Bacillus amyloliquefaciens bkm b-2714d strain having apparent antagonism in relation to salmonella typhi, staphylococcus aureus, listeria monocytogenes and resistance to tetracycline and trimethoprim. RU patent 2509148.
 Dry chromogenic feed medium for detection of coliform bacteria and e.coli (versions) / 2508400Invention can be used for detection of coliform bacteria and E.coli in specimens of food products and water at performance of bacteriological tests. Feed medium includes a nitrogen source represented by meat peptone or pancreatic hydrolysate of fish flour, sodium chloride, dibasic sodium phosphate, potassium monophosphate, sodium pyruvate, L-tryptophane, sodium dodecyl sulphate, 6-chloro-3-indolyl-β-D-galactopyranoside (Salmon - GAL), 5-bromine-4-chloro-3-indolyl-β-D-glucoronide-(X-GLUC), isopropyl- β-D1-tiogalactopyranoside (IPTG) and microbiological agar in the specified ratio. |
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Dry differential diagnostics feed medium for detection and consideration of e.coli and coliform bacteria / 2508399 Invention can be used for detection and considering of E.coli and coliform bacteria in water, food products, clinic material, etc. Feed medium contains pancreatic hydrolysate of fish flour dried with Tergitol 7 on the bases of 0.1 g of Tergitol 7 per 6 g of dry pancreatic hydrolysate of fish flour, yeast extract, 1-water D (+) lactose, bromthymol blue, sodium dodecyl sulphate, 2,3,5-triphenyltetrazolium chloride, sodium carbonate and microbiological agar in the specified ratio. |
 Strain of chlorella vulgaris microalgae for obtaining lipids as raw material for production of motor fuel / 2508398Invention propose a strain of Chlorella vulgaris IPPAS C-616 microalgae for obtaining lipids as raw material for production of motor fuel. |
 N-glycan from c jejuni presented in salmonella enterica and its derivatives / 2507253Invention refers to genetically modified bacteria Salmonella enterica, which contain at least one operon pgI from Campylobacter jejuni or its functional derivative and refers to presentation of at least one N-glycan from Campylobacter jejuni or derivative of this N-glycan on their cell surface. In bacteria one or several genes for biosynthesis of bacillozamine are inactivated by mutation and/or partial or complete deletion of genes pgID, E, F, G. Invention provides method for obtainment of genetically modified bacteria Salmonella enterica. Invention is directed to application of modified bacteria in pharmacological compositions of medical and veterinary purpose and methods for treatment and/or prevention of infections Campylobacter and, not obligatory, Salmonella. |
 Glarea lozoyensis mutant strain and its application / 2507252Mutant strain is obtained by impact on Glarea lozoyensis ATCC 20957 strain by nitrosoguanidine and it is deposited in CGMCC with number CGMCC 2933. |
 Method for obtaining biomass of green microalgae, enriched with fatty acids / 2507251Inoculum of microalgae Desmodesmus sp. strain 2C166E is introduced into mineral medium BG-11 till final concentration of chlorophyll in mixture of 4-6 mcg/ml. Incubation is performed under constant lighting and environment barbotage by atmospheric air during 12-16 days under temperature of 25-27°C with consequent separation of microalgae biomass from nutritional medium with obtainment of microalgae biomass containing 33-35% of fatty acids from dry weight of cells. |
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Method of identifying vibrio bacteria / 2506313 Invention relates to microbiology and biotechnology. Material to be investigated - pure culture of rod-like, gram-negative, glucose-fermenting, oxidase-positive or oxidase-negaive bacteria - is collected first. The investigated daily bacterial culture is seeded on the surface of nonselective nutrient agar (GRM-agar) with 1% sodium chloride. A paper disc is then placed seeded surface, said disc containing vibriostatic substance niclosamide (2,5-dichloro-4-nitrosalicylanilide) in amount of 10 mcg or 16 mcg per disc. The seeded material is incubated in aerobic conditions at 35°C for 24 hours. Vibrio bacteria are indicated a zone of inhibited bacterial growth around the disc. |
 Consortium of probiotic strains of lactobacillus rhamnosus and lactobacillus plantarum for producing bacterial preparation and direct administration ferment for producing fermented milk and fermented beet juice / 2506308Invention relates to microbiology and biotechnology. Disclosed is a consortium of Lactobacillus rhamnosus VKM B-2726D and Lactobacillus plantarum VKM B-2725D strains. |
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Starter material of pure growth of lactate microorganisms for preparation of cultured milk products / 2505601 Strain Lactococcus casei VKPM V-8730 is obtained at available growth-supporting media and is used as starter material in cultured milk products production. |
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Starter material of pure growth of lactate microorganisms for preparation of cultured milk products / 2505600 Strain Enterococcus durans VKPM B-8731 is obtained at available growth-supporting medium and is used as starter material in cultured milk products production. |
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Dry chromogenic feed medium for detection of coliform bacteria and e.coli (versions) / 2508400 Invention can be used for detection of coliform bacteria and E.coli in specimens of food products and water at performance of bacteriological tests. Feed medium includes a nitrogen source represented by meat peptone or pancreatic hydrolysate of fish flour, sodium chloride, dibasic sodium phosphate, potassium monophosphate, sodium pyruvate, L-tryptophane, sodium dodecyl sulphate, 6-chloro-3-indolyl-β-D-galactopyranoside (Salmon - GAL), 5-bromine-4-chloro-3-indolyl-β-D-glucoronide-(X-GLUC), isopropyl- β-D1-tiogalactopyranoside (IPTG) and microbiological agar in the specified ratio. |
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Dry differential diagnostics feed medium for detection and consideration of e.coli and coliform bacteria / 2508399 Invention can be used for detection and considering of E.coli and coliform bacteria in water, food products, clinic material, etc. Feed medium contains pancreatic hydrolysate of fish flour dried with Tergitol 7 on the bases of 0.1 g of Tergitol 7 per 6 g of dry pancreatic hydrolysate of fish flour, yeast extract, 1-water D (+) lactose, bromthymol blue, sodium dodecyl sulphate, 2,3,5-triphenyltetrazolium chloride, sodium carbonate and microbiological agar in the specified ratio. |
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Method for obtaining actinobacillus pleuropneumonia apxi or apxiii toxins in liquid culture media supplemented by air enriched with carbon dioxide / 2507267 Invention relates to method for obtaining of RTX-toxins Apxl or ApxIII by culturing of Actinobacillus pleuropneumoniae bacteria in liquid culture media. Characterised method consists in the following: during exponential growth phase of bacteria and production of RTX-toxins air passes through the medium, carbon dioxide content in air is above normal atmospheric level and is up to 10 % vol. |
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N-glycan from c jejuni presented in salmonella enterica and its derivatives / 2507253 Invention refers to genetically modified bacteria Salmonella enterica, which contain at least one operon pgI from Campylobacter jejuni or its functional derivative and refers to presentation of at least one N-glycan from Campylobacter jejuni or derivative of this N-glycan on their cell surface. In bacteria one or several genes for biosynthesis of bacillozamine are inactivated by mutation and/or partial or complete deletion of genes pgID, E, F, G. Invention provides method for obtainment of genetically modified bacteria Salmonella enterica. Invention is directed to application of modified bacteria in pharmacological compositions of medical and veterinary purpose and methods for treatment and/or prevention of infections Campylobacter and, not obligatory, Salmonella. |
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Consortium of probiotic strains of lactobacillus rhamnosus and lactobacillus plantarum for producing bacterial preparation and direct administration ferment for producing fermented milk and fermented beet juice / 2506308 Invention relates to microbiology and biotechnology. Disclosed is a consortium of Lactobacillus rhamnosus VKM B-2726D and Lactobacillus plantarum VKM B-2725D strains. |
 Bacillus subtilis bacteria strain having high level of production of phytase (versions), animal feed composition and method of feeding animals / 2506307Invention relates to biotechnology. Disclosed are Bacillus subtilis DSM 19467, Bacillus subtilis DSM 19489 and Bacillus subtilis DSM 19466 strains which produce high levels of phytase. An animal feed composition is obtained based on any of said strains coupled with other suitable additives. Also disclosed is a method of feeding animals, which involves serving the composition to animals along with other feed ingredients. |
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Starter material of pure growth of lactate microorganisms for preparation of cultured milk products / 2505601 Strain Lactococcus casei VKPM V-8730 is obtained at available growth-supporting media and is used as starter material in cultured milk products production. |
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Starter material of pure growth of lactate microorganisms for preparation of cultured milk products / 2505600 Strain Enterococcus durans VKPM B-8731 is obtained at available growth-supporting medium and is used as starter material in cultured milk products production. |
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Application of l casei ssp paracasei as antimycotic agent / 2505599 Invention pertains to the use of L. casei ssp. paracasei CNCM 1-1518 strain for giving antimycotic properties to fermented dairy product. Inhibiting of mould development of ascomycete class in the said product is made by dairy substrate fermentation in presence of this strain for, at least, 5 hours. Thallome diameter after 7 days of incubation amounts 8.8±1 mm. |
 Probiotic strains lactobacillus and their consortium for prophylaxis and treatment of urogenital infectious diseases of females / 2504580Group of inventions refers to microbiology and biotechnology. Proposed strains Lactobacillus crispatus "ВКМ" B-2727D, Lactobacillus gasseri "ВКМ" B-2728D and Lactobacillus plantarum "ВКМ" B-2731D have antagonistic activity in relation to pathogenic and semi-pathogenic microorganisms. Besides, the invention proposes a consortium based on the above strains for production of a bacterial preparation, a biologically active additive, and direct inoculation for obtaining fermented milk product of functional nutrition. |
 Strain of lactobacilli lactobacillus paracasei cncm i-2116 (ncc 2461) eliciting ability to prevent intestine colonization with pathogenic microorganisms causing diarrhea, supernatant of its culture and oral agent for prophylaxis and/or treatment of diarrhea-associated disorders / 2243779Invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea. |
FIELD: biotechnologies.
SUBSTANCE: bacillus amyloliquefaciens BKM B-2714D strain has apparent antagonism in relation to Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and resistance to tetracycline and trimethoprim antibiotics. It is deposited in the All-Russia Collection of Microorganisms of the Institute of Biochemistry and Physiology of Microorganisms Named after G.K. Skryabin of the Russian Academy of Science (IBFM RAN) and has the following registration number: BKM B-2714D, and can be used at production of probiotic bacterial preparations that can be used in veterinary medicine.
EFFECT: invention allows increasing protease and xylanase activities.
1 tbl, 3 ex
The invention refers to biotechnology, namely to the probiotic bacterial drugs, which may find application in veterinary medicine.
Probiotic are bacterial preparations based on strains of microorganisms possessing antagonistic activity to enteropathogenic bacteria.
Classical studies Iaeneea these properties were studied on the example of lactic acid bacteria. Then the circle of producers of probiotic preparations have been expanded.
Probiotics based strains of spore-forming bacteria relatively recently started to be used in veterinary medicine. Antagonistic properties of these bacteria attracted the attention of many developers.
The complexity of the treatment of infectious diseases is the emergence of pathogenic microorganisms of polirezistentnami to antibiotics. Moreover, antibiotics often had an adverse effect on the organism of animals and cause intestinal dysbacteriosis, aggravating the course of the basic disease. This causes the need for new approaches to solving this problem.
Drugs-probiotics by bacteria of the genus Bacillus that suppress the development of pathogenic microorganisms are widely used in agriculture for the suppression of phytopathogenic bacteria and fungi in medicine, veterinary medicine for treatment of various diseases of birds and farm animals [1].
Currently, preparations, probiotics are bacteria of the genus Bacillus, are widely used in veterinary medicine for treatment of various diseases of birds and farm animals. Proposed numerous variants of probiotics of spore-forming bacteria: backspin [5], bacilloscopy [6], sporoloth [7], sporobacterin [8], subalin [9]. The basis of the above drugs - probiotics are various strains of bacteria of the genus Bacillus with antagonism against pathogenic bacteria.
The overwhelming majority of representatives of the genus Bacillus is harmless to mammals, are antagonistic activity towards a wide spectrum of pathogenic and conditionally pathogenic microorganisms resistant to lytic enzymes, technological production, stable in storage, is ecologically safe.
Given the possibility of the use of antibiotics in feed production, a very promising application in structure of probiotic preparations strains with as high antagonistic activity against enteropathogen and MDR to antibiotics.
Besides preparations based on Bacillus subtillis and Bacillus licheniformis developed preparations based on Bacillus amiloliquefaciens.
As a prototype selected strain of Bacillus amiloliquefaciens SISTER-5950, which is the basis of the drug ESOPS 10].
The object of the present invention to provide a strain Bacillus amiloliquefaciens, which has antagonistic activity towards a wide spectrum of pathogenic test-organisms, as well as the resistance to common antibiotics.
Technical effect that can be obtained by using the obtained strain, is to enhance the effectiveness of treatment, due to the combined use of bacterial medication with antibiotics.
To solve this problem, a strain of Bacillus amiloliquefaciens VKM B-2714D. Strain deposited at the all-Russian collection of microorganisms.
Identification of the strains was performed by «the Determinant of microorganisms» Berga (1997).
The original soil isolates selected in the Kirov region. Soil isolate was subjected mutagenic effect of nitrosoguanidine (dose of 40 to 100 mcg/ml) with subsequent screening on selective media.
Cultural morphological and microscopic features of strain : rod-shaped cells direct the size of 0,5-0,7 x 2.0-3.0 mm, form a heat-resistant endospores, no more than 1 in cell-sporangium, the sporulation is not inhibited by exposure to air, the location is Central. Cell mobile. Standard dense nutrient media (MPA, LB, RM-1) forms white crater colony. Catalase: polozitelnye, aerobe. Used sources of carbon for aerobic growth: glucose, arabinose, xylose, lures, dextrose, casein, starch. The optimum pH for the growth of 6.0-8.0, the temperature is from +4 C to +45 C the Resistance to antibiotics: tetracycline - 20 mcg/ml trimethoprim - 100 ug/ml.
The invention is illustrated by the following examples.
Example 1. The strain is characterized by high antagonistic activity against pathogenic and conditionally pathogenic microorganisms. Antagonistic activity was examined by the method of deferred antagonism.
For this suspension cells washed with agar medium, sown stroke in diameter Petri dishes with agar medium Gause N 2. The crops were incubated in thermostat at 37 C for 48 hours Then grown culture was podesavati stroke test-organisms (500 - million suspension subsistence crops physiological solution). Records of the results of conducted through 18-20 hours of incubation at 37 C largest areas absence of growth of the test cultures. The control of the growth of the test cultures served parallel growing them on cups with agar medium Gause N 2 without the studied culture. The results are shown in table.
Stunting test cultures, mm
E.coliS.typhi-murium
S.typhiPs.aeruginosa
St.aureus
L.monocytogenes
0 0 14 0 9 13Example 2. They studied the resistance of the following antibiotics: kanamycin, streptomycin, gentamicin, trimethoprim, nalidixic acid, Cefazolin, ampicillin and benzylpenicillin sodium salt. The test was carried out at the deep cultivation of a producer on glucose peptone medium with yeast extract at various concentrations of antibiotics. It was found that the strain has resistance to antibiotics: tetracycline - 20 mcg/ml trimethoprim - 100 ug/ml.
Example 3.
The enzyme activity in culture fluid defined in submerged cultivation of a producer on glucose peptone medium with yeast extract in two days. The following were identified values of the enzymatic activity:
α-amylase activity - 0.09 IU/ ml of medium at pH 6.0, xilanaza activity - 2.0 IU/ml of medium at pH 7.0, proteaza activity - 2.143 IU/ml of medium with pH 7.0.
It should be noted high protease and xylanase activity strain Bacillus amiloliquefaciens VKM B-2714D.
Sources of information
Now CENTURIES, Reznik S.R., Vyunitsky V.O. Modern understanding of the mechanisms of therapeutic-and-prophylactic action of the probiotic bacteria of the genus Bacillus. // Microbiol. Ukr. - 1993. - №4. - pp.92-112.
2. Bakulina L. F., Timofeyev I.V., N.G. Perminova, Polushkina A.F., Pechurkina N.I. Probiotics by the spore-forming microorganisms of the genus Bacillus and their use in veterinary medicine. // Biotechnology. 2001, №2, p.48-56.
3. Hong NA, Due L.H., Cutting with The use of bacterial spore formers as probiotics .// FEMS Microbiol.Rev. - 2005. - Vol.29, N 4. - P. 813-835.
4. Probiotics Microbes: The Scientific Basis / R.J. Collier, et al. // A Report from the American Academy of Microbiology Washington, 2006. - 28 p.
5. RF patent 2130316. Therapeutic and preventive biological product, backspin. A61K 35/74, BI № 14, 1999, prior. 23.01.97.
6. Patent USSR 1710575. A strain of bacteria Bacillus sp - component therapeutic-preventive medication against dysbacteriosis and allergies, 12 N1/20, BI №5, 1992.
7. RF patent 2035186. Preventive biological sporoloth. A61K 35/66, BI №14, 1995.
8. Aleshkin, A.V., Afanasiev, S., Davydkin, VY, Manko, I.E., Davydkin, YOU Bacterial preparation of new generation «Sporobacterin». // Honey. card index. - 2003. - №4
9. RF patent №2035185 Preventive biological preparation of Subalin. -1995 Prior. 31.01.1992.
10. Haipeng Cao, Shan't, Ruopeng Wei, Marek Diong, Liqun Lu. Safety and efficacy of Ecobiol (Bacillus amiloliquefaciens) as feed additive for chickens for fattening. The EFSA Journal (2008) 773, 1-13.
The strain of Bacillus amyloliquefaciens VKM B-2714D, with the strong antagonism against Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and resistance to antibiotics tetracycline and trimethoprim.