Method for determining adhesive properties of bacteria of enterococcus family by means of caco-2 cell line
SUBSTANCE: method involves preparation of bacterial suspension, separation of the obtained biomass of bacteria by centrifugation, dilution of the obtained biomass in physiological solution, preparation of monolayer of CaCo-2 cells, introduction of bacterial culture, cultivation of cells, flushing with physical solution, removal of monolayer with bacterial cells and counting of the amount of bacterial cells related to 1000 cells of CaCo-2; with that, bacteria refer to highly-adhesive ones if the number of bonded cells is 1010 to 3000, to average-adhesive ones of 210 to 1000, and to low-adhesive ones of 0 to 200.
EFFECT: improvement of the method.
The invention relates to the field of Microbiology and biotechnology and can be used in food biotechnology. The method is used to determine the adhesive properties of bacteria using the cell line of SASO-2 (epileptologie cells adenocarcinoma colon man in the monolayer.
A known method of determining the adhesion of microbes to the epithelial cells of the intestine of rats or mice, are described in the THROES 4.2.2602-10. 4.2. Control methods. Biological and microbiological factors. System pre-clinical studies the safety of drugs. The selection, validation and storage of industrial strains used in the production of probiotics. Methodical instructions. For staging techniques adhesion use daily culture strains. The daily culture of the strain grown on stubble adequate nutrient medium wash phosphate buffered saline (SFR) and then washed twice SFR with a pH of 7.2 by centrifugation (1500 rpm, 10 min). In addition, culture can be grown on solid medium, covered with cellophane, at 37°C for 16-18 hours Preparing a suspension containing 2×109CFU/ml Adhesiveness of the strains studied in the in vitro system in the epithelial cells of the intestine and colon of rats Fischer. Rats or mice hammer carbon dioxide from the intestines secrete pieces size ×5 mm, carefully washed from mucus and content in SFR. Epithelial cells derived from pieces of the intestinal mucosa on the vibrator model runway.
Cells are washed twice with cooled SFR (pH of 7.2) by centrifugation at 800 rpm for 10 min. After determining the concentration of suspended cells with the camera Goryaeva under a light microscope (bring it up to 2×106cells/ml of Preparing a suspension of the tested culture concentration of 2×109CFU/ml To 0.5 ml suspension of epithelial cells are 0.5 ml of bacterial suspension culture. The cell is a bacterial mixture incubated in a thermostat at (37±1)°C for 30 min, shaking occasionally. The mixture is then washed three times SFR from unattached bacteria at 600 rpm for 10 minutes All manipulations carried out in the cold. To the precipitate add 1-2 drops SFR and prepare smears on the glass. Drugs record 96° alcohol and stained by Romanovsky-Giemsa. Under light microscope, count the average number of bacteria adhering to 25 enterocytes or colonocytes. When assessing the adhesion of each strain of microorganism experience repeated at least 3 times. Calculate the average adhesion (SPA), the number of microbes on 1 cell - microbe/cell 10 fields of view, taking into account the results of all experiments.
The level of adhesion of certain bacteria strains conditionally separated into four degrees:
- non-adhesive (When A = 0);
- slaboagressivnoy (SPA = 1-5);
- sredneagressivnyh (SPA = 5-10);
- high adhesive (SPA above 10).
The described method is a placeholder.
The disadvantage of the above method is that this method uses animal cells (cells of rats and mice). Testing on animals used for food borne pathogens has long been known. But currently, the development of cell science is allowed to conduct research directly on the epithelial cells of the person. It plays an important role, especially in the field of food biotechnology.
The technical result is to increase the accuracy of detection of the adhesive activity of probiotic bacteria of the genus Enterococcus was developed by way of definition on the epithelial cells of the colon of a person line SASO-2 monolayer.
The task of the invention is directed to the development of a method for determining the adhesion properties of the probiotic bacteria of the genus
Enterococus. The method uses a cell line SASO-2 monolayer, which is obtained from the colon of a person.
The problem is solved developed a method for determining the adhesive properties of bacteria of the genus Enterococus using cell lines SASO-2, including the preparation of bacterial suspensions: the cultivation of bacterial strains on the corresponding liquid medium for 18 h; branch is derived biomass from the culture medium by centrifugation, for this purpose, 20 ml of culture suspension of each strain centrifuged at minus 4°C with a frequency of 4000 rpm for 15 min; the breeding of obtained biomass in saline to a concentration of 4×107CFU/cm3; preparation of monolayer cells SASO-2: growing monolayer cells SASO-2 in culture medium DMEM with 15% serum, fetal bovine, 5% of the antibiotic penicillin-streptomycin, 6-hole tablet for 5 days; removing the formed monolayer nutrient laboratory aspirator; rinsing the monolayer cells twice with saline solution; determination of adhesive properties of bacteria in each well of the tablet to form a monolayer of cells SASO-2 contribute 0.5 ml of bacterial culture and 2 ml of DMEM medium with 15% serum fetal bovine; cultured for 1 h at 37 ° °C; remove nutrient medium laboratory aspirator and washed the monolayer cells twice with saline solution, removing the monolayer with related bacterial cells carry out the solution of Versene 0.02% and trypsin 0,25% in the ratio of 3:1 in each well of a 6-hole tablet add 1 ml of solution for counting related bacterial cells use dilution method for growing cells on a corresponding dense environment, the adhesive properties of microorganisms is determined by the number of the bacteria, contacting 1000 cells SASO-2: from 1010 to 3000 bacterial cells - high adhesive strain; from 210 to 1000 - sredneetazhnye; from 0 to 200 - riskadvisory.
The method of determining the adhesion properties of the probiotic strains of bacteria of the genus Enterococus using lines of epithelial human cells based on the interaction of bacteria with a cell wall epithelial cells of colon carcinoma human colon, SASO-2 monolayer, and the counting of bound peroxidase bacteria monolayer.
The advantages of the proposed method are that the method uses directly the human cell SASO-2 to determine the adhesive properties of bacteria, which is one of the important properties of probiotic strains used in the food industry.
The method allows to study analyzed the properties directly on the epithelial cells of the human, which provides a basis for extrapolating the results to humans in General. With this method cells are used in the form of a monolayer, it allows to determine the amount of adhesion of bacteria with greater accuracy.
To calculate related bacterial cells were used dilution method for growing cells in a dense environment MRS. Adhesive properties of microorganisms is determined by the number of bacteria that communicates with 1000 cells SASO-2: from 1010 to 3000 smear the territorial cells high adhesive strain; from 210 to 1000 - sredneetazhnye; from 0 to 200 - riskadvisory. The gap between the results given by the error in the count.
A specific example of the proposed method
Determination of adhesive properties of strains of Enterococcus thailandicus PBC-2 (VKPM B-10684) and Enterococcus faecalis 55 (VKPM B-8652).
Carry out the preparation of bacterial suspensions: cultivated strains of Enterococcus thailandicus PBC-2 and Enterococcus faecalis 55 on the respective liquid medium for 18 h; separate the produced biomass from the culture medium by centrifugation, for this purpose, 20 ml of culture suspension of each strain centrifuged at minus 4°C with a frequency of 4000 rpm for 15 min, diluted obtained biomass in saline to a concentration of 4×107CFU/cm3. Growing monolayer cells SASO-2 in culture medium DMEM with 15% serum, fetal bovine, 5% of the antibiotic penicillin-streptomycin, 6-hole tablet for 5 days; then remove from the formed monolayer culture medium laboratory aspirator; washed the monolayer cells twice with saline. To determine the adhesive properties of bacteria in each well of the tablet to form a monolayer of cells SASO-2 contribute 0.5 ml of bacterial suspension and 2 ml of DMEM medium with 15% serum fetal bovine; cultivate the for 1 h at 37°C; remove culture medium laboratory aspirator and washed the monolayer cells twice with saline. Removing the monolayer with related bacterial cells carry out the solution of Versene 0.02% and trypsin 0,25% in the ratio of 3:1 in each well of a 6-hole tablet add 1 ml of solution.
To calculate related bacterial cells use dilution method for growing cells on a corresponding dense environment, the adhesive properties of microorganisms was determined by the number of bacteria by contacting 1000 cells SASO-2: from 1010 to 3000 bacterial cells - high adhesive strain; from 210 to 1000 - sredneetazhnye; from 0 to 200 - riskadvisory.
The results showed that with 1000 cells SASO-2 contacts 2×103the cells of E. thailandicus PBC-2, 109×103the cells of E. faecalis 55. This explores what it Is. thailandicus PBC-2 and E. faecalis 55 are characterized by high adhesive properties.
Grown on tablets monolayer cells SASO-2 can be considered as a model of the intestinal epithelium, therefore the use of this method improves the accuracy of detection of the adhesive activity of probiotic microorganisms used in the food industry.
The method for determining the adhesive properties of bacteria of the genus Enterococcus using cell lines SASO-2, including the preparation of bacterial suspension, the Department received b is MASSY at 4000 rpm for 15 min, breeding the obtained biomass in saline to a concentration of 4×107CFU/cm3growing monolayer cells SASO-2, removing the formed monolayer nutrient laboratory aspirator, washing of the monolayer cells twice with saline solution and determining the adhesive properties of bacteria in each well of the tablet to form a monolayer of cells SASO-2 contribute 0.5 ml of bacterial culture and 2 ml of DMEM medium with 15% serum, fetal bovine, cultured for 1 h at 37°C, remove the culture medium laboratory aspirator and washed the monolayer cells twice with saline solution, removing the monolayer with related bacterial cells carry out the solution of Versene 0.02% and trypsin 0,25% in the ratio 3:1, to count related bacterial cells use dilution method for growing cells on a corresponding dense MRS adhesive properties of bacteria was determined by the number of bacteria by contacting 1000 cells SASO-2: 0-200 - riskadvisory strain, from 210 to 1000 - sredneetazhnye, from 1010 to 3000 bacterial cells - high adhesive.
SUBSTANCE: aggravated chronic obstructive bronchitis in the females suffering A(H3N2) influenza on their second trimester of gestation is analysed by paired blood serums. The first blood serum (titre) (A) is analysed for the content of anti-influenza antibodies; there are also measured the differences of maximal temperature at the height of the disease on the 2nd-4th day and maximal body temperature on the 6th-8th day of the diseases (B) and the variations of the average-molecular peptide concentrations at the height of the disease on the 2nd-4th day and on the 6th-8th day of the diseases (optical density units) (C). A discriminant equation is used to calculate a discriminant function D: D=-0.063×A+2.386×B+294.340×C. If D is equal to 4.54 or more, the absence of the intrauterine foetus infection is predicted, while D less than 4.54 enables predicting the intrauterine foetus infection in the females diagnosed with the above.
EFFECT: using the method provides a probability of the correct prediction, enables the well-timed therapeutic measures aiming at the prevention of the intrauterine infection in aggravated chronic obstructive bronchitis in the females with A influenza on their second trimester of gestation.
SUBSTANCE: substance of the declared method consists in determining the genotypes and alleles of polymorphous variants of folate metabolism genes MTHFR C677T and A1298C, MTRR G66A by polymerase chain DNA synthesis with restriction analysis: If observing a combination of the allele T of the MTHFR C677T promoter polymorphism, the allele C of the MTHFR A1298C promoter polymorphism, the allele A of the MTRR G66A promoter polymorphism, the development of food intolerance is predicted in children.
EFFECT: using the method enables planning the preventive measures and therapeutic approach to an infant, ensures preventing the development of food-related aggravations and complications, and providing the maximum improvement of quality of life and working ability.
SUBSTANCE: children's blood serum is examined for lymphocyte count expressing CD3+; CD4+; CD8+, interferon γ (INFγ) and interleukin 4 (IL-4) on their surface. Reducing CD3+ in 1.5 sigmal interval to 51.7-59.9%; CD4+ - to 20.3-31.1%; CD8+ - to 13.8-19.6% and increasing IL-4 in CD4+ cell populations in 1.5 sigmal interval to 22.3-27.7%, IL-4 in CD8+ cells - to 27.4-31.6%, and INFγ in CD8+ cells - to 13.2-18.0% enable diagnosing acute burn toxaemia in children.
EFFECT: technique is easily reproducible, requires no expenses, providing early diagnosis of the diseases and enables diagnosing acute burn toxaemia in children.
2 tbl, 2 ex
SUBSTANCE: method consists in measuring the information values: newborn's birth weight, erythrocyte count, hemoglobin and haptoglobin levels to calculate a diagnostic index by formula: DI=0.004×BW+0.971×Er-0.013×Hem-0.418×Hapt-4.344, wherein BW is newborn's birth weight (gram), Er is the venous blood erythrocyte count (×109/l), Hem is the venous blood hemoglobin level (g/l), Hapt is the umbilical blood serum haptoglobin concentration (mcg/dl). If DI is more than 0, the viability of the newborn with ELBW is considered to be high, while DI less than 0 enables stating the viability of the premature newborn with ELBW to be low.
EFFECT: method enables the objective assessment of the newborn's state, viability potential and the follow-up approach correction.
SUBSTANCE: cervical mucus MIP-1α is determined in the pregnant women diagnosed with an urogenital infection on their 1st trimester. If the cervical mucus MIP-1α value is more than 36.9 pg/ml, a high risk of the 1st trimester miscarriage is predicted.
EFFECT: more accurate prediction of the threatening miscarriage accompanying the urogenital infection for the purpose of prescribing the current therapeutic measures.
SUBSTANCE: invention can be used in laboratory diagnostics for determination of blood atherogenicity as per level of content of minimum modified lipoproteins of low density (MM-LPLD) in serum or plasma of human blood. MM-LPLD is aggregated from blood serum or plasma by treatment with buffer containing polyvinyl pyrrolidone (PVP) with molar weight of 12600±2700 at final PVP concentration of 11.3% to 14.2% in the sample. After incubation during 10 minutes, light absorption in test and check samples is measured, difference is calculated and at the difference value of more than 10 U blood atherogenicity of the examined person is stated due to increased MM-LPLD level.
EFFECT: use of easy, available and easy-to-implement method of determination of MM-LPLD in human blood allows performing screening investigations in order to determine availability of an atherosclerotic process at a pre-clinical stage and clinical examination of population.
3 tbl, 3 ex
SUBSTANCE: what is involved is the immunopathological examination using indirect immunofluorescence. At the first stage, patient's serum is incubated on monkey's renal tissue section; at the second stage; it is incubated with goat anti-species fluorescent serum. The reaction is recorded by fluorescence microscopy by specifying the number of renal tubules with a fluorescent basal membrane and expressing the measurement as a percentage. If the derived value exceeds 15%, a considerable increase of the probability of the non-surgical treatment failure is predicted.
EFFECT: method is easy to implement; it takes 2 hours to be implemented and requires no expensive chemicals to be used.
SUBSTANCE: liposomes are used as a matrix for activated ferment - horseradish peroxidase. To 5 mg of horseradish peroxidase oxygenated with a periodate method there added is 1 ml of liposome suspension in 0.01 M solution of carbonate-bicarbonate buffer at pH 9.5, and subjected to ultrasonic treatment during 1 min. Then it is incubated for 1 hour; immobilised with immunoglobulins in concentration of 5 mg during 2 hours at the temperature of 22±4°C; stabilised with 5 mg of sodium borane with further gel-chromatographic cleaning.
EFFECT: invention allows obtaining liposomal-immunoperoxidase conjugate for indication of infectious agents in an immunoenzymometric analysis and increasing service life of a preparation up to 6 years.
1 tbl, 3 ex
SUBSTANCE: invention refers to biotechnology, namely to a method for Mycobacterium leprae antibody test. The method involves the immunoenzyme blood serum test for Mycobacterium leprae antibodies. A test antigen in the immunoenzyme test is an aqueous suspension of Mycobacterium lufu cultured in a thermostate on the Lowenstein-Jensen medium for 7 days at temperature 37°C, heated at 100°C for 1.5 hours on a water bath.
EFFECT: presented invention enables simplifying the method for Mycobacterium leprae antibody test.
SUBSTANCE: versions of an antibody or its fragment, which are specific in relation to β-amyloid protein, are proposed. Each version is characterised by the fact that it includes H- and L-chains, or areas VH and VL, each of which contains three corresponding CDR. The following is described: polypeptide VL, polypeptide VH, as well as coding nucleic acid, expression vector containing it and a cell carrying the vector, which are used for obtaining an antibody or its functional fragment. The following is proposed; a test kit, versions of pharmaceutical composition, a mixture to be used as a medicine based on the antibody or its functional fragment. Versions of the method used for production of an antibody are described: using a cell, nucleic acid or a vector. A composite preparation method, as well as an in vitro amyloid disease diagnostics method, a method for determination of a degree of loading with in vitro amyloidogenic patches, a method for curing or relief of actions of amyloid disease, which use an antibody or its functional fragment, are described. Inventions can be used in therapy and diagnostics of Alzheimer disease and other enlisted amyloid diseases.
EFFECT: proposed inventions provide new antibodies that bind the epitope contained in the area of 12-23 protein αβ1-42; with that, residues 15-20 have a fundamental importance.
44 cl, 18 dwg, 9 tbl, 16 ex
SUBSTANCE: preparation containing biodestructor of oil contamination represents centrate of cultural liquid of microbial mass of consortium of oil-oxidising microorganisms immobilised on peat carrier. Consortium composition includes the following: Rhodococcus eqvi SRI MCC B-1115 bacteria strain, Rhodotorula glutinis SRI MCC Y-1113 yeast and Rhodotorula glutinis ARSRI MCC Y-1114 yeast strain in the ratio of 1:1:1 respectively, which have been grown jointly.
EFFECT: invention allows improving quality of soil cleaning from oil and oil products under conditions with reduced temperatures.
SUBSTANCE: proposed chimeric protein with SEQ ID NO:02 is fluorescent biosensor, built on the basis of HyPer protein and mutant of PH-domain of Btk tyrosine kinase.
EFFECT: proposed inventions allow performing simultaneous monitoring of product of hydrogen peroxide and phosphatidyl inositol-3,4,5-triphosphate in a living cell.
4 cl, 4 dwg, 3 ex
SUBSTANCE: 1% sterile solution of glucose is prepared on the basis of a physiological solution, which is used as a nutrient medium. An absorber by Zaytsev is connected to an aspirator of "Briz-1" grade, into the flask of which 10 ml of the prepared 1% glucose solution is placed. The device is placed into the investigated room, and the aspirator is switched on for 15 min. Microorganisms that are in air go through the glucose solution and are retained in it. The solution is placed into a test tube and thermostatted at 37°C for 2 hours. Solution electroconductivity is measured with the help of a sensor KDS-1038. The number of microorganisms in air is determined according to the curve of empirical dependence of solution electroconductivity on the number of microbes, which is built in accordance with the produced values.
EFFECT: invention makes it possible to reduce time for determination of microorganism number in the air of the working area down to 2 hours and 20 min.
SUBSTANCE: method to protect yeast Saccharomyces cerevisiae against oxidising stress as a result of exposure to hydrogen peroxide includes growing of yeast culture under standard conditions till the end of the logarithmic or the start of the stationary phase of growth, incubation with a protective agent, exposure to hydrogen peroxide with subsequent determination of the number of survived cells. The protective agent is represented by soy inhibitors of proteases in concentration of 0.1-2.0 g/l. The time of incubation with the protective agent makes 5-6 hr, concentration of hydrogen peroxide is 700-750 mM. The invention provides for high extent of yeast cells production with no depressant action.
EFFECT: share of survived yeast cells during method realisation increases in comparison with the reference one 1,2-5,4 times.
SUBSTANCE: composition for production of organosilicon sol-gel matrix for immobilisation of microorganisms in biosensor analysers is provided. The composition consists of 20% polyethyleneglycol solution in phosphate buffer solution, Tetraethoxysilane, and 0.2 mol/dm3 catalyst solution NaF, an additionally introduced hydrophobic additive - methyltriethoxysilane. The components are taken at a volume ratio of PEG:TES:MTES: NaF 4:(18-3.4) (2-16.6):1.
EFFECT: reducing the toxic effect of the matrix on the biomaterial and increase in its mechanical strength.
1 tbl, 1 ex
SUBSTANCE: present invention relates to microbiology and a method of detecting and counting viable Legionella pneumophila microorganisms in a sample. The described method involves: (1) contacting said microorganisms of said sample with at least one reducing compound which contains glutamate and pyruvate, and a culture medium which is a buffered charcoal yeast (BCYE) or GVPC agar culture medium, (2) incubating the product of step (1), and (3) detecting and determining the number of viable microorganisms. The reducing compound used directly or indirectly affects metabolism, reducing oxidative stress of the microorganism by reducing formation and/or breaking down reactive forms of oxygen.
EFFECT: disclosed method enables to accurately count Legionella pneumophila microorganisms in a stressed condition.
7 cl, 5 dwg, 5 ex
SUBSTANCE: method of qualitative assessment of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys in operation of spacecrafts and the suspension of spore materials fungi of implementation of the said method is proposed. The test and control samples of aluminium and magnesium alloys are prepared. The prepared samples are dried, sterilised. Fungal cultures - strains of microorganisms Paecilomyces variotii Bainier of All-Russian collection of microorganisms F-4039D, Ulocladium botrytis Preuss of All-Russian collection of microorganisms F-4032D, Penicillium chrysogenum Thorn of All-Russian collection of microorganisms F-4034D, Aspergillus sydowii (Bainier et Sartory) Thorn et Church of All-Russian collection of microorganisms F-4037D, Cladosporium sphaerospermum Penz. of All-Russian collection of microorganisms F-4041D are inoculated for growing spores into tubes with a sloped agar medium of Czapek-Dox. The tubes are thermostated at a temperature of (29+2)°C for 14-28 days until appearance of mature spores. Spore materials suspensions of individual cultures of the said fungi are prepared, which are then mixed in equal proportions. And the concentration of spores of each fungi species in the suspension should be in the range of 1-2 mln/cm3. The resulting suspension is applied to the sterilised test samples. Then they are dried and placed as one sample in each Petri dish on the surface of agar medium of Czapek-Dox. The control samples not treated by the spore material are also placed on the surface of the agar medium of Czapek-Dox in the Petri dishes. The Petri dishes with the test and control samples are placed in different desiccators at the bottom of which water is poured to maintain the humidity more than 90%. The desiccators are closed and incubated in the thermostat at a temperature of (29±2)°C. The exposition of test and control samples is carried out for 40, 120 and 180 days. Then the samples are removed from the Petri dishes, and the corrosion products and mycelium are removed from the surface of the samples washing them in running water. The samples are soaked in 70% ethyl alcohol for 30 minutes, then they are washed with detergent and dried. Then, using a scanning electron microscope a qualitative assessment of biocorrosion damages is carried out. According to the change in the appearance of the surface of the samples the initial stages of biocorrosion and its type is evaluated, as well as the distribution of corrosion damage on the surface of the samples, and the dependence of the biocorrosion process on time is determined.
EFFECT: inventions enable to carry out the qualitative assessment of the initial stages of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys with the thickness of more than 2 mm.
2 cl, 9 tbl
SUBSTANCE: invention relates to means of controlling quality of organic and inorganic products, and can be used to evaluate safety of food and fodder products, natural water and waste water, ground, soil, determining the maximum allowable concentration of contaminants, as well as the impact of human activities on the environment, including oil extraction and refining products. The apparatus consists of a computer with a software system and a biodetector. The biodetector has a housing 1 inside of which there is a displacement controller for a board 2 with containers 3 for test objects, a light source - light-emitting diodes 4, an optical system with a television camera 5 and a lens 6 mounted on a support 7. The apparatus is provided with a light-proof casing 8 for closing the top of the board with containers for test objects, the inner surface of which is coated with white paint. The optical system with a television camera 5 and a lens 6 is mounted on the support 7 by a turning mechanism 9 in form of a ball head. Inside the housing 1 there is a stepper motor 10 for rotating the board 2 with containers 3.
EFFECT: invention increases measurement accuracy while reducing the measuring time, simplifying the design and reducing the size of the apparatus.
2 cl, 2 dwg
SUBSTANCE: device comprises a through cell equipped with holes, where at least one hole represents an inlet hole for intake of fluid medium from the specified technological flow, and at least one hole is an outlet hole for discharge of fluid medium from the specified through cell. To one of specified holes an RK probe is attached, possibly, an OVP probe, a cleaning accessory. The first pipeline is connected to the inlet hole. Possibly, the second pipeline is connected to the outlet hole. A valve is connected to the specified through cell. With the help of the specified devices and methods they measure volume microbiological activity and surface microbiological activity in a process flow of water by means of measurement of dissolved oxygen concentration.
EFFECT: method improvement.
45 cl, 10 dwg, 2 tbl, 4 ex
SUBSTANCE: method includes the following stages: contact of a sample with a source of nutrition for cells, containing antioxidant, representing pyroracemic acid or its salt, and an inhibitor of cell proliferation, which is selected from ciprofloxacin and cefalexin; contact of the specified sample with fluorescent-marked oligonucleotide probes, capable of specific hybridisation at least with one section of ribosomal nucleic acids, which belong to microorganisms of Legionella pneumophila kind and type; and detection and quantitative determination of a fluorescent signal.
EFFECT: provided method and set make it possible to more accurately and reliably detect and calculate viable microorganisms of Legionella pneumophila type, having excluded natural fluorescence of microorganisms from calculation.
6 cl, 2 dwg, 6 tbl, 2 ex
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.