Patent Description:
Treatment and prevention of bacterial infections are a global problem of modern medicine.

Bacterial infections are not only an important public health issue, but also they are a significant and increasing problem for veterinary medicine, environmental safety and economics. According to the data of World Health Organization (WHO), European Food Safety Authority (EFSA) and European Centre for Disease Prevention and Control (ECDC), public health risks related to anthroponotic bacterial diseases, including salmonellosis, escherichiosis, shigellosis, Staphylococcus, Streptococcus, Pseudomonas infections - are increasing each year. Animals or fowls, affected with bacterial infections, may become a source of infection for humans; animal and poultry products used as food - such as meat, egg - contaminated with microorganisms (Salmonelleae, Shigellae, enteropathogenic Escherichia coli, Proteae, enterotoxigenic strains of Staphylococcus spp. and others), may cause toxicoinfections, intoxications and infectious diseases as well. Furthermore, the sources of infection during the secondary contamination of animal/poultry meat and products thereof, are environmental objects (soil, water, transport etc.), as well as infected people or asymptomatic carriers.

According to the experts of European Food Safety Authority (EFSA), economic burden of anthroponotic diseases may exceed <NUM> billion euros per year in Europe. Continuous increase in anthroponotic bacterial infections in various countries of the world, growth in the number of bacterial agents (their serovars) found in humans, animals and poultry, considerable microbial contamination of food products, environmental objects and economic burden therefrom, make these infections not only medical, but also ecological, veterinary and social problem.

According to WHO, bacterial infections are distinguished by severe disease progression and difficulty in their ellimination. One of the reasons, leading to these problems, is a large number of pathogenic microorganisms (their serotypes). The most noteworthy issue is increasing antibiotic resistance of infection-causing bacterial agents and global spread of their multidrug-resistant strains. One more challenge, facing physicians, is bacterial carriage too, not only in humans, but also in animals and poultry; for instance, carriage prevalence of Salmonelleae is up to <NUM>%.

Generally, antibiotic resistant microorganisms play a key role in the development of bacterial purulent-inflammatory and intestinal infections. Throughout the years, widespread consumption of antibiotics without clinical urgency and strict monitoring for treatment of bacterial infections conditioned emergence and ubiquity of antibiotic-resistant bacterial strains, which is a logical consequence of evolutionary process of microbial development. Due to irrational consumption of antibiotics, we may appear as vulnerable to infections, as we were before the discovery of penicillin, - as stated by World Health Organization. Considering multidrug-resistance of bacterial pathogens, contraindications and complications of antibiotics (allergic reactions, toxic, immunosuppressive, embryotoxic, teratogenic effects, disruption of normal gut microflora - dysbiotic changes and etc.), the issue of treatment and prevention of bacterial infections is still of great importance in practical medicine, needs new methods of approach and development of optimal treatment plans.

Strictly specific, harmless, safe, effective biopreparations - bacteriophages, having the ability to intentionally eliminate conditionally-pathogenic microorganisms - one of the alternatives to antibiotics - are recognized as one of the most biological preparations for treatment and prevention of purulent-inflammatory and intestinal infections (used in humans, animals and poultry, as well as in environmental sanitation).

Bacteriophages or simply phages are bacterial viruses, which cause specific lysis of microorganisms, conditioning bacterial infections - they selectively disrupt bacterial cells. Bacteriophage adsorbs to the membrane of a homologous bacterial cell, disrupts its integrity, penetrates inside, multiplies and causes its lysis, along with releasing of a new population of phage progeny. (<IMG> (<NUM>) <IMG>; <IMG> (<NUM>) <IMG>; <IMG> B. (<NUM>) <IMG>, <IMG>, Nº7, c. <NUM>-<NUM>; <IMG> B. , <IMG>, <NUM>, <IMG>, <IMG>; <NPL>). Re-derived phage progeny continues infecting new microbial cells until infection liquidation - exactly the above-mentioned process is a basis of the successful phage therapy and prophylaxis. Nowadays, up to <NUM> bacteriophage isolates, specific to approximately <NUM> bacterial genera are studied.

Phage therapy was developed shortly after the discovery of bacteriophages (<NUM>. Felix d'Herelle). In the <NUM>, D'Herelle worked out monocomponent dysentery-phage, as well as polycomponent intesti- and pyobacteriophage preparations for treatment and prevention of intestinal and purulent-surgical infections. Eliava together with D'Herelle founded Institute of Bacteriology, Microbiology and Virology (IBMV) in Georgia. A lot of phages, specific to various bacterial pathogens, were isolated and studied in Georgia. The similar activities also took place in Russia, France and Poland. In the <NUM>, shortly after beginning of the antibiotic era, phage therapy was forgotten in Western Europe. However, after the <NUM>, increase in life threatening, antibiotic resistant bacterial infections and spread of them, as well as the possibility of deriving their new forms, rekindled interest in phage preparation consumption (as an alternative remedy to antibiotics) for treatment and prevention of bacterial infections. (<NPL> ; <NPL>). Nowadays, too much attention is focused on this field in the West - phage therapeutic companies are founded, and bacteriophages are actively promoted and advertised. Scientific-clinical researches, which have been conducted for many years, reveal a number of significant factors, determining phage advantages over antibiotics and other chemical antibacterial preparations. These advantages are as follows:.

Thus, consumption of therapeutic phage preparations - alternative to antibiotics - is of great importance for practical medicine to treat and prevent various bacterial infections. Over the years, recommendations regarding the use of phage preparations for treatment and prevention of bacterial infections have been based on the analysis of scientific-clinical researches, clearly certifying the high efficacy of phage therapy and prophylaxis without any contraindications and complications.

Numerous clinical trials corroborated that application of phage preparations, in case of its administration at the onset of the disease and susceptibility of the pathogenic bacteria to the bacteriophage, improves a patient's condition after <NUM> hours in <NUM>%, and <NUM> hours in <NUM>-<NUM>%.

It has to be mentioned that substitution of antibiotics with phage preparations for treatment of various bacterial infections, cause developing of antibiotic-susceptible cells in bacterial (natural and clinical) populations. There are some clinical cases described, where antibiotic resistant bacterial strains were replaced with antibiotic sensitive ones after treating patients with phage preparations. ,<IMG> <IMG>, <NUM>, <IMG>, Nº7, c. <NUM>-<NUM>).

It is worth noting, that application of therapeutic bacteriophage preparations during bacterial infections is recommended as follows:.

High therapeutic potential of phage preparations, their multi-species pattern and polyvalent composition, strict specificity and complete safety - assigns a high priority to consumption of therapeutic bacteriophage preparations in practical medicine as one of the alternatives to antibiotics for treatment and prevention of bacterial infections. (<IMG>, <IMG>, <IMG>, <NUM>, <IMG>. <NUM>-<NUM>; <IMG> A. ,<IMG> <IMG>- <IMG>, <NUM>, Nº6, c. <NUM>-<NUM>; <IMG> E. <IMG> <IMG> <IMG> <IMG>, <NUM>; <IMG> A. ,<IMG> <IMG>, <NUM>, <IMG>, c. <NUM>-<NUM>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>;<NPL>; <NPL>; <NPL>;<NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>).

Therapeutic and prophylactic bacteriophage preparations may be monocomponent (monovalent or polyvalent) or polycomponent (combined polyvalent). The monocomponent preparations comprise one or more true virulent bacteriophages, homologous with a single species of bacteria (serotype, serovar). The examples of the monocomponent preparations are: Staphylococcus phage (Phagestaph), Pseudomonas aeruginosa phage (Phagepy), Salmonella phage (Phagesal, polyvalent), dysentery phage (Phagedys), Klebsiella, Vibrio cholerae, Serratia bacteriophages, etc. (<NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>; <NPL>).

Polycomponent preparations comprise multiple true virulent bacteriophages, homologous with various species (two or more) of bacteria (serotype, serovariant).

According to Prior Art, there are known a number of polycomponent preparations, some of which are described in the following sources:<NPL>; <NPL>.

Even though there are a lot of phage containing preparations nowadays, development of a highly efficient medication for the infectious diseases, caused by various microbes, is still extremely important. The fact that a plurality of diseases, conditioned by multidrug-resistant microbial associations, has been noticeably increased - makes the above-mentioned issue more significant. Therefore, treatment of such diseases requires polycomponent preparations, comprising true virulent bacteriophages, which have the ability to jointly, thoroughly cover the whole specter of disease-causing microbe.

One particular embodiment of the invention relates to an antimicrobial composition against gastrointestinal infections, which comprises:.

In another embodiment of the invention, the above-mentioned composition may have a form, which will be selected from the following group: liquid formulation, spray, tablet, powder, capsule, cream and suppository.

In another embodiment of the invention, the above-mentioned composition is used for treatment and/or prevention of intestinal infections.

In another embodiment of the invention, the above-mentioned composition is used for treatment and/or prevention of intestinal infections in humans.

In another embodiment of the invention, the above-mentioned composition is used for treatment and/or prevention ofintestinal infections in animals and poultry.

In another embodiment of the invention, the above-mentioned composition is used for treatment and/or prevention of dysentery (shigellosis), salmonellosis, escherichiosis (E. coli-infection), dyspepsia, dysbiosis, foodbome toxicoinfections, enteritis, gastroenteritis, colitis, enterocolitis and gastroenterocolitis in humans.

Another embodiment of the invention is defined as isolated bacteriophage strains, constituents of the above-mentioned composition, which have a lytic activity against Shigella Pexneri and are selected from the following group: DSM <NUM> and DSM <NUM>. All the above-named strains are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as isolated bacteriophage strains, constituents of the above-mentioned composition, which have a lytic activity against Shigella sonnei and are selected from the following group: DSM <NUM> and DSM <NUM>. All the above-named strains are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as an isolated bacteriophage strain DSM <NUM>, which is constituent of the above-mentioned composition, has a lytic activity against Salmonella choleraesuis and is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as an isolated bacteriophage strain DSM <NUM>, which is constituent of the above-mentioned composition, has a lytic activity against Salmonella newport and is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as an isolated bacteriophage strain DSM <NUM>, which is constituent of the above-mentioned composition, has a lytic activity against Salmonella paratyphi A and is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as an isolated bacteriophage strain DSM <NUM>, which is constituent of the above-mentioned composition, has a lytic activity against Salmonella typhimurium and is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as an isolated bacteriophage strain DSM <NUM>, which is constituent of the above-mentioned composition, has a lytic activity against Salmonella paratyphi B and is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as an isolated bacteriophage strain DSM <NUM>, which is constituent of the above-mentioned composition, has a lytic activity against Salmonella heidelberg and is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Another embodiment of the invention is defined as isolated bacteriophage strains, constituents of the above-mentioned composition, which have a lytic activity against Enterococcus and are selected from the following group: DSM <NUM> and DSM <NUM>. All the above-named strains are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

According to one of the preferred embodiments of the invention, there is provided an antimicrobial composition, which contains:.

According to the invention, a phage, which is a component of the composition, refers to the deposited bacteriophage and progeny derived therefrom, the genetic profile of which is substantially equivalent to the profile of said bacteriophage.

Based on the data of World Health Organization, <NUM>% of the gastrointestinal infections are caused by pathogenic and conditionally pathogenic bacteria: Salmonella, Shigella, E. coli, Proteus, Pseudomonas aeruginisa, Staphylococcus, Enterococcus.

Shigella flexneri and Shigella sonnei cause shigellosis. Shigellosis accounts for a large number of acute intestinal infections. Worldwide, Shigella causes <NUM> million cases of disease annually, of which <NUM> are fatal. Children are affected in <NUM>-<NUM>% of all shigellosis cases. Shigella infection is the most common in children between the ages of <NUM>-<NUM>. Complications of severe dysentery in younger children may include nephritis, toxic and hypovolemic shock, rectal prolapse, bleeding, invagination or peritonitis. Bronchopneumonia, stomatitis, pyoderma, furuncolosis, osteomyelitis, urinary tract infections, hemolytic-uremic syndrome with acute kidney failure, non-purulent arthritis and Reyter's syndrome may also occur; Shigella-induced bacteremia causes death in <NUM>% of patients.

The main diseases, caused by Salmonelleae, may be divided into three groups: typhoid and paratyphoid fever, gastroenteritis and septicemia.

Typhoid and paratyphoid fever - (typhus abdominalis, S. paratyphi A and S. paratyphi B infections) - belong to the group of acute infections, caused by Salmonella typhi, Salmonella Paratyphi A, Salmonella schottmulleri. These diseases are characterized by fever, general intoxication, bacteremia, hepatosplenomegaly, enteritis and specific injury of intestinal lymphatic system. Salmonelleae release endotoxin when they are disrupted, which induces symptoms of general intoxication and plays an important role in development of small intestinal ulcers, leukopenia and potentially toxic shock syndrome (TSS).

Salmonellosis is an acute infectious disease, which is caused by Salmonelleae and characterized by a wide range of clinical manifestations, including asymptomatic carriage or even septic forms. It mainly affects gastrointestinal tract (gastroenteritis, colitis). The leading causes of salmonellosis in children are: S. typhimurium, S. enteritidis, S. infantis, S. heidelberg and others. Septic form of salmonellosis is characterized by especially severe course of the disease. The illness does not respond to antibiotic-therapy well. Secondary purulent foci are frequently formed in the musculoskeletal system (osteomyelitis, arthritis, spondylitis). Sometimes salmonellosis-related infective endocarditis, aortitis with development of late aortic aneurysm, purulent meningitis; rarely - liver abscess, purulent strumitis and infection of the ovarian cyst may also occur. Severe forms of the disease may cause dehydration, as well as toxic shock syndrome (TSS). Salmonella is estimated to cause up to <NUM>. <NUM> food poisoning cases, <NUM> hospitalizations and <NUM> deaths in the United States every year.

Esherichia coli causes enteritis, urethritis, cystitis, pyelonephritis, cholecystitis, peritonitis, septicemia, meningitis in children, wound infections etc. E. coli is a main causative microorganism of acute intestinal infections. Escherichiosis (synonym - intestinal E. coli infection) elicits the symptoms of gastroenteritis and gastroenterocolitis.

Diarrheagenic E. coli is divided into <NUM> categories: enterotoxigenic (ETEC), enteroinvasive (EIEC), enteropathogenic (EPEC), enterohemorrhagic (EHEC) and enteroadherent (EAEC). Enterotoxigenic E. coli is a main causative agent of traveler's diarrhea. It also is a common cause of gastroenteritis in young children. Enterohemorrhagic E. coli causes hemorrhagic colitis. Except for diarrhea, enterohemorrhagic E. coli causes developing of hemolytic-uremic syndrome (HUS). Enterohemorrhagic strains of E. coli is distinguished by high virulence and pathogenicity.

The most common etiologic agent of urogenital disorders during pregnancy (<NUM>-<NUM>%) is E.

Proteus vulgaris causes urinary tract infections (UTIs), abscesses, meningitis, gastroenteritis, burn wound infections, as well as secondary septic lesions after surgical procedures and burn injuries.

Staphylococcus aureus is a major pathogen responsible for neonatal nosocomial infections (<NUM>%). It is a main causative agent of pneumonia (<NUM>%), sepsis (<NUM>-<NUM>%), meningitis, skin infections (<NUM>%) and bacterial sinusitis (<NUM>-<NUM>%). Sputum culture of the patients, having cystic fibrosis (mucoviscidosis), which has been a global issue over the past <NUM> years, most commonly reveals S. aureus (<NUM>-<NUM>%) in young age groups. Staphylococcus aureus is detected in <NUM>,<NUM>-<NUM>,<NUM>% of urogenital disorders during pregnancy, such as: pyelonephritis, cystitis, urethritis, bacteriuria, colpitis, endocervicitis, endometritis, salpingo-oophoritis.

Staphylococcal enterocolitis may be the primary manifestation of infection or develop secondary to sepsis or other underlying conditions (pneumonia, meningitis, omphalitis etc.). The disease most frequently occurs in weak, premature babies up to <NUM> months of age, with different accompanying diseases.

Pseudomonas aeruginosa causes <NUM>-<NUM>% of nosocomial infections, <NUM>-<NUM>% of hospital-acquired pneumonia (HAP) and <NUM>-<NUM>% of purulent-surgical infections. One-third of urogential tract infections (UTIs) is also resulted from P. aeruginosa. aeruginosa is a causative agent of keratitis, endocarditis, enteritis, conjunctivitis, otitis, meningitis, bacteremia/septicemia, perirectal and rectal abscesses, osteomielitis and arthritis. Mortality rate for bacteremia-septicemia is <NUM>-<NUM>%. The most common pathogens detected with a sputum culture of the patients, having cystic fibrosis (mucoviscidosis) are mucoid (virulent), multidrug-resistant P. aeruginosa strains. <NUM>% of chronic infections conditioned by the above-mentioned microorganism is fatal.

Enterococci (Enterococcus faecium and Enterococcus faecalis) cause enteritis, colitis, urinary tract infections (UTIs), endocarditis, bacteremia-septicemia, neonatal sepsis. Most of the enterococcal infections are considered as nosocomial. <NUM>% of the strains, isolated from hospitals, is vancomycin-resistant Enterococcus faecium.

The aforementioned data of World Health Organization (WHO) and Georgian National Center for Disease Control and Public Health (NCDC), as well as the fact that an infection is rarely caused by a single microbial species, led to the development of an effective antimicrobial composition. True virulent bacteriophages, constituents of the composition, result in lysis of the main aforementioned pathogens and are highly efficient in treating and preventing the infections, caused by microbial associations. It is worth mentioning, that different types of bacteriophages, constituents of the composition, do not reveal antagonistic action to each other, which proves their strict specificity once more.

In a preferred embodiment of the invention, titer of the phages and phage concentrates in the composition are as follows:.

According to the invention, the composition contains bacteriophages and optionally a carrier. The composition may have a liquid or lyophilized powder formulation. The preparation, produced on the basis of the composition, may have a form of injection, infusion, spray, tablet, capsule, cream or suppository. The preparation should preferably comprise a pharmaceutically acceptable carrier.

In order to produce the needed form (e.g. solution for injection, nasal spray etc.) or before using it, the composition of lyophilized powder formulation may be suspended in water for injection, buffered solution, <NUM>% glucose solution, glycerine, dextran, polyethylene glycol, sorbitol or any kind of solution, that maintain phage viability and are not toxic to humans.

According to the invention, sachets, tablets and capsules may be produced on the basis of the powder composition using the well-known pharmaceutical technologies. The above-mentioned preparations may contain stabilizers, conservatives, additional active ingredients, e.g. antibiotics. Tablets and capsules may be produced by the technology, which will ensure the release of the active ingredient in the intestines.

The preparation, made on the basis of the composition, may have a liquid form for oral administration, e.g. suspensions, solutions, emulsions or syrups. Such formulations may contain suspending agents, emulsifiers, conservatives, aromatizing agents, sweeteners etc..

On the basis of the composition topical preparations may also be developed. Such preparations include creams and suppositories. They contain bases, which are well-known in the pharmaceutical industry, maintain phage viability and are not toxic to humans.

The composition comprises isolated bacteriophages. Cultivation of the isolated bacteriophages is carried out separately from the environment. Thus, each strain of the isolated bacteriophage is pure and practically does not include other bacteriophage mixtures.

Deposited bacteriophages, constituent of the composition, are specific to the targeted bacteria and have the ability to lyse them.

According to the invention, the concept of a bacteriophage, constituent of the composition, includes the deposited bacteriophages, as well as progeny thereof, genetic profile of which are essentially equivalent to the profile of said bacteriophages, thus, they fully maintain specificity to the targeted bacteria. The above-mentioned phage progeny may have some genetic variations, the limits of which corresponds with the standards for "closely related microorganisms", developed by Tenover (<NPL>). Bacteriophages, needed to produce the composition, are cultivated by the method, technique and materials of which are well-known for the specialists of the above-mentioned field. More specifically, each production strain of the targeted bacteria, susceptible to the deposited bacteriophages, is cultivated in a culture medium and then inoculated with its specific bacteriophages (deposited bacteriophages which are specific to the aforementioned bacteria) at the pre-determined optimal multiplicity of infection (MOI). After incubation and bacterial lysis, bacteriophages are collected, purified and concentrated, which result in obtaining the phages, necessary for the composition production. The steps of purification and concentration include various systems of filtration and centrifugation, which are well-known for the field specialists (<NPL>).

Determination of a particular bacteriophage concentration is carried out by phage titration assay. If higher concentration of the bacteriophages - specific to the particular bacterial strain - is needed, concentrating is carried out by filtration and centrifugation, and if less concentration is required, it is suspended in water or buffer. Finally, in order to produce the composition, progeny of each deposited bacteriophage strain, obtained by this method, is combined into a single phage cocktail.

The composition contains the following bacteriophage strains, specific to Shigella flexneri: DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

The optimal conditions for propagation of the above-mentioned bacteriophages are: pH - <NUM>-<NUM>,<NUM>; temperature - <NUM>-<NUM>; culture media: pancreatic hydrolizate of fish meal or soya broth; multiplicity of infection (MOI) - <NUM>,<NUM>; cultivation duration - <NUM>-<NUM>.

Plaque morphology and lytic activity of these bacteriophages were determined. Morphology of the bacteriophages - DSM <NUM> and DSM <NUM> were investigated with the aid of electron microscope. The interaction between the above-mentioned phages and their host cells, particularly, adsorbtion rate of bacteriophages onto the host cells in short period of time (<NUM>), as well as adsorbtion rate constant - K were also studied.

The data of the above-noted assays are shown in table <NUM> (the table shows bacterial strains of the collection, maintained by JSC "Biochimpharm"), while <FIG> depict electron-microscopic images of the above-mentioned bacteriophages.

Genomes of the bacteriophages - DSM <NUM> and DSM <NUM>, particularly, restriction fragment length polymorphism profiles (RFLP) were also studied. In order to determine RFLP profiles, restriction enzyme - Afl II was used.

RFLP profiles of the above-mentioned bacteriophages are shown in <FIG>.

The composition contains the following bacteriophage strains, specific to Shigella sonnei: DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Plaque morphology and lytic activity of these bacteriophages were determined. Morphology of the bacteriophages - DSM <NUM> and DSM <NUM> were investigated with the aid of electron microscope. The interaction between the above-mentioned phages and their host cells, particularly, adsorbtion rate of the bacteriophages onto the host cells in short period of time (<NUM>), as well as adsorbtion rate constant - K were also studied.

The composition contains the following bacteriophage strain, specific to Salmonella choleraesuis: DSM <NUM> - which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

The optimal conditions for propagation of the above-mentioned bacteriophage are: pH - <NUM>-<NUM>,<NUM>; temperature - <NUM>-<NUM>; culture media: pancreatic hydrolizate of fish meal or soya broth; multiplicity of infection (MOI) - <NUM>,<NUM>; cultivation duration - <NUM>-<NUM>.

Plaque morphology and lytic activity of this bacteriophage were determined. Morphology of the bacteriophage - DSM <NUM> was investigated with the aid of electron microscope. The interaction between the above-mentioned phage and its host cell, particularly, adsorbtion rate of the bacteriophage onto the host cell in short period of time (<NUM>), as well as adsorbtion rate constant - K were also studied.

The data of the above-noted assays are shown in table <NUM> (the table shows bacterial strains of the collection, maintained by JSC "Biochimpharm"), while <FIG> depicts electron-microscopic image of the above-mentioned bacteriophage.

Genome of the bacteriophage - DSM <NUM>, particularly, restriction fragment length polymorphism profile (RFLP) was also studied. In order to determine RFLP profile, restriction enzyme - Hind III was used.

The composition contains the following bacteriophage strain, specific to Salmonella newport DSM <NUM> - which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

The composition contains the following bacteriophage strain, specific to Salmonella paratyphi A: DSM <NUM> - which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

The composition contains the following bacteriophage strain, specific to Salmonella typhimurium: DSM <NUM> - which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Genome of the bacteriophage - DSM <NUM>, particularly, restriction fragment length polymorphism profile (RFLP) was also studied. In order to determine RFLP profile, restriction enzyme - Spe I was used.

The composition contains the following bacteriophage strain, specific to Salmonella paratyphi B: DSM <NUM> - which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

The composition contains the following bacteriophage strain, specific to Salmonella heidelberg. DSM <NUM> - which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

The composition contains the following bacteriophage strains, specific to Escherichia coli: DSM <NUM>, DSM <NUM> and DSM <NUM> - which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Plaque morphology and lytic activity of these bacteriophages were determined. Morphology of the bacteriophages - DSM <NUM>, DSM <NUM> and DSM <NUM> - was investigated with the aid of electron microscope. The interaction between the above-mentioned phages and their host cells, particularly, adsorbtion rate of the bacteriophages onto the host cells in short period of time (<NUM>), as well as adsorbtion rate constant - K were also studied.

Genomes of the bacteriophages - DSM <NUM>, DSM <NUM> and DSM <NUM>, particularly, restriction fragment length polymorphism profiles (RFLP) were also studied. In order to determine RFLP profiles, restriction enzymes: EcoR V and Afl II were used.

The composition contains the following bacteriophage strains, specific to Proteus vulgaris: DSM <NUM>, DSM <NUM> and DSM <NUM> - which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Genomes of the bacteriophages - DSM <NUM>, DSM <NUM> and DSM <NUM>, particularly, restriction fragment length polymorphism profiles (RFLP) were also studied. In order to determine RFLP profiles, restriction enzymes: Hind III and Afl II were used.

The composition contains the following bacteriophage strains, specific to Staphylococcus aureus DSM <NUM>, DSM <NUM> and DSM <NUM> - which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Genomes of the bacteriophages - DSM <NUM>, DSM <NUM> and DSM <NUM>, particularly, restriction fragment length polymorphism profiles (RFLP) were also studied. In order to determine RFLP profiles, restriction enzymes: EcoR I, EcoR V, Hind III, Spe I, Afl II were used.

The composition contains the following bacteriophage strains, specific to Pseudomonas aeruginosa: DSM <NUM>, DSM <NUM> and DSM <NUM>- which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Plaque morphology and lytic activity of these bacteriophages were determined. Morphology of the bacteriophages - DSM <NUM>, DSM <NUM> and DSM <NUM>- was investigated with the aid of electron microscope. The interaction between the above-mentioned phages and their host cells, particularly, adsorbtion rate of the bacteriophages onto the host cells in short period of time (<NUM>), as well as adsorbtion rate constant - K were also studied.

Genomes of the bacteriophages - DSM <NUM>, DSM <NUM> and DSM <NUM>, particularly, restriction fragment length polymorphism profiles (RFLP) were also studied. In order to determine RFLP profiles, restriction enzymes: EcoR V and Hind III were used.

The composition contains the following bacteriophage strains, specific to Enterococcus: DSM <NUM> and DSM <NUM> - which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ).

Plaque morphology and lytic activity of these bacteriophages were determined. Morphology of the bacteriophages - DSM <NUM> and DSM <NUM> - was investigated with the aid of electron microscope. The interaction between the above-mentioned phages and their host cells, particularly, adsorbtion rate of the bacteriophages onto the host cells in short period of time (<NUM>), as well as adsorbtion rate constant - K were also studied.

Genomes of the bacteriophages - DSM <NUM> and DSM <NUM>, particularly, restriction fragment length polymorphism profiles (RFLP) were also studied. In order to determine RFLP profiles, restriction enzyme Hind III were used.

In vitro lytic activity of bacteriophages, which were specific to <NUM> bacterial strains, constituents of the composition, were also studied. Particularly, lytic activity of bacteriophages, specific to <NUM> bacterial strains of the collection, maintained by JSC "Biochimpharm", as well as <NUM> bacterial strains of international collections and various countries (Spain, Germany, Australia, etc.) were investigated.

The results of phage lytic activity against bacterial strains of the collection, maintained by JSC "Biochimpharm" are shown in table <NUM>. According to the table, in vitro efficiency - spectrum of lytic activity of bacteriophages, which are constituents of the above-mentioned composition, against their homologous bacterial strains are as follows: Shigella (<NUM> strains) - <NUM>-<NUM>%; Salmonella (<NUM> strains) - <NUM>-<NUM>%; E. coli (<NUM> strains) - <NUM>,<NUM>-<NUM>,<NUM>%; Proteus (<NUM> strains) - <NUM>%; S. aureus (<NUM> strains) - <NUM>,<NUM>-<NUM>,<NUM>%; P. aeruginosa (<NUM> strains) - <NUM>,<NUM>-<NUM>,<NUM>%; Enterococcus (<NUM> strains) - <NUM>-<NUM>,<NUM>%. Screening analysis showed that spectrum of the lytic activity of these <NUM> bacteriophages, constituents of the composition, overlaps each other; thus, in vitro efficiency - spectrum of the lytic activity of the batceriophages, constituents of the composition, against <NUM> bacterial strains of the collection, maintained by JSC "Biochimpharm", is <NUM>%.

Lytic activity of bacteriophages, constituents of the composition, against bacterial strains of international collections and various countries (Spain, Germany, Australia, USA) is as follows: against Staphylococcus aureus strains - <NUM>,<NUM>%; against E. coli strains - <NUM> %; against Proteus strains -<NUM>%; against P. aeruginosa strains - <NUM>%; against Enterococcus strains - <NUM>%; against Salmonella strains - <NUM>%; against Shigella strains - <NUM>%.

The composition is used for treatment and prevention.

The composition is indicated to be used for treatment of the following diseases: dysentery (shigellosis), salmonellosis, escherichiosis (E. coli infection), dyspepsia, dysbiosis, foodbome toxicoinfections, enteritis, gastroenteritis, colitis, enterocolitis, gastroenterocolitis.

The composition is recommended to be used for prevention during outbreaks of intestinal infections in food establishments, places of mass gatherings, closed groups (kindergartens, schools, hospitals, etc.), military units (under field conditions and during military operations), and during natural disasters.

Except the above-mentioned conditions, the composition may also be used in phage diagnostics, phage indication and phage prophylaxis. Its application can be directed towards normalizing the ecological condition, environmental sanitation, agricultural and aquaculture products.

For reduction, elimination and prevention of colonization with pathogenic bacteria, food products may be processed by the composition, proposed by the invention. The preparations, produced on the basis of the composition, may be used in manufacturing facilities, retirement homes, kindergartens, etc. for the purpose of environmental sanitation and prevention of bacterial colonization.

The composition may be used as an additive in cosmetic products (creams, lotions, gels, etc.).

Besides, in vivo therapeutic efficiency of the composition was also studied. Over the course of phage therapy (<NUM>-<NUM> days) with the composition in children with intestinal infections of mild and medium severity, complete elimination of pathogenic and conditionally pathogenic microbes: hemolytic E. coli strains (in <NUM>% of the patients) and S. aureus (in <NUM>%), as well as partial eradication of Proteus vulgaris, Klebsiella, Serratia - were achieved. Yet it is noteworthy that in <NUM>% of patients, who did not undergo anti-fungal therapy, revealed elimination of Candida spp. infections.

The clinical trials studying the composition (<NUM> patients with the diagnosis of acute bacterial intestinal infection of medium severity, etiologic agents: S. enteritidis, S. typhimurium, S. flexner) revealed significant improvement in <NUM> hours after treatment initiation in <NUM>% of patients (reduction in the intensity and frequency of intoxication, abdominal pain, lack of appetite, malaise, diarrhea). After a short-course monotherapy with the composition (<NUM>-<NUM> days), full recovery of patients, proven by clinical and laboratory findings, was achieved.

Moreover, efficiency of the composition was compared with the effectiveness of antibiotics, Sextaphage (RU2410084 (FEDERAL NOE GUP NPOB MED IMMUNOBIOLOGICHESKIM PREPARATAM MIKROGEN MIN ZDRAVOOKHRANENIJA RF) <NUM>. <NUM>) and Pyophage (RU2036232 (UFIM Nil VAKTSIN I SYVOROTOK) <NUM>. Efficiency of bacteriophages, constituents of the composition, during intestinal infections, which were caused by antibiotic-resistant bacterial strains, is <NUM>-<NUM>%, particularly: <NUM>,<NUM>% against Staphylococcus aureus strains, <NUM>% against E. coli strains, <NUM>% against Proteus strains, <NUM>% against P. aeruginosa strains, <NUM>% against Enterococcus strains, <NUM>% against Salmonella strains, <NUM>% against Shigella strains. Meanwhile, effectiveness of Sextaphage and Pyophage is <NUM>-<NUM>%, particularly, <NUM>-<NUM>% against Staphyloccccus aureus strains, <NUM>-<NUM>% against E. coli strains, <NUM>-<NUM>% against Proteus strains, <NUM>-<NUM>,<NUM>% against P. aeruginosa strains. As for antibiotic therapy, its efficacy does not exceed <NUM>% (<NPL>).

Hence, the composition, proposed by the invention, is highly efficient medium to treat and prevent diseases, caused by various microbes, especially, microbial associations.

Claim 1:
The antimicrobial composition comprising:
a) Bacteriophage strains, specific to Shigella flexneri. DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
b) Bacteriophage strains, specific to Shigella sonnei. DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
c) Bacteriophage strain, specific to Salmonella choleraesuis. DSM <NUM>, which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
d) Bacteriophage strain, specific to Salmonella newport DSM <NUM>, which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
e) Bacteriophage strain, specific to Salmonella paratyphi A: DSM <NUM>, which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
f) Bacteriophage strain, specific to Salmonella typhimunum. DSM <NUM>, which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
g) Bacteriophage strain, specific to Salmonella paratyphi B: DSM <NUM>, which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
h) Bacteriophage strain, specific to Salmonella heidelberg DSM <NUM>, which is deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
i) Bacteriophage strains, specific to Escherichia coll. DSM <NUM>, DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
j) Bacteriophage strains, specific to Proteus vulgaris. DSM <NUM>, DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
k) Bacteriophage strains, specific to Staphylococcus aureus. DSM <NUM>, DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
l) Bacteriophage strains, specific to Pseudomonas aeruginosa:. DSM <NUM>, DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ),
m) Bacteriophage strains, specific to Enterococcus. DSM <NUM> and DSM <NUM>, which are deposited in German Collection of Microorganisms and Cell Cultures (DSMZ)
and optionally a pharmaceutically acceptable excipient.