RU2326692C2 - Probiotic medicinal agent for agricultural animals and birds - Google Patents

Abstract

FIELD: medicine; veterinary science.

SUBSTANCE: agent contains mixed suspensions of bacteria strain and protective medium. As protective medium, agent contains saccharose-gelatin composition with the following ratio of mixture (mass%): saccharose - 15, gelatin - 5, 9% sodium chloride solution - 80. As suspensions of bacteria strain, agent contains suspension of the following strains (mass%): Azomonas agilisc B-2586 - 8, Azotobacter chroococcum B-3162 - 7, Azotobacter vinelandii B-2587 - 7, Bacillus subtillis B-5225 - 7, Bacillus subtillis B-4828 - 9, Bifidum bacterium globosum B-2584 - 10, Escherichia coli B-4412 - 8, Enterococcus faecium B-2898 - 7, Lactobacillus acidophilus B-3488 - 7, Lactobacillus acidophilus B-2585- 7, Propionibacte-rium freudenreichii B-6561- 7, Pseudomonas sp. B-3908 - 7, Pseudomonas sp. B-2589- 7, Saccharomyces cerevisiae Y-365 - 2.

EFFECT: agent enables to normalize bowel microbiocenose of healthy animals and birds; reduce mortality and to provide mass increase of young animals.

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Description

FIELD OF THE INVENTION

The invention relates to biotechnology and microbiology, in particular to probiotic drugs for farm animals and poultry, and can be used in veterinary medicine for the prevention and treatment of diseases, reduce mortality and increase weight gain in young animals.

State of the art

A probiotic medicinal product for veterinary medicine is known, including Bacillus subtilis 12B-DEPT biomass and filler in the following ratio of components, wt.%: Bacillus subtilis 12B-DEPT (living microbial cells) biomass - 1-2 × 10 ⁹ in 1 ml, solvent 92- 95%, filler 5-8%. The biological product inhibits the development of pathogenic microorganisms of the genera: Streptococcus, Esherichia coli, Pseudomonas, Sthaphilococcus, Proteus, Shigella, has broad antibiotic resistance and treatment effectiveness (RF patent No. 2172175, CL A61K 35/74, C12N 1/20, publ. .2001).

The disadvantage of this biological product is the limited species composition and insufficient therapeutic effect.

Known probiotic drugs of various species composition intended for the prevention of gastrointestinal diseases of young animals and birds:

- “Apinik”, developed by VGNKI - NPO “CMEI”, which includes L. acidophilus, Saccharomyces cerivisiae yeast extract;

- “Bifidumbacterin”, the developer of which is “Partner” JSC, Moscow, consisting of Bif.adolescentis, Bif.bifidum, Bif.longum;

- “Bifacidobacterin”, developed by MGTSPB - LLC “M-TECH”, which includes L.acidophilus, Bif.adolescentis;

- "Bifinorm", the developer of the Bryansk Agricultural Institute, which includes Bif.adolescentis.

The disadvantage of these biological products is the limited species composition (Malik NI, Panin AN Veterinary, 2001, No. 1).

The closest in technical essence and the achieved positive effect adopted by the authors for the prototype is the veterinary probiotic drug "Bactoneotim", which includes L. acidophilus, Str.faecium, Bif.adolescentis, Bif.bacterium globosum (developer of LLC "Veda", Pushchino) (Malik N.I., Panin A.N. Veterinary Medicine, 2001, No. 1).

Significant disadvantages of this biological product are the limited species composition (4 strains of microorganisms belonging to 3 types of bacteria), as well as the use of strains isolated from the human intestine or taken from the collection of strains for food biotechnology. This applies to strains of L. acidophilus, Bif.adolescentis (Malik N.I., Panin A.N. Veterinary science, 2001, No. 1).

Disclosure of invention

The objective of the invention is the development of a probiotic drug from effective microorganisms for farm animals and poultry, having a multi-species composition, which includes a mixture of suspensions of living effective microorganisms belonging to different genera and species, corresponding to the normal intestinal microbiocenosis of healthy animals and birds, allowing to reduce the incidence of , mortality and increase body weight gain in young animals.

The technical result that can be achieved using the present invention is to normalize the microbiocenosis of the intestines of healthy animals and birds, reduce the incidence of purulent-inflammatory and dyspepsia and death, increase body weight gain in young animals.

The technical result is achieved using a probiotic drug from a mixture of suspensions of effective microorganisms for farm animals and poultry containing a strain of bacteria L.acidophilus and a protective medium, while the strain L.acidophilus is taken B-2585, and as a protective medium using a sucrose-gelatin mixture in the ratio: sucrose: gelatin: 9% sodium chloride solution (wt.%) 15: 5: 80, and it additionally contains a mixture of suspensions of strains of Azomonas agilisc B-2586; Azotobacter chroococcum B-3162; Azotobacter vinelandii B-2587; Bacillus subtilis B-5225; Bacillus subtilis B-4828; Bifidum bacterium globosum B-2584; Escherichia coli B-4412; Enterococcus faecium B-2898; Lactobacillus acidophilus B-3488; Propionibacterium freudenreichii B-6561; Pseudomonas B-2589; Pseudomonas B-3908; Saccharomyces cerevisiae Y-365, isolated from healthy productive farm animals, poultry and most fully corresponding to their normal microbiocenosis, with the following effective composition and ratio of components, wt.%:

Azomonas agilisc B-2586 - 8;

Azotobacter chroococcum B-3162-7;

Azotobacter vinelandii B -2587-7;

Bacillus subtilis B-5225-7;

Bacillus subtilis B-4828-9;

Bifidum bacterium globosum B-2584-10;

Escherichia coli B-4412-8;

Enterococcus faecium B-2898-7;

Lactobacillus acidophilus B-3488-7;

Lactobacillus acidophilus B-2585-7;

Propionibacterium freudenreichii B-6561-7;

Pseudomonas B-3908 - 7;

Pseudomonas B-2589-7;

Saccharomyces cerevisiae Y-365 - 2 (table).

Thus, the macroorganism is closely associated with microflora, the composition of which is normally characterized by the constancy and balance of the populations of its main representatives. Without normal microflora, complete digestion and assimilation of food, maintaining the constancy of the internal environment of the body, and its protection from pathogenic microflora are impossible.

In a sick body, the quantitative ratios of microbial populations, their distribution in the intestinal tract, upper respiratory tract, on the skin, as well as their biological activity, are very often violated. The emergence of powerful antibiotics and the deterioration of environmental conditions led to disruption of normal microbiocenoses.

The widespread production of harmful chemical products that enter the environment has changed the immunobiological reactivity of animals, especially young animals. All this leads to disorders of the basic regulatory systems of the body, contributing to a massive increase in the incidence of disease, negative genetic and other changes.

Overuse of antibiotics has even more sad consequences. Today, there are many antibiotic-resistant forms of pathogenic microorganisms that reduce the effectiveness of the antibiotics used.

In violation of the conditions of keeping and feeding animals, especially young animals, succession processes of intestinal microorganisms change in a negative direction. At the same time, a high concentration of staphylococci, protea, yeast-like fungi and other microorganisms was established in the composition of the intestinal biocenosis, the population level of bifidobacteria and lactic acid bacteria was reduced, and the structure of the population of Escherichia changes in the direction of decreasing enzymatic activity.

World experience shows that in the prevention and treatment of gastrointestinal diseases of young animals, the importance of substitution therapy aimed at restoring the intestinal biocenosis through the regular introduction of live bacteria - representatives of normal intestinal microflora (Antipov V.A. Agriculture Abroad, 1981, No. 2) ; (Tarakanov B.V., Nikolicheva T.A. Veterinary, 2000, No. 7); (Gilliland S.R., Speck M.L. J. Food Pretect, 1977, No. 40).

The main function of friendly microbes is protective: they suppress pathogenic and conditionally pathogenic bacteria. Soon after taking effective microorganisms (EM), biologically active substances begin to stand out and microbial cell systems function, having both a direct effect on pathogenic and conditionally pathogenic microorganisms, and indirectly by activating specific body defense systems. In addition, being biocatalysts of many vital processes in the digestive tract, they actively produce enzymes, amino acids, antibiotics and other physiologically active substrates.

The essence of the invention consists in compiling a probiotic drug from a mixture of suspensions of live strains of effective microorganisms (EM) in a specific species composition and ratio. The strains that make up a single bioculture, isolated from healthy productive farm animals and birds, belong to different genera and species, differ in biological properties and direction of action, which makes it possible to reduce morbidity and mortality when a drug is added to food or water in certain doses and increase weight gain in young animals.

The implementation of the invention

Examples of specific performance of obtaining a probiotic drug from effective microorganisms for livestock and poultry:

Example 1. Prepare a series of probiotic drugs of the following composition and quantitative ratio, wt.%: 1) Azomonas agilisc B-2586 - 10%; 2) Azotobacter chroococcum B-3162 - 8%; 3) Bacillus subtilis B-4828 - 20%; 4) Escherichia coli B-4412 - 20%; 5) Lactobacillus acidophilus B-2585 - 20%; 6) Pseudomonas B-3908 - 20%; 7) Saccharomyces cerevisiae Y-365 - 2%. For this, bacterial strains are grown separately on a nutrient medium that is optimal for each strain for (36 ± 2) hours at a temperature of 37 ° C. The grown bacterial cultures are washed off with a 9% sodium chloride solution, and the number of microbial cells in 1 ml of suspension is calculated according to the optical turbidity standard (TOC 45-28-59-85-P). Then, a suspension of each strain is prepared in a protective medium, for example, in a sucrose-gelatin mixture, bringing the concentration of microbial cells of each culture to 1.0-1.5 × 10 ⁹ MK / ml of this protective medium. Then prepare a mixture of suspension of strains in a certain ratio.

In order to prevent gastrointestinal diseases, calves in the experimental group received an EM preparation daily mixed with milk: calves under 20 days of age received 5 ml of EM preparation per day, with an age of 20 days to 3 months - 20 ml per the head.

As a result of observations, it was found that no calves were observed in the experimental group of diseases. In the control group that did not receive the drug, 5 calves were ill with enteritis, however, after a single application of an EM drug at a therapeutic dose of 20 ml per head per day, diarrhea in calves stopped after 16 hours. The appearance of the calves of the experimental group was noticeably different from the type of calves of the control group - the coat was shiny, and the calves of the experimental group showed good appetite. The ration of feeding in the conditions of the experimental and control groups (per 1 head per day): 5-10 kg of green mass; 0.5 kg of concentrated feed: 0.01 kg of salt; 5 kg of milk. The duration of the experiment is 20 days. During the experiment and after its completion, the animals of the experimental and control groups were weighed.

As a result, in the experimental group, the total weight gain (25 goals) per day was 12.5 kg (0.5 kg per 1 head), and for 18 days - 225 kg. In the control group, the weight gain per day (for 25 animals) was 11.25 kg (0.45 kg per animal), for 18 days - 202.5 kg. The difference in daily calf gain for the experimental and control groups was 1.25 kg, and for the entire duration of the experiment - 22.5 kg.

In the experimental group of piglets treated with an EM preparation at a dose of 5 ml per head per day, there were no cases of dysbacteriosis. In the control group, symptoms of dysbiosis (diarrhea) were observed in 6 piglets. After a single use of an EM preparation of 5 ml per head, after 16 hours, the signs of the disease disappeared.

An EM preparation was also included in the diet of 2 pregnant sows 3 days before farrowing (dose of the preparation was 30 ml per day). The newborn piglets did not have diarrhea, their general appearance and appetite were good, their behavior was active, there was no death.

The daily diet of one piglet was: 0.2 kg of concentrated feed; 0.2 kg of milk; 0.3 kg of green mass. The duration of the experiment was 18 days. During the experiment and after its completion, the animals of the experimental and control groups were weighed.

In the experimental group, the total weight gain (per 20 goals) per day was 3.7 kg (0.185 kg per head), and for 18 days - 66.6 kg. In the control group (20 goals), the total weight gain per day was 2.4 kg (0.12 kg per head), and for 18 days - 43.2 kg. The difference in the daily gain of piglets for the experimental and control groups was 1.3 kg, and for the entire experiment - 23.4 kg.

In experiments on 30 thousand (15 thousand - the experimental group, 15 thousand - the control group) of chickens of approximately the same weight, an additive to the feed, consisting of the above consortium of microorganisms, was tested. The calculation of the required amount of EM-preparation per 1 bird’s head depended on its age: 0.3-0.5 ml per 10 heads for chickens under 10 days old; 10-20 days - 0.7-0.8 ml; 20-30 days - 0.8-1.0 ml; 30-40 days - 1.2-1.3; 40-50 days - 1.3-1.5 ml. The additive was used in a mixture with food or water 1 time per day. Strictly observed food and temperature conditions. To avoid crowding, the chicks observed the rules of zootechnical planting. The bird was monitored daily for 2 months, counting the number of dead in both the experimental and control groups, control weighing after 4-5 days and the behavioral status of the chickens.

The use of the above EM preparation increased the overall weight gain of the bird by 19.8% and reduced the mortality rate from 10% to 2.8% compared with the control. In addition, it was noted that the poultry stock was healthy, active, willingly eating food treated with a suspension of living microorganisms.

Example 2. Prepare a series of probiotic drugs of the following composition and quantitative ratio, wt.%: 1) Azomonas agilisc B-2586 - 20%; 2) Azotobacter vinelandii B-2587 - 10%; 3) Bacillus subtilis B-5225 - 10%; 4) Bifidum bacterium globosum B-2584 - 18%; 5) Enterococcus faecium B-2898 - 10%; 6) Lactobacillus acidophilus B-3488 - 10%; 7) Propionibacterium freudenreichii B-6561 - 10%; 8) Pseudomonas B-2589 - 10%; 9) Saccharomyces cerevisiae Y-365 - 2%, using the manipulations described in example 1.

After daily administration of the drug for 18 days in the experimental group of calves, the total weight gain (25 goals) per day was 13.5 kg (0.54 kg per 1 head), and for 18 days - 243 kg. In the control group, the weight gain per day (for 25 animals) was 11.25 kg (0.45 kg per animal), for 18 days - 202.5 kg. The difference in daily gain of calves for the experimental and control groups was 2.25 kg, and for the entire experiment - 40.5 kg.

In the experimental group of piglets treated with this biological product, the total weight gain (per 20 goals) per day was 3.9 kg (0.195 kg per head), and for 18 days - 70.2 kg. In the control group (20 goals), the total weight gain per day was 2.4 kg (0.12 kg per head), and for 18 days - 43.2 kg. The difference in the daily gain of piglets for the experimental and control groups was 1.5 kg, and for the entire duration of the experiment - 27 kg.

The use of the above EM preparation increased the overall weight gain of the bird by 20.7% and reduced the mortality rate from 10% to 3.2% compared with the control. In addition, it was noted that the poultry stock was healthy, active, willingly eating food treated with a suspension of living microorganisms.

Example 3. Prepare a series of biological product of the following composition and quantitative ratio, wt.%: 1) Azomonas agilisc B-2586 - 8%; 2) Azotobacter chroococcum B-3162 - 7%; 3) Azotobacter vinelandii B-2587 - 7%; 4) Bacillus subtilis B-5225 - 7%; 5) Bacillus subtilis B-4828 - 9%; 6) Bifidum bacterium globosum B-2584 - 10%; 7) Escherichia coli B-4412 - 8%; 8) Enterococcus faecium B-2898 - 7%; 9) Lactobacillus acidophilus B-3488 - 7%; 10) Lactobacillus acidophilus B-2585 - 7%; 11) Propionibacterium freudenreichii B-6561 - 7%; 12) Pseudomonas B-3908 - 7%; 13) Pseudomonas B-2589 - 7%; 14) Saccharomyces cerevisiae Y-365 - 2%, using the manipulations described in example 1.

After daily administration of the drug for 18 days in the experimental group of calves, the total weight gain (25 goals) per day was 15 kg (0.6 kg per 1 head), and for 18 days - 270 kg. In the control group, the weight gain per day (for 25 animals) was 11.25 kg (0.45 kg per animal), for 18 days - 202.5 kg. The difference in daily gain of calves for the experimental and control groups was 3.7 kg, and for the entire duration of the experiment - 67.5 kg.

In the experimental group of piglets treated with this biological product, the total weight gain (per 20 goals) per day was 5.5 kg (0.275 kg per head), and 99 kg in 18 days. In the control group (20 goals), the total weight gain per day was 2.4 kg (0.12 kg per head), and for 18 days - 43.2 kg. The difference in the daily gain of piglets for the experimental and control groups was 3.1 kg, and for the entire experiment - 55.8 kg.

The use of the above EM preparation increased the total weight gain of the bird by 25% and reduced the mortality rate from 10% to 4% compared with the control. In addition, it was noted that the poultry stock was healthy, active, willingly eating food treated with a suspension of living microorganisms.

As shown by experimental tests of probiotic drugs on farm animals and poultry, the composition and mixture of suspensions of the following effective microorganisms are optimal in composition and percentage: Azomonas agilisc B-2586 - 8%; Azotobacter chroococcum B-3162 - 7%; Azotobacter vinelandii B-2587 - 7%; Bacillus subtilis B-5225 - 7%; Bacillus subtilis B-4828 - 9%; Bifidum bacterium globosum B-2584 - 10%; Escherichia coli B-4412 - 8%; Enterococcus faecium B-2898 - 7%; Lactobacillus acidophilus B-3488 - 7%; Lactobacillus acidophilus B-2585 - 7%; Propionibacterium freudenreichii B-6561 - 7%; Pseudomonas B-3908 - 7%; Pseudomonas B-2589 - 7%; Saccharomyces cerevisiae Y-365 - 2%.

The advantage and positive effect is due to the fact that, in contrast to the prototype, the proposed probiotic drug, due to its specific composition of effective directed microorganisms, provides the colonization of the gastrointestinal tract of animals and birds with normal microflora, which suppresses the pathogenic flora, increases immunity, normalizes the intestines, improves them Well-being, increases appetite, improves the digestibility of feed, which increases the gain and leads to the improvement of agricultural governmental animals and birds.

Table No pp The name of the microorganism Strain number Strain properties Selection source one. Azomonas agilisc B-2586 Stimulates the growth of animals and fish Gastrointestinal tract 2. Azotobacter chroococcum B-3162 Stimulates the growth of animals and fish Gastrointestinal tract 3. Azotobacter vinelandii B-2587 Stimulates the growth of agricultural animals GIT of cattle four. Bacillus subtilis B-5225 Pathogenic Klebsiel Antagonist From the colon of a healthy 10-day-old calf 5. Bacillus subtilis B-4828 Antagonist of pathogenic microflora. Prevention of the disease of newborn calves with diarrhea Large intestine calf 6. Bifidum bacterium globosum B-2584 Antagonist of pathogenic and conditionally pathogenic microflora GIT of cattle 7. Escherichia coli B-4412 Stimulates the growth of animals and fish GIT of cattle 8. Enterococcus faecium B-2898 Feed fortification The intestinal tract of carp 9. Lactobacillus acidophilus B-3488 Suppresses the growth of pathogenic E. coli Goose litter 10. Lactobacillus acidophilus B-2585 Treatment and prevention of diarrhea and dysbiosis Ram scar eleven. Propionibacterium freudenreichii B-6561 Producer of B vitamins Bull scar 12. Pseudomonas B-3908 Staphilicoccus growth inhibitor producer 13. Pseudomonas B-2589 Staphilicoccus growth inhibitor producer fourteen. Saccharomyces cerevisiae U-365 Enzyme producer, interferon inducer

Claims (1)

A probiotic drug for farm animals and poultry containing a mixture of suspensions of bacterial strains and a protective medium, characterized in that it contains a sucrose-gelatin mixture as a protective medium in the following ratio, wt.%: Sucrose 15; gelatin 5; 9% solution of sodium chloride 80, and as a mixture of suspensions of strains contains suspensions of the following strains, wt.%: Azomonas agilisc B-2586 8 Azotobacter chroococcum B-3162 7 Azotobacter vinelandii rekc B-2587 7 Bacillus subtilis B-5225 7 Bacillus subtilis B-4828 9 Bifidum bacterium globosum B-2584 10 Escherichia coli B-4412 8 Enterococcus faecium B-2898 7 Lactobacillus acidophilus B-3488 7 Lactobacillus acidophilus B-2585 7 Propionibacterium freudenreichii B-6561 7 Pseudomonas sp. B-3908 7 Pseudomonas sp. B-2589 7 Saccharomyces cerevisiae y-365 2