RU2628312C2 - Antibacterial composition for prevention or treatment of hospital infections (versions), bacteriophage stamps used to obtain such composition - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: composition includes 7 strains of bacteriophage, each represented by a combination of bacterial phagolysates filtrates with lytic activity of at least 10^-4 according to Appelman concerning test strains and isolated bacterial isolates, as well as target additives in an amount of 0.01±99.99 wt % of composition weight. At that, Staphylococcus aureus phagolysate filtrate obtained using the Staphylococcus aureus SCH1 bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-105, Staphylococcus aureus phagolysate filtrate obtained using the Staphylococcus aureus SCH111 bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-95, Klebsiella pneumoniae phagolysate filtrate, obtained using the Klebsiella pneumoniae KPV15 bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-90, Klebsiella pneumoniae phagolysate filtrate, obtained using the Klebsiella pneumoniae KPV811 bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-91, Pseudomonas aeruginosa phagolysate filtrate, obtained using the PA5 Pseudomonas aeruginosa bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-88, Pseudomonas aeruginosa phagolysate filtrate, obtained using the Pseudomonas aeruginosa PA10 bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-89, Acinetobacter baumannii, phagolysate filtrate, obtained using the Acinetobacter Baumannii AM24 bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-106. The composition version also contains Acinetobacter baumannii phagolysate filtrate, obtained using the Acinetobacter baumannii AP22 bacteriophage strain, deposited in the collection of FBIS SSC PVB of Rospotrebnadzor under number Ph-42. The appropriate bacteriophage strains are also proposed. Nucleic acid molecules corresponding to the genome of the said bacteriophages are also proposed.

EFFECT: expanded range of products containing highly selective natural antibacterial components as the main active ingredient, ensured biological stability and activity of bacteriophages in the agent composition, the composition has a low risk of toxic and side effects, allows to expand versions and methods of practical application of the agent containing bacteriophages, provides storage stability and application efficiency over a wide temperature range.

28 cl, 1 dwg, 4 tbl, 24 ex

Description

The invention relates to biotechnology and can be used in the manufacture of therapeutic products based on bacteriophages.

The proliferation of antibiotic-resistant bacteria, as well as the discovery of the role of antibiotic-resistant biofilms in the development of chronic infections, revived interest in the use of bacteriophages.

Currently, antibiotics are widely used both for treatment and for the prevention of various diseases. However, the frequency and severity of the course of purulent-septic infections do not tend to decrease. One of the reasons for this is the increase in the etiological role of microorganisms that are resistant to most antibiotics. The use of antibiotics is accompanied by a violation of the composition of normal microflora, allergization and the rapid adaptation of microorganisms to the environment.

According to the US Centers for Disease Control and Prevention (CDC), 1 out of every 20 hospitalized patients has a hospital infection. This corresponds to approximately 1.7 million hospital infections per year and causes 99,000 deaths per year. CDC estimates show that the direct costs of inflation-adjusted nosocomial infections exceed $ 30 billion annually.

About 16% of hospital infections are associated with multidrug resistance of pathogens:

methicillin-resistant Staphylococcus aureus (8%)

vancomycin-resistant Enterococcifaecium (4%)

carbapenem-resistant P. aeruginosa (2%)

cephalosporin-resistant K. pneumonia (1%)

carbapenem-resistant A. baumannii (0.5%)

Methicillin-resistant Staphylococcus aureus (MRSA) causes a range of diseases: from skin and wound infections to pneumonia and bloodstream infections, which can lead to sepsis and death. Staphylococcal bacteria, including MRSA, are one of the most common causes of human infectious diseases. Resistance to methicillin and similar antibiotics (nafcillin, oxacillin) and cephalosporin resistance according to CDC estimates in 2011 led to 80,461 cases of the disease caused by MRSA and 11,285 deaths.

K. pneumonia is an important cause of nosocomial infections. In 1998, K. pneumonia and K. oxytoca accounted for 8% of nosocomial bacterial infections in the United States and Europe, including urinary tract infections, pneumonia, septicemia, and soft tissue infections. K. pneumonia is considered a hospital opportunistic pathogen that usually affects patients with implanted medical devices. K. pneumonia produces an expanded spectrum of β-lactamases and the infections caused by it cannot be treated with third-generation cephalosporins. This severely limits therapeutic options.

P. aeruginosa is often the cause of infection, especially in patients with compromised immune defenses, including pneumonia, bloodstream infections, urinary tract infections, and surgical infections. This is the most common pathogen isolated from patients hospitalized for more than 1 week. Some strains of P. aeruginosa are resistant to almost all antibiotics, including aminoglycosides, cephalosporins, fluoroquinolones, carbapenems.

In connection with the foregoing promising direction in improving the prevention and treatment of infections, including purulent-septic, is the use of therapeutic bacteriophages, which can also affect bacterial strains resistant to antibiotics. They themselves or in combination with other antibacterial drugs increase the effectiveness of etiotropic therapy. In addition, preparations of bacteriophages are not toxic, do not cause the development of dysbiosis and other side effects.

The advantages of bacteriophages over antibiotics are quite obvious and are as follows: bacteriophages are able to destroy bacteria resistant to antibiotics, because they act only on certain bacteria; freely penetrate into the tissues of the human and animal body, without disturbing the balance of the higher organism; practically do not cause side effects; do not inhibit the growth of normoflora; do not weaken the immune system; do not develop bacterial resistance; combined with any medications; have an immunostimulating effect.

Many specialists include bacteriophages in the complex of therapeutic measures and are convinced of their high effectiveness both with local and oral administration. In some cases, bacteriophages were used without antibiotics. It was noted that clinical improvement occurred on average 3-6 days faster than in the control group. Significantly reduced the length of stay of patients in a bed.

It was established that, despite the method of application (local or general), phages penetrate the blood and lymph and enter the focus of inflammation, having a positive effect on the immune status. Bacteriophages are able to quickly penetrate the blood and lymph and excreted through the kidneys with urine. The activity of therapeutic and prophylactic bacteriophages against pathogens of purulent-septic and enteric diseases is quite high - from 72 to 90%, including and in respect of strains of hospital origin characterized by multiple antibiotic resistance. The correspondence of bacteriophage preparations to the modern etiological structure of pathogens is achieved due to their constant adaptation to circulating strains by updating phage races and bacterial production strains. This feature distinguishes phages from other antimicrobial agents - antibiotics, eubiotics or vaccines, where production strains (producer strains) or a synthesized substance are not subject to any modifications. Under the influence of phage, phagocytosis is first activated, the activity of neutrophils and their metabolic activity increase, which prevents the recurrence of infection and the chronicity of the inflammatory process. A decrease in the number of leukocytes and neutrophils was noted, and the level of lymphocytes increased in the treatment of chronic inflammatory diseases against the background of immunosuppressive conditions, bacterial carriage.

Modern therapeutic and prophylactic preparations of bacteriophages are composed of polyclonal virulent bacteriophages of a wide range of action, which are also active against bacteria resistant to antibiotics.

Currently, in Russia for phagotherapy and phagoprophylaxis of hospital infections, the following are produced and used:

- polyvalent salmonella bacteriophage;

- monovalent bacteriophages - typhoid, dysentery, protea, Pseudomonas aeruginosa, cholera, staphylococcal, streptococcal, coliphage (Escherichia coli);

- combined preparations of polyvalent bacteriophages, among which the most famous are:

- Combined pyobacteriophage (Pyopolifag) is able to lyse staphilo-, streptococci (including enterococci), protea, Pseudomonas aeruginosa and Escherichia coli.

Designed for the prevention and treatment of purulent-inflammatory diseases of the ear, throat, nose, sinuses, respiratory tract, lungs; surgical infections (suppuration, abscess, phlegmon, osteomyelitis, peritonitis), urogenital infections (urethritis, cystitis, pyelonephritis); gynecological infections (colpitis, endometritis, salpingo-oophoritis); enteric infections (gastroenterocolitis, cholecystitis, dysbiosis); purulent-septic diseases of the newborn.

- Polyvalent pyobacteriophage (Sextafag) has the ability to specifically lyse staphylococcus aureus, streptococci (including enterococci), Escherichia coli, Proteus, Pseudomonas aeruginosa and Klebsiellupneumonia. Designed for the prevention and treatment of various forms of purulent-inflammatory and enteric diseases: purulent-inflammatory diseases of the ear, throat, nose, respiratory tract, lungs and pleura; surgical, urogenital and enteric infections; generalized septic diseases, including purulent-septic diseases of newborns and infants.

- The complex liquid pyobacteriophage is a mixture of staphilo-, strepto-, strepto-, enterococci, E. coli, Proteus (mirabilis and vulgaris), Pseudomonas aeruginosa and Klebsiella (pneumonia and oxytoca) phagolysates. Used for the treatment and prevention of purulent-inflammatory and intestinal diseases: diseases of the ear, throat, nose, respiratory tract and lungs; surgical, urogenital, enteric infections; generalized and septic diseases; purulent-inflammatory diseases of the newborn, for the treatment of postoperative and freshly infected wounds.

The disadvantage of these complex preparations is the fact that the bacteriophages that make up their composition as an active principle, as well as the collection strains of sensitive microorganisms on which they are cultivated, do not have a complete biological and molecular genetic characteristics. The collection of strains of sensitive microorganisms is replenished and constantly updated with circulating pathogens (Funkner EV Microbiological and technological aspects of the development of a complex preparation of bacteriophages. Abstract of dissertation for the degree of candidate of medical sciences. - Perm, 2007. - 24 p.). As a result, with regulated passages and updates of bacteriophage strains, moderate bacteriophages can appear in the preparations, potentially lysogenic bacteria can be included in the collection, which together affects the safety and effectiveness of these drugs.

In addition, none of the above preparations contains bacteriophage strains against nosocomial infections caused by A. baumannii, which is the most clinically significant species of the genus Acinetobacter, which causes 2-10% of gram-negative infections in Europe and the USA, up to 1% of all nosocomial infections. The patent literature describes a strain of bacteriophage A. baumannii deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number AP22, which is used to identify microorganisms of this type and to develop complex therapeutic drugs against nosocomial infections, but in practice these preparations you are not used (RF patent №2439151).

As general risk factors for infections caused by A. baumannii, there are: male gender; elderly age; the presence of concomitant diseases (malignant blood diseases, cardiovascular or respiratory failure, disseminated intravascular coagulation); the duration of the use of invasive methods of treatment and monitoring (mechanical ventilation for more than 3 days; inhalation administration of drugs; administration of a nasogastric tube; tracheostomy; catheterization of the bladder, central vein, artery, surgery); long stay in a hospital or intensive care unit (ICU); previous antibiotic therapy using cephalosporins, fluoroquinolones or carbapenems.

Prior hospitalization in ICU surgery increases the risk of infection by about 5 times.

A. baumannii in most cases causes disease in seriously ill immunocompromised patients. This microorganism can cause respiratory tract infections (sinusitis, tracheobronchitis, pneumonia), blood flow (sepsis, endocarditis of natural and artificial valves), urinary tract infections, wound and surgical infections, skin and soft tissue infections (including necrotizing fasciitis), and the nervous system (meningitis , ventriculitis, brain abscess), intra-abdominal (abscesses of various localization, peritonitis), musculoskeletal system (osteomyelitis, arthritis).

The main task solved by the invention is to expand the spectrum of the specific activity of the composition of therapeutic and prophylactic purposes based on bacteriophages by including in its composition the active ingredients of virulent bacteriophages with a non-overlapping or partially overlapping spectrum of lytic activity and the selection of targeted additives that stabilize the activity of bacteriophages that are part of composition and its purpose, depending on the dosage form used (hereinafter in the description, formulas and the abstract of the invention - target additives).

The task is achieved due to the fact that the antibacterial composition includes a combination of bacterial phagolysate filtrates with lytic activity of at least 10–4 according to Appelman in relation to test strains and bacterial isolates isolated from the clinical material: Staphylococcus aureus phagolysate filtrate obtained using the Staphylococcus aureus bacteriophage strain SCH1 deposited in the collection of the Museum of Microorganisms of the Federal Budgetary Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" Federal State Supervision of Consumer Rights Protection and Human Well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-105, the filtrate of the phagolysate Staphylococcus aureus, obtained using the bacteriophage strain Staphylococcus aureus SCH111, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State Scientific Center for Applied Microbiology and Biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-95, filtrate fa olysate Klebsiella pneumoniae obtained using the bacteriophage strain Klebsiella pneumoniae KPV15 deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer protection and human well-being (FBSI SSC PMB Rospotrebnad) Ph-90, Klebsiella pneumoniae phagolysate filtrate obtained using the bacteriophage strain Klebsiella pneumoniae KPV811, deposited in the collection of the Museum of microorganisms F. State Research Center for Applied Microbiology and Biotechnology, Federal Budgetary Institution of Science, Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-91, Pseudomonas aeruginosa phagolysate filtrate obtained using the Pseudomonas aeruginosa bacteriophage strain deposited in the collection of the Museum of Microorganisms of the Federal Budgetary Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-88, the filtrate of the phagolysate Pseudomonas aeruginosa, obtained using the bacteriophage strain Pseudomonas aeruginosa RA10, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science “State scientific and research center and Biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-89, Acinetobac phagolysate filtrate ter baumannii, obtained using the bacteriophage strain Acinetobacter baumannii AM24, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) Ph-106 and / or Acinetobacter baumannii phagolysate filtrate obtained using the Acinetobacter baumannii AP22 bacteriophage strain deposited in the collection of the Fede Museum of Microorganisms cial budgetary institution of science "State Scientific Center of Applied Microbiology and Biotechnology" Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor FBUN SRCAMB) under the Ph-42 number. The composition may include targeted additives in an amount of 0.01 ÷ 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected depending on the dosage form used. This composition due to the sufficient spectrum of stable lytic activity can be used for the prevention and treatment of hospital infections and is presented in the form of various dosage forms: suspension, ointment, suppository, powder, tablets, capsules, gel, spray, etc.

Two variants of the antibacterial composition are claimed using 7 or 8 bacteriophages as the active principle of the filtrates of the phagolysates.

The selection of 8 strains of virulent bacteriophages was carried out in the course of work with 40 strains of Staphylococcus aureus, 46 strains of Pseudomonas aeruginosa), 40 strains of Klebsiella pneumoniae and 80 strains of Acinetobacter baumannii.

Bacterial culture media and conditions

Bacterial cultures were grown on solid and liquid nutrient media: GRM-broth - based on the drug “nutrient broth for the cultivation of dry microorganisms (GRM-broth)” (FBUN SSC PMB, Obolensk), GRM-agar based on the preparation “nutrient agar for the cultivation of microorganisms dry (timing-agar) ”(FGUN SSC PMB, Obolensk).

Assessment of the lytic activity of bacteriophages.

The spectrum of lytic activity of bacteriophages was evaluated in several ways:

a) Micromethod. Serial ten-fold dilutions of the phagolysates were prepared in SM buffer in a 96-well plate. Aliquots of the dilutions were transferred with a replicator to the lawns of bacterial strains applied to the surface of nutrient agar in semi-liquid (0.6%) agarose. The results were taken into account after incubation of the dishes at 37 ° C. overnight. Strains in which negative zones were observed in several dilutions were considered sensitive. Strains in which lysis zones were observed upon application of the native phage preparation (not diluted) were classified as conditionally sensitive. Strains with no lysis zones were considered resistant. In this way, the lytic activity of several bacteriophages was evaluated simultaneously on one lawn.

b) 10 μl of successive ten-fold dilutions of the phagolysate was applied to the lawn of the bacterial strain in semi-liquid (0.6%) agarose. The results were taken into account after incubation of the dishes at 37 ° C. overnight. The interpretation of the results is similar to that indicated in the first method. This method allows you to evaluate not only the spectrum of lytic activity of the phage, but also to calculate the number of plaque forming units per unit volume of the phagolysate.

Strains of virulent bacteriophages used to obtain filtrates of phagolysates

1. The strain of the bacteriophage Staphylococcus aureus SCH1 was isolated from clinical samples and dressings of hospitals in Moscow and Chelyabinsk using a bacterial culture of the strain Staphylococcus aureus 2004 and deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for supervision in the field of supervision consumer protection and human welfare (FBUN SSC PMB Rospotrebnadzor) under the number Ph-105.

The strain of the bacteriophage Staphylococcus aureus SCH1 is characterized by the following properties: on the lawn of a sensitive strain, phages form small negative transparent colonies with a diameter of 1-2 mm with smooth edges without a halo.

The nucleotide sequence of the genome of the strain is presented in the graphic part.

2. The strain of the bacteriophage Staphylococcus aureus SCH111 was isolated from clinical samples and dressings of hospitals in Moscow and Chelyabinsk using a bacterial culture of the strain Staphylococcus aureus 2004 and deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal surveillance service consumer protection and human welfare (FBUN SSC PMB Rospotrebnadzor) under the number Ph-95.

The strain of the bacteriophage Staphylococcus aureus SCH111 is characterized by the following properties: on the lawn of a sensitive strain, phages form small negative transparent colonies with a diameter of 1-2 mm with smooth edges without a halo.

The nucleotide sequence of the genome of the strain is presented in the graphic part.

The assessment of the spectrum of lytic properties of phages was determined on 40 indicator clinical strains of S. aureus using the spot test method. The accumulated phagolysates were sterilized with chloroform and applied to freshly planted lawns of S. aureus. The sensitivity of S. aureus strains to phages was judged by the appearance of a sterile spot on the lawn of the indicator strain grown after 16-18 hours. The results of studying the spectrum of the lytic action of staphylococcal bacteriophages are presented in table. one.

Table 1. The spectrum of lytic action of staphylococcal bacteriophages in relation to clinical strains of S. aureusino (results of a spot test).

As follows from the data obtained, the bacteriophage SCH1 possesses the broadest spectrum of lytic action against indicator cultures: it inhibits the growth of 92.5% of staphylococcal strains; bacteriophage bacteriophage SCH111 is less active (72%). It should be noted that on sensitive cultures of S. aureus in the spot test, the bacteriophages SCH1 and SCH111 formed sterile lysis spots when diluted 10,000 times.

Two bacteriophages of S. aureus are resistant to chloroform: their titer in phagolysates sterilized by filtration through a filter with a pore diameter of 0.45 μm did not exceed the titer in similar phagolysates treated with chloroform.

When studying the viability of staphylococcal phages, their sensitivity to relatively low temperatures was revealed: heating bacteriophage preparations to a temperature above 56 ° C completely inactivates them in 15 minutes. The optimal temperature for maintaining the initial phage titer for several months is from 4 to 10 ° C.

3. The strain of the bacteriophage Klebsiella pneumoniae KPV15 was isolated from samples of Moscow wastewater on a bacterial culture of the strain Klebsiella pneumoniae B-716 and deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-90.

The strain of the bacteriophage Klebsiella pneumoniae KPV15 is characterized by the following properties: it forms small opaque negative colonies with a diameter of 1-2 mm with a weak halo (1-2 mm) or without it.

The nucleotide sequence of the genome of the strain is presented in the graphic part.

4. The strain of the bacteriophage Klebsiella pneumoniae KPV811 was isolated from Moscow wastewater samples from a bacterial culture of the strain Klebsiella pneumoniae B-811 and deposited in the collection of the Museum of microorganisms of the Federal State Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for the Supervision of Consumer Protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-91.

The strain of the bacteriophage Klebsiella pneumoniae KPV811 is characterized by the following properties: it forms small opaque negative colonies with a diameter of 1-2 mm with a weak halo (1-2 mm) or without it.

The nucleotide sequence of the genome of the strain is presented in the graphic part.

Both phages are resistant to chloroform: their titers in phagolysates sterilized by filtration through a filter with a pore diameter of 0.45 μm did not exceed the titer in similar phagolysates sterilized with chloroform. When the bacteriophage preparations KPV15 and KPV811 were heated to a temperature above 60 ° C for 30 minutes, they were completely inactivated. The optimum temperature to maintain the initial phage titer for four months (observation period) is from 4 to 10 ° C.

The lytic spectrum of bacteriophages KPV15 and KPV811 in relation to strains of K. pneumoniae isolated in clinical institutions in the last 5 years are presented in table. 2.

As follows from the data in Table 2, the bacteriophage KPV15 actively multiplies and lyses 14 of the 40 clinical indicator strains of K. pneumoniae: cultures of K. pneumonia sensitive to it are lysed when the phagolysate is diluted to a titer of 1 × 10 ^-7 - 1 × 10 ^-8 . On the lawn of sensitive crops, the KPV15 phage forms clear plaques. In addition, the zero dilution phagolysate of the bacteriophage KPV15 lyses another 20 indicator K. pneumoniae strains, forming cloudy spots on their lawn. Thus, out of 40 indicator strains, the bacteriophage KPV15 with different degrees of activity lyses 34 strains (85%).

The bacteriophage KPV811 actively multiplies (plaque formation occurs when the phagolysate is diluted to 10-7-10-8) and lyses 7 out of 40 indicator cultures of K. pneumoniae, and the bacteriophage is replicated on those strains on which the bacteriophage KPV15 is not replicated, i.e. two bacteriophages, KPV15 and KPV811, are lysed with varying degrees of activity of 92.5% of the clinical strains of K. pneumoniae. It should be noted that the set of indicator strains includes cultures of K. pneumoniae, isolated mainly in 2012-2014 in large clinical centers in Moscow.

5. The strain of the bacteriophage Pseudomonas aeruginosa RA5 was isolated from wastewater from Moscow and Serpukhov in a bacterial culture of the strain Pseudomonas aeruginosa B-1304 and deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of the protection of rights consumer and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-88.

The strain of the bacteriophage Pseudomonas aeruginosa PA5 is characterized by the following properties: The bacteriophage PA5 on the lawn of the strain P. aeruginosa B1304 forms transparent negative colonies with a diameter of 1-2 mm.

The nucleotide sequence of the genome of the strain is presented in the graphic part.

6. The strain of the bacteriophage Pseudomonas aeruginosa PA10 was isolated from wastewater from Moscow and Serpukhov in a bacterial culture of the strain Pseudomonas aeruginosa 176 and deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-89.

The strain of the bacteriophage Pseudomonas aeruginosa PA10 is characterized by the following properties: the bacteriophage PA10 on the host strain P. aeruginosa 176 forms colonies with a diameter of 4-5 mm with a transparent center and a halo at the periphery.

The nucleotide sequence of the genome of the strain is presented in the graphic part.

Both bacteriophages are resistant to chloroform and sensitive to heat: at a temperature of 55 ° C they are inactivated for 15 minutes. They are well preserved in freeze-dried state at a temperature of 4-8 ° C. The maximum titer of bacteriophages PA5 and PA10 when they multiply in liquid and solid nutrient media is about 10 ¹⁰ phage particles in 1 ml of phagolysate.

The spectrum of lytic activity of pseudomonas bacteriophages in relation to clinical strains of P. aeruginosa is presented in table. 3.

Table 3. The spectrum of lytic activity of pseudomonas phages in relation to clinical P. aeruginosa strains isolated in Serpukhov, Chelyabinsk, Vologda and Moscow.

As follows from the table, the isolated bacteriophages have a relatively narrow spectrum of lytic activity: for phages PA5, PA10 it is 27, 29%, respectively. However, a mixture of these phages causes the lysis of 52% of indicator P. aeruginosa strains.

7. The strain of the bacteriophage Acinetobacter baumannii AP22 was isolated from samples of clinical material from Moscow hospitals using a bacterial culture of the strain Acinetobacter baumannii 1053 and deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of the protection of rights consumer and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-42. The strain is protected by RF patent No. 2439151.

The strain of the bacteriophage Acinetobacter baumannii AP22 is characterized by the following properties: The bacteriophage AP22 on the lawn of the sensitive strain Acinetobacter baumannii 1053 forms round transparent negative colonies with a diameter of about 2-3 mm with smooth edges surrounded by an opaque halo.

8. The strain of the bacteriophage Acinetobacter baumannii AM24 was isolated from samples of clinical material from Moscow hospitals using a bacterial culture of the Acinetobacter baumannii B-05 strain and deposited in the collection of the Museum of microorganisms of the Federal State Institution of Science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for Surveillance in the Field of consumer protection and human welfare (FBUN SSC PMB Rospotrebnadzor) under the number Ph-106.

The strain of the bacteriophage Acinetobacter baumannii AM24 is characterized by the following properties: The bacteriophage AM24 on the lawn of the sensitive strain B-05 forms round transparent negative colonies with smooth edges with a diameter of about 1 mm, surrounded by an opaque halo. The diameter of the halo is from 2 to 6 mm.

The nucleotide sequence of the genome of the strain is presented in the graphic part.

Both bacteriophages are resistant to chloroform and sensitive to heat: at a temperature of 55 ° C they are inactivated for 15 minutes. They are well preserved in freeze-dried state at a temperature of 4-8 ° C. The maximum titer of the AP22 bacteriophage when it is multiplied in liquid and solid nutrient media is 5 × 10 ¹⁰ PFU / ml, the titer of the bacteriophage AM24 does not exceed 7 × 10 ⁹ PFU / ml. The spectrum of lytic activity of acinetobacter bacteriophages against 80 clinical strains of Acinetobacter baumannii is presented in table. four.

The claimed invention is based on a new combination of original distinctive features that provides a solution to the problem: for the first time, it is proposed to use two variants of compositions of seven or eight bacteriophage strains that complement the spectrum of lytic activity in relation to test strains and isolates of bacteria that cause hospital infections during the first time a selected mixture of targeted additives that stabilize the activity of bacteriophages when used in various dosage forms.

The invention is illustrated by the following examples.

Example 1. Obtaining individual phagolysates and their combinations

The bacterial culture of the host strain in a titer of 10 ⁸ -10 ⁹ CFU / ml is sown in a vessel for cultivation - a glass microbiological mattress on a beveled dense nutrient medium with a layer thickness of 10 mm to 25 mm, cultivated for 3-3.5 hours at optimal temperature for the growth of the culture of the host strain, then the uterine bacteriophage in the titer of 10 ⁵ -10 ⁶ PFU / ml is sown on the obtained grass lawn of the culture of the host strain, the culture vessel is sealed, cultured for 13-15 hours at the optimum temperature for growth of the strain culture ba of the cteriophage and the thickness of the air layer above the surface of the dense nutrient medium from 25 mm to 40 mm, a phagolysate is obtained by suspending the bacteriophage from the surface of the dense nutrient medium with saline or buffer solution with a pH of 7.0-7.2 in an amount of 0.04-0.045 ml per 1 cm ^{2 of the} surface of a dense nutrient medium, the phagolysate is suctioned into a sterile container, chloroform is added, it is kept for 30-45 minutes with continuous shuttling, centrifuged for 30-45 minutes at 5000-6000 rpm, the supernatant is sterilized by filtration Through a filter with a pore diameter of 0.2-0.22 microns and pass the obtained filtrate through a column containing an agent affinity for endotoxin. After centrifugation, supernatants containing bacteriophages (7 or 8) are mixed, then the mixture of supernatants is sterilized by filtration through a filter with a pore diameter of 0.2-0.22 microns and the filtrate obtained is passed through a column containing an endotoxin affinity agent.

Example 2. The obtained antibacterial composition for the prevention or treatment of hospital infections (option 1) with lytic activity not lower than 10-4 according to Appelman in relation to test strains and bacterial isolates isolated from the human body, including the staphylococcus aureus phagolysate filtrate obtained using a bacteriophage strain Staphylococcus aureus SCH1, deposited in the collection of the Museum of Microorganisms of the Federal Budgetary Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for to the Consumer Protection and Human Welfare Supervision Act (FBSI SSC PMB Rospotrebnadzor) under the number Ph-105, the staphylococcus aureus phagolysate filtrate obtained using the bacteriophage strain Staphylococcus aureus SCH111, deposited in the collection of the Museum of Microorganisms of the Federal Budget Institution of Science “The State Scientific and Scientific Research Center and Biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-95, filtrate of the phagolysate Klebsiella pne umoniae obtained using the bacteriophage strain Klebsiella pneumoniae KPV15, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph -90, Klebsiella pneumoniae phagolysate filtrate obtained using the Klebsiella pneumoniae KPV811 bacteriophage strain deposited in the collection of the Museum of Microorganisms of the Federal Budget State Research Center for Applied Microbiology and Biotechnology of the Federal Service for Supervision of Consumer Rights Protection and Human Wellbeing (Federal State Institution of Science and Medicine of the Russian Federal Public Health Service) under the number Ph-91, the filtrate of the phagolysate Pseudomonas aeruginosa obtained using the bacteriophage strain Pseudomonas aeruginosa RA deposited in the collection of the Museum of Microorganisms of the Federal Budgetary Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for Supervision in the Field of protection of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-88, the filtrate of the phagolysate Pseudomonas aeruginosa obtained using the bacteriophage strain Pseudomonas aeruginosa PA10, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center of applied microbiology" The Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-89, and the filtrate of the phagolysate Acinetobacter baumannii, obtained identified using a bacteriophage strain Acinetobacter baumannii AM24 deposited in the collection of the Museum of microorganisms of the Federal budget institution of science “State scientific center for applied microbiology and biotechnology” of the Federal service for the supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-106 . The composition includes target additives in an amount of 0.01 ÷ 99.99 wt. % by weight of the composition as a mixture of ingredients.

The stability studies of the polyvalent activity of individual strains of bacteriophages included in the composition of the claimed composition in the samples of the composition stored for a year, and testing of all bacteriophages of the composition for lysogenesis were carried out. The lytic activity of all strains of bacteriophage composition was not less than 10-4 according to Appelman. Tests for lysogenesis were negative. To test the bioavailability of the obtained composition, its sample weighing 0.6 g was introduced into medium 199. It was noted that the preparation was uniformly dispersed in an aqueous medium and that the solution retained at least 99% of the lytic activity of the composition.

Example 3. The obtained antibacterial composition for the prevention or treatment of hospital infections (option 2) with lytic activity not lower than 10-4 according to Appelman in relation to test strains and bacterial isolates isolated from the human body, including the staphylococcus aureus phagolysate filtrate obtained using a bacteriophage strain Staphylococcus aureus SCH1, deposited in the collection of the Museum of Microorganisms of the Federal Budgetary Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for to the Consumer Protection and Human Welfare Supervision Act (FBSI SSC PMB Rospotrebnadzor) under the number Ph-105, the staphylococcus aureus phagolysate filtrate obtained using the bacteriophage strain Staphylococcus aureus SCH111, deposited in the collection of the Museum of Microorganisms of the Federal Budget Institution of Science “The State Scientific and Scientific Research Center and Biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-95, filtrate of the phagolysate Klebsiella pne umoniae obtained using the bacteriophage strain Klebsiella pneumoniae KPV15, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph -90, Klebsiella pneumoniae phagolysate filtrate obtained using the Klebsiella pneumoniae KPV811 bacteriophage strain deposited in the collection of the Museum of Microorganisms of the Federal Budget State Research Center for Applied Microbiology and Biotechnology of the Federal Service for Supervision of Consumer Rights Protection and Human Wellbeing (Federal State Institution of Science and Medicine of the Russian Federal Public Health Service) under the number Ph-91, the filtrate of the phagolysate Pseudomonas aeruginosa obtained using the bacteriophage strain Pseudomonas aeruginosa RA deposited in the collection of the Museum of Microorganisms of the Federal Budgetary Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for Supervision in the Field of protection of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-88, the filtrate of the phagolysate Pseudomonas aeruginosa obtained using the bacteriophage strain Pseudomonas aeruginosa PA10, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology" Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-89, Acinetobacter baumannii phagolysate filtrate, obtained using the strain of the bacteriophage Acinetobacter baumannii AP22 deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-42 , and the Acinetobacter baumannii phagolysate filtrate obtained using the bacteriophage strain Acinetobacter baumannii AM24, deposited in the collection of the Museum of microorganisms of the Federal budget rezhdeniya science "State Scientific Center of Applied Microbiology and Biotechnology" Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor FBUN SRCAMB) under the Ph-106 number. The composition includes target additives in an amount of 0.01 ÷ 99.99 wt. % by weight of the composition as a mixture of ingredients.

The stability studies of the polyvalent activity of individual strains of bacteriophages included in the composition of the claimed composition in the samples of the composition stored for a year, and testing of all bacteriophages of the composition for lysogenesis were carried out. The lytic activity of all strains of bacteriophage composition was not less than 10-4 according to Appelman. Tests for lysogenesis were negative. To test the bioavailability of the obtained composition, its sample weighing 0.6 g was introduced into medium 199. It was noted that the preparation was uniformly dispersed in an aqueous medium and that the solution retained at least 99% of the lytic activity of the composition.

Example 4. The study of toxicity, prophylactic and therapeutic efficacy of the claimed composition in experiments on laboratory animals

The study of acute toxicity of the claimed antibacterial composition (AK) was performed on 10 experimental and 5 control outbred mice, females, live weight of 18-20 g. Mice of the experimental group were once administered intraperitoneally with 0.5 ml of AK with a concentration of phage particles of 1 × 108 PFU. The control animals were injected in the same way with the buffer used to prepare AK. Animals of both groups were monitored for 7 days. In animals, external signs of intoxication, activity, appetite, coat condition were recorded, and changes in live weight were monitored. 7 days after the administration of AK, peripheral blood was taken from the animals of the experimental and control groups, in which the total leukocyte count was examined.

The criterion for the harmlessness of the drug for mice are: the absence of death, the absence of signs of disease, the absence of weight loss in animals, good appetite and good condition of the coat. The condition of the experimental (treated AK) animals should not differ from that of the control animals. The number of leukocytes in the blood of experimental mice should not differ significantly from the number of leukocytes in the blood of control animals.

After blood sampling, animals of both groups were euthanized, opened and a macroscopic picture of internal organs was described in them. In experimental (treated AK) animals, this picture should not differ from that of the control.

The study of chronic AK toxicity was performed on outbred (outbred) white mice, females (or males) with a live weight of 18-20 g.

Two groups of mice were used in the experiments: experimental and control, 10 mice each. An experimental group of animals daily for 20 days once a day received AKperos in a volume of 0.5 ml with a phage concentration of 1 × 10 ⁸ PFU, the maximum equivalent to a single dose for humans in terms of unit body weight. A group of control animals instead of AK once a day for 20 days received peros in 0.5 ml of buffer solution, which is used in the preparation of AK. The animals of both groups were observed during the entire period of receiving AK (20 days) and in the next 7 days after drug withdrawal. Observations were carried out similar to those described above for groups of animals in which acute toxicity of AK was experienced. On the next and 7th day after the end of the drug administration in animals (males and females), blood was taken from the experimental and control groups for hematological analysis, paying attention to the total number of leukocytes. After blood sampling, animals of both groups were euthanized and material was taken for histological studies (pieces of spleen, liver, mesenteric lymph nodes, lungs, heart, tongue root, brain, thin and thick stomach), on the basis of which a possible damaging effect of AK on the body of mice was judged .

Chronic toxicity of AK was evaluated by observing the behavior and health of animals, by hematological and histological studies (in comparison with similar studies in the control group of animals).

Study of the prophylactic efficacy of AK in Klebsiella infection in mice.

An experiment to evaluate the prophylactic effectiveness of AK was performed on 4 experimental and 2 control groups of outbred white mice (females or males) with a live weight of 18-20 g (10 animals in each group). The animals of the experimental groups were injected with the preparation AK (intraperitoneally, once at a dose of 1 × 10 ⁸ PFU in a volume of 0.5 ml), respectively, 24, 12, 6 and 1 hour before infection with cells of the strain K. pheumoniae 9. Animals of the experimental groups (treated LPP) and one control (group No. 5, not treated with a bacteriophage) were simultaneously infected with a strain of K. pneumoniae 9 subcutaneously at a dose of 1 × 10 ⁴ PFU in a volume of 0.5 ml, which corresponds to approximately 600 LD ₅₀ . The sixth control group (No. 6) was not infected: she was once administered the drug AK (toxicity control) at a maximum dose (1 × 10 ⁸ PFU). Infected experimental animals were monitored for 14 days, recording their behavior, coat condition, and appetite. After 14 days, the animals were euthanized and opened. Seeds were made from blood, spleen, lungs, and liver for contamination by the culture of the infecting strain of K. pneumoniae 9 with the calculation of the CFU of the pathogen in 1 g of the test sample. The effectiveness of prophylactic treatment with AK of infected animals was evaluated by their condition during the entire observation period, based on the percentage of surviving animals treated after infection with AK, as well as by the degree of elimination of the pathogen from parenchymal organs of treated AK animals

Study of the therapeutic efficacy of AK in Klebsiella infection in mice.

The therapeutic efficacy of AK in acute Klebsiella infection in outbred mice was evaluated in 8 groups of animals, 10 animals each: six experimental and two control. Animals of six experimental groups and one control at the same time were infected with a strain of K. pneumoniae 9 subcutaneously at a dose of 1 × 10 ⁴ CFU (600 LD ₅₀ ) in a volume of 0.5 ml, the eighth group of animals was not infected (control of the toxicity of the bacteriophage preparation). The animals of the first four experimental groups, respectively, 1, 6, 12, and 24 hours after infection, were treated with AK at a dose of 1 × 10 ⁸ PFU in a volume of 0.5 ml intraperitoneally once a day for 10 days. The animals of the fifth and sixth groups were treated with the antibiotic ciprofloxacin 12 hours and 24 hours after infection with the strain K. pneumoniae 9, respectively, by administering the drug intragastrically at a dose of 2 mg / animal twice a day (after 12 hours) for five days. Seventh group - control of the effectiveness of infection: animals were infected subcutaneously with 1 × 10 ⁴ CFU of K. pneumoniae 9 cells in a volume of 0.5 ml, without treatment. The eighth group is the control of toxicity of LPP: animals were injected intraperitoneally with a dose of 1 × 10 ⁸ PFU once every ten days. During the course of treatment and within 14 days after the end of therapy, animals were monitored, recording their behavior, condition of the coat and appetite. In the experimental and control groups of animals recorded the death of animals. Fallen animals were opened, parenchymal organs were sown on nutrient media for subsequent assessment of the contamination of the cells of the infecting strain of K. pneumoniae 9 with their cells. The surviving white mice were euthanized, and their organs were examined for the presence of the pathogen - bacteria of the strain K. pneumoniae 9 with a CFU count of 1 g of the test sample. The efficacy of treating acute Klebsiella infection in mice with the use of a bacteriophage preparation and an antibiotic was evaluated by the life expectancy of the treated animals, by the behavior of the experimental animals during the entire observation period compared with the control groups, by the percentage of survivors after infection with the bacteria treated by the bacteriophage, and by the degree of elimination of the pathogen from parenchymal organs of animals treated with phage.

The effectiveness of phage therapy for acute Klebsiella infection in outbred white mice is shown in Fig. 1. However, there were no significant differences in the effectiveness of the two tested antibacterial combinations.

Example 5. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a suspension containing the target additives in an amount of 0.01-95.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: licorice root syrup, apple pectin, sodium phosphate buffer, HEPES buffer, chinosol. The lysogenesis test was negative. To test the bioavailability of the resulting suspension, its sample weighing 0.6 g was added to medium 199. It was noted that the suspension was uniformly dispersed in the aqueous medium and that the solution retained at least 99% of the lytic activity of the composition.

Example 6. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a suppository containing target additives in an amount of 50.0-95.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: glycine, cysteine, histidine, albumin, castor oil, lanolin, confectionery fat, edible fat, spermaceti, petrolatum, glucose, solid fats (SolPro), T-2 emulsifier, paraffin , cocoa butter, water, vitebsol, quinosol, gelatin, glycerin, calcium gluconate, polyethylene oxide 400, polyethylene oxide 1500 and salts capable of forming a phosphate or acetate or tris buffer solution. The lysogenesis test was negative. To test the bioavailability of the obtained suppository, its sample weighing 0.6 g was added to medium 199. It was noted that the suspension was uniformly dispersed in an aqueous medium and that the composition retained at least 99% of the lytic activity.

Example 7. The obtained antibacterial composition according to Example 2 or Example 3 in the form of an ointment containing the target additives in an amount of 50.0-95.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: salts forming a buffer solution, glycine, cysteine, histidine, chinosol, polyglucin, wax, salomas, kombizhir, lanolin, spermaceti, petrolatum, glycerin, ceresin, liquid paraffin, artificial petrolatum , palmitin, hydrogenated fat, confectionery fat, palm olein, palm stearin, polyethylene oxide 1500, polyethylene oxide 400, cocoa butter, castor oil, peach oil, linseed oil, hemp oil, propylene glycol, hexylene glycol. The lysogenesis test was negative.

Example 8. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a cosmetic gel containing target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: carbapol ETD, calendula extract, lavender essential oil, nipazole (propyl paraben), NaOH, RD brand, purified water.

The drug is indicated for the prevention or treatment of purulent-inflammatory lesions of the skin and subcutaneous tissue arising from various injuries of the skin: burns, wounds, abrasions, etc. The gel is recommended as an effective cosmetic product for skin care after laser cosmetic procedures, as well as a prophylactic for various violations of the integrity of the skin. The drug is used in combination with other antibacterial agents, and also as monotherapy for patients who are intolerant of antibiotic therapy and for antibiotic resistance of the pathogen strains. The drug is used 1-2 times a day in the form of applications on a bandage or on an open surface. For prevention, the drug is applied to clean skin for 15-20 minutes.

Example 9. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a gel containing target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: povidone, PEG 40 hydrogenated castor oil, laureth-9, sodium hydroxide, sodium benzoate, potassium sorbate, methyl paraben, water, magnesium lactate, D-panthenol, benzocaine, menthol, propylene glycol , ASD fraction 2.

Appointment: for use as an auxiliary, additional tool in the complex therapy of burns of any origin (temperature, radiation, chemical). The complex of bacteriophages provides protection (in outpatient and clinical use) from infection of the inflamed area with the main pathogens of purulent bacterial infections, as well as the sanitation of an infected wound. Helps reduce the manifestations of acidosis, sensitization and toxicity of damaged tissues, stabilizes blood microcirculation. It has an anti-inflammatory and mild analgesic effect.

Method of application: at the initial stages (necrosis, acute inflammation), apply to wound dressings until they are moistened and soaked 3-4 times a day, as well as a field for changing dressings. In subsequent stages (scarring), apply directly to the affected area at least 3 times a day. Duration of the course according to indications.

Example 10. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a spray for the oral cavity containing the target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: povidone, PEG 40 hydrogenated castor oil, sodium hydroxide, sodium benzoate, potassium sorbate, water, fructooligosaccharide, lactulose, potassium lactate, D-panthenol, flavor. Appointment: for hygienic, prophylactic and improving care of the oral cavity, predisposed to various manifestations of dysbiosis - unpleasant odor and / or taste, gingivitis, periodontitis, stomatitis, caries. To reduce the risk of various inflammations and infectious complications after treatment and other intensive procedures in the oral cavity.

Method of application: irrigate the oral cavity, directing the spray to problem areas. Do not rinse with water or drink for 30 minutes. Use 3-5 times as needed, but at least 3 times a day. The duration of the course is from 2 to 5 weeks.

Example 11. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a gel for applications containing target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: hypromellose, PEG 40 hydrogenated castor oil, sodium benzoate, potassium sorbate, phosphate buffer, water, perfume, arginine, menthol, propylene glycol. It is applied to a bandage consisting of a textile base (polyamide mesh) and is used for local treatment of wounds, burns and skin diseases.

Example 12. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a spray for disinfection of indoor air and surfaces, containing target additives in the amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: carbomer, triethanolamine, benzyl alcohol, methylchloroisothiazolinone, methylisothiazolinone, PEG 12 dimethicone, chinosole, perfume water.

Appointment: for hygienic, sanitary and disinfection treatment of air and surfaces in residential, domestic and industrial premises, in places of animals, in medical and veterinary institutions, including for sanitizing antibacterial treatment of air, indoor surfaces from pathogens of nosocomial infections and instruments.

Method of application: by aerosol spraying, spray the product in the room air at the rate of 0.3-0.5 cm ^{3 of the} product per m ^{3 of the} air space of the room. To apply the product on surfaces with a high probability of seeding with bacterial pathogenic microflora and on surfaces that require special purity and additional protection against contamination (at the rate of 0.2-0.3 cm ³ per m ^{2 of the} treated surface).

Example 13. Obtaining lyophilized dried filtrates of bacteriophage phagolysates used to prepare the composition according to Example 2 or 3 in the form of lyophilized powder, tablets and capsules.

Protective media and the freeze-drying regime of phages and bacteria on freeze-drying EPSILON 2-60 (CHRIST, Germany) were tested.

In a protective medium for bacterial cultures (sucrose 10%, gelatin 1%), a suspension of bacteria was prepared with a concentration of 1 × 10 ⁹ m.k. / ml. A protective medium for bacteriophages (sucrose 20%, gelatin 2%) was added to the phagolysate in an equal volume. The prepared suspensions of phages and host bacterial cultures were poured into 0.5 ml vials and dried for 24 hours at a temperature of -85 ° C.

As a result of the work carried out, preparations of eight bacteriophages and ten bacterial strains were lyophilized dried and characterized (by concentration).

Example 14. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a lyophilized powder containing target additives in an amount of 30.0-94.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: aerosil, sucrose, lactose, glucose, maltose, beckoning, fructose, ascorbic acid, citric acid, succinic acid, glycine, thiourea, polyglucin, pectin, dextrin, albumin, gelatin, buffer solution salts, calcium stearate, titanium dioxide, lactulose, polyvinylpyrrolidone, microcrystalline cellulose, starch, talc, cysteine, histidine, chinosol. The lysogenesis test was negative.

Example 15. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a tablet with enteric coating containing target additives in an amount of 50.0-94.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: aerosil, sucrose, lactose, glucose, maltose, beckoning, fructose, ascorbic acid, citric acid, succinic acid, glycine, thiourea, polyglucin, pectin, dextrin, albumin, gelatin, buffer salts, calcium stearate, titanium dioxide, lactulose, polyvinylpyrrolidone, microcrystalline cellulose, starch, talc, cellulose, methyl cellulose, acetate phthalate cellulose, sodium carboxymethyl cellulose, croscarmellose sodium, sodium glycolate in, histidine, cysteine, quinosol. The lysogenesis test was negative.

Example 16. The obtained antibacterial composition according to Example 2 or Example 3 in the form of a capsule containing target additives in an amount of 30.0-94.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: aerosil, sucrose, lactose, glucose, maltose, beckoning, fructose, ascorbic acid, citric acid, succinic acid, glycine, thiourea, polyglucin, pectin, dextrin, albumin, gelatin, buffer solution salts, calcium stearate, titanium dioxide, lactulose, polyvinylpyrrolidone, microcrystalline cellulose, starch, talc, cellulose, methyl cellulose. cellulose acetate phthalate, crospovidone, chinosol.

Example 17. In a patient hospitalized in a neurosurgical clinic with a traumatic brain injury, after successfully performing an operation in the intensive care unit and intensive care unit (ICU), Klebsiellapneumoniae and Pseudomonasaeruginosa с identified as strains were isolated on the 3rd day after connecting to the ventilator for bacteriological analysis of endotracheal aspirate multiple antibiotic resistance. The aerosol use of a spray containing an antibacterial composition based on a cocktail of bacteriophages twice a day for 2 days made it possible to confirm the eradication of these microorganisms in an endotracheal aspirate with repeated bacteriological analysis.

Example 18. In a patient hospitalized in a neurosurgical clinic with hemorrhagic stroke, after a successful operation in the intensive care unit (ICU) on the 5th day after connecting to the ventilator for bacteriological analysis of endotracheal aspirate, and Klebsiella pneumoniae were isolated after two days and urine and Acinetobacter baumannii, identified as strains with multiple antibiotic resistance. The use of a suspension containing an antibacterial composition based on a cocktail of bacteriophages twice a day for 3 days for rinsing the oral cavity and treating the endotracheal tube made it possible to confirm the eradication of these microorganisms during repeated bacteriological analysis of endotracheal aspirate and urine.

Example 19. An outbreak of nosocomial infection caused by a pan-resistant strain of Klebsiella pneumoniae occurred in a surgical hospital in ICU. Before the operation, enteric capsules containing lyophilized substance on the basis of a cocktail of bacteriophages were prescribed prophylactically for 2 days twice a day twice a day before surgery to the unit # 1 of ICU. Patients who entered the ICU unit No. 2 after surgery did not receive a bacteriophage preparation. As a result of a comparative study for 30 days, only 5% of patients from block No. 1 during bacteriological analysis of urine and endotracheal aspirate were seeded with a pan-resistant strain of Klebsiella pneumoniae, while in block No. 2 of ICU the frequency of nosocomial infection exceeded 50%.

Example 20. In a patient hospitalized with burns of 18% of the body surface, on day 10 with bacterial monitoring of wound surfaces from the wound discharge MRSA strains were seeded. The use of an ointment based on a cocktail of bacteriophages for 14 days allowed the eradication of these microorganisms from the wound discharge of the patient.

Thus, the claimed invention allows to expand the range of products containing highly selective natural antibacterial components (bacteriophages) as the main active principle, to ensure biological stability and activity of bacteriophages in the composition of the product. In addition, the composition has a low risk of toxic and side effects, allows you to expand options and methods for the practical use of agents containing bacteriophages, provides storage stability and efficiency in a wide temperature range. A list of nucleotide sequences according to the WIPO standard is presented for seven strains of bacteriophages.

It was found that the nucleotide sequence of each individual strain encoding the bacteriophage genome is 90-100% identical to the corresponding sequence presented in the list of nucleotide sequences, namely identical: SEQ ID NO: 1 by 93%, 96%, 99% or one hundred%; SEQ ID NO: 2 93%, 96%, 99% or 100%; SEQ ID NO: 3 at 90%, 93%, 96%, 99% or 100%; SEQ ID NO: 4 at 90%, 93%, 96%, 99% or 100%; SEQ ID NO: 5 at 99% or 100%; SEQ ID NO: 6 100%; SEQ ID NO: 7 at 90%, 93%, 96%, 99% or 100%. The above nucleotide sequences can be transformed into a sensitive bacterial cell, followed by bacteriophage. The term "transformation" means the process of absorption by a cell of a free DNA molecule from the environment. According to the prior art, a method is known for transforming bacteriophage DNA into a sensitive bacterial cell with subsequent development of a productive phage infection (L. Planelles, C. Maranon, JM Requena, M.S. Lopez, Phage recovery by electroporation of naked DNA into host cells avoids the use of packaging extracts. Analytical biochemistry, 1999, Vol. 267, No. 1, p. 234-235).

Example 21. A DNA molecule representing a nucleotide sequence identical to SEQ ID NO: 1 by 93% was transformed into a suspension of electrocompetent cells of S. aureus strain by electroporation. The culture of S. aureus strain after transformation was incubated in meat and peptone broth at 37 ° C for 18 hours, then filtered through a filter with a pore diameter of 0.22 μm. The obtained filtrate contained a staphylococcal bacteriophage in a titer of 5 × 105 PFU / ml.

Example 22. A DNA molecule representing a nucleotide sequence identical to SEQ ID NO: 3 by 90% was transformed into a suspension of chemically competent cells of the strain K. pneumoniae by a chemical method in the presence of calcium chloride. The culture of the strain K. pneumoniae after transformation was incubated in meat and peptone broth at 37 ° C for 12 hours, then filtered through a filter with a pore diameter of 0.45 μm. The obtained filtrate contained Klebsiella bacteriophage in a titer of 2 × 104 PFU / ml.

Example 23. A DNA molecule representing a nucleotide sequence identical to SEQ ID NO: 5 by 99% was transformed into a suspension of chemically competent cells of P. aeruginosa strain by a chemical method in the presence of calcium chloride. The culture of P. aeruginosa strain after transformation was incubated in meat and peptone broth at 37 ° C for 10 hours, then filtered through a filter with a pore diameter of 0.45 μm. The obtained filtrate contained a pseudomonas bacteriophage in a titer of 3 × 105 PFU / ml.

Example 24. A DNA molecule representing a nucleotide sequence identical to SEQ ID NO: 7 by 100% was transformed into a suspension of electrocompetent cells of A. baumanii strain by electroporation. The culture of A. baumanii strain after transformation was incubated in meat and peptone broth at 37 ° C for 18 hours, then filtered through a filter with a pore diameter of 0.45 μm. The obtained filtrate contained the bacteriophage A. baumanii in a titer of 1 × 104 PFU / ml.

Claims (28)

1. An antibacterial composition for the prevention or treatment of hospital infections, including a combination of bacterial phagolysate filtrates, characterized in that it has an Appelman lytic activity of at least 10-4 for test strains and bacterial isolates isolated from the clinical material and includes a Staphylococcus aureus phagolysate filtrate obtained using a strain of the bacteriophage Staphylococcus aureus SCH1 deposited in the collection of the museum of microorganisms of the Federal budget institution of science "State scientific center of microbiology and biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-105, the filtrate of the phagolysate Staphylococcus aureus, obtained using the bacteriophage strain Staphylococcus aureus SCH111, deposited in the museum’s collection of the microorganism museum institutions of science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBUN SSC PMB of Rospotrebnadzor) under the number Ph-95, the Klebsiella pneumoniae phagolysate filtrate obtained using the bacteriophage strain Klebsiella pneumoniae KPV15, deposited in the collection of the Museum of microorganisms of the Federal State Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for the Supervision of Consumer Rights Protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-90, the filtrate of the phagolysate Klebsiella pneumoniae obtained using the bacteriophage strain Klebsiella pneumoniae KPV811, de The State Scientific Center for Applied Microbiology and Biotechnology, Federal State Institution of Science, collection of the Museum of Microorganisms, Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-91, Pseudomonas aeruginosa phagolysate filtrate obtained using strain of bacteriophage Pseudomonas aeruginosa RA5 deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center of applied micro of biology and biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-88, the filtrate of the phagolysate Pseudomonas aeruginosa obtained using the bacteriophage strain Pseudomonas aeruginosa PA10, deposited in the collection of the Museum of microorganisms of the Federal microorganisms Science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBUN SSC PMB Rospotre supervision) under the number Ph-89, the Acinetobacter baumannii phagolysate filtrate obtained using the Acinetobacter baumannii AM24 bacteriophage strain, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for the Protection of Consumer Rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-106, and target additives in an amount of 0.01 ± 99.99 wt. % by weight of the composition. 2. An antibacterial composition for the prevention or treatment of hospital infections, comprising a combination of bacterial phagolysate filtrates, characterized in that it has an Appelman lytic activity of at least 10-4 for test strains and bacterial isolates isolated from the clinical material and includes a Staphylococcus aureus phagolysate filtrate obtained using a strain of the bacteriophage Staphylococcus aureus SCH1 deposited in the collection of the museum of microorganisms of the Federal budget institution of science "State scientific center of microbiology and biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-105, the filtrate of the phagolysate Staphylococcus aureus, obtained using the bacteriophage strain Staphylococcus aureus SCH111, deposited in the museum’s collection of the microorganism museum institutions of science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBUN SSC PMB of Rospotrebnadzor) under the number Ph-95, the Klebsiella pneumoniae phagolysate filtrate obtained using the bacteriophage strain Klebsiella pneumoniae KPV15, deposited in the collection of the Museum of microorganisms of the Federal State Institution of Science "State Scientific Center for Applied Microbiology and Biotechnology" of the Federal Service for the Supervision of Consumer Rights Protection and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-90, the filtrate of the phagolysate Klebsiella pneumoniae obtained using the bacteriophage strain Klebsiella pneumoniae KPV811, de The State Scientific Center for Applied Microbiology and Biotechnology, Federal State Institution of Science, collected in the collection of the Museum of Microorganisms of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-91, the Pseudomonas aeruginosa phagolysate filtrate obtained using strain of bacteriophage Pseudomonas aeruginosa RA5, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center of applied mik of biology and biotechnology »of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-88, the filtrate of the phagolysate Pseudomonas aeruginosa obtained using the bacteriophage strain Pseudomonas aeruginosa PA10, deposited in the collection of the Museum of microorganisms of the Federal microorganisms Science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for Supervision of Consumer Rights Protection and Human Well-being ) under the number Ph-89, the Acinetobacter baumannii phagolysate filtrate obtained using the Acinetobacter baumannii AM24 bacteriophage strain deposited in the collection of the Museum of microorganisms of the Federal budget institution of science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for the Protection of Consumer Rights and human well-being (FBSI SSC PMB Rospotrebnadzor) under the number Ph-106, the filtrate of the Acinetobacter baumannii phagolysate obtained using the Acinetobacter baumannii AP22 bacteriophage strain, deposited in the collection of the Museum of Microorganisms of the Federal Budgetary Institution of Science “State Scientific Center for Applied Microbiology and Biotechnology” of the Federal Service for Supervision of Consumer Rights Protection and Human Well-Being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-42, and target additives in the amount of 0.01 ± 99.99 wt. % by weight of the composition. 3. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 0.01-95.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: licorice root syrup, apple pectin, sodium phosphate buffer, HEPES buffer, chinosol, and is presented as a suspension. 4. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 50.0-95.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: glycine, cysteine, histidine, albumin, castor oil, lanolin, confectionery fat, edible fat, spermaceti, petrolatum, glucose, solid fats (SolPro), T-2 emulsifier, paraffin , cocoa butter, water, vitebsol, quinosol, gelatin, glycerin, calcium gluconate, polyethylene oxide 400, polyethylene oxide 1500 and salts capable of forming a phosphate, or acetate, or tris-buffer solution, and is presented in the form of a suppository. 5. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 50.0-95.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: salts forming a buffer solution, glycine, cysteine, histidine, chinosol, polyglucin, wax, salomas, kombizhir, lanolin, spermaceti, petrolatum, glycerin, ceresin, liquid paraffin, artificial petrolatum , palmitin, hydrogenated fat, confectionery fat, palm olein, palm stearin, polyethylene oxide 1500, polyethylene oxide 400, cocoa butter, castor oil, peach oil, linseed oil, hemp oil, propylene glycol, hexylene glycol, and is presented as an ointment. 6. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: carbapol ETD, calendula extract, lavender essential oil, nipazole (propylparaben), NaOH, RD brand, purified water, and presented as a cosmetic gel. 7. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: povidone, PEG 40 hydrogenated castor oil, laureth-9, sodium hydroxide, sodium benzoate, potassium sorbate, methyl paraben, water, magnesium lactate, D-panthenol, benzocaine, menthol, propylene glycol , ASD fraction 2, and is presented as a gel for burns. 8. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: povidone, PEG 40 hydrogenated castor oil, sodium hydroxide, sodium benzoate, potassium sorbate, water, fructooligosaccharide, lactulose, potassium lactate, D-panthenol, flavor, and is presented as a spray for the oral cavity. 9. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: hypromellose, PEG 40 hydrogenated castor oil, sodium benzoate, potassium sorbate, phosphate buffer, water, fragrance, arginine, menthol, propylene glycol, and is presented as a gel for dressings. 10. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it contains target additives in an amount of 99.99 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: carbomer, triethanolamine, benzyl alcohol, methylchloroisothiazolinone, methylisothiazolinone, PEG 12 dimethicone, chinosole, water, fragrance, and is presented as a disinfectant. 11. The antibacterial composition for the prevention or treatment of hospital infections according to claim 1 or 2, characterized in that it comprises a combination of bacterial phagolysates filtrates in the form of dry biomass. 12. The antibacterial composition for the prevention or treatment of hospital infections according to claim 11, characterized in that it contains target additives in an amount of 30.0-94.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: aerosil, sucrose, lactose, glucose, maltose, beckoning, fructose, ascorbic acid, citric acid, succinic acid, glycine, thiourea, polyglucin, pectin, dextrin, albumin, gelatin, the salts forming the buffer solution, calcium stearate, titanium dioxide, lactulose, polyvinylpyrrolidone, microcrystalline cellulose, starch, talc, cysteine, histidine, chinosol and are presented in the form of lyophilized powder. 13. The antibacterial composition for the prevention or treatment of hospital infections according to claim 11, characterized in that it contains target additives in an amount of 50.0-94.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: aerosil, sucrose, lactose, glucose, maltose, beckoning, fructose, ascorbic acid, citric acid, succinic acid, glycine, thiourea, polyglucin, pectin, dextrin, albumin, gelatin, buffer salt, calcium stearate, titanium dioxide, lactulose, polyvinylpyrrolidone, microcrystalline cellulose, starch, talc, cellulose, methyl cellulose, acetate phthalate cellulose, sodium carboxymethyl cellulose, croscarmellose sodium, sodium glycolate in, histidine, cysteine, hinozol, and is presented in a tablet with an enteric coating. 14. The antibacterial composition for the prevention or treatment of hospital infections according to claim 11, characterized in that it contains target additives in an amount of 30.0-94.0 wt. % by weight of the composition in the form of a mixture of ingredients selected from the series: aerosil, sucrose, lactose, glucose, maltose, beckoning, fructose, ascorbic acid, citric acid, succinic acid, glycine, thiourea, polyglucin, pectin, dextrin, albumin, gelatin, salts of the buffer solution, calcium stearate, titanium dioxide, lactulose, polyvinylpyrrolidone, microcrystalline cellulose, starch, talc, cellulose, methyl cellulose, cellulose acetate, crospovidone, chinosol, and are presented in capsule form. 15. The strain of the bacteriophage Staphylococcus aureus SCH1, used to obtain the antibacterial composition according to claim 1 or 2, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-105. 16. The nucleic acid molecule corresponding to the bacteriophage genome according to claim 15, presented by SEQ ID NO: 1. 17. The strain of the bacteriophage Staphylococcus aureus SCH111, used to obtain the antibacterial composition according to claim 1 or 2, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-95. 18. The nucleic acid molecule corresponding to the bacteriophage genome according to claim 17, presented by SEQ ID NO: 2. 19. The strain of the bacteriophage Klebsiella pneumoniae KPV15, used to obtain the antibacterial composition according to claim 1 or 2, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-90.  20. The nucleic acid molecule corresponding to the bacteriophage genome according to claim 19, presented by SEQ ID NO: 3. 21. The strain of the bacteriophage Klebsiella pneumoniae KPV811, used to obtain the antibacterial composition according to claim 1 or 2, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-91. 22. The nucleic acid molecule corresponding to the bacteriophage genome according to claim 21, presented by SEQ ID NO: 4. 23. The strain of the bacteriophage Pseudomonas aeruginosa RA5, used to obtain the antibacterial composition according to claim 1 or 2, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center of applied microbiology and biotechnology" of the Federal service for supervision of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-88. 24. The nucleic acid molecule corresponding to the bacteriophage genome according to claim 23, presented by SEQ ID NO: 5. 25. The strain of the bacteriophage Pseudomonas aeruginosa PA10, used to obtain the antibacterial composition according to claim 1 or 2, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center of applied microbiology and biotechnology" of the Federal service for supervision of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-89. 26. The nucleic acid molecule corresponding to the bacteriophage genome according to claim 25, presented by SEQ ID NO: 6. 27. The strain of the bacteriophage Acinetobacter baumannii AM24, used to obtain the antibacterial composition according to claim 1 or 2, deposited in the collection of the Museum of microorganisms of the Federal budget institution of science "State scientific center for applied microbiology and biotechnology" of the Federal service for the supervision of consumer rights and human well-being (FBUN SSC PMB Rospotrebnadzor) under the number Ph-106. 28. The nucleic acid molecule corresponding to the bacteriophage genome according to claim 27, presented by SEQ ID NO: 7.

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