-compared with a yearly mortality of 15,000 in the early 1990s.

Ninety percent (90%) of staphylococci --- the pathogens responsible for fifteen percent (15%) of all bacterial infections-are penicillin resistant, and forty percent (40%) are resistant to methycillin.

Physicians are resorting to extreme measures:
Sixty-three percent (63%) of US vancomycin prescriptions violate the Centers for Disease Control and Prevention (US CDC) guidelines and, thereby, accelerate the formation of bacterial resistance.

The medical community has thus inadvertently entered the "post-antibiotic" era, with no conventional remedy in sight. The indiscriminate over-use of antibiotics has succeeded in eradicating only the antibiotic-susceptible infectious strains while empowering highly resistant "super bugs." Furthermore, without a significant overhaul in antibiotic-use policy, history suggests that most or all chemical antimicrobials in the development pipeline will also trigger rapid evolution of target-bacterium resistance

The Unconventional Solution
A valid, proven, and practical alternative to the chemical-antibiotic treatment of bacterial infections has been successfully practiced in Eastern Europe from as early as the late 1930s.

Phage therapy is a method of antibacterial treatment that harnesses the bacteria-killing properties of otherwise harmless viruses. Phage therapy is practiced routinely in the former Soviet Union as an alternative, combinatory, and complimentary form of treatment in conjunction with, or in lieu of, antibiotics. This time-proven Eastern European practice has received surprisingly little exposure in the West, and, consequently, it has failed to win its due recognition in the West.

Standardized phage medicines in numerous forms are being produced in several locations in Russia and Georgia. These concentrated, polyvalent phage preparations are typically comprised of mixtures of different phages of wide host range that infect and kill many bacterial species and strains, including:

• Brucellae,
• Enterococci,
• Pathogenic strains of Escherichia coli (e.g., O157:H7),
• Klebsiellae (atypical pneumonia),
• Mycobacterium tuberculosis,
• Protei (nosocomial urinary tract infections),
• Pseudomonas aeruginosa,
• Salmonellae (typhoid fever and food poisoning),
• Serratia spp,
• Shigella spp. (bacillary dysentery),
• Staphylococci (skin abscesses, food poisoning, toxic shock syndrome),
• Streptococci (strep throat),
• Vibrio cholerae (cholera), and
• Yersinia spp (plague/black death).

In large-scale clinical trials of various phage-therapy preparations and techniques conducted in Poland in the mid-1980s, a decisive recovery rate of ninety-two percent (92%) was achieved. (Slopek et al, 1983, 1985, 1987).

Western medical culture has been unaware of phage therapy's considerable achievements in the former Soviet Union owing to a variety of historical, political, and bureaucratic circumstances. Hardly any of the numerous Russian-language publications have been translated into English, let alone reviewed.

What are Lytic Bacteriophages?
Lytic bacteriophages ("phages" for short) are bacteria-specialized viruses, and they are among the simplest and most abundant organisms on earth.

Typically comprised of a head filled with genetic material, a syringe-shaped tail, and several fibers that selectively attach themselves to specific receptors on the host bacterial surface, phages bore into their respective host bacteria and inject them with the phage's own genetic material. Within infected bacteria, phage DNA is replicated and then incorporated into bacteria-infectious particles that are manufactured from chemical components stolen from the bacterial host.

These "virions" are then released from their parent bacterial cells via a process known as "lysis," which kills bacterial cell. The production and subsequent release of phage particles allows subsequent phage infection of additional bacteria in a rapid, exponential pattern. (In contrast to lytic phages, temperate phages, which can bolster their bacterial host's virulence, resilience, and general capacity to proliferate are generally unsuitable for therapeutic applications.)

The Phage Advantage

1. Phages thrive in the presence of bacteria, and die out in their absence.
2. Because of phages' extreme specificity and chemically large nature, they have induced neither side nor adverse effects when used as therapy in clinical practice.

3. Phages do not cause allergies or affect the body's natural immune system.
4. Phages generally display a low chemotherapeutic index, particularly upon primary administration systemically or upon topical administration, and they are vastly more diverse in their potential to overcome bacterial resistance than known antibiotics also displaying comparatively low chemotherapeutic indices.

5. Phages support and enhance vital microflora (in contrast with the indiscriminate action of wide-spectrum antibiotics, which can decimate the body's protective normal microflora).

6. Phages may be used both prophylactically and in the treatment of ongoing infections.
7. Phages constantly evolve and can adapt in situ to resistant bacteria strains.
8. Phages may also be more effectively targeted to growing bacteria in local infections.
9. Phages are administered in a limited number of small doses over a short period of time.
10. Phages eliminate pathogens more rapidly and effectively than standard antibiotics.
11. Phage medicines have a long shelf life (up to 2 years).
12. Production costs of phages are low.
13. Phage therapy is consistent with "green-natural-alternative" ideology, and its production is environment-friendly.

14. Phages present the only viable alternative and, potentially, the last resort for the treatment of antibiotic-resistant pathogens.

Historical Background and Context
The biologic phenomena known as "bacteriophages" were discovered during the mid-1910s. Phage-therapy experiments during the 1920s and 1930s yielded inconclusive results (mostly owing to a lack of knowledge about phages' high specificity). Phage therapy, as a clinical method, was rejected altogether in the West upon the discovery, immediate popularization, and wide-scale dissemination of penicillin in the early 1940s.

However, in the USSR, interest and research in phages persisted.
The G. Eliava Institute of Bacteriophage, Microbiology, and Virology, founded in Tbilisi in 1934 - and generously funded and staffed on Stalin's directive - engaged in the laborious process of sampling, cultivating, matching, and producing phage preparations against most known pathogens.

In the late 1980s, when the institute was at the height of its scientific achievement, 1,200 scientists were employed in R&D, maintaining the extensive phage collections and producing two tons of phage preparations per day. Unfortunately, the Georgian civil war of the early 1990s instigated the demise of the institute, the loss of most of its phage collections, the collapse of its infrastructure, and, subsequently, the relocation of phage research and production to Russia.

Prior to the dismantlement of the Soviet Union, phage preparations had been produced in most of the main metropolitan centers. Phage therapy had been spurred by the chronic shortage of adequate and sufficient antibiotics. The phage industry has been adversely affected by the general collapse of the post-Soviet economy, and only a handful of producers remain. Presently, demand for phage therapeutics greatly exceeds supply. ...

Unable to refute its validity, doctors will soon face an adamant demand from patients to provide them with the phage alternative. ...

Possible Problems and Resistance
The introduction of Phage Therapy is expected to encounter the following obstacles:

1. Pervasive fixation on chemical antibiotics within the clinical establishment;
2. Resistance of the pharmaceutical sector, which is heavily invested in chemical antibiotics;
3. Physicians' reluctance to forego wide-spectrum antibiotics in favor of the highly specific phages;
4. Structural and functional reorganization required for a coordinated and responsive diagnosis-production-administration chain;

5. Pervasive aversion within the biotech research establishment to revert to "archaic" microbiology;
6. Need to constantly adapt and refresh phage preparations in response to pathogen evolution;
7. Delay between clinical presentation and antibacterial administration;
8. General disregard for Russian research and clinical practices;
9. Concerns with bacterial evolution of resistance to phages; and
10. Lack of a regulatory reference basis.

3

http://my.webmd.com/content/article/14/1668_50850
Germ Warfare: How Viruses May Help.
By Mitchell Leslie
WebMD Feature
April 30, 2000

Deep in the wilds of your local sewer system, a microscopic drama is unfolding. Invisible to the naked eye, a virus with a bulbous head, spindly neck, and spidery legs glides toward a plump bacterial cell. After alighting, the bacteriophage punctures the cell's membrane and injects its own genes, which force the cell to mass produce viruses. In less than an hour, the victim cell explodes, scattering a brood of 200 newborn viruses. Each of them immediately begins prowling the sewage for more prey.

Bacteriophages, or phages for short, do nothing but attack and destroy bacteria. They thrive anywhere bacteria are abundant -- in sewage, on food, in water, even in your body -- and they've honed their killing technique for more than a billion years. ...

.. each kind of phage usually attacks only one species of bacterium.
That means that phages are extremely unlikely to turn on us -- they don't have a taste for human cells -- and they won't mow down the helpful bacteria that live in our intestines, as antibiotics often do. This pickiness also explains why the phages within your body don't automatically kill off invading bacteria before you get sick. With so many kinds of phages around, you probably don't have the right kind to fight that particular bug.

Finally, phages can evolve along with the bacteria, so that the bacteria can't develop permanent resistance to them as they can to antibiotics. ...

Along with all these benefits come some risks.
When doctors first tried giving phages to patients, they sometimes accidentally included poisons from the bacteria in the medicine, making patients sicker. In other cases, the phages may have done their work too fast, bursting too many bacteria at once, and releasing an overwhelming dose of poison from the bacterial cells. As a result, many patients given phage therapy died. So, except for occasional instances of "compassionate use" for dying patients, phage therapy has not been tried in the West for 60 years.

... researchers in the Soviet Republic of Georgia kept working to overcome the dangers. Millions of patients in the USSR were treated with phage therapy for everything from diarrhea and burns to lung infections.

In one instance, workers building a stretch of railroad through Siberia in 1975 fell prey to a virulent strain of staphylococcus bacteria. Infections that began as skin lesions on the malnourished workers were invading their lungs, then spreading throughout their bodies. David Shrayer, MD, then a young microbiologist at the Gamaleya Institute in Moscow, was called in. Finding antibiotics useless, he arranged for the workers to receive phage therapy. Shrayer, now a Brown University oncologist, says they were quickly cured.

Phage preparations are still available today in Georgia and Russia. "I like to emphasize their safety," says Alexander Sulakvelidze, PhD, the former head of the state microbiology lab in the Republic of Georgia. ....

4

http://www.hhmi.org/research/investigators/jacobs.html
Functional and Genomic Gold from Dirt.
Unveiling the Survival Strategies of the World’s Most Effective Pathogen, Mycobacterium tuberculosis.
by William R. Jacobs Jr.
September 27, 2005

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), has evolved to become the world's most successful pathogen. Because of its highly infectious nature and its ability to establish persistent infections in individuals with healthy immune systems, M. tuberculosis has infected more than one-third of the world's population, according to World Health Organization (WHO) estimates. ...

The vast majority of infected individuals are subclinically infected, with no symptoms. However, when these people suffer some compromise in their immune systems, they often develop TB, and this promotes the global increase in TB. Combined with these increases, strains of M. tuberculosis have emerged that are resistant to two or more antituberculosis drugs, typically called multidrug-resistant tuberculosis (MDR-TB). The combination of increasing incidence of TB due to increasing numbers of HIV infections and the emergence of MDR-TB prompted WHO to declare TB a global health emergency in 1993, a distinction never accorded to another disease. ...

Mycobacteriophages: Functional and Genomic Gold from Dirt
Viruses that infect mycobacteria (mycobacteriophages) were used by our lab to develop the first gene transfer systems for mycobacteria. ...

Mycobacteriophages are a unique and novel set of unexplored evolutionary material. In collaboration with Graham Hatfull (HHMI Professor, University of Pittsburgh), we have isolated, characterized, and determined the DNA sequences of 14 independent mycobacteriophages. Most of these phages were discovered by our labs in soil samples from such diverse places as backyards, barnyards, or the Bronx Zoo. Several phages were isolated by high school students working in each lab. In collaboration with Reid Schwebach (Albert Einstein College of Medicine), we have developed Phagefinders, a summer research program for high school students. ...

... the phage Bxz1, isolated from a zebra field at the Bronx Zoo, possesses a homolog of the human gene that encodes the protein Ro, to which autoantibodies are generated with the disease lupus. The discovery in Bxz1 may lead to understanding of the function of human Ro. ... it is estimated that 1031 phage particles exist on planet Earth, ...

Growth and Multiplication in Mammalian Hosts:
Overcoming Host Innate and Adaptive Immune Responses M. tuberculosis has evolved the functions required to infect, replicate, and multiply in mammalian cells in the face of innate immune responses. Moreover, the tubercle bacillus has evolved strategies to survive an adaptive immune response. ... M. tuberculosis must make most amino acids in order to replicate in mammalian cells, thus defining intracellular growth requirements. ...

... M. tuberculosis can secrete both offensive and defensive weapons that protect it from innate immune responses. Genes required for the synthesis and export of unique lipids of M. tuberculosis, such as phthiocerol dimycocerosate (PDIM), are essential for lung growth. ... Two of the proteins that are not secreted include catalase peroxidase and superoxide dismutase, proteins that cells use to protect themselves from oxidative killing mechanisms. This secretion system appears to be common to all gram-positive pathogens. Thus, secreted lipids and secreted proteins appear to play roles for survival in the face of innate immune responses. ....

5

http://runews.rockefeller.edu/index.php?page=engine&id=309
Novel Way to Kill Streptococci Bacteria.
for Vincent Fischetti, Ph.D.
Laboratory of Bacterial Pathogenesis,
Rockefeller University,
March 20, 2001

... a powerful new way to destroy on contact the bacteria that cause strep throat, flesh-eating disease and a variety of other infections. The technique, which may not cause the bacteria to evolve resistant strains as antibiotics do, also could have applications for many other bacterial diseases. ...

The new method uses enzymes produced by bacteriophages, tiny viruses that infect bacterial cells, make copies of themselves and then exit to infect other cells. The bacteriophages (or "phages") produce the enzymes after they have finished replication and need to dissolve the bacterial cell wall in order to escape. ...

We can take 10 million organisms in a test tube, add a very small bit of enzyme, and five seconds later, they are all dead. ...

... the phage enzymes also are highly specific.
Fischetti says every type of bacteria has a corresponding phage that infects it. An enzyme made by a streptococcus phage will, when purified, kill only certain streptococci when applied to the microbes on mucous membranes, leaving harmless bacteria alone. "It’s what we call targeted killing, in which we kill only the disease bacteria without disturbing the normal bacteria needed for health, unlike antibiotics which kill everything," he says.

[In 2001], the lab harvests the enzymes from phages that have infected bacteria, but they have the ability to make it artificially if necessary. Fischetti says that eventually production costs could be as low as 10 cents per dose.

"The phage enzymes will not likely cure an infection–its importance lies in lowering the chance that strep will cause infection in the first place," Fischetti says. "The enzymes would be used to eliminate the source of the disease bacteria, which in most cases are the human mucous membranes. The organisms are generally spread from an infected or colonized individual through contaminated saliva. The enzyme could be given in the form of a spray, administered at frequent intervals–such as once or twice a day–to maximize effectiveness." ...

After phages were discovered in 1917, researchers initially thought they would provide an effective way to kill bacteria. They soon learned, however, that phages must bind to specific receptors on the surface of bacteria before injecting their DNA; as bacteria evolve, they change their receptors and shut out the phages. Scientists would constantly have to develop new phages in order for them to be effective. Because of this drawback, phage therapy waned as a technology in most countries. ...

... phage toxins are also released when the bacteria burst, and these agents cause human tissue to be more susceptible to infection. ....

6

http://colloidalsilverresearch.com/watertreat.htm
Non-specific title.
Colloidal Silver research findings.
by multiple researchers.
Water Science and Technology Vol 35 No 11-12 pp 87-93
IWA Publishing 1997

... using H2O2 [hydrogen peroxide] with colloidal silver as an additive to stabilize the Cs and to assist the antibacterial activity. H2O2 is a natural weapon used by most of your cells to oxidize pathogens and the combination has recently been well researched as a sterilizer for potable water systems. ...

Ag+ [silver] was able to inactivate target bacteria (E. coli-B, E. coli-K12), while H2O2 was more effective than Ag+ against MS-2 phages. Copper (250 g/l) had no bactericidal effect but possessed an appreciable viricidal effect. When hydrogen peroxide and copper were combined, a pronounced increase in both bactericidal and viricidal effects was obtained. ...

Silver and hydrogen peroxide (HP) acted synergistically on the viability of E. coli K-12. ...

It is not a question of whether using copper and silver or using chlorine is the better way to keep a swimming pool sanitized and algae free. A good case can be made for either methods. Studies have shown that an ideal method is to use a combination of all three elements.

G.R. Taylor, at Surrey University, proposed a dual disinfection method after his tests showed that

"Two different chemicals of metals added together may allow more efficient disinfection kinetics to be achieved. One substance targets the surface of a micro-organism killing and injuring the cells while a second substance targets the nucleic acid of the micro-organism destroying the remaining injured micro-organisms. By using this method of dual disinfection, reduced levels of both substances may be more effective than much higher levels of either individual substance."

[Copper and Silver are both] highly effective in the control and killing of both bacteria and algae. Together they give superior results. Copper has the ability to pierce the protective outer membrane of a cell and disrupt enzyme balance. Silver is effective because of its capabilities of interfering with DNA production and accelerating the death phase. This method has the advantage over chlorine of remaining very stable in swimming pools. The pool will stay sanitized for days or weeks with the system turned off and no additional copper or silver being added.

Copper and silver are not absorbed through the skin and, therefore, are not carried out of the pool by swimmers. They are also not affected by sunlight and actually become slightly more effective as the water is heated or the pH increases.

... effectiveness of copper and silver in killing E. Coli bacteria.
Many other tests have shown that silver also kills viruses and other types of bacteria. Coleman reported that herpes simplex virus (HSV) type I was quite sensitive to silver. Richards reported that only 3 ug/1 silver was necessary to prevent the growth of pseudomonas. Moroz reported that silver kills salmonella and E. Coli and can kill bacteria highly resistant to antibiotics. ...

Since metallic silver is a catalyst for the breakdown of H2O2 to water and oxygen gas, it can provide the intersticial force, from bubble formation, to break the weak crystals down to possibly an atomic size. ...

Ionic Colloidal Silver has a distinct bitter taste while metallic silver colloids have a distinct metallic taste, as would be expected. It is a simple test to see if you bought a semi-useless metallic silver colloid.

Others have reported on the increased effectiveness of the combination of H2O2 and ionic Cs on patients and much has been written about "most infections start in the ears and can also be treated/cured there" with Colloidal Silver.

7

http://www.biomedcentral.com/1741-7015/2/32
BMC Medicine, BioMed Central
Emergence of new Salmonella Enteritidis phage types in Europe?
by Karin Nygård1, Birgitta de Jong, Philippe J Guerin1,
Yvonne Andersson, Agneta Olsson and Johan Giesecke
April 19, 2004

... Among human Salmonella Enteritidis infections, phage type 4 has been the dominant phage type in most countries in Western Europe during the last years. This is reflected in Salmonella infections among Swedish travellers returning from abroad. ...

Salmonella Enteritidis is the most common serovar causing food-borne salmonellosis in humans, causing approximately 80% of salmonellosis cases reported in Europe [1]. During the 80s and early 90s, a steady increase in S. Enteritidis infections was reported in Europe and North America. ... Approximately 70% of outbreaks caused by S. Enteritidis in Europe during the 90s, were related to eggs and egg products. ...

Of 13,271 cases of S.Enteritidis infections notified during 1997–2002, 11,570 cases (87%) were reported as infected abroad, ... Eighty-six percent (10,049) of the isolates from 1997 to 2001 were phage-typed, increasing from 75% in 1997 to 95% in 2001. ...

People returning from travel abroad may have a higher tendency to seek medical care and have a stool sample taken if an imported infection is suspected. In addition, visitors may be more susceptible to pathogens circulating in the community than the local inhabitants. ...

If proper investigations, and appropriate prevention and control measures are to be implemented in the countries visited, it is important that the surveillance information compiled from the traveller's home countries is rapidly communicated to the affected countries.

8

http://www.sciencentral.com/articles/view.php3?
article_id=218391794&cat=2_2
Exploding Anthrax.
by Joyce Gramza
August 21, 2002

Lysins to Kill
... showed that lysin can kill anthrax in laboratory tests and in living animals, they also demonstrated that anthrax is likely to have a hard time developing resistance to lysins.

... The research is funded by the Defense Advanced Research Projects Agency. Fischetti says development of lysin as an anti-anthrax treatment is being fast-tracked and could be completed within three years.