RU2526806C2 - Method for prevention of developing bacterial intestinal infection - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, namely to medical microbiology, and can be used for treating intestinal yersiniosis, or pseudotuberculosis, or salmonellosis experimentally. That is ensured by simulating one of the above pathologies in experimental animals followed by oral administration of therapeutic preparations in therapeutically effective doses with controlling the content of pathological microflora for a period of time providing elimination of the disease. Stimbifid or a culture fluid of lactic acid bacilli is administered orally as the therapeutic preparation into each animal for the first 2 hours after infection once a day as a monotherapeutic preparation promoting the colonisation resistance of the intestinal mucosa to an agent. Stimbifid is administered into each animal in a dose of 13 mg for 5 days from the moment of the infection, while the culture fluid is administered in an amount of 0.2 ml throughout 7 days from the moment of the injection. What is also presented is a method for preparing an agent for treating intestinal yersiniosis, or pseudotuberculosis, or salmonellosis experimentally.

EFFECT: group of inventions provides a high percentage of pathology elimination by using the prebiotic preparation and the liquid probiotic complex with no accompanying negative consequences.

4 cl, 3 tbl, 3 ex

Description

The invention relates to the field of medicine, infectology and medical microbiology and can be used to simulate conditions when selecting means to prevent the development of a sensitive intestinal infection caused by pathogenic bacteria.

Man and animals are born sterile, already in the first hours and days after birth, their skin and mucous membranes are colonized by microorganisms, the number and species composition of which are determined by the conditions of birth, the state of the environment and the type of feeding [Shenderov B.A. Normal microflora and its role in maintaining human health. // Grew. journal gastroenterol., hepatol., coloproctol. - 1998. - No. 1. - S. 61-65; Shenderov B.A. Medical microbial ecology and functional nutrition: in 3 volumes. Volume 1: The microflora of humans and animals and its functions. - M .: Publishing house GRANT, 1998. - 288 s]. The adult organism is abundantly populated with microorganisms, of which more than 60% colonize the intestines [Salminen S., Isolauri E., Onela T. Gut flora in normal and disordered states // Chemotherapy. - 1995 .-- Vol.41 (Suppl.l). - P. 5-15; Ardatskaya M.D. Intestinal dysbiosis: concept, diagnosis, principles of therapeutic correction // Consilium medicum. - 2008. - T. 10, No. 8. - S.86-92].

At the present stage of the study of microecological disturbances in the intestine and in general the infectious process, enough data has been accumulated that testify to the ability of individual representatives of their own intestinal microflora, which have certain healing mechanisms, to cause disease with a decrease in immunity. In this regard, some representatives of the intestinal microflora, causing purulent-inflammatory diseases, bacteremia of the newborn, local or generalized uroinfection, arthritis and other autoimmune diseases, are classified as opportunistic microorganisms against the background of decreased immunity.

It is possible to prevent the translocation of opportunistic microorganisms through the intestinal wall and their further dissemination through the sensitive organism by affecting the colonization resistance of the intestinal mucosa [Glushanova N.A., Shenderov B.A. Relationships of probiotic and indigenous host lactobacilli under co-culture in vitro. Zhurn. microbiol. - 2005. - No. 2. - 56-61]. Normal intestinal microflora, along with other functions, supports the colonization resistance of the intestinal mucosa, and with it the inhibition of adhesion of pathogenic microorganisms to the epithelium and their reproduction. The main mechanism of colonization resistance is the activation of the immune system. At the same time, the state of colonization resistance is associated with the biological properties of microorganisms of the intestinal microflora, pathogenic, conditionally pathogenic, as well as probiotic microorganisms in case of prescribing the latter for prophylactic or therapeutic purposes. The relationship between these groups of microorganisms can be antagonistic, synergistic or indifferent in nature, which makes them competitive or compatible. Thus, maintaining or increasing colonization resistance, providing an indigenous intestinal microflora with the help of modern drugs, a microecological advantage, it is possible to influence the infectious process caused not only by conditionally pathogenic, but also highly virulent microorganisms capable of initiating severe diseases of the gastrointestinal tract with severe violations of the species and the quantitative composition of intestinal microflora.

A complex method of treating acute intestinal infections using a diet, etiotropic and pathogenetic therapy, taking into account the severity and form of the disease, is known. All patients with abdominal and generalized forms of intestinal diseases are subject to hospitalization. The purpose of the appointment of antibacterial drugs is to stop the acute manifestations of the disease and prevent the generalization of infection, exacerbations, relapses, prolonged and chronic course. Antibiotics and chemotherapeutic drugs are the main means of combating infectious diseases of a bacterial nature [Bondareva TA, Kalininsky VB, Borisevich IV, Bondarev VP, Fomenkov O.O. Current status and prospects for solving the problem of increasing the effectiveness of emergency prevention and treatment of systemic bacterial infections // Molecular Medicine. - 2009. - No. 5. - S.21-25). However, the abundance of these drugs is often accompanied by their unsystematic use, which not only reduces the effectiveness of the use of antimicrobial drugs of various classes, but also leads to a number of side effects [Belmer S.V. Intestinal dysbiosis as a complication of antibiotic therapy // Children's infections. - 2007. - T.6, No. 2. - S. 44-48]. The latter include toxic reactions, damage to the liver parenchyma, damage to the kidneys, damage to the peripheral nervous system and hemopoietic organs, dysbiosis (dysbiosis), allergic effect and immunosuppressive conditions.

Even the safest antibacterial drugs have a negative effect on the patient’s body and can cause the death of not only pathogenic microorganisms in natural biotopes, but also microorganisms that make up the normal human microflora [B. Shenderov Normal microflora and some issues of microecological toxicology. // Antibiotics and honey. biotechnol. - 1987. - T. 32, No. 2. - S. 18-241.

Therefore, new means and methods of treating intestinal infections are needed. When developing such tools and methods, an important place is given to their modeling in the experiment [Guide to the experimental (preclinical) study of new pharmacological substances. / Ed. R.Yu. Khabrieva. - M .: Medicine, 2005. - 832 p.].

Closest to the claimed invention both in the part "Method of treatment ..." and in the part "Method of obtaining funds ..." is a technical solution known from the article [Darmov IV, Marakulin IV, Pogorelsky I.P. ., Novikova O.D., Portnyagina O.Yu., Solovyova T.F. Prevention of experimental pseudotuberculosis infection by immunization with porin from Yersinia pseudotuberculosis. // Bull. experimental biology and medicine. - 1999. - T.127, No. 2. - S.221-223].

However, the known method cannot be used if there is an infectious agent in the animal’s body, but it requires preliminary immunization of healthy individuals.

The technical result of the invention both in the part “Method of treatment ...” and in the part “Method of obtaining funds ...” is the development of such a method of treatment and a method of obtaining funds for its implementation, which would allow them to be used in animals already infected with an infectious agent, providing this is a high percentage of elimination of pathology without associated negative consequences.

This technical result in the part “Method of treatment ...” is achieved by the fact that in the inventive method of treating intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in an experiment by forming in animals an experimental pathology corresponding to the etiological agent followed by oral administration of therapeutic drugs in therapeutically effective doses under controlled maintenance pathogenic microflora during the period ensuring the elimination of the disease, as a therapeutic agent orally in the first 2 hours after To infect each animal, once a day, either Steambifide or lactobacillus culture fluid is administered as a monotherapy that provides colonization resistance of the intestinal mucosa to the pathogen, while each animal is injected with Steambifid at a dose of 13 mg for at least 5 days, and culture fluid a volume of 0.2 ml for at least 7 days.

Colonization resistance of the intestinal mucosa is judged by the absence of dysbiotic changes in the composition of the intestinal microflora, determined by counting the total number of microorganisms of the intestinal microflora and its individual representatives (CFU · g ^-1 ) taking into account the grown colonies on selective nutrient media at a temperature of 37 ° C for 48 hours.

The formation of experimental intestinal pathology in animals is carried out by oral administration of pathogens or intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in an amount of 10 LD ₅₀ .

This technical result in the part "Method of obtaining funds ..." for the implementation of the treatment method is achieved by the fact that in the claimed method for the treatment of intestinal yersiniosis or pseudotuberculosis or salmonellosis in an experiment containing the products of cultivation of lactobacilli, lactobacteria are isolated from the drug Lactobacterin, they are cultured in a liquid nutrient medium at 37 ° C for 72 hours to a concentration of (2.8-3.0) x 10 ⁹ CFUs · ml ^-1, and then the culture liquid was centrifuged in sealed test tubes to remove Ia therefrom cells or their fragments to give the desired product as a supernatant.

A method of treating intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in an experiment is as follows.

1. In a living organism infected with an intestinal infection pathogen, a biologically active complex is introduced at a certain dose through the mouth (per os), creating priority conditions for the propagation of microorganisms of their own intestinal microflora, stabilizing the barrier properties of the intestinal mucosa, stimulating the immune system and producing secretory immunoglobulins that inhibit growth pathogenic microflora and its displacement from the body, maintaining ionic homeostasis of the intestinal epithelial zone, primarily bifidoba bacteria, as well as lactobacilli, Escherichia.

2. Diagnose the disease using the bacteriological method and the determination of clinically significant symptoms of the disease.

3. A prebiotic or microbial exometabolite is administered until a therapeutic effect is achieved - normalization of intestinal functions and stabilization of the intestinal microflora, determined by the bacteriological method, for which a biologically active complex (action of a positive exogenous factor) is determined, the oral administration of which should stop the generalization of intestinal infection.

4. Determine the total number of viable microorganisms of the intestinal microflora and its individual representatives in experimental animals before infection with the causative agent of intestinal infection and at the end of observation (on day 21) or at the time of death of the animal from generalization of intestinal infection.

5. Calculate the difference in the numbers of intestinal microflora and its individual representatives at two time points (before infection with the causative agent of intestinal infection and on day 21 of observation or at the time of death of the animal from generalization of intestinal infection) to assess the microecological state of intestinal microflora.

6. Determine the presence and number of bacteria of intestinal infection in the intestinal contents of dead experimental animals as a result of generalization of intestinal infection, as well as in surviving animals on the 21st day of observation.

7. By the number of surviving individuals in the experimental group of infected animals that received an orally selected biologically active complex, as well as by the total number of microorganisms of the intestinal microflora and its individual representatives, before infection and at the end of observation on day 21 they made a judgment about the protective effectiveness of the biologically active complex.

8. A judgment is made on the effectiveness of the oral administration of a biologically active complex into an infected organism based on the improvement of well-being and recovery, as well as determination of the total number of microorganisms of the intestinal microflora in 1 g of feces, indicating the absence of dysbiotic changes in the composition of the intestinal microflora.

9. The following can be used as a biologically active complex: the prebiotic preparation Steambifid, manufactured by V-MIN LLC / MedStar LLC, Russia. The drug is based on fructooligo and fructopolysaccharides, contains the premix vitamin-mineral Immunity and excipients (sodium bicarbonate, lactose, calcium stearate).

Description of the Method of obtaining funds for implementing the method of treatment of intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in the experiment.

1. Lactobacilli is isolated from the drug Lactobacterin, for which, under aseptic conditions, a bottle containing a lyophilized culture of lactobacilli is opened, 5.0 ml of sterile sodium chloride solution is added to the bottle to rehydrate the culture of lactobacilli and obtain a homogeneous suspension.

2. Select 2.0 ml of rehydrated culture of lactobacilli and transfer into 100 ml of liquid culture medium in a bottle with a cotton-gauze stopper.

3. Lactobacilli are cultured in a liquid nutrient medium under microaerophilic conditions at a temperature of 37 ° C for 72 hours to a concentration of (2.8-3.0) · 10 ⁹ CFU · ml ^-1 , after which the native culture is centrifuged (3000g, 15 min) in closed tubes until the cells or their debris are removed from it and the target product is obtained in the form of a supernatant.

4. The supernatant (liquid probiotic complex), obtained from the native culture of probiotic lactobacilli, freed by centrifugation of the cellular elements of lactobacilli and containing low molecular weight exometabolites that accumulate in the culture medium during the growth and reproduction of lactobacilli under optimal microaerophilic conditions, is used for oral white administration .

The common feature that combines the prebiotic preparation Steambifid and the liquid probiotic complex in the supernatant of the native culture of lactobacilli isolated from the probiotic preparation Lactobacterin is the multidirectionality of the prebiotic action, which exclusively stimulates the reproduction of bifidobacteria and other representatives of the normal intestinal microflora, which inhibits the growth of pathogenic microflora , increasing the rate of restoration of normal intestinal microflora, stabilizing arernye properties of the intestinal mucosa, which stimulates the production of secretory immunoglobulin in the lumen of the gut and enhances the phagocytic activity of the liver cells.

Modeling of the disease is as follows.

1. Select a specific causative agent of intestinal disease, assess the pathogenicity (as such can be selected:

the causative agent of pseudotuberculosis, causing a serious infectious disease, with a septic form which mortality can reach 25-50%;

the causative agent of intestinal yersiniosis - zoonotic bacterial infection with severe polymorphism of clinical manifestations and damage to the gastrointestinal tract;

the causative agent of salmonellosis is an acute zoonotic-anthroponous bacterial infection, which is characterized by intoxication and predominant damage to the gastrointestinal tract).

2. Evaluation of the pathogenicity of the causative agent of intestinal disease is carried out with the enteral (oral) route of entry of the pathogen into the body of a laboratory animal, natural for intestinal yersiniosis, or pseudotuberculosis, or salmonellosis. Taking into account the results of oral infection in laboratory animals, the degree of pathogenicity of the pathogen of intestinal infection (virulence) is calculated as an indicator of the average lethal dose LD ₅₀ [Ashmarin I.P., Vorobyov A.A. Statistical methods in microbiological research. - L .: Medgiz, 1962. - 280 p.]. The average lethal dose indices determined for conventional white mice (LD ₅₀ ) were: for the causative agent of intestinal yersiniosis, 7.4 · 10 ⁷ CFU; for the causative agent of pseudotuberculosis - 8.5 · 10 ⁴ CFU; for the pathogen of salmonellosis - 3.1 · 10 ⁸ CFU.

Example 1

A pseudotuberculosis microbe culture is grown at a temperature of 5 ± 3 ° C for 3 days, after which a bacterial suspension is prepared on an isotonic sodium chloride solution based on the calculation of the concentration of 10 LD ₅₀ microbial cells in the injected volume of the suspension for oral infection of animals. The indicated concentration of bacteria in the suspension, equal to 10 average lethal doses of the pseudotuberculosis microbe, ensures the guaranteed development of the infectious process with the oral administration of the bacterial suspension.

Take 10 acclimatized conventional white mice weighing 18-20 g and sit in a ditch with a lid. Fecal masses were taken from each animal at the beginning of the study to calculate the total number of microorganisms of the intestinal microflora and its individual representatives (CFU · g ^-1 ), taking into account the grown colonies on selective solid nutrient media at a temperature of 37 ° C for 48 hours.

.A suspension of pseudotuberculosis microbe bacteria in a volume of 0.2 ml containing 10 LD _{50 of the} pathogen was orally administered to conventional white mice, animals were observed, noting the outcome of the infection. Table 1 shows the result of infection of conventional white mice with the causative agent of pseudotuberculosis $$(\overline{X}\pm I_{95},n=10)$$

.

The data presented in table 1 indicate that as a result of infection by the causative agent of pseudotuberculosis, all experimental animals of the control group died within 5 to 13 days from the start of infection. A pure culture of a pseudotuberculosis microbe was isolated from all dead animals: in 1 g of feces, the number of bacteria is 8.6 · 10 ⁷ -1.2 · 10 ⁸ CFU · g ^-1 . The decrease in the total number of normal intestinal microflora is 3.7 · 10 ⁵ times, bifidobacteria - 5.9 · 10 ⁴ times, lactobacilli -2.2 · 10 ⁶ times, Escherichia - 1.4 · 10 ³ times.

The results obtained indicate the generalization of infection, the death of animals from pseudotuberculosis and the development of deep dysbiotic changes in the composition of the intestinal microflora.

Example 2

A culture of a pseudotuberculosis microbe is grown according to the method described in Example 1. For subsequent infection of experimental animals, a suspension of bacteria in an isotonic sodium chloride solution is used, 0.2 ml of which contains a bacterial count of 10 LD ₅₀ when administered orally to animals.

Take 10 acclimatized conventional white mice weighing 18-20 g and sit in a ditch with a lid. Fecal masses are taken from each animal at the beginning of the study and at the end of the observation period to calculate the total number of microorganisms of the intestinal microflora and its individual representatives (CFU · g ^-1 ) taking into account the grown colonies on selective solid nutrient media at a temperature of 37 ° C for 48 hours .

A suspension of pseudotuberculosis microbe bacteria in a volume of 0.2 ml containing 10 LD _{50 of the} pathogen is orally administered to conventional white mice.

A suspension of the prebiotic Steambifide is prepared in distilled water, 0.2 ml of which contains 13 mg of the drug. The indicated dose in the volume of 0.2 ml of the suspension corresponds to the conversion factor of the daily dose of the stebifid prebiotic for humans. The daily dose of the steambifid prebiotic is administered orally to conventional white mice 2 hours after oral infection of the experimental animals, and the suspension of the steambifide prebiotic is continued to be administered to the infected animals for 5 days after their infection. The animals are monitored for 21 days, the outcome of infection is noted (table 2).

.Table 2 shows the protective efficacy of the prebiotic Steambifide for oral infection of conventional white mice with the causative agent of pseudotuberculosis $$(\overline{X}\pm I_{95},n=10)$$

.

Prebiotic Steambifide, as follows from the results presented in Table 2, completely prevents the multiplication of the causative agent of pseudotuberculosis on the surface of the intestinal mucosa, its translocation through the intestinal wall, entry into the bloodstream and dissemination throughout the body. None of the animals infected with the causative agent of pseudotuberculosis, on the background of oral administration of the prebiotic Steambifid, did not get sick and did not die. At the same time, bacteriological examination of animal feces did not reveal any dysbiotic changes in the intestinal microflora, and the number of bifidobacteria in the intestinal contents even increased.

The results obtained indicate that the prebiotic Steambifid provides the formation in conventional white mice of a high level of colonization resistance of the intestinal mucosa, which does not allow adhesion of colonization, translocation of the causative agent of pseudotuberculosis and the generation of the infectious process.

Example 3

Lactobacillus strain L. plantarum 8P-A3, grown from the Lactobacillus probiotic preparation, is grown in a liquid nutrient medium at a temperature of 37 ° C under microaerophilic conditions for 72 hours to a concentration of (2.8-3.0) · 10 ⁹ CFU · ml ^{- 1} . The resulting native culture is freed from microbial cells by centrifugation in lockable centrifuge tubes. The supernatant of the native culture of lactobacilli is used for oral administration to conventional white mice.

A culture of a pseudotuberculosis microbe is grown according to the method described in Example 1. For subsequent infection of experimental animals, a suspension of bacteria in an isotonic sodium chloride solution is used, 0.2 ml of which contains the amount of bacteria equal to 10 LD ₅₀ when administered orally to animals.

Take 10 acclimatized conventional white mice weighing 18-20 g and sit in a ditch with a lid. Fecal masses are taken from each animal at the beginning of the study and at the end of the observation period to calculate the total number of microorganisms of the intestinal microflora and its individual representatives (CFU · g ^-1 ) taking into account the grown colonies on selective solid nutrient media at a temperature of 37 ° C for 48 hours .

A suspension of pseudotuberculosis microbe bacteria in a volume of 0.2 ml containing 10 LD _{50 of the} pathogen is orally administered to conventional white mice.

Orally-infected conventional white mice are injected with a supernatant of a native culture of lactobacilli in a volume of 0.2 ml 2 hours after oral administration of a suspension of a pseudotuberculosis microbe to animals. The introduction of the supernatant of the native culture of lactobacilli to experimental animals is carried out for 7 days after their infection. The animals are observed for 21 days, the outcome of infection is noted. Table 3 shows the protective efficacy of the supernatant of a native culture of lactobacilli containing low molecular weight microbial exometabolites during oral infection of conventional white mice with the causative agent of pseudotuberculosis (X ± I ₉₅ , n = 10).

The results shown in table 3 indicate that the supernatant of the native culture of lactobacilli of the strain L. plantarum 8P-A3, containing low molecular weight microbial exometabolites that are formed during the growth and reproduction of lactobacilli, effectively protects against the introduction of infected conventional white mice from pseudotuberculosis. The prevention of colonization of the intestinal mucosa by bacteria of the pseudotuberculosis microbe and their elimination from the intestines of animals occurs due to the maintenance of the natural microbial community of normal intestinal microflora, which is facilitated by low molecular weight microbial exometabolites introduced into experimental animals with the supernatant of the native culture of lactobacilli. The preservation of the microbial community in the intestinal contents is indicated by the figures of the total number of microorganisms of the intestinal microflora and its individual representatives - bifidobacteria, lactobacilli and Escherichia.

Preservation of numerical values of the total amount of normal intestinal microflora and its individual representatives should be associated with the prebiotic effect of low molecular weight microbial exometabolites, ensuring the preservation of a high level of colonization resistance of the intestinal mucosa.

The presented results confirm that an important characteristic of intestinal microbiocenosis is the quantitative ratio of microflora species. The microbial community is dominated by anaerobic and microaerophilic groups of microorganisms. They constitute the “species core” of the microbiocenosis, creating a habitat for the entire community. However, rare and small species are also important for the life of the microbial community: they create its species diversity, increase the number of biocenotic connections, serve as a reserve for filling and replacing the dominant species, i.e. give microbiocenosis stability under different conditions [Vorobev A.A., Abramov N.A., Bondarenko V.M., Shenderov B.A. Dysbacteriosis is an urgent problem of medicine. // Bulletin of RAMS. - 1997. - No. 3. - C.4-7; Bondarenko V.M., Petrovskaya V.G. The early stages of the development of the infectious process. // Bulletin of RAMS. - 1997. - No. 3. - S.7-10; Pokrovsky V.I. Man and microorganisms. Health and disease. // Bulletin of RAMS. - 2000. - No. 11. - C.3-6]. The species composition of the microbial community and the number of individual types of intestinal microbiocenosis are determined by the conditions of the closed space of the intestine, in which very specific conditions are created in an adult. And the more specific the conditions, the poorer the species composition of microorganisms and the higher the number of individual species. In various diseases of the gastrointestinal tract, such as decreased gastric secretion, impaired function or resection of the ileocecal valve, impaired intestinal digestion and absorption, bowel obstruction, structural disorders of the intestinal wall (diverticulosis), the effects of surgery, etc., there is an excess of bacteria in small intestine, favorable conditions are created for the proliferation of bacteria and stasis in the intestine [Minushkin ON, Ardatskaya MD, Zverkov IV, Chicherin I.Yu. Dysbacteriosis (concept, diagnosis, principles of medical correction). Modern possibilities of prebiotic therapy: a training manual for doctors and cadets of the cycle of improvement of doctors. M .: FGU "Educational Scientific Medical Center" of the Office of the President of the Russian Federation. 2010. - 50 s].

The claimed method takes into account this theoretical information and allows you to save and even increase the level of colonization resistance of the intestinal mucosa, which together with a number of protective factors of the intestine ensures the immunity of the whole organism to the causative agent of intestinal infection.

Table 1 A method for the treatment of intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in an experiment and a method for producing a means for its implementation Number of animals

(X_min-X_max)The terms of death, day, $$\overline{X}$$

(X _min -X _max ) The content of live bacteria in 1 g of feces on ... the day of the experiment, CFU · g ^-1 experienced fallen from pseudotuberculosis isolated microorganisms start of experiment on the day of death 10 10 8.8 (5-13) Total amount (6.0 ± 0.7) · 10 ⁹ (1.6 ± 0.6) · 10 ⁴ Bifidobacteria (6.5 ± 0.6) · 10 ⁶ (1.1 ± 0.4) · 10 ⁴ Lactobacilli (2.8 ± 0.6) · 10 ⁸ (1.3 ± 0.5) · 10 ² Esherichia (1.6 ± 0.5) · 10 ⁴ (1.1 ± 0.3) · 10 ¹

table 2 Number of animals Deadlines The content of live bacteria in 1 g of feces on ... the day of the experiment, CFU · g ^-1 experienced fallen from pseudotuberculosis isolated microorganisms start of experiment on the day of death 10 0 - Total amount (6.1 ± 0.8) · 10 ⁹ (5.8 ± 0.6) · 10 ⁹ Bifidobacteria (6.3 ± 0.7) · 10 ⁶ (5.1 ± 0.7) · 10 ⁷ Lactobacilli (2.5 ± 0.7) · 10 ⁸ (4.0 ± 0.5) · 10 ⁸ Esherichia (1.7 ± 0.6) · 10 ⁴ (3.9 ± 0.7) · 10 ⁴

Table 3 Number of animals Deadlines, days The content of live bacteria in 1 g of feces on ... the day of the experiment, CFU · g ^-1 experienced fallen from pseudotuberculosis isolated microorganisms start of experiment on the day of death 10 0 - Total amount (6.5 ± 0.8) · 10 ⁹ (8.1 ± 0.6) · 10 ⁹ Bifidobacteria (6.4 ± 0.6) · 10 ⁶ (6.8 ± 0.7) · 10 ⁷ Lactobacilli (2.8 ± 0.7) · 10 ⁸ (5.3 ± 0.6) · 10 ⁸ Esherichia (2.0 ± 0.5) · 10 ⁴ (1.5 ± 0.5) · 10 ⁴

Claims (4)

1. A method for the treatment of intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in an experiment by forming in animals an experimental pathology corresponding to the etiological agent, followed by oral administration of therapeutic drugs in therapeutically effective doses under the control of the content of pathogenic microflora for a period providing elimination of the disease, characterized in that as a therapeutic agent, they are orally administered in the first 2 hours after infection to each animal 1 time per day or Steambi feed, or culture fluid of lactobacilli as a monotherapy, providing colonization resistance of the intestinal mucosa in relation to the pathogen, while each animal is injected with a dose of 13 mg for 5 days from the moment of infection of the animal, and culture fluid in a volume of 0.2 ml per within 7 days from the moment of infection of the animal. 2. The method according to claim 1. characterized in that the colonization resistance of the intestinal mucosa is judged by the absence of dysbiotic changes in the composition of the intestinal microflora by counting the total number of microorganisms of the intestinal microflora and its individual representatives (CFU · g ^-1 ) taking into account the grown colonies on selective nutrient media at a temperature of 37 ° C in within 48 hours. 3. The method according to claim 1, characterized in that the formation of experimental intestinal pathology in animals is carried out by oral administration of pathogens or intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in an amount of 10 LD ₅₀ . 4. A method of obtaining funds for the treatment of intestinal yersiniosis, or pseudotuberculosis, or salmonellosis in an experiment containing products of the cultivation of lactobacilli, characterized in that lactobacteria are isolated from the drug Lactobacterin, they are cultured in a liquid nutrient medium at a temperature of 37 ° C for 72 hours before concentration (2.8-3.0) · 109 CFU · ml ^-1 , after which the culture fluid is centrifuged in closed tubes until the cells or their debris are removed from it and the target product is obtained in the form of a supernatant.