Bacillus licheniformis strain having apparent antagonism in relation to salmonella typhi, staphyloccus aureus, listeria monocytogenes and resistance to streptomycin and nalidixic acid. RU patent 2501851.
 Lactobacillus paracasei subspecies paracasei strain having antimicrobial and immunomodulating properties, and food product on its basis / 2501850Strain is deposited in CNCM and has number 1-3689. Strain has ability to inhibit the growth of pathogenic microorganisms in culture, and namely Escherichia coli, Salmonella enteritidis and Listeria monocytogenes. Strain has anti-inflammatory properties at the ratio of IL-10/IL-12 equal to 11.8. |
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Bacillus licheniformis vkm b-2713d strain having apparent antagonism in relation to salmonella typhi, staphyloccus aureus, listeria monocytogenes and resistance to streptomycin and nalidixic acid / 2501849 Bacillus licheniformis VKM B-2713D strain has apparent antagonism in relation to Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes. Strain has resistance to antibiotics at streptomycin concentration of 20 mcg/ml and at concentration of nalidixic acid of 20 mcg/ml. Strain has α-amylase activity of 0.015 U/ml of the medium at pH 6.0, protease activity of 2.34 U/ml of the medium at pH 7.0. |
 Method for obtaining recombinant core protein of hepatitis e virus and recombinant vaccine for prophylaxis of hepatitis e virus / 2501809Invention proposes a method for obtaining recombinant core protein of hepatitis E virus (rtHEV-ORF2) and recombinant vaccine for prophylaxis of hepatitis E virus. Core protein is obtained by cultivation of recombinant yeast strain Hansenula polymorpha "КБТ"-11/pHEV-001, which contains DNA sequence integrated into genom of yeast cell and coding the fragment of amino-acid sequence from position 86 to 607 of core protein of hepatitis E virus of genotype 3 (rtHEV-ORF2) under control of promoter of MOX gene. The method allows obtaining immunogenic antigene of hepatitis E virus, which has properties of natural protein. Based on the obtained antigene there created is recombinant vaccine for prophylaxis of hepatitis E virus. Vaccine includes effective amount of rtHEV-ORF2 protein, adjuvant and a physically acceptable diluter. |
 Recombinant bacillus licheniformis bacteria strain-producer of heat-resistant lipase / 2500812Invention represents Bacillus licheniformis bacteria strain All-Russian collection of industrial microorganisms B-11302 - producer of heat-resistant lipase. At fermentation of the obtained strain in 3 l of a laboratory fermenter the ferment activity in culture liquid reaches the level of 500 U/ml. |
 Recombinant bacillus subtilis bacteria strain-producer of specific phospholipase / 2500811At fermentation of obtained strain in 3 l of a laboratory fermenter in minimal medium at the temperature of 30°C, ferment activity in culture liquid reaches the level of 210 U/ml. |
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Production method of keratinase from penicillium citrinum / 2499833 Production method of keratinase provides for directed adaptation of strain Penicillium citrinum PC-54-91"ВИЛАР" by three-time subcultivation on Czapek agar medium containing 2% of hair keratin as a carbon source. Cultivation under deep conditions on Czapek medium containing 10% of hair keratin and 0.5% of saccharose during 6 days. Separation of biomass from culture fluid with further extraction of target product by two-stage chromatographic cleaning involving gel filtration on TSK-Gel TOYOPEARL HW-40 and affine chromatography on protein A to sepharose CL-4B with further lyophilisation. |
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Production method of complex feed preparation with lysin producer based on corynebacterium glutamicum / 2499829 Liquid culture of strain Corynebacterium glutamicum All-Russian collection of industrial microorganisms B-1959 - lysine producer by means of a deep method and liquid cultures of strains Bacillus subtilis All-Russian collection of industrial microorganisms B-8130, Bacillus subtilis All-Russian collection of industrial microorganisms B-2984, Bacillus subtilis All-Russian collection of industrial microorganisms B-4099 and Bacillus licheniformis All-Russian collection of industrial microorganisms B-4162 are prepared. To liquid culture of strain Corynebacterium glutamicum All-Russian collection of industrial microorganisms B-1959 there added in quantity of 30 l is 35 l of the mixture consisting of liquid cultures Bacillus subtilis All-Russian collection of industrial microorganisms B-8130, Bacillus subtilis All-Russian collection of industrial microorganisms B-2984 and Bacillus subtilis All-Russian collection of industrial microorganisms B-4099, which have been taken in the ratio of 6:6:1 respectively. Common mixture of liquid cultures is applied onto the carrier prepared in advance - sterile exhausted beet pulp treated with cellulolytic ferment and enriched with fermentolysate of Saccharomyces cerevisiae yeast. Then, it is mixed and exposed; after that, solid-phase fermentation is carried out under the specified conditions of restricted oxygen access. Liquid culture Bacillus licheniformis All-Russian collection of industrial microorganisms B-4162 containing at least 5.6×108 CFU/g is added in the amount of 65 l. The mixture is mixed and dried to the humidity of 8-10%. Dry powders of purple echinacea and fruits of holy thistle are added in terms of 20-50 g per 1 kg of the end product. The obtained mixture is stirred and subject to crushing till indiscrete mass is obtained. |
 Strain of micromycete aspergillus oryzae-producer of proteolytic and amylolytic ferments to be used in food industry / 2499039Strain Aspergillus oryzae RCAM01135 is a producer of proteolytic and amylolytic ferments, which has ability to produce proteolytic and amylolytic ferments. It was deposited at GNU VNIISKHM with the following registration number: RCAM01135. It can be used at production of different food products (fermented spices, additives, and drinks). |
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Strain of unicellular algae dunaliella salina ippas d-295-producer of biologically active substances having antioxidant activity / 2497945 Strain of unicellular algae Dunaliella salina IPPAS D-295-producer of biologically active substances having antioxidant activity was deposited in the culture collection of microalgae of the Institute of Plant Physiology n.a. K.A.Timiryazev RAS (IPP RAS (IPPAS)) under registration number IPPAS D-295 and can be used to produce biologically active substances having antioxidant activity and antagonistic activity against opportunistic pathogenic bacteria. |
 Strain of bacteriophage escherichia coli ecd4, having lytic activity in respect to bacteria escherichia coli of serotype o104:h4 / 2496874Invention relates to a strain of the bacteriophage Escherichia coli ECD4. The strain of the bacteriophage Escherichia coli ECD4 is isolated from faeces of broiler chickens on the culture of the bacteria of the strain Escherichia coli O104.H4 RK1No.112027 and is deposited in the State Collection of Pathogenic Microorganisms and Cell Cultures "GKPM-Obolensk" under the number Ph63. The proposed strain of the bacteriophage has lytic activity in respect to the bacteria of the strain Escherichia coli O104:H4 RKINo.112027, lyses Escherichia of the serotype 0157:H7, does not suppress growth of cells Escherichia coli M-17, has lytic activity in respect to several other clinically significant serotypes of Escherichia, and also to several types of Shigella. The strain of the bacteriophage Escherichia coli ECD4 propagates on the laboratory non-pathogenic strain Escherichia coli K-12 C600F. |
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Lactobacillus paracasei subspecies paracasei strain having antimicrobial and immunomodulating properties, and food product on its basis / 2501850 Strain is deposited in CNCM and has number 1-3689. Strain has ability to inhibit the growth of pathogenic microorganisms in culture, and namely Escherichia coli, Salmonella enteritidis and Listeria monocytogenes. Strain has anti-inflammatory properties at the ratio of IL-10/IL-12 equal to 11.8. |
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Bacillus licheniformis vkm b-2713d strain having apparent antagonism in relation to salmonella typhi, staphyloccus aureus, listeria monocytogenes and resistance to streptomycin and nalidixic acid / 2501849 Bacillus licheniformis VKM B-2713D strain has apparent antagonism in relation to Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes. Strain has resistance to antibiotics at streptomycin concentration of 20 mcg/ml and at concentration of nalidixic acid of 20 mcg/ml. Strain has α-amylase activity of 0.015 U/ml of the medium at pH 6.0, protease activity of 2.34 U/ml of the medium at pH 7.0. |
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Cleaning method of water solution containing copper salt from copper ions / 2501745 Cleaning method of water solution containing copper salt from copper ions is proposed. The method provides for contact of water solution containing 20 mg of Cu2+/l with 0.2 g of suspension of homogenised culture per 1 l of the solution. The suspension contains the following composition of dominant types of cyanobacteria: Phormidium ambiguum (Jom.), Phormidium boryanum (Kutz.), Leptolyngbya foveolarum (Rabenhor stex Gom), and Plectonema boryanum (Gom.f.boryanum). Contact is performed during 1 to 3 hours. |
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Nutritional composition promoting normal development and growth / 2501553 Invention refers to pharmaceutics and represents a nutritional composition for an infant or: : a child containing lipid or fat, a protein source, a source of long-chain polysaturated fatty acids which contains docosahexaenoic acid; a supplementary calcium source to 2.5 wt % with at least 20% of the supplementary calcium source representing calcium gluconate, and PDGF-β 0.015 to 0.1 ppm (pg/mcg). |
 Immunomodulatory extracts of bacteria lactobacillus and methods for preparing and using them / 2500412Invention refers to pharmaceutical industry, namely to an extract of one or more bacterial strains Lactobacillus. The extract of one or more bacterial strains Lactobacillus representing a soluble extract, wherein the extract contains chemically modified bacterial molecules prepared by the action of an alkaline medium on one or more bacterial strains Lactobacillus; the extract is effective in treating diseases associated with the anti-inflammatory cytokine production imbalance. A method for preparing the extract of one or more bacterial strains Lactobacillus. A pharmaceutical composition effective for reducing at least one symptom associated with at least one condition specified in a respiratory disorder, an allergic condition, an urinary disorder and a gastric disorder, containing the extract. A nutritional composition. A pharmacological composition effective in treating the diseases associated with the anti-inflammatory cytokine production imbalance, containing the extract. A method of relieving the above symptoms. The extract prepared by the above method. |
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Production method of complex feed preparation with lysin producer based on corynebacterium glutamicum / 2499829 Liquid culture of strain Corynebacterium glutamicum All-Russian collection of industrial microorganisms B-1959 - lysine producer by means of a deep method and liquid cultures of strains Bacillus subtilis All-Russian collection of industrial microorganisms B-8130, Bacillus subtilis All-Russian collection of industrial microorganisms B-2984, Bacillus subtilis All-Russian collection of industrial microorganisms B-4099 and Bacillus licheniformis All-Russian collection of industrial microorganisms B-4162 are prepared. To liquid culture of strain Corynebacterium glutamicum All-Russian collection of industrial microorganisms B-1959 there added in quantity of 30 l is 35 l of the mixture consisting of liquid cultures Bacillus subtilis All-Russian collection of industrial microorganisms B-8130, Bacillus subtilis All-Russian collection of industrial microorganisms B-2984 and Bacillus subtilis All-Russian collection of industrial microorganisms B-4099, which have been taken in the ratio of 6:6:1 respectively. Common mixture of liquid cultures is applied onto the carrier prepared in advance - sterile exhausted beet pulp treated with cellulolytic ferment and enriched with fermentolysate of Saccharomyces cerevisiae yeast. Then, it is mixed and exposed; after that, solid-phase fermentation is carried out under the specified conditions of restricted oxygen access. Liquid culture Bacillus licheniformis All-Russian collection of industrial microorganisms B-4162 containing at least 5.6×108 CFU/g is added in the amount of 65 l. The mixture is mixed and dried to the humidity of 8-10%. Dry powders of purple echinacea and fruits of holy thistle are added in terms of 20-50 g per 1 kg of the end product. The obtained mixture is stirred and subject to crushing till indiscrete mass is obtained. |
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Method for isotope enrichment of e.coli cells / 2499042 Method for enrichment of E.coli cells with magnesium isotopes provides for cultivation of E.coli cells during 10-16 h at the temperature of 37°C in water solution enriched with magnesium isotope 24Mg or 25Mg or 26Mg. Water solution includes NH4Cl, glucose, Na2HPO4, KH2PO4, NaCl and MgSO4 at the following component content per litre of distilled water: NH4Cl - 2 g, MgSO4 - 200-260 mg, glucose (dry) - 8-10 g, Na2HPO4 - 11.9-12.1 g, KH2PO4 - 5.95-6.05 g, and NaCl - 0.98-1.02 g. |
 Improvement of growth of bifidus bacteria in fermented dairy products / 2499041Production method of a fermented product involves inoculation of milk with bacteria, milk fermentation and packaging of the ready product. Inoculation is performed using Bifidobacterium animalis lactis BB-12© DSM 15954 strain, Lactobacillus bulgaricus bacteria and Streptococcus thermophilus ST6008 DSM 18111 strain. The invention proposes a starter culture containing 105 to 1012 CFU/g of Bifidobacterium animalis lactis BB-12© DSM 15954 strain, 105 to 1012 CFU/g of Streptococcus thermophilus ST6008 DSM 18111 strain and Lactobacillus bulgaricus bacteria. |
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Method of growing acidophilic lactic acid microorganisms / 2497946 Method comprises mixing a decoction of marine invertebrates and/or fish with protein product of animal origin (wastes of fish and/or seafood processing) crushed into particles of size 5 mm, and/or protein product of plant origin (soyabean cake) in a predetermined ratio to produce nutrient medium. The resulting nutrient medium is pasteurised with stirring and cooled. The cooled nutrient medium at a temperature of about 38-40°C is inoculated with acidophilic lactic acid microorganisms followed by culturing at a temperature of 51-52°C for 6-6.5 hours with stirring to obtain an additive. |
 Method of biodegradation of drotaverine hydrochloride / 2496866Method is proposed for biodegradation of drotaverine hydrochloride (spasmolytic NOSHPA). The method provides for the process of reaction of drotaverine hydrochloride with cells of the strain Rhodococcus rhodochrous, Institute of Ecology and Genetics of Microorganisms 608. The process is performed under aerobic conditions in the medium containing sources of carbon, nitrogen, phosphorus, mineral salts. At the same time the additional source of carbon in the nutrient medium may be glucose. The cells of the strain Rhodococcus rhodochrous, Institute of Ecology and Genetics of Microorganisms, 608, may be used in the immobilised form. The strain is stored in the Regional Profiled Collection of Alkanotroph Microorganisms (acronym of the collection of the Institute of Ecology and Genetics of Microorganisms). Duration of biodegradation of drotaverine hydrochloride makes at least 80% for 25 days. |
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Lactobacillus paracasei subspecies paracasei strain having antimicrobial and immunomodulating properties, and food product on its basis / 2501850 Strain is deposited in CNCM and has number 1-3689. Strain has ability to inhibit the growth of pathogenic microorganisms in culture, and namely Escherichia coli, Salmonella enteritidis and Listeria monocytogenes. Strain has anti-inflammatory properties at the ratio of IL-10/IL-12 equal to 11.8. |
FIELD: biotechnologies.
SUBSTANCE: Bacillus licheniformis strain has antagonism in relation to Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes and resistance to the following antibiotics: streptomycin and nalidixic acid. The strain is deposited with number BKM B-2712D in the All- Russian collection of microorganisms of the Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Science (IBFM RAN).
EFFECT: strain can be used for creation of probiotic bacterial preparations in veterinary.
The invention refers to biotechnology, namely to the probiotic bacterial drugs that can find application in veterinary medicine.
Probiotic bacterial preparations based on strains of microorganisms possessing antagonistic activity to bacteria.
Classical studies I.A. Mechnikov these properties were studied on the example of lactic acid bacteria. Then the circle of producers of probiotic preparations was expanded.
Probiotics based on strains of spore-forming bacteria relatively recently started to be used in veterinary medicine. Antagonistic properties of these bacteria have drawn attention of many developers
The complexity of the treatment of infectious diseases consists of the emergence of pathogenic microorganisms to antibiotics. Moreover, antibiotics often have spillover effects on the organism of the animals and cause intestinal dysbacteriosis course of the basic disease. It causes the necessity of search of new approaches to solving this problem.
Drugs-probiotics on the basis of bacteria of the genus Bacillus, which suppress the development of pathogenic microorganisms are widely used in agriculture for the suppression of phytopathogenic bacteria and fungi, medicine, veterinary medicine for treatment of different diseases of birds and farm animals [1].
Currently drugs-probiotics from bacteria of the genus Bacillus, are widely used in veterinary medicine for treatment of various diseases of birds and farm animals. Offered numerous variants of probiotics of spore-forming bacteria: [5], [6], [7], sporobacterin [8], [9]. The basis of the above drugs-probiotics are different strains of bacteria of the genus Bacillus, having antagonism against pathogenic bacteria.
To increase the efficiency of the prevention and treatment of disbacteriosis offer drugs of two strains of bacteria of the genus Bacillus Bacillus subtilis and Bacillus licheniformis. It is believed that the type of bacteria Bacillus subtilis - strict aerobes, while representatives of Bacillus licheniformis can grow in conditions. It is also believed that these properties of the two types strengthen the effectiveness of probiotic treatment [2-4]. The vast majority of the members of the genus Bacillus harmless for warm-blooded animals, are antagonistic activity towards a wide spectrum of pathogenic and conditionally pathogenic microorganisms resistant to political enzymes and their application in production, stable in storage, is ecologically safe.
Given the possibility of the use of antibiotics in feed production, is very promising use of probiotic preparations strains possessing both high antagonistic activity against and MDR antibiotics.
As a prototype can be seen strain of Bacillus licheniformis PMBC-2336, part of a veterinary drug [11].
Object of the present invention is to obtain a strain of Bacillus licheniformis having antagonistic activity towards a wide spectrum of pathogenic test-organisms, as well as resistance to the most commonly used antibiotics.
Technical effect which can be obtained by using the resulting strain, is to enhance the effectiveness of treatment due to the possibility of a simultaneous application of bacterial preparation of antibiotics.
To solve this problem, a strain of Bacillus licheniformis VKM B-2712D. Strain deposited in the national collection of microorganisms.
Identification strain was carried out by « microorganisms» Berga (1997).
The original soil isolates selected in the Kirov region. Soil isolate was subjected to impact nitrozoguanidine mutagenesis (dose of 40 to 100 mcg/ml) with subsequent screening on selective media.
Culturally-morphological and microscopic features of strain: the rod-shaped cells direct, size: 0,5-0,7 x to 2.0-3.0 mm, heat-resistant forms endospores, no more than 1 in a cage sporangia, , not suppressed, by exposure to air, the location is Central. Cells are immobile. The standard nutritionally dense environments (MPA, LB, GRM-1) forms small transparent convex colonies. Colonies grow into the agar. Catalase: _, optional . Sources of carbon 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 degrees C to +45 C.
Antibiotic resistance: streptomycin -10 mcg/ml, nalidiksova acid -20 mcg/ml.
The invention is illustrated by the following examples.
Example 1. Strain is characterized by high antagonistic activity against pathogenic and conditionally pathogenic microorganisms. Antagonistic activity tested method of deferred antagonism.
For this suspension cells, washed out from the agar medium, plant touch on diameter of a Petri dish with agar medium Gause N 2. Crops were incubated in thermostat at 37 C for 48 H. Then, which has grown culture stroke test-organisms (500-million suspension subsistence crops in the physiological solution). Records of the results of conducted through 18-20 hours of incubation at 37 C largest areas lack of growth in the test cultures. The control of the growth of test cultures served parallel growing them in cups with an agar medium Gause N 2 without the studied culture. The results are shown in the table.
Study of antagonistic properties culture
Growth inhibition test cultures, mm
E. coliS. typhimurium
S. typhiPs. aeruginosa
St. aureus
L. monocytogenes
0 0 8 0 13 15Example 2. It studied the resistance to the following antibiotics: kanamycin, streptomycin, gentamicin, trimethoprim, nalidixic acid, Cefazolin, ampicillin, benzylpenicillin sodium salt. A check was carried out at the deep cultivation producer on glucose peptone medium with yeast extract at various concentrations of antibiotics. It was found that the strain has to antibiotic resistance: streptomycin - 10 mcg/ml, nalidiksova acid - 20 mcg/ml
Example 3.
The enzyme activity in the cultural liquid determined in submerged cultivation producer on glucose peptone medium with yeast extract in two days. The following were identified values of enzymatic activity:
α activity - 0.1 U/ml of the medium at pH 6.0, activity - 1,728 U/ml of the medium with pH 7.0.
It should be noted high activity strain Bacillus licheniformis VKM B-2712D.
Literature
1. Smirnov V.V., Reznik S.R., V.O. Modern understanding of the mechanisms of therapeutic and prophylactic action of probiotics from bacteria of the genus Bacillus // . Journe. - 1993. - №4. - Pp.92-112.
2. Bakulina L. F., Timofeyev I.V., N.G. Perminova, Polushkina A.F., NI Probiotics on the basis of spore-forming microorganisms of the genus Bacillus and their use in veterinary medicine // Biotechnology. 2001, №2, P.48-56.
3. Hong, N.A., 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 / RJ. Collier, et al. // A Report from the American Academy of Microbiology Washington, 2006. - 28 p.
5. Patent of the Russian Federation 2130316. Medical and prophylactic preparation . A61K 35/74, BI №14,1999, prior. 23.01.97.
6. USSR patent 1710575. A strain of bacteria Bacillus sp - component therapeutic and preventive preparation against dysbacteriosis and allergies, C12N 1/20, BI №5,1992.
7. Patent of the Russian Federation 2035186. Preventive biopreparation . A61K 35/66, BI №14, 1995.
8. Aleshkin, A.V., Afanasiev, S., , V.Y, Manko, so-CALLED, , I.YU Bacterial preparation of new generation «Sporobacterin» // Honey. card index. - 2003. - №4
9. Patent of Russian Federation №2035185 Preventive biopreparation . -1995 Prior. 31.01.1992.
10. Patent of the Russian Federation 2115725. Strain of Bacillus subtilis, resistant to rifampicin, ampicillin, and streptomycin, has activity against pathogenic species of microorganisms. Prior. 23.07.1997.
11. Gataullin A.G. Biological properties of strains of Bacillus subtilis looking to create a new probiotics // Diss. kand. Biol. of Sciences, Moscow, 2005
Strain of Bacillus licheniformis BKM B-2712D, has expressed antagonism towards Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and resistance to antibiotics streptomycin and nalidixic acid.