RU2723411C2 - Bacillus cereus strain rcam04578, a biologically active compounds producer having probiotic properties - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.SUBSTANCE: invention relates to a Bacillus cereus bacteria strain for producing a probiotic preparation. Strain of Bacillus cereus bacteria (DPRK-17) is deposited in the Departmental collection of useful microorganisms for agricultural purposes (RCAM) under number RCAM04578.EFFECT: invention can be used for production of probiotic preparations for prevention and treatment of gastrointestinal diseases, dysbacteriosis, which can be used together with antibiotics.1 cl, 1 dwg, 5 ex

Description

The invention relates to biotechnology, pharmacology, cosmetology, nutrition, microbiology, agriculture, aquaculture, veterinary medicine and medicine. The bacterial strain Bacillus cereus, deposited in the departmental collection of beneficial microorganisms for agricultural purposes (RCAM) under registration number RCAM04578, is a new non-pathogenic strain of spore-forming bacilli, producer of polysaccharides, a complex of antioxidant enzymes (catalase, superoxide dismutase), digestive enzymes, amylases, lipases (amylases) antibiotic-resistant group of beta-lactams and cephalosporins. The strain can be used as a production strain for the production of probiotic preparations for the prophylaxis and treatment of diseases of the gastrointestinal tract, dysbiosis, which can be used in conjunction with antibiotics.

A strain of microorganisms with beneficial properties has been identified. The non-pathogenic avirulent strain of Bacillus cereus RCAM04578 (DPRK-17) was isolated from cow's milk. The strain is cultivated on nutrient media: nutrient and meat and peptone broth and its various modifications. The strain can be used for fermentation of milk and raw materials to obtain fermented preparations, probiotic preparations and biologically active additives. The invention provides for the production of fermented therapeutic, prophylactic, probiotic and other drugs. The probiotic properties of this strain are to suppress the growth of pathogenic enterobacteria, including enterotoxigenic Escherichia coli and increase the number of lactobacilli and bifidobacteria - representatives of healthy resident intestinal microflora, as well as the immunomodulatory effect and increase growth in farm animals.

The technical result consists in expanding the arsenal of special-purpose drugs (creating fermented preparations and products, immunostimulating drugs, food additives, cosmetics, feed additives, probiotic preparations), in increasing the biological value of the resulting product, as well as in expanding the range of products for dietary and medical preventive nutrition, including products used as feed, feed additives, treats or veterinary preparations for domestic and farm animals (cattle, poultry), aquaculture (fish and crustaceans).

The intestinal bacterial flora of domestic animals plays an important role in the digestion and absorption of animal feed. She takes part in the metabolism of nutrients such as carbohydrates, proteins, lipids and minerals, as well as in the biosynthesis of vitamins. In addition to these physiological effects on nutrition, the bacterial microflora provides protection against certain diseases and infections by inhibiting the growth of pathogenic organisms in the intestines.

Enterobacteria are closely associated with the development of conditions such as colibacteriosis, salmonellosis and clostridial enteritis in domestic animals and birds. A healthy intestinal microflora provides animals with the synthesis of vitamins, help in the absorption and absorption of nutrients, resistance to infections and antagonism towards harmful microorganisms. On the other hand, pathogenic intestinal microflora produce toxic metabolites, cause rotting processes, gas formation and toxinogenesis, which inhibit animal growth, reproductive ability, reduce the nutritional value of feeds due to lack of absorption, and also reduce the ability of animals to resist infection. When the balance of the intestinal bacterial flora is disturbed as a result of stressful conditions, abnormal growth of pathogenic microorganisms begins, which are rare in the normal microflora, and this causes diarrhea and enteritis. Such painful conditions lead to infections with harmful effects on pets, in farm animals productivity and product quality are reduced.

In light of the relationship between the bacterial flora of the intestine and the host animal, maintaining a good balance of the intestinal flora is important for the health of pets and birds, whether it is pets (cats, dogs) or farm animals.

In livestock production in recent years, animals and birds are often bred and maintained in tight colonies or herds to increase economic efficiency. But this often leads to poorer feeding conditions, inadequate nutrition and poor hygiene management due to lack of labor. In addition, this is combined with many other stress factors, which ultimately leads to an imbalance in the intestinal bacterial flora. This is due to the current use of a large number of antimicrobial feed additives, such as antibiotics and synthetic antimicrobial agents to suppress or eliminate harmful microorganisms.

But as a result, there were problems in the field of public health in connection with the presence of antibacterial drugs used in meat, etc., as well as the problem of pathogenic bacterial strains multi-resistant to antibiotics. In addition, it is known that prolonged administration of an antibiotic creates a unique microflora in which the proportion of bacteria resistant to antibiotics increases, and such microflora causes new diseases. In the light of these circumstances, the use of antibiotics to improve animal growth has been banned completely since January 2006 in Europe. Other countries are also seeking to ban or limit the use of antibiotics, and in this regard, safe drugs are being developed that can replace antibiotics or reduce their use.

In some countries of Europe and America, as well as in Japan, probiotics (preparations containing live bacteria) have been used for many years as medicines to normalize the intestines and to treat diarrhea and enteritis in humans. Given their effectiveness, probiotics have been created for the prevention and treatment of diarrhea and enteritis in domestic animals. Several drugs originally intended for humans have been approved and have become commercially available as veterinary drugs.

The active ingredients of probiotics are new bacterial strains that are isolated in nature and selected because of their positive effect on the growth and health of pets. In addition to lactic acid bacteria, there is a group of spore probiotic bacteria that naturally live in soil, on moist grass, and on plant roots, protecting them from infection by pathogenic microorganisms - for example, Bacillus subtilis and Bacillus cereus. They are able to perform a similar function in humans and animals, producing substances in the gastrointestinal tract that inhibit the growth of pathogenic microflora.

The advantages of the bacterial spores that form these bacteria are their resistance to heat, allowing them to be stored at room temperature and in dry form. In addition, these bacteria are able to reach the intestines, since they survive at a low pH of the stomach. The use of probiotic bacterial spores ranges from nutritional supplements to growth stimulants and is used in aquaculture (e.g. shrimp). Probiotic preparations involving some Bacillus species are recommended for use with antibiotics, as these strains are resistant to them (e.g. B. cereus). Bacillus coagulans have been used as adjunctive therapy to relieve rheumatoid arthritis. B. subtilis is associated with a Japanese product known as Natto. The consumption of this product can have a stimulating effect on the immune system and helps to reduce blood coagulation. The secretion of antimicrobials, such as coagulin, amikoumacin and subtilisin, is also found in bacteria of the genus Bacillus. The mechanisms of the probiotic effects of bacteria of the genus Bacillus are based, among other things, on immunomodulation, which occurs by stimulation of the intestinal lymphoid tissue, through the production of cytokines, the competitive exclusion of gastrointestinal pathogens (for example, competition for adhesion sites) and the secretion of antimicrobial substances. Several studies have been conducted on the safety of Bacillus species using animal and vitro models to evaluate the toxicity or side effects of strains. Use of B. subtilis is approved for use as a dietary supplement in Italy and the UK (Olmos 2014).

To this we can add that spore bacteria belonging to the genus Bacillus, such as Bacillus cereus or Bacillus subtilis, are the most powerful producers of digestive enzymes (amylolytic, lipolytic, antithiolytic), bacteriocins and other substances that inhibit the growth of pathogenic enterobacteria and fungi, due to which, the fermented product obtained with their participation will contribute to more efficient digestion of food, normalization of the microflora of the gastrointestinal tract and elimination of representatives of pathogenic microflora, protection against intestinal toxicoinfections, normalization of digestion and stool (Kozasa 1989; Soccol 2010; Olmos 2014).

In Japan, the most widespread among probiotics for farm animals and poultry is the spore probiotic Toyotserin (Toyocerin, Toyo Jozo, Japan), containing spores of non-pathogenic bacteria Bacillus cereus. This microorganism, according to the literature, is referred to as transient microflora and is able to effectively prevent and treat enterocolitis and stimulate the growth of farm animals. A single dose of Bacillus cereus bacteria causes a significant decrease in the number of enterotoxigenic strains of E. coli in the intestines of pigs suffering from diarrhea. At the same time, it increases the amount of healthy resident microflora, such as representatives of the genus Lactobacillus, while increasing body weight and feeding efficiency. In rabbits, Bacillus cereus inhibits the proliferation of pathogenic E. coli variants in the intestines, preventing diarrhea, improves weight gain and improves survival. In an experiment where hens caused heat stress, there was a significant decrease in the number of lactobacilli in the gastrointestinal tract and, as a result, intestinal dysbiosis. Oral administration of Bacillus cereus bacteria as a probiotic prevented a decrease in the number of lactobacilli and stably maintained the intestinal microflora in a normal state even under conditions of heat stress. In addition, it was shown that Bacillus cereus produces various digestive enzymes, such as amylases and proteases, and is also able to synthesize B vitamins. Also, Bacillus cereus led to a decrease in the concentration of ammonia in the intestines and blood. When used as a probiotic for farm animals, Bacillus cereus increased the content of volatile fatty acids, such as propionic acid in ruminal juice, maintained a normal pH of gastric juice, and contributed to the normalization of ruminal function (Kozasa 1989).

Bacillus cereus has also been proposed as a probiotic for fish, including Nile tilapia (Garcia-Marengoni et al 2015). Bacillus cereus improved growth parameters and reduced mortality due to infections caused by Vibrio vulnificus and V. parahaemolyticus in T. trichopterus fish (Subharanjani et al. 2015).

A known strain of Bacillus subtilis 534, which is resistant to polymyxin and is used to obtain the drug "Sporobacterin" for the prevention and treatment of diseases of the gastrointestinal tract, dysbiosis (USSR Patent 1723116, 1988).

However, this strain is sensitive to beta-lactams. This limits the scope of the drug for severe forms of purulent-inflammatory diseases, the treatment of which requires the joint administration of a bacterial drug and an antibiotic.

A known strain of Bacillus subtilis 3H (GISK 248), which has the property of antibiotic resistance to polymyxin and rifampicin and used to obtain the probiotic drug "Baktisporin", which is used in conjunction with antibiotics for the treatment and prevention of dysbiosis, digestive enzyme deficiency, purulent infections, food allergies ( 2067616).

The disadvantage of this strain is the lack of resistance to beta-lactams and cephalosporins, as one of the most commonly prescribed groups of antibiotics.

In the present invention, the isolated bacterial strain Bacillus cereus RCAM04578 (DPRK-17) is natural and non-pathogenic, does not require gene modification. It is safe for animals, as demonstrated by tests on mice and chickens. This strain can be used as an oral probiotic supplement that normalizes the intestinal microflora (reduces the number of enteropathogenic microflora and increases the number of healthy microflora representatives - bifidobacteria and lactobacilli). It has an immunomodulatory effect. Due to the resistance of the strain to antibiotics of the beta-lactam, cephalosporins and carbapenems group, this strain can be used as a probiotic supplement during antibiotic therapy for the prevention or treatment of dysbiosis.

The objective of the invention is to obtain a non-pathogenic strain of bacteria Bacillus cereus RCAM04578 (DPRK-17) with useful properties that is resistant to beta-lactams and cephalosporins and produces a complex of biologically active compounds in the nutrient medium and is intended to create various therapeutic, prophylactic drugs or additives (immunostimulating , probiotic and other drugs) and fermented products.

The strain was identified using 16S rDNA sequencing on an AE3000 automatic sequencer, a comparison of sequences and the construction of kinship trees based on GosNIIgenetika. When sequencing the variable regions of 16S rDNA, the following assembled nucleotide sequence was obtained for the studied strain:

Initial screening using the GenBank and RDP-II database showed that the strain studied belongs to the following systematic Bacteria groups; Firmicutes Bacilli; Bacillales; Bacillaceae; Bacillus, moreover, the homology with some species of the genus Bacillus is 99%. The sequences were aligned with the corresponding sequences of the nearest bacterial species available from the GenBank database.

The results of processing sequences using a computer program located on the RDB II website (Ribosomal Database Project II), designed to determine the affinity of microorganisms and build phylogenetic trees, are presented in graphical form. Further analysis by the RDP II 16S rRNA database showed homology with the same bacterial species.

According to the analysis, a phylogenetic tree was constructed (see Fig. 1) with homologous strains.

S000005079 0.984 0.980 1395 Bacillus thuringiensis; WS 2617; Z84585 S002291145 0.985 0.984 1465 Bacillus cereus (T); ATCC 14579; AE016877 S004061999 0.985 0.963 1465 Bacillus toyonensis (T); BCT-7112; CP006863 S000006577 0.985 0.981 1395 Bacillus thuringiensis; WS 2626; Z84588 S000008663 0.985 0.978 1397 Bacillus anthracis; Sterne; AF290552 S000012518 0.985 0.981 1397 Bacillus anthracis; Vollum; AF290553 S000015508 0.984 0.980 1404 Bacillus cereus; ATCC14579; AF290547

Where unknown is the test strain

The criterion for classifying a microorganism to a particular species is considered to be at least 97% homology. According to this criterion, the studied strain can be attributed to the species: Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis.

An analysis of phylogenetic kinship constructed using typical strains of closely related bacteria showed that the species Bacillus cereus is the closest to the studied strain.

Additionally, identification was carried out by sequencing the sequence of the ITS region (Lasseter et al., 2009; Pelucchi et al., 2012), as well as biochemical and morphological parameters (Yoshino et al., 2010; Lancefield, 1934).

According to the results of the analysis of sequencing of variable regions of genes encoding 16S rRNA, the test strain is closest to the species Bacillus cereus (99%).

Sequenced nucleotide sequence of the ITS region:

The result of comparing the obtained nucleotide sequence with the ITS region of typical strains (for species B. thuringiensis and B. cereus):

B. thuringiensis type strain ATCC 10792T

Query cover - 98%

Identity - 99.7%

B. cereus type strain ATCC 14579T

Query cover - 98%

Identity - 99.3%

When culturing strain RCAM04578 (DPRK-17) on MPA for 24 hours, the formation of grayish-whitish, flat, fine-tuberous, waxy colonies with a wavy edge with a diameter of 1.5-5 mm was noted. Light microscopy showed the presence in the culture of rod-shaped cells (1-1.5 × 3-4 μm), single or located in pairs and chains, motile, forming endospores located in the center of the cells. The morphological features of cells and colonies of strain RCAM04578 (DPRK-17) indicate its belonging to the species B. thuringiensis or B. cereus. Comparison of the ITS region of strain RCAM04578 (DPRK-17) with homologous sequences of the typical B. thuringiensis and B. cereus strains (99.7 and 99.3% similarity, respectively) suggested that the studied strain is closer to the B. thuringiensis species. However, taking into account the results of studying the 16S rRNA gene and the absence of endotoxin crystals in the cytoplasm that are characteristic of B. thuringiensis, strain RCAM04578 (DPRK-17) can be designated as Bacillus cereus.

The strain of Bacillus cereus RCAM04578 (DPRK-17) has the following characteristics:

1. Cultural and morphological:

- optional anaerobic;

- gram-positive sticks;

- length 600-800 nm;

- mobile;

- single or arranged in chains;

- form an endospore located in the center of the cell;

The study of the cultural or morphological properties of the selected strain was carried out on a solid nutrient medium - nutrient agar. On the surface of dense nutrient media after 24 hours at a temperature of 37 ° C form: a grayish-whitish colony, flat, finely tuberous, waxy with a wavy edge, with a diameter of 1.5-5 mm.

2. Physiological and biochemical:

- the strain ferments glucose.

- grows at a temperature of 37 ° C;

- grows in meat and peptone broth at pH 7.3;

- the strain is resistant to oxacillin (resistance to beta-lactams, cephalosporins, carbopenems). Sensitive to ciprofloxacin, tetracycline, erythromycin, azithromycin, vancomycin, clindamycin, amikacin, nitrofurantoin. It is sensitive to antibiotic groups tetracyclines, macrolides, fluorchionols, lincosamides, aminoglycosides, nitrofurans.

- produces enzymes of the antioxidant system: catalase, superoxide dismutase, peroxidase, reduced glutathione;

- produces proteolytic, amylolytic, lipolytic enzymes;

- produces lipopolysaccharides (LPS).

The method, conditions and composition of the media for long-term storage of the strain - lyophilization; in test tubes with nutrient broth at a temperature of from -15 to -200 ° C for long-term storage; culture tubes stored at a temperature of + 4-8 ° C for 3 months.

The method, conditions and composition of the media for the propagation of the strain at a temperature of 37 ° C; environments: various options of nutritious broth.

The proposed new strain, easy to cultivate and store, is not pathogenic to humans, animals and plants.

Example 1. Culture passage: Bacillus cereus strain RCAM04578 (DPRK-17) was introduced into tubes with nutrient broth and placed in a thermostat at 37 g. for 24 hours until a precipitate forms. At the end of the fermentation process, the nutrient medium is saturated with biologically active metabolic products and spores of this bacterial strain. The quantification of the enzyme antioxidant enzymes (catalase and superoxide dismutase) was carried out according to the methods (Morozov et al., 2018).

Example 2. Determination of the parameters LD50, acute and chronic toxicity: Determination of the LD50 for the strain of Bacillus cereus RCAM04578 (DPRK-17) was carried out according to the method of Reed-Mench on 200 white mice. The LD50 when administered orally to white mice is 8.95 * 10 ¹⁵ CFU / kg, which indicates the safety of this strain for humans and animals when administered orally at the safety level of probiotic strains of lactobacilli and bifidobacteria. To study acute and chronic toxicity, a preparation containing the Bacillus cereus strain RCAM04578 (DPRK-17) was used. To study acute toxicity, a preparation with a concentration of microbial bodies in 1 ml of -1.7 × 10 ⁶ CFU was used. administered to white mice orally with a gastric tube in a volume of 0.3-, 0 ml.

In an experiment to determine acute toxicity, the finished drug was administered intragastrically, taking into account the maximum volume of a single injection. The maximum attainable dose at a single dose was 1.7 * 10 ⁷ CFU / kg, and with a triple fractional dose it was 5.1 * 10 ⁷ CFU / kg. For the experimental determination of acute toxicity, nonlinear mice and males were used, with an initial weight of 24-30 g; in total, 130 laboratory animals were used in the experiment. Laboratory animals were divided into 7 groups, ten animals in each group, and 50 animals were used for preliminary selection of doses. And 10 goals control group.

The chronic toxicity of a preparation containing a strain of Bacillus cereus RCAM04578 (DPRK-17) was studied on outbred mice, males, with an initial weight of 13-16 g, the total number of animals in this series is 40. The preparation is intragastric, the duration of administration is 3 months or 90 days.

During the experiments, their behavior, appearance, physical activity, and reaction to external stimuli were monitored. The clinical picture of "acute" poisoning of laboratory animals was recorded with the introduction of the studied drugs in the highest achievable doses.

The following parameters of the functional state of laboratory animals were recorded daily: Activity - lethargy, normal activity, increased activity. Movement - impaired coordination, muscle tone, tremor.

Appearance: Condition - obesity, normal, exhaustion, cachexia. Wool - shiny, dull, smooth, sticking out, hair loss. Eyes - lacrimation, inflammation, clouding of the cornea, adhesion. Ears - color - normal, pallor, redness; inflammation, secretion, scab formation, twitching. Teeth - color, breakage, loss. Extremities - color, swelling.

Physiological functions: Respiration - rapid, normal slow, wheezing, wheezing, labored. Salivation - insufficient or excessive. Saliva is watery or clammy. Urine is the color. Excretion - color, quality.

Weighed animals, which allowed us to determine the dynamics of body weight of laboratory animals in the experiment. Hematological and biochemical parameters were also determined.

With oral administration of the test samples for 24, 48 hours, 14 days, the death of animals in all groups was not observed. If it is not possible to determine the LD50, the maximum dose of the test drug is administered to animals in our studies, 112,000 mg / kg. According to the toxicity tabulation, for single use according to Hodge and Sterner, 1943, this indicator corresponds to class VI - is relatively harmless.

In accordance with GOST 12.1.007-76 with the subcutaneous and intraperitoneal method, it belongs to hazard class 4 - low toxicity.

Considering that the threshold dose (single action threshold) is the smallest amount of a substance that causes, in a single exposure, such changes in the body that are detected by biochemical or physiological tests in the absence of external signs of poisoning, in our studies was 45,000 mg / kg - the drug, according to the given classification can be attributed to class VI - relatively harmless.

Example 3. The effect on the intestinal microflora. The use of the Streptosubtil preparation containing Bacillus cereus RCAM04578 (DPRK-17) has a positive effect on the intestinal microbiocenosis of broiler chickens, increasing the amount of beneficial microflora and reducing the level of conditionally pathogenic. When using the drug with food, the number of representatives of the genus Bifidobacterium increases by 144 times compared with the control. When drinking the drug through the drinking system, the number of representatives of the genus Bifidobacterium increased by 9.5 times, and microorganisms of the genus Lactobacillus - by 4.3 times compared with the control. Also in the experimental groups, a decrease in the level of conditionally pathogenic microflora belonging to the group of Escherichia coli (BHEC) was noted - in the first experimental (with food) by 1.84 times, in the second (through the drinking system) - by 3.72 times compared with control.

Example 4. The effect on weight gain of animals. The use of the Streptosubtil preparation containing Bacillus cereus RCAM04578 (DPRK-17) at a dose of 10 ml / kg live weight for 40 days has a positive effect on the increase in live weight of broiler chickens. When using this drug with food, the average daily gain in live weight when used with food was 58.57 grams, which is 5.68% higher than the control value. With the introduction of the drug into the drinking system from the first day of life of chickens, this figure was 57.94 g. (2.85% higher than control). Thus, the use of a preparation containing Bacillus cereus RCAM04578 (DPRK-17) has a positive effect on the increase in live weight of broiler chickens. A positive effect is observed both when using this drug with food, and when introducing it into the drinking system from the first day of life of chickens.

Example 5. Immunostimulating effect: In animals treated with Streptosubtil containing Bacillus cereus RCAM04578 (DPRK-17), in a dose of 28,000 to 112,000 mg / kg once, for 14 days the number of leukocytes in the circulating blood is higher than in the control group due to the lymphocytic fraction, which indicates a stimulating effect on leukopoiesis, lymphopoiesis.

Literature:

1. USSR patent 1723116, 1988

2. RU 2067616. STRAIN OF BACTERIUS BACILLUS SUBTILIS, BEARING THE PROPERTIES OF ANTIBIOTIC RESISTANCE USED TO OBTAIN THE PRODUCT "BAKTISPORIN"

3. Garcia-Marengoni N., et al. Use of probiotic Bacillus cereus var. toyoi and Bacillus subtilis C-3102 in the diet of juvenile Nile tilapia cultured in cages \\ Lat. Am. J. Aquat. Res. 2015.43 (3). Pp. 601-606.

4. Kozasa Probiotics for animal use in Japan \\ Rev. sci. tech. Off. int Epiz., 1989, 8 (2), 517-531.

5. Subharanjani S. Supplementation of B. cereus as probiotic in fish feed of Trichogaster trichopterus (Blue Gourami) and calculating its growth and survival \\ Int.j. Curr.Microbiol.App.Sci. 2015.4 (12). Pp. 744-751.

6. Carlos Ricardo Soccol, Luciana Porto de Souza Vandenberghe, Michele Rigon Spier, Adriane Bianchi Pedroni Medeiros, Caroline Tiemi Yamaguishi, Juliano De Dea Lindner, Ashok Pandey and Vanete Thomaz-Soccol The Potential of Probiotics: A Review \\ C.R. SOCCOL et al .: The Potential of Probiotics, Food Technol. Biotechnol. 48 (4) 413-434 (2010)

7. Jorge Olmos and J Paniagua-Michel Bacillus subtilis A Potential Probiotic Bacterium to Formulate Functional Feeds for Aquaculture \\ J Microb Biochem Technol 6: 361-365. doi: 10.4172 / 1948-5948.1000169

8. Morozov, A.A., Chuiko, G.M., Yurchenko, V.V. Annual variations in hepatic antioxidant defenses and lipid peroxidation in a temperate fish, common bream Abramis brama (L.) \\ International Aquatic Research Volume 9, Issue 3, 1 September 2017, Pages 249-257

9. Report: Identification of strain DPRK-17 to a species using 16S RNA analysis. Date: 05/04/2017. All-Russian collection of industrial microorganisms FSBI "GosNIIgenetika" Ministry of Science. Director of the BRC VKPM, Professor Sineokiy S.P.

10. Report on the identification of the microorganism strain Bacillus sp. using the sequencing method of the ITS region under the contract No. 27/2017 / 10n of 06/19/2017. FEDERAL AGENCY OF SCIENTIFIC ORGANIZATIONS Federal State Budget Scientific Institution "All-Russian Scientific Research Institute of Agricultural Microbiology" (FGBNU VNIISCHM). Responsible Contractor: Head of Laboratory No. 10, Ph.D. IN AND. Safronova.

11. Report on the second stage of research work under the contract No. 10-779 / 17-N on the topic: To study the pathogenic properties of Bacillus cereus DPRK-17. INSTITUTE OF EXPERIMENTAL VETERINARY OF SIBERIA AND THE FAR EAST SIBERIAN FEDERAL SCIENTIFIC CENTER FOR AGRIBIOTECHNOLOGIES OF THE RUSSIAN ACADEMY OF SCIENCES. Responsible Executive: Senior Researcher laboratories of diseases of young animals, candidate of veterinary sciences V.Yu. Koptev. Novosibirsk 2017

12. Report on research work under the contract No. 10-779 / 17-Н on the topic: Conduct preclinical trials and study the effect of Streptocvasha preparation on the body of chickens. INSTITUTE OF EXPERIMENTAL VETERINARY OF SIBERIA AND THE FAR EAST SIBERIAN FEDERAL SCIENTIFIC CENTER FOR AGRIBIOTECHNOLOGIES OF THE RUSSIAN ACADEMY OF SCIENCES. Responsible Executive: Senior Researcher laboratories of diseases of young animals, candidate of veterinary sciences V.Yu. Koptev. Novosibirsk 2018

Claims (1)

The bacterial strain Bacillus cereus RCAM04578 (DPRK-17), deposited in the "Departmental collection of beneficial microorganisms for agricultural purposes" (RCAM) - designed to obtain a probiotic preparation.

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