Strain of bacteria bacillus thuringihnsis bios-1, possessing insectoacaricidal activity
SUBSTANCE: strain of bacteria Bacillus thuringiensis BIOS-1 VKPM B-10709, possessing insectoacaricidal activity against representatives of leaf-eating and sucking pests, such as representatives of orders Lepidoptera, Coleoptera, Homoptera, Thysanoptera and Acariformes, doing harm to crops, is deposited in All-Russian collection of industrial microorganisms (VKPM), Federal State Unitary Enterprise GosNIIgenetika under number B-10709.
EFFECT: invention makes it possible to increase mortality of representatives of leaf-eating and sucking pests, doing harm to crops.
6 tbl, 2 ex
The invention relates to the microbiological industry, Microbiology and biotechnology, namely the production of bacterial biological products intended for the control of pests of agricultural crops, which are the representatives of the orders Lepidoptera, Coleoptera, Homoptera, Thysanoptera and Acariformes.
Effective pest control plays an important role in maintaining and increasing yield, because agricultural production losses associated with the activity of insect pests, up to 30% of the expected harvest.
At the present time to control pests of agricultural crops is widely used chemicals. Prolonged use of the latter led to the formation of resistant to these drugs pest populations . In addition, chemicals toxic to warm-blooded animals and beneficial insects, as well as able to penetrate into plants, resulting in poor sanitary and hygienic conditions of agricultural production. The most widely used of chemicals is karbofos.
An increasingly important role in protecting plants from pests play a biological control with the use of products derived from viral, bacterial and fungal pathogens or their products synthesized. The advantage of b is listwa known microbiological plant protection products compared to chemical - they are environmentally friendly.
The most widely as microbiological plant protection products on the basis of different strains of the entomopathogenic bacteria Bacillus thuringiensis. These bacteria during sporulation synthesize crystalline proteins, or, as they are called Delta-endotoxins, which differ among themselves on the spectrum of entomopathogenic actions . Delta-endotoxins have a selective toxic effect only in relation to insects.
Some strains of Bacillus thuringiensis produce more and thermostable exotoxin, which is in contrast to endotoxin does not have the selectivity and toxicity manifests in relation to a very wide range of invertebrates and vertebrates, causing allergic reactions in individuals working with these drugs .
Known strains of Bacillus thuringiensis, which possess insecticidal activity against representatives of the order of Coleoptera and Lepidoptera [4, 5]. However, they are not toxic to the representatives of the orders Homoptera, Thysanoptera (insecticidal activity) and Acariformes (acaricide activity).
Known bacterial strain Bacillus thuringiensis ssp.kurstaki VKPM B-8715 with insecticide and acaricide activity against the representatives of the orders Lepidoptera, Coleoptera, Homoptera, Thysanoptera and Acariformes . The strain chosen as a prototype.
The problem is obtained of the invention - is to teach the bacterial strain of Bacillus thuringiensis, with increased microbiological productivity and insecticide and acaricide activity.
The problem is solved by breeding strains of bacteria Bacillus thuringiensis ssp.kurstaki VKPM B-8715 on laboratory and industrial nutrient media. When mnogochislennyh the subcultures and rasiah to individual colonies was obtained strain of Bacillus thuringiensis BIOS-1, with increased insecticide and acaricide activity against listography and sucking pests.
The resulting strain deposited in Russian national collection of industrial microorganisms (VKPM) how Bacillus thuringiensis BIOS-1 VKPM B-10709.
The strain VKPM B-10709 has the following chacteristic:
Cultural and morphological characteristics.
Gram-negative motile Bacillus size of 4.1-1.6 ám. In the stationary growth phase to form crystals of endotoxins and oval spores.
Grows well in hydrolysates of casein (pancreatic hydrolysate of casein - 1, yeast extract and 0.2, MnSO4, - 0.5, and MgSO4- 3, KH2PO4- 0,1, glucose - 0,6, water - the rest, the pH of the medium to 7.4 and 7.5, medium composition in wt.%) at 27-29°C and a pH of 6.8 to 7.5. On agar medium casein (pancreatic hydrolysate of casein - 1, yeast extract - 0.5, and sodium chloride - 0.05, the agar - 1,5, water - the rest, the pH of the medium to 7.4 and 7.5, medium composition in wt.%) 10 days at 27-29°C forms a homogeneous colonies 3-5 mm in diameter, light gray in color with a slightly jagged edge. The structure of the ur fine.
On the H-antigen refers to serovariants III (IV).
Under cultivation of strain both in liquid and on solid nutrient media output phage does not occur.
Physiological and biochemical characteristics.
The sources of carbon assimilates with the formation of acid, glucose, maltose, corn and potato starch, soy and corn flour. Not absorb lactose, xylose, arabinose, lecithin. Peptonized milk, hydrolyzes starch, gelatine not thins.
The sources of nitrogen: restores nitrates, assimilates ammonium forms of nitrogen.
Relationship to temperature: grows well at 27-29°C, the optimum temperature of 28°C.
Relation to the pH of the medium: grows well when 6,8-7,5, optimum pH of 7.2. Relation to oxygen: aerobe.
Relationship to phage: sustainable for the main industrial phages P., 22, s/t, And-77, 25-16.
Beta-exotoxin not synthesize.
Not pathogenic for humans, domestic and wild animals, beneficial insects, birds and fish.
Not a genetically modified strain.
Claimed properties: synthesizes components with insecticide and acaricide activity against the representatives of the orders Lepidoptera, Coleoptera, Homoptera, Thysanoptera and Acariformes.
Marker signs: activity against the representatives of the orders Lepidoptera, Coleoptera, Homoptera, Thysanoptera and Acariformes, the lack of ability to synthesize beta-Exo is oxen.
Storage of strain: strain is stored in the vials after lyophilization paracrystalline mixture in the usual way.
The invention is illustrated by the following examples.
EXAMPLE 1. Microbiological yield was obtained strain.
Assessment of microbiological productivity of the obtained strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 on laboratory culture media was carried out according to two main parameters, namely the title dispute and concentration of the main insecticidal protein (Delta-endotoxin) in comparison with the prototype strain of Bacillus thuringiensis ssp.kurstaki VKPM B-8715.
For this purpose, the strains of Bacillus thuringiensis were grown in flasks with a capacity of 750 ml in volume environment growing 200 ml to stage full of sporulation on a circular shaker at 28°C and shaking 250 rpm/min For growing used medium of the following composition, wt.%:
1. Wednesday 1:
|Pancreatic hydrolysate of casein||1|
|the pH of the medium||7,4-7,5|
2. Wednesday 2:
|Trypticase casein hydrolysate||1|
|the pH of the medium||7,4-7,5|
3. Wednesday 3:
|the pH of the medium||7,4-7,5|
The grown culture was separated by centrifugation and washed 1M NaCl in the presence M EDTA. Crystal proteins from paracrystalline mixture was dissolved in carbonate buffer pH 11.0 in an hour, undissolved precipitate was separated by centrifugation, the pH of the supernatant was made up to 10.0. The protein concentration was determined by Bradford method , using bovine serum albumin as standard. Complete dissolution was controlled by re-extraction in the same buffer. If re-extraction was dissolved not more than 7% protein. Determination of the titer of viable spores was carried out on Petri dishes with agar medium of the following composition, wt.%:
|Pancreatic hydrolysate of casein||1|
|the pH of the medium||7,47,5|
The research results are summarized in table 1:
|Nutrient medium||Wednesday 1||Wednesday 2||Wednesday 3|
|The strains of Bacillus thuringiensis||PMBC|
|The title dispute, ×108per ml||4,5||6||1,8||2,6||3,5||5,0|
|Protein concentration (mg/ml)||0,25||0,4||0,35||0,5||0,25||0,4|
As follows from table 1, the resulting strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 on the three studied laboratory nutrient medium is superior in microbial productivity (titer dispute, conc is ntrace main insecticidal protein, Delta-endotoxin) strain of Bacillus thuringiensis ssp.kurstaki VKMV-8715 (prototype) by 25-30%.
Evaluation of the obtained strain on industrial nutrient medium was performed according to the setting of microbiological productivity as a title dispute.
For preparation of inoculum the resulting strain and the prototype were grown in 2 ml of medium 1 for 12 hours at 28°C as described above. Then 1 ml of microbial cultures seeded in flasks with a capacity of 750 ml volume of medium growing 50 ml-driven polysaccharide environment (DPS) of the following composition, wt.%: yeast - 36, corn flour and 26.8, soy flour to 5.0, chalk - 3,0, water - the rest, the pH of the medium was 7.2-7.5 and cultivated on a pie rocking chair to the stage full of sporulation. Determination of the titer of viable spores was carried out on Petri dishes with agar medium as described above. The results of the experiment are shown in table 2.
|Microbiological production(billion JV./ml)||4,3||6,0|
From the results shown in the tables is 2, it follows that the productivity of the obtained strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 industrial environment, higher productivity of the strain Bacillus thuringiensis ssp.kurstaki VKPM B-8715 (prototype).
Microbial productivity (title dispute, the concentration of the main insecticidal protein, Delta-endotoxin) of the obtained strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 on laboratory culture media and industrial nutrient medium is higher than that of strain-prototype (Bacillus thuringiensis ssp.kurstaki VKPM B-8715).
EXAMPLE 2. Evaluation of insecticide and acaricide activity of the obtained strain.
Determination of insecticide and acaricide activity of the obtained strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 and prototype Bacillus thwingiensis ssp.kurstaki VKPM B-8715, Baloo conducted by representatives of the orders Lepidoptera, Coleoptera, Homoptera, Thysanoptera and Acariformes in the laboratory.
The resulting strain VKPM B-10709 and prototype VKPM B-8715 grew to the stage full of sporulation as in example 1 in environment 1.
Evaluation of insecticidal activity for representatives of Lepidoptera were conducted at two sites: the caterpillars of the Gypsy moth Lymantria dispar 2 age and caterpillars mill the liquidation Galleria mellonella age 7.
Caterpillars of the moth grew stored in the refrigerator at a temperature of +4°C egg in a medium of the following composition, wt.%:
|Haricot beans are swollen||72|
|The dry yeast||of 5.4|
|Paper filter finely chopped||2|
|Potassium phosphoric acid disubstituted||1,4|
|Benzoic acid + alcohol||3,5|
|The oak leaf dry (powdered)||1|
When biotesting 3 g medium was mixed with 1 ml of the test suspension of spore-crystal mixture (twice washed with saline) and well rubbed the mixture in a porcelain mortar with a pestle and mortar. The resulting mixture with a spatula transfer on the cover of the Petri dishes and placed in the form of a ring with a diameter of about 3, see Inside of this ring was planted on ten tracks. At the bottom of the Petri dishes placed is whether a disk of filter paper. Each sample suspension was tested in 3 parallel replications. The experiment was carried out at room temperature and natural light. The number of dead insects was counted on day 6. The results of the experiment are shown in table 3.
|The strains of Bacillus thuringiensis||Mortality (%) Gypsy moth (Lepidoptera, moth Lymantria dispar)|
The resulting strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 on insecticidal activity against moth exceeds the prototype of Bacillus thuringiensis ssp.kurstaki VKPM B-8715 25%.
Evaluation of insecticidal activity against the mill the liquidation (Galleria mellonella) 7 age was performed by means of grafting larvae mill the liquidation in a Petri dish filled with food (Merv + wheat bran).
Food was Packed up in Petri dishes (9 g each), which were introduced into 3 ml of the drug.
In each Petri dish (after drying of feed) was infused for 10 caterpillars of the liquidation 7 age (weight 75±5 mg) in three replications. Petri dishes with the tracks contained on the lab tables at temperature 25±20°C and relative humidity of the air is 60-80%. Mortality of larvae was calculated for 6 days. The results of the experiment are shown in table 4.
|The strains of Bacillus thuringiensis||Mortality (%) mill the liquidation (Lepidoptera, Galleria mellonella)|
The resulting strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 on insecticidal activity against mill the liquidation exceeds the prototype of Bacillus thuringiensis ssp.kurstaki VKPM B-8715 30%.
Evaluation of the insecticidal activity of the obtained strain VKPM B-8715 for members of the order of Coleoptera were performed on larvae of the Colorado potato beetle Leptinotarsa decemlineata.
Colorado potato beetle larvae younger children (3rd and early 4th instar) were placed in Petri dishes with potato leaves treated by immersion in solutions of drugs. Table 5 shows the results of the death of the larvae of the Colorado potato beetle on the 5th day.
|The strains of Bacillus thuringiensis||Mortality (%) Colorado potato beetle (Coleoptera, Leptinotarsa decemlineata)|
The resulting strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 on insecticidal activity against larvae of the Colorado potato beetle exceeds the prototype of Bacillus thuringiensis ssp.kurstaki VKPM B-8715 25%.
A comparative assessment of insecticide and acaricide activity of the obtained strain against the representatives of the orders Homoptera, Thysanoptera and Acariformes conducted at three sites: bocheva aphid, Western flower thrips and spider mite.
For test objects in the laboratory used the method of immersion of host plants in solution culture fluid with spores and crystals of the investigated strains obtained after completion of sporulation as in example 1 on the environment of growing 1.
For melon aphid (Aphis gossypii Glov., Homoptera) cut the leaves of cucumber were immersed in solutions of the test samples for 6 seconds, after which they were placed on moist filter paper in a Petri dish and was inhabited by the female aphid.
For Western flower thrips (Frankliniella occidentalis Pergande, Thysanoptera) used dvuhlitrovye bean plant. 7 days prior experience on plants of the female thrips were laying eggs. After hatching the larvae of the plants were cut and the leaves were immersed in solutions of the test samples for 6 seconds, after which he set the glasses of water, substituting the petioles plastic mugs, almasan the e on the edges with petroleum jelly.
In the case of the common spider mite (Tetranychus urticae Koch, Acariformes) cut dvuhlitrovye bean plants the day before the experience was inhabited by the female mite. The plants were immersed in solutions of drugs for 6 seconds and was installed in vessels of water.
The results of the death of the test object on the 5th day are shown in table 6.
|The strains of Bacillus thuringiensis||VKPM B-8715||VKPM B-10709|
|Mortality (%) melon aphid (Homoptera, Aphis gossypii Glov.,)||55||69|
|Mortality (%) spider mites (Acariformes, Tetranychus urticae Koch,)||79||95|
|Mortality (%) Western flower thrips (Thysanoptera, Frankliniella occidentalis Pergande,)||49||61|
As follows from the results presented in table 6, the resulting strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 on insecticide and acaricide activity against the representatives of the orders Homoptera, Thysanoptera and Acariformes exceeds the prototype of Bacillus thuringiensis ssp.kurstaki VKPM B-8715 20-25%.
Insecticide and acaricide activity of the obtained strain of Bacillus thuringiensis BIOS-1 VKPM B-10709 n is otiv of representatives of the orders Lepidoptera, Coleoptera, Homoptera, Thysanoptera and Acariformes exceeds the prototype of Bacillus thuringiensis ssp.kurstaki VKPM B-8715 20-30%.
Sources of information
1. National Research Council. 1986. Pesticide reistance: strategies and tactics for management. National Academy of Sciences, Washington, D.C.
2. Crickmore, N., Zeigler, D.R., Feitelson, J., Schenepf, E., van Rie, J., Lereclus, D., Baum, J. and Dean, D.H. 1998. Revision of the nomenclature for the Bacillus thuringienis pesticidal crystal proteins. Environ. Molec. Biol. Rev. 62:807-813.
3. McConnell, E. and Richards, A.G. 1959. The production by Bacilllus thuringiensis Berliner of a heat-stable exotoxin toxic to insects. Can. J. Environ. 5: 161-168.
4. USA patent 6,156,308.
5. RF patent №2125091.
6. RF patent №2278159.
7. Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analyt. Biochem. 72: 248-254.
The bacterial strain Bacillus thuringiensis BIOS-1 VKPM B-10709 with insecticide and acaricide activity against members of the leaf-eating and sucking pests.
SUBSTANCE: there is offered a method of preparing the antibiotic Cephalosporin C. A new antibiotic producer Acremonium chrysogenum strain, RNCM No. F-4081D is used. Acremonium chrysogenum, RNCM No.F-4081D is cultivated on an enzymatic nutrient medium containing carbohydrate sources - glucose, corn starch, dextrin, nitrogen sources - a corn extract, Pharmamedia, ammonium sulphate, and also inorganic salts - potassium phosphate, potassium sulphate, chalk, copper sulphate, zinc sulphate, manganese sulphate, ferrous sulphate, and as vegetable fat - rapeseed oil and in addition a phosphatidylcholine and/or sitosterol.
EFFECT: twice increased antibiotic production level, reduced fermentation time.
1 tbl, 7 ex
SUBSTANCE: invention can be used in producing nutrient mediums which create the optimum conditions of legionella activity. The nutrient medium contains a enzymatic hydrolyzate of a pig's lung, a enzymatic hydrolyzate of peanut, 1-substituted potassium phosphate, 2-substituted 3-aqueous potassium phosphate, activated carbon, L-cysteine hydrochloride, microbiological agar and distilled water.
EFFECT: invention allows reducing time for legionella detection.
SUBSTANCE: avirulent Vibrio cholerae strain of biovar eltor of serovar Ogawa - a producer of a major protective O1 antigen of serovar Ogawa is produced in a simulation experiment of virulent natural V.cholerae M569 Ogawa strain by spontaneous loss of "СТХφ" prophage at cholera agent continuance in sterile river water. The strain is deposited in the State Collection of pathogenic bacteria "Microbe" Registration No. 262 of the Collection of Miscroorganisms. A feature of the strain is the absence of genes of a core region of "СТХφ" prophage. The strain is avirulent for suckling rabbits, agglutinated by cholera serums O1 and Ogawa in dilution exceeding a diagnostic titre.
EFFECT: use of the invention allows producing a vaccine of O1 protective antigen of serovar Ogawa providing immunity formation in cholera.
SUBSTANCE: avirulent Vibrio cholerae KM 263 strain of biovar eltor of serovar Inaba - a producer of a protective O1 antigen is produced in a simulation experiment of virulent natural V.cholerae M569 Inaba strain by spontaneous loss of "СТХφ" prophage at cholera agent continuance in sterile river water. The strain is characterised by a high production level of the major protective O1 antigen of serovar Inaba.
EFFECT: invention aims at preparing chemical cholera vaccines by formation of antibacterial cholera immunity, and producing purified preparation of the O1 antigen of serovar Inaba for a diagnostic serum preparation .
SUBSTANCE: blood serum is diluted with physiologic saline in the ratio 1:2, 1:5, 1:10, 1:20 and 1:50, and tracheobronchial aspirates are diluted with physiologic saline in the ratio 1:2, 1:4, 1:8, 1:16 and 1:32. A solid nutrient medium of the following composition is prepared: a nutrient microbiological dry agar 26.5 g, a nutrient yeast extract 1.22 g, lactose 10.7 g, disodium phosphate 0.48 g, anhydrous sodium sulphite 0.83 g, sodium carbonate 0.03 g, sodium hydroxide 5.0 g, distilled water 1000 ml, pH 8.0 which contains the test strain Micrococcus lysodeiktikus 2665 in the concentration 50 million microbial cells in 1 ml of the medium. Simultaneously with the material being analysed, a reference concentration 5 mcg/ml is introduced in the wells of the diameter 8 mm on each Petri dish. In 18 hours of the incubation procedure at 37°C, a diameter of test strain growth retardation zone surrounding the wells are measured. The vancomycin concentration, mcg/ml is determined by a calibration curve of growth retardation zone diameter to the reference vancomycin concentration.
EFFECT: use of the declared method allows determining the vancomycin concentration in the biological fluids and ensuring higher clinical effectiveness.
SUBSTANCE: method involves cultivation of recombinant koji mycelial fungi in a fluid medium, collection of recombinant protein from the prepared product. The culture medium contains as a raw substance, at least one selected from a group consisting of barley and wheat surface of which is completely or partially coated at least in the concentration 1 to 20% (weight/volume) and as nutrients, at least one inorganic salt. Recombinant koji mycelial fungi is produced by a procedure consisting in the fact that a gene coding a target protein is ligatured below a promotor of a gene coding an enzyme exposed to catabolite repression due to concentration of the nutrients, such as saccharides and amino acids, for preparing thereby the ligation product, and the ligation product is introduced in koji mycelial fungi as a host.
EFFECT: invention allows higher yield protein.
4 cl, 3 tbl, 1 ex
SUBSTANCE: invention refers to a method of producing a mutant lactobacillus Streptococcus thermophilus, to a milk ferment, a method of producing a fermented milk product and to the fermented milk product. The offered invention can be used for producing the fermented milk products with improved storage characteristics. A method of producing the mutant lactobacillus Streptococcus thermophilus characterised by weaker postoxidation, than a parent strain is implemented by introducing in DNA a genome of said parent strain of mutation codon 552 coding histidine, of teh domain HA of lactose permease. Said mutation induces replacement of said histidine by amino acid which is distinct from serine, tyrosine, histidine and threonine.
EFFECT: invention allows producing the mutant lactobacillus Streptococcus thermophilus characterised by weaker postoxidation and suitable particularly for producing the fermented milk products.
10 cl, 5 dwg, 3 tbl, 2 ex
SUBSTANCE: Streptomyces caespitosus 3810/OE strain is a mitomycin C producer prepared from a stock strain recovered from soil by processing with chemical mutagens followed by stage selection, and deposited in the Collection of Cultures of State Institution G.F.Gauze Research Institute for Investigation of New Antibiotics of the Russian Academy of Medical Science, No. INA 01082. For producing mitomycin C, the strain is grown on a sowing medium of the following compoisiton (%): glucose - 0.6-1.4, starch - 0.6-1.4, soya flour - 2-5, corn steep - 0.6-1.0, KH2PO4 - 0.01-0.03, chalk - 0.05-0.12, (NH4)2SO4 - 0.06-0.14, main water, pH before sterilisation 6.8-7.2 and then is transferred to an enzyme medium of the following composition (%): starch - 0.6-1.4, sucrose - 1-3, soya flour - 2-4, corn steep - 0.6-1.0, CoCl2•6H2O - 0.007-0.012, KH2PO4 - 0.02, chalk - 0.05-0.12, (NH4)2SO4 - 0/06-0.04, FeSO4•7H2O - 0.01-0.02, MgSO4•7H2O - 0.01-0.03, NaCl - 0.2-0.6, main water, pH before sterilisation 6.8-7.2. The strain is grown in an aeration environment for 100-120 h, then mycelium is removed by centrifugation or filtration, and a supernatant containing an end product is extracted by mixed acetonitrile and methyl or ethyl alcohol. A level of strain produced mitomycin C is 85 to 100 mcg/ml.
EFFECT: improved method of producing antibiotic mitomycin.
2 cl, 1 ex
SUBSTANCE: method provides introducing a preparation of active dry yeast Saccharomyces cerevisiae in water in the ratio 1:10. Then the mixture is exposed for 4-6 h at temperature 23-27°C. The produced suspension is layered to reactivated yeast and a supernatant. The supernatant is removed.
EFFECT: method allows producing reactivated yeast with more complete living function recovery.
2 tbl, 3 ex
FIELD: food industry.
SUBSTANCE: nutrient medium contains soya flour and distilled water at preset quantitative ratios.
EFFECT: Deinococcus radiodurans yield increase.
2 tbl, 8 ex
SUBSTANCE: strain of Bifidobacterium longum is separated from bowels content of healthy baby and is deposited in Government collection of microorganisms of normal microflora (GKNM) FSIS "MNIIEM named after G.N. Gabrichevskiy Rospotrebnadzor " under registration number 230. Strain multiplies in various nutrition mediua with accumulation of production biomass with high concentration of bifidobacteria.
EFFECT: Bifidobacterium longum strain has acid-forming activity, antagonistic activity with respect to pathogenic and opportunistic microorganisms, ferments milk, which makes it possible to apply it for obtaining bifidocontaining production.
2 tbl, 5 ex
SUBSTANCE: method of indicating hospital strains of Pseudomonas aeruginosa, Staphylococcus aureus, Enterobacter cloacae involves identification of an isolated pure culture of bacteria, inoculating the identified bacteria on blood-meat infusion agar while washing accumulated bacteria - S.aureus - after 12 hours, and Exloacae and P.aeruginosa - after 8 hours with sodium chloride solution. Electrical resistance of the suspension of bacteria in the sodium chloride solution is determined at bacteria concentration of 500000 in 1 ml in a dc electric field with 2.8 V across the electrodes. If electrical resistance for P.aeruginosa bacteria ranges from 579 to 674 kom, S.aureus 545-642 kom, Exloacae 452-584 kom and epidemiological data on recording five or more disease cases from one infection source, the isolated infectious agent is considered a hospital strain.
EFFECT: invention cuts time for conducting laboratory investigations and detecting circulation of hospital strains in hospitals, use of the invention does not require intraspecific identification of bacteria.
2 tbl, 3 ex
SUBSTANCE: nutrient medium contains enzymeatic hydrolyzate of beef meat, sodium chloride, sodium phosphate bisubstituted, sodium sulfite, Karpuzidi stimulant and purified water in specified component ratio.
EFFECT: invention makes it possible to obtain quality nutrient medium for culturing microbial cells of plague microbe of vaccine strain EB in sufficient quantity during 48 h.
1 tbl, 3 ex
SUBSTANCE: method involves treating a water-based medium containing sulphate-reducing bacteria SRB in industrial aqueous systems of chemical production and oil refining. Inhibition of production of biogenic sulphide with SRB takes place as a result of synergetic action of a biocide component in a first concentration and a metabolic inhibitor in a second concentration. The biocide immediately destroys the first portion of SRB. The biocide component is selected from a group comprising aldehydes, amine-type compounds, halogenated compounds, sulphur compounds, salts of quaternary phosphonium and/or combinations thereof. The metabolic inhibitor inhibits growth of a second portion of SRB without its direct destruction. The metabolic inhibitor component is selected from a group comprising nitrite, molybdate, tungstate, selenate, anthraquinone and/or combinations thereof. Contact between the SRB and the biocide and metabolic inhibitor can take place continuously, intermittently or simultaneously.
EFFECT: method ensures efficient inhibition of production of biogenic sulphide with SRB during combined use of components in considerably lower concentrations than if the biocide or metabolic inhibitor was used separately.
25 cl, 2 dwg, 1 ex
SUBSTANCE: method to produce a probiotic preparation based on sporogenous strains Bac.subtilis and Bac.licheniformis includes their cultivation, mixing of bacterial cells biomasses with protector of microbial cells and subsequent sterile dehydration. For this purpose the method applies the strain Bac.subtilis VKM V-2287 D and the strain Bac.licheniformis VKM V-2414D. Strain biomasses are mixed at the ratio of 1:1. A protector of microbial cells is lactose in amount that provides for cell titre in the finished product after drying of at least 1X1011 CFU/g.
EFFECT: method makes it possible to produce a pluripotential preparation, which may be used to prevent gastrointestinal diseases in farm animals and birds, prevention of stress actions, correction of microflora in intestine in case of digestion processes disturbance.
6 tbl, 2 ex
SUBSTANCE: method to produce a probiotic preparation based on sporogenous strains Bac.subtilis and Bac.licheniformis includes their cultivation, mixing of bacterial cells biomasses with protector of microbial cells and subsequent sterile dehydration. At the same time the method uses the strain Bac. subtilis VKPM V-10172 and the strain licheniformis VPKM V-10135. Biomasses of strains are mixed at the ratio of 1:1. A protector of microbial cells is a gelatine-saccharose protective medium in amount that provides for cell titre in the finished product after drying of at least 2X1012 CFU/g.
EFFECT: method makes it possible to produce a preparation with high efficiency of probiotic component, which ensures higher digestibility of fodders and increases weight gain of farm animals and birds.
3 tbl, 1 ex
SUBSTANCE: invention refers to biotechnology, and can be used for creation of vaccine preparations for colibacillosis in animals. A method for producing colibacillosis anatoxin provides sampling of epizootic Escherichia coli strains able to produce a thermolabile, thermostable and shiga like toxin, separate cultivation on a nutritious broth for 6-7 days, culture inactivation by adding formalin to the concentration 0.3-0.4 % at temperature 37°C for 14 days and mixing in equal volumes, and separation of a bacterial mass by means of sterilisation filtration.
EFFECT: invention provides more efficient prevention of colibacillosis in animals.
1 tbl, 1 ex
SUBSTANCE: method involves Aeromonas sobria vaccine strain seeding, cultivation and inactivation. Aeromonads are cultivated in a liquid nutrient medium of a Hottinger broth in a fermenter for 4-6 hours at temperature 26±1°C, immediately after seeding, an oxidation-reduction potential of the cultural liquid is reduced to (-40) - (-100) mV. Thereafter, before termination of a cultivation process, pO2 in the cultural fluid is kept at 20-30 % from air oxygen saturation, pH of the cultural fluid is regulated at 7.1-7.3 pH units, and fractional glucose supply is carried out in doses to the concentration 0.25-0.35 % with glucose limitation of aeromonad growth.
EFFECT: method allows higher yield of the vaccine with reduced time of preparation.
4 tbl, 5 ex
SUBSTANCE: according to the invention, versions of an immunobiological antiallergic agent contain Lactobacillus acidophilus 100 "аш" PA strain, collection No. 207, is stored in the State collection of microorganisms of normal microflora Federal State Research Institution Gabrichevskiy Moscow Research Institution of Epidemiology and Microbiology of Federal Service for Supervision of Consumer Rights and Human Welfare in a culture medium in amount 106-1010 CFU/ml. The immunobiological antiallergic agent additionally contains species-specific virulent bacteriophages and bacteriophages with induced virulence, a biomass of Lactobacillus acidophilus "КзШз4" strain in a culture medium in amount 106-1010 CFU/ml, a biomass of Lactobacillus acidophilus NKi strain in the culture medium in amount 106-1010 CFU/ml and a biomass of Bifidobacterium bacteria in the culture medium in amount 106-1010 CFU/ml. The immunobiological antiallergic agent additionally contains target additives in amount 0.01-95.0 wt % from mass of the agent. The additives are selected from a number of: glycine, cysteine, copper sulphate, copper gluconate, quinosol, chitosan, licorice root extract, hips extract, origanum extract, cranberry extract and a flavouring agent. The immunobiological antiallergic agent is presented in the form of a suspension. The Lactobacillus acidophilus 100 "аш" PA strain is produced by means of a sequence of multiple passages through a nutrient medium MPC with added sources of histamine, Cu1+, Cu2+, Co2+, Ni2+ ions and at culture pH 6.2-7.4. The agent is used for prevention and treatment of allergic or pseudo-allergic disease without an accompanying pathology or in a combination with an infectious disease, intestinal dysbacteriosis and psychological adaptation disorders.
EFFECT: more efficient antiallergic action of the immunobiological agent on the basis of lactobacilli.
10 cl, 3 tbl
SUBSTANCE: invention refers to a mutant bacterium producing higher levels of intracellular and/or extracellular folate as compared with a wild type which is sensitive to methotrexate, and has growth rate at least 0,1 µ/hour when grown on a medium which contains methotrexate 1.25 mg/l and no folate-dependent metabolites. Also, the invention refers to a composition for foodstuff or food additive enrichment containing said bacterium, and to a method of sampling said bacteria.
EFFECT: invention allows producing high-folate bacteria.
13 cl, 3 dwg, 2 tbl
SUBSTANCE: agent is a strain of a ectomycorrhiza-forming fungus Suillus sibiricus (6)No.LE(BIN)2178. The strain is deposited in the Collection of basidium fungi cultures LE(BIN) of the Institution of the Russian Academy of Science of the Botanical Institute named after V.L. Komarov, RAS, with appointment of a registration number No.LE(BIN)2178.
EFFECT: invention makes it possible to eliminate difficulties related to heterovegetative reproduction and re-planting of precious breeds of coniferous trees, to increase establishment of conifers at woodless areas and urbanised lands, on depleted soils in landscape and gardening works, to strengthen dams and to arrange green spaces of coal quarries.
6 dwg, 5 tbl