Nutrient medium for extraction of bacteria of shigella kind from water objects |
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IPC classes for russian patent Nutrient medium for extraction of bacteria of shigella kind from water objects (RU 2517750):
 Method to prepare medicated product from live strains and microorganisms of lactobacilli and bifidobacteria "lb-complex plus" / 2517734
Method provides for cultivation at 37±1°C of strains of lactobacilli and bifidobacteria in the medium and packing of liquid product with account of daily dose necessary for patients. The medium contains components in the following quantities: caseine hydrolysate dissolved by distilled water, 0.33-0.4 g/l, sodium chloride 5 g/l, fructose 10 g/l, peptone 2 g/l, agar-agar 0.75 g/l for lactobacilli, for bifidobacteria - 1.0 g/l, ascorbic acid 0.25 g/l, distilled water 0.67-0.6 g/l. Strains-producers are Lactobacillus plantarum 8 RA-3, Lactobacillus fermentum 39, Lactobacillus fermentum 90 TC-4, Bifidobacterium bifidum 791, Bifidobacterium longum 379, Bifidobacterium bifidum 1. For lactobacilli and bifidobacteria they prepare accordingly media with different content of agar-agar, amine nitrogen in caseine hydrolysate 160-170 and 180-200 mg% and medium pH 7.8-8.0 and 8.5-8.6. Starting from the first generation the bifidobacteria and lactobacilli are cultivated for 24±1 hours, two strains together, one separately. The produced biomass is mixed with the fresh nutrient medium at the ratio of 1:1000, and strains of lactobacilli are cultivated 24±1 hours, strains of bifidobacteria - 48±1 hours. Upon completion of cultivation of biomass of strains they are mixed at the ratio of 2:1:2:1.
 Nourishing composition, including probiotics and improving sleep pattern / 2517616
Invention relates to application of probiotic bacterial strain for production of a probiotic composition for reducing sleep disorders and/or improvement of sleep quality in people and animals. As a bacterial strain used is Lactobacillus reuteri DSM 17938 or Bifidobacterium longum NCC 3001 (ATCC BAA-999).
Preparation for purification of water and soil from oil pollution and method of obtaining thereof / 2516412
Group of inventions relates to field of biochemistry, ecology, protection of the environment. Claimed is preparation for purification of water and soil from oil pollution, which contains microorganisms, oil destructors, sorbent, cryoprotector - glycerol, microfertilisers - sodium nitrate 0.5% and potassium phosphate 0.5%. As oil destructor, it contains association of oil-oxidising microorganisms: Bacillus subtilis VKM B-81, Pseudomonas spp. VKM B-892, Pseudomonas putida VKM B-1301, Rhodococcus sp. VKM Ac-950, Mycobacterium flavescens VKM Ac-1415 in amount 75-85% of the total number of cells, as well as soil bacteria Agrobacteium radiobacter VKM B-1219 in amount 15-25% of the total number of cells. Sorbent in preparation represents finely-dispersed dehydrated zeolite with size of granules 0.1-0.5 mm, powdered with Aerosil A-300 nanoparticles. Ratio of components in preparation (wt %) is the following: zeolite - 94±1, Aerosil A-300 - 3±0.5, glycerol - 1±0.2, sodium nitrate - 0.5±0.2, potassium phosphate- 0.5±0.2, association of oil-oxidising microorganisms with Agrobacteium radiobacter in efficient quantity 2-3*108 kl/g - 1±0.5. Also claimed is method of preparation obtaining. Assocoation of oil-oxidising bacteria and fraction of soil bacteria are grown separately. After that, two obtained culture liquids are mixed in ratio 75-85% of association of oil-oxidising bacteria of the total quantity of cells and 15-25% of soil bacteria of the total quantity of cells. Suspension is concentrated to concentration 2*1011 kl/ml. Zeolite is preliminarily crushed to granules with size 0.1-0.5 mm, kept in furnace at temperature 250°C to the stage of swelling, cooled to temperature 20°C at humidity 10-12% and mixed with Aerosil A-300. Glycerol, sodium nitrate, potassium phosphate are introduced into concentrated suspension, and after that mixed with zeolite. Obtained zeolite and concentrated suspension are mixed with ratio 9:1. After that process of contact-chemical dehydration is carried out.
 Hybrid protein having prolonged action, based on recombinant human interferon alpha-2 (variants), method of its production and strain of saccharomyces cerevisiae for implementing this method (versions) / 2515913
Hybrid proteins GFN80 and GFN100 are formed based on recombinant human interferon alpha-2 fused on the N-terminus with the amino acid sequence of polypeptide S(G4S)16 or S(G4S)20, respectively. The strains of producer Saccharomyces cerevisiae RNCIM Y-3927 and Saccharomyces cerevisiae RNCIM Y-3928 are produced by recombinant method. The strains are used in the method of production of the hybrid protein GFN80 and GFN100, which comprises culturing under suitable conditions of yeast cells transformed by the expression vector, which contains the region of replication initiation of endogenous 2-micron plasmid of yeast Saccharomyces cerevisiae, and the promoter of yeast GAL1 controlling the expression of the gene comprising the DNA sequence SEQ ID NO:1 or SEQ ID NO:2, respectively, followed by isolation of the hybrid protein from the culture fluid.
Strain of fungus stagonospora cirsii davis having herbicidal activity against canada thistle / 2515899
Strain of the fungus Stagonospora cirsii Davis of All-Union Research Institute of Plant Protection 1.42 has herbicidal activity against canada thistle and species closely related to it - S. Cirsii. It was deposited in the State Collection of Microorganisms of All-Union Research Institute of Plant Protection under the collection number of Stagonospora cirsii Davis of All-Union Research Institute of Plant Protection 1.42 and can be used for biological control of canada thistle.
 Medication (versions), composition (versions) and application of medication (versions) for reduction of halitosis / 2515113
Invention relates to versions of medication for reduction of halitosis, versions of compositions based on said medications and versions of application of said medications. Device for reduction of halitosis represents strains of microorganisms Lactobacillus acidophilus, selected from group Lactobacillus acidophilus DSM 19825, Lactobacillus acidophilus DSM 19826 and Lactobacillus acidophilus DSM 19827. Version of said medication represents culture supernatant of said strains. Also claimed are compositions, which contain said microorganisms, and their application for reduction of halitosis.
 Method of differentiating bacillus anthracis from other closely related species of genus bacillus based on determining differences in structure of chromosomal genes / 2514663
Method includes sample preparation, DNA isolation, PCR statement. At that in carrying out PCR, the oligonucleotide primers are used, which are complementary to sequences of the chromosomal genes fliC and hom2, having the following sequences: fliC-F: 5'-TGGAGCAGTAACAATTGG-3', fliC-R: 5'-GCACCACTGATAGAAATGTTAG-3', hom2-F: 5'-GACGTGTTAAAAGAAGCCCA-3', hom2-R: 5'-CACCAATTTCGTCTTTTACA-3', followed by electrophoretic analysis of the amplification products, when the formation of the amplification product is 153 bps in size it is indicative of belonging of the strain under study to the species B.anthracis, formation of the amplification product with size of 550 bps is indicative of belonging of the strain under study to the other species of the genus Bacillus.
Strain of filamentous fungus aspergillus oryzae - producer of maltogenic alpha-amylase / 2514224
Strain of fungus Aspergillus oryzae Amy T-52-3-21 produces maltogenic α-amylase, and it is deposited in the All-Russian Collection of Microorganisms Institute of Biochemistry and Physiology of Microorganisms n.a. GK Scriabin RAS under the number F-4476D. The strain is made on the basis of strain Aspergillus oryzae of All-Russian Collection of Microorganisms F-3927D using the genetic engineering methods. Activity of α-amylase at 120 h of growth of the strain is 600-640 units/ml.
Strain of bacteria bacillus vallismortis - destructor of oil and oil products / 2513702
Strain of bacteria Bacillus vallismortis VKPM V-11017 is proposed - destructor of oil and oil products. Strain may within short period of time in the wide range of temperatures from +8 to +37°C degrade oil by 78.3%.
Method to clean permafrost soils and water environment by spore-forming bacteria bacillus atrophaeus vkpm v-10592 / 2513699
Strain of bacteria Bacillus atrophaeus VKPM V-10592 is grown, and suspension is prepared from it, which is introduced into permafrost soil and water environment. Maintained at the specified parameters from 7 to 60 days, and then they determine quantity content of oil and oil products in permafrost soil and water environment.
Method of differentiating bacillus anthracis from other closely related species of genus bacillus based on determining differences in structure of chromosomal genes / 2514663
Method includes sample preparation, DNA isolation, PCR statement. At that in carrying out PCR, the oligonucleotide primers are used, which are complementary to sequences of the chromosomal genes fliC and hom2, having the following sequences: fliC-F: 5'-TGGAGCAGTAACAATTGG-3', fliC-R: 5'-GCACCACTGATAGAAATGTTAG-3', hom2-F: 5'-GACGTGTTAAAAGAAGCCCA-3', hom2-R: 5'-CACCAATTTCGTCTTTTACA-3', followed by electrophoretic analysis of the amplification products, when the formation of the amplification product is 153 bps in size it is indicative of belonging of the strain under study to the species B.anthracis, formation of the amplification product with size of 550 bps is indicative of belonging of the strain under study to the other species of the genus Bacillus.
 Differential diagnostic nutrient medium for identification of yersinia bacterium / 2511436
Invention refers to biotechnology, microbiology, and concerns the recovery and identification of pseudotuberculosis and intestinal yersiniosis agents (Y. Pseudotuberculosis and Y. Enterocolytica). A nutrient medium contains microbiological agar (dry), lactose, glucose, urea, calcium chloride, 1% alcoholic phenol red, 1% alcoholic methylene blue and distilled water in specific proportions.
Method for preparing microgravimetric immunosensor / 2510830
Invention refers to biotechnology and microbiology. What is presented is a method for preparing a microgravimetric immunosensor. A surface of a quartz crystal resonator is activated first by plasma spray coating with polyethylene imine of molecular weight less than 10000 Da for 10 s in a vacuum unit at UHF field power 30 Wt. Thereafter, immunoglobulins from a solution with the protein concentration of 0.125 mg/ml are immobilised on the activated surface of the quartz crystal resonator. That is followed by washing in distilled water and drying in an air flow.
 Method for quantitative assessment of bactericidal activity of disinfectants / 2510610
Invention refers to microbiology and biotechnology. Analysed bacterial strains are inoculated on a dense nutrient medium. Paper disks impregnated with a disinfectant are applied. They are incubated in a temperature chamber until the bacteria start growing. The bacterial growth inhibition areas are measured. A quantity of the grown colonies is counted to construct a dependence diagram of the bacterial growth inhibition area and the quantity of the grown colonies after the disinfectant reaction. The diagram and Shughart inspection sheet are used to assess the disinfectant activity on specific types of the microorganisms. The disinfectants with mean measurements of the bacterial growth inhibition area are above an upper control limit of the Shughart inspection sheet are considered to be high bactericidal activity agents. The disinfectants with mean measurements of the bacterial growth inhibition area are below a lower control limit of the Shughart inspection sheet are considered to be low bactericidal activity agents. The disinfectants with mean measurements of the bacterial growth inhibition area between the control limits of the Shughart inspection sheet are considered to be mean bactericidal activity agents in relation to all analysed agents.
 Method of species identification of l.casei/paracasei, l.fermentum, l.plantarum and l.rhamnosus / 2508406
Invention refers to a method of species identification of L.casei/paracasei, L.fermentum, L.plantarum, L.rhamnosus lactobacilli. The proposed method involves performance of a PNR reaction with species-specific primers; besides, primers are built, which are specific to the first gene of operon FIFO ATP of synthase (a gene of subunit a) and a gene of uracylphosphoribosyltransferase, which precedes it, for L.casei/paracasei and L.rhamnosus and a gene of uracyltransport protein for L.plantarum and L. fermentum.
 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.
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.
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.
 Method for quick growth, detection and identification or counting of microcolonies of microorganisms at early stage / 2505607
Invention pertains to the method for quick growth, detection and identification or counting of microcolonies of microorganisms at early stage. The described method includes the following stages: obtaining the container with medium with porous element located above or under the top surface of the medium, note that the medium has nutritious substances for quick growth of microcolonies of microorganisms and porous element has pores from 1000 to 107 Da; pouring the liquid sample without serial dilution into container to the area above porous element; capturing the microorganisms in porous element or at the medium above porous element; incubation of container for the time sufficient for quick growth of microcolony at an early stage; transfer of porous element and any medium above it from container to the secondary medium for evaluation, detection and identification of microorganisms; and microcolonies research relatively the growth, detection, identification or counting of microorganisms. The said method for growth, detection and identification or counting of microorganisms takes not more than approximately six hours.
Method for increasing biocidal and therapeutic action of suspension-cream with linco-spectin / 2505285
Invention represents a method for increasing biocidal and therapeutic action of a suspension-cream with linco-spectin consisting in detoxification and polymerisation of linco-spectin 100 g in water 300 ml by 0.15±0.05% glutaric aldehyde 0.15±0.05% alkyldimethyl benzylammonium chloride at 38-40°C for 2-3 days.
Method to prepare medicated product from live strains and microorganisms of lactobacilli and bifidobacteria "lb-complex plus" / 2517734
Method provides for cultivation at 37±1°C of strains of lactobacilli and bifidobacteria in the medium and packing of liquid product with account of daily dose necessary for patients. The medium contains components in the following quantities: caseine hydrolysate dissolved by distilled water, 0.33-0.4 g/l, sodium chloride 5 g/l, fructose 10 g/l, peptone 2 g/l, agar-agar 0.75 g/l for lactobacilli, for bifidobacteria - 1.0 g/l, ascorbic acid 0.25 g/l, distilled water 0.67-0.6 g/l. Strains-producers are Lactobacillus plantarum 8 RA-3, Lactobacillus fermentum 39, Lactobacillus fermentum 90 TC-4, Bifidobacterium bifidum 791, Bifidobacterium longum 379, Bifidobacterium bifidum 1. For lactobacilli and bifidobacteria they prepare accordingly media with different content of agar-agar, amine nitrogen in caseine hydrolysate 160-170 and 180-200 mg% and medium pH 7.8-8.0 and 8.5-8.6. Starting from the first generation the bifidobacteria and lactobacilli are cultivated for 24±1 hours, two strains together, one separately. The produced biomass is mixed with the fresh nutrient medium at the ratio of 1:1000, and strains of lactobacilli are cultivated 24±1 hours, strains of bifidobacteria - 48±1 hours. Upon completion of cultivation of biomass of strains they are mixed at the ratio of 2:1:2:1.
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FIELD: biotechnologies. SUBSTANCE: nutrient medium is proposed for extraction of Shigella kind bacteria from water objects. The nutrient medium contains the following components: 0.1% alcohol solution of bromcresol green - 9.0 ml; 0.2% alcohol solution of methyl red - 3.0 ml; fermentative peptone - 0.5 g; extract of fodder yeast - 4.5 g; potassium dihydrophosphate - 8.7 g; caustic soda - 1.4 g; sodium chloride - 5.0 g; distilled water - up to 1000 ml; medium pH is 6.70-6.85. EFFECT: invention makes it possible to increase accumulating properties of nutrient medium and to simplify its composition. 3 tbl, 6 ex
The invention relates to sanitary bacteriology and can be used by the institutions of the state sanitary-epidemiological service, oversight and monitoring of water quality in surface water bodies, areas of recreation, sewage and drinking water. No special culture media for isolation of Shigella from water bodies complicates their inoculation, which does not allow to assess the epidemiological security of water sources. Known nutrient medium for the accumulation of bacteria of the family Enterobacteriaceae containing as a source of nitrogen nutrition pancreatic hydrolysate fish and fodder yeast extract, glucose as carbon source of power, a buffer salt such as sodium phosphate and potassium dihydrophosphate, sodium chloride, giving the environment isotonic properties, selective agents - purified bile and brilliant green (RU 2267530 C2). This medium provides an inhibitory effect on representatives of gram-positive microorganisms, but it has the same degree of accumulation for all Enterobacteriaceae, including E. coli, which at high content in the studied sites have an antagonistic effect on Shigella. Therefore, this environment may not be used for isolation of Shigella when the research is of water bodies, obviously containing E. coli in large quantities. In practical laboratories for isolation of Shigella from environmental objects, including water, used Selenite environment (Guide to medical Microbiology. Private Microbiology and etiological diagnosis of infections. Book II/Ed. Labinsky A.S., Kostyukova N.N., Ivanova CM. - Moscow: BINOM, 2010. - S), selected as a prototype. Selenite environment included in HOWTO MU 2.1.5. Methods of sanitary-microbiological analysis of the coastal waters of the seas in places of water use of the population" (Ministry of health of Russia, Moscow, 2011) as a medium for isolation of Shigella. This environment has the following composition:
The environment is prepared from d the two solutions. Solution 1. Pre-determine the proportion of phosphates in one sample peptone and sodium hydrosalinity to ready medium had a pH of 7.0±0,1. After determining the ratio of phosphate to the solution add peptone and lactose. Poured into vials 50,0 ml and sterilized fluid steam 2 days for 30 minutes. Solution 2. a 10% solution of sodium hydrosalinity cookex temporesterile distilled water. Before working in a bottle with a 50.0 ml solution of 1 aseptically add 2.0 ml of solution 2. The finished medium is aseptically poured into 5,0-7,0 ml in sterile tubes and tightly stoppered. Further sterilization is not required. The disadvantage of environment is the fact that along with the concomitant inhibition of microorganisms, including E. coli, there is a simultaneous inhibition of the growth of Shigella. The aim of the invention is a liquid nutrient medium for isolation of Shigella from the water, providing efficiency savings of Shigella. The medium contains peptone is an enzymatic source of nitrogenous food, fodder yeast extract as a growth promoter, potassium dihydrophosphate and caustic soda as a buffer mixture and a source of mineral nutrition, sodium chloride to give the isotonic properties of the environment, bromocresol green and methyl red in kachestvennyie growth concomitant microflora in the following amounts:
Environment prepare three solutions. Solution 1. Sample peptone is an enzymatic extract of fodder yeast, potassium dihydrophosphate, caustic soda, sodium chloride is dissolved in a small amount of distilled water and bring to 1000 ml Heat the solution to dissolve the ingredients. Sterilized at 112°C (0.5 kgf/m2within 30 minutes Solution 2. 0,1 alcohol solution of bromocresol green cook ex tempore20% solution of ethanol: 0.1 g of bromocresol green is dissolved in 99.9 g of 20% ethanol. Solution 3. 0,2% alcohol solution of methyl red cookex tempore60% ethanol: 0.2 g of methyl red was dissolved in 99.9 g of 60% ethanol. Before you begin to one liter of prepared nutrient medium add 9 ml of 0.1% alcohol solution of bromocresol green and 3 ml of 0.2% alcohol solution of methyl red. Solutions 2 and 3 are used only freshly prepared. This method is illustrated by the following examples. Example No. 1. 0,1% alcohol solution of bromocresol green - 5,0 ml of 0.2% methyl alcohol solution of red - 2.5 ml, peptone 0.3 g EKD - 4.0 g, potassium dihydrophosphate - 8.0 g, sodium hydroxide, 1.2 g, sodium chloride 3.0 g distilled water to 1000 ml, pH of 6.3. Further, the method is carried out as described above. Example No. 2. 0,1% alcohol solution of bromocresol green, and 9.0 ml of 0.2% methyl alcohol solution of red - 3,0 ml peptone - 0.5 g, EKD - 4.5 g, potassium dihydrophosphate - 8.7 g, caustic soda 1.4 g sodium chloride 5.0 g distilled water to 1000 ml, pH of 6.7. Further, the method is carried out as described above. Example No. 3. 0,1% alcohol solution of bromocresol green - 10.0 ml, and 0.2% methyl alcohol solution of red - 4,0 ml peptone - 1.0 g, EKD - 5.0 g, Kali is the dihydrophosphate - 9.2 grams, sodium hydroxide and 1.5 g of sodium chloride 5.5 g, distilled water to 1000 ml, pH of 7.1. Further, the method is carried out as described above. Data on cumulative environment properties are presented in table 1, which shows that the growth of S.sonnei registered before dilution 10-7on the environment with the minimum and the optimum content of the components. On the environment with the maximum content of the components of growth S.sonnei recorded breeding 10-6. Growth S.flexneri was absent in dilutions of 10-6, 10-7on the environment with minimal amount of components and in dilutions of 10-7on the environment with the maximum content of components, while in the medium with the optimum content of the components of the growth of the test strain was observed in the dilution 10-7. The growth of E. coli (estimated accompanying microflora) before dilution 10-7on the environment with minimal amount of components, environment optimum, and maximum content in the dilution 10-5. Thus, the optimum proportion of the ingredients for the growth of bacteria of the genus Shigella and suppression of concomitant microflora is:
Example No. 4. The sensitivity of the environment was determined by the highest dilution, which in the subculture on solid nutrient medium was observed the growth of the test strains. Test strains - S.flexneri a, S.sonnei, E. coli 3912/41 obtained from the collection of gisk named after. L.A. Tarasevich. Comparative characteristics of sensitivity of the two environments accumulation are presented in table 2. The table shows that the experimental environment is more sensitive than Selenite (Wednesday comparisons), as the growth of the test strains was observed before dilution 10-7, while Selenite on the growth environment S.flexneri was absent in the breeding of 10-5 growth S.sonnei was observed only up to a dilution of 10-6. The growth of E. coli was absent in both environments accumulation in dilution 10-5. Example No. 5. Efficiency savings of Shigella in two environments was investigated on the example of the test strains: S.flexneri and S.sonnei. Seeding of microorganisms was carried out in Selenite and experienced environment in the form of microbial suspensions prepared according to the standard sample turbidity TOC No. 517-1V 2010 issue of dilutions from 10-4up to 10-7subsequent reseeding on mastopathy agar. The results of the comparison of the efficiency savings of Shigella grown during incubation on experienced and Selenite environments, are presented in table 3. These data indicate that the efficiency of accumulation of test strain S.sonnei on the experimental environment in 107times higher than Selenite, a S.flexneri - 106times, respectively. Example No. 6. Explored the practical application environment for the accumulation and excretion of Shigella from surface water reservoirs with different degree of pollution. Quantitative determination of Shigella were carried out by seeding the water in two parallel rows in the volume: 100 ml, 10 ml, 1 ml and so on, depending on the degree of biological contamination of water object. Just was studied 28 samples R. don. With the help of experienced environment selected 3 strain S.flexneri, when using Selenite environment is not what was allocated a single strain of Shigella. Thus, the proposed nutrient medium provides the increase of the cumulative properties of the nutrient medium for the reduction in inhibitory effects on the growth of Shigella and significant inhibition of growth of associated microorganisms, including E. coli.
Nutrient medium for isolation of bacteria of the genus Shigella from water bodies containing peptone is an enzymatic, distilled water, characterized in that it contains the extract of fodder yeast, potassium dihydrophosphate, caustic soda, sodium chloride, bromocresol green and methyl red in the following amounts:
pH 6,70-6,85.
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