Differential diagnostic nutrient medium for identification of yersinia bacterium

FIELD: medicine, pharmaceutics.

SUBSTANCE: 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.

EFFECT: invention enables reducing the length of studies.

3 dwg, 2 tbl

 

The invention relates to medicine, namely to Microbiology, and for the isolation and identification of pathogens pseudotuberculosis and intestinal yersiniosis (K. pseudotuberculosis and Y. enterocolytica).

Pseudotuberculosis and intestinal yersiniosis are infectious diseases caused by Yersinia spp., transmitted through nutritional and characterized by polymorphism of clinical manifestations and a variety of clinical forms. On the territory of the Russian Federation and in the world are registered each year group and sporadic cases of yersiniosis, resulting in preschool institutions, schools, orphanages, country groups, military units, enterprises and educational institutions United by a common power source.

The most reliable results, confirming the diagnosis, get a microbiological isolation of Yersinia from pathological material ("gold standard" of diagnosis). However, isolation and identification of Yersinia associated with certain difficulties and primarily with the duration of the process. This leads to the need to improve methods of isolation and identification of Y. pseudotuberculosis and Y. enterocolytica by creating effective differential diagnostic of nutrient media.

Scheme microbiological release of Yersinia adopted at the present time, consists of after ovately steps: enrichment primary cultures on Petri dishes with nutrient media (such environments Serov, No. 67, Endo, Levin, Maurice and others), secondary screening with solid nutrient medium in a test tube with one of the differential media containing urea, in the form of a beveled column (Kligler, Altnickol and others). In these environments, Yersinia grow in the form of a dry matte coating on the beveled surface and in the course of injection over the entire height of the column with the decomposition of urea and formation of alkali (column after 24 hours incubation acquires crimson color). Cultures are then used for further identification by biochemical tests and serological tipirovanii (Fig.1).

Known nutrient medium for the diagnosis of bacteria of the genus YERSINIA.

1)the Differential diagnosis of combined nutrient medium for isolation of Yersinia (SBTS), manufactured by state research Institute of applied Microbiology (Obolensk, Moscow region).

The composition of the medium: nutrient agar pH of 7.8; for cultivation of microorganisms - 35,0 g

the purified bile - 6.0 g

glucose - 10, 0 g

urea-5.0 g

bronchiology blue water-soluble - 0,128 g

disubstituted phosphate potassium (Csnrt) - 1,0,

Method of preparation: components contribute in 1.0 l of distilled water, heated to boiling and boiled for 5 minutes on slow fire. Then filtered through a cotton-gauze filter is, again boil for 1-2 minutes, cool to 45-50°C and pour into Petri dishes. Keep the environment within 7-10 days at 4-7°C.

2) Medium with urea on Preus (modification of TZNIUS them. I.I. Mechnikov) indicator bratisalava blue and Christensen indicator bromophenol red.

The use of these environments has made more reliable determination of urease activity of bacteria, which in parallel with sowing in a combined environment do the planting in a test tube environment Christensen (prepa).

3) Environment Christensen with urea, composition:

Solution I. Peptone - 1.0 g

Sodium chloride - 5.0 g

Glucose - 1.0 g

Potassium phosphate one-deputizing - 2.0 g

Phenol red (solution 1:500) to 6.0 ml

Urea - 20,0 g

Distilled water to 100.0 ml

Solution II.

Agar-agar - 15.0 g

Distilled water - 900,0 ml

Sterilized with 0.5 ATM for 15 minutes.

The cooking. To a cooled to 50-55°C. the solution II was added 100 ml of solution I, stirred, poured aseptically into sterile tubes, the medium is cooled in an inclined position for receiving the column and the jamb. Prepared medium yellow color.

The course of the study.

Make massive sowing across the surface of the casing. Incubated at 37°C. are counted after 2, 4, 6 hours and the next day. In some cases, with negative results to see 4-7 days (voznagrazhdenie reaction). Positive result - red color.

However, the use of additional media with urea extends the detection and identification of Yersinia another 1-2 days.

Thus, microbiological identification of Yersinia takes 5 days, which is unacceptably long for making clinical. In addition, significantly increases the cost of bacteriological tests.

As a prototype, the applicant has chosen a well-known and widely used 3-sugar environment Altnickol and its modification. The use of this nutrient medium is regulated by the appropriate methodical documents (Epidemiology, laboratory diagnosis of yersiniosis, organization and carrying out of preventive and antiepidemic measures. The statement, approved. The USSR Ministry of health 30.10.1990, No. 15-6/42; Prevention of yersiniosis. Sanitary and epidemiological regulations SP 3 3.1.7.2617-10, appr. the resolution of the Chief state sanitary inspector of the RF dated 26.04.2010, No. 37; Epidemiological surveillance and prevention of pseudotuberculosis and intestinal yersiniosis. Methodical instructions. MU 3.1.12438-2009).

The composition of the medium, Altnickol classic and modified variants registered in the Ministry of health of the USSR №535-84, 1)

Table 1
The composition of the medium Oltenitza what about the classic and modified versions
IngredientsClassicModified
Nutrient agar, pH 7,2-7,4, ml1000
Lactose, g1,0
Sucrose, g10,010,0
Glucose, g1,01,1
Urea, g10,010,0
Sodium thiosulfate (Hypo), g0,31,0
Ammonium iron (II) sulfate (Salt Mora)0,20,2
Phenol red, 0,4% wadnr-R, ml4,0
The environment of GIS with lactose and Indyk. (BP), g45,0
Distilled water mlto 1000,0

The medium is sterilized at 0.5 ATM 20 is minutes or flowing steam 3 days in a row in 20 minutes Environment cool in slanted position for receiving the column height of 2.5 cm and jamb length 4 see the Finished medium rose-pink.

The course of study

Sow a stroke on the beveled surface and injected into the column. Incubated at 37°C 18-24 hours. Kislotoobrazovanie causes the appearance of a yellow color, the decomposition of urea (increase pH), reddening.

The disadvantage is that the main purpose of this environment is the identification of bacteria of the family Enterobacteriacea, which includes 12 genera and over 300 species of microorganisms. Therefore, the environment Altnickol does not solve the problem of differential-diagnostic for the identification of bacteria of the genus Yersinia (figure 2).

The objective of the invention is to provide a simple differential diagnostic of nutrient medium with high resolution, which allows to determine whether the selected culture to the genus Yersinia without the use of additional environments, resulting in a significant reduction in terms of research.

The problem is solved by making the differential diagnosis of nutrient medium for the identification of bacteria of the genus Yersinia, containing microbiological agar (dry), lactose, glucose, urea, dye phenol red distilled water, according to the invention additional nutrient medium which contains dye methylene blue and calcium chloride in the following ratio of components, g/100 ml:

Dry agar-3,0

Lactose - 1,0

Glucose -0,15

Urea - 0,3

Calcium chloride is 0.2

Phenol red (1% alcoholic aq) - 0.2 ml

Methylene blue (1% alcoholic aq) 0.1 ml

Distilled water up to 100 ml

a pH of 7.0 to 7.2.

As a solvent for the dye phenol red and methylene blue uses ethyl alcohol.

The development of the proposed nutrient medium is taken into account and used biochemical peculiarities of the genus Yersinia and individual species of this genus.

Simultaneous use in the claimed composition of the nutrient medium alcohol solution of methylene blue and phenol red allows you to get specific staining (bar raspberry, mowed the purple part) is specific to bacteria of the genus Yersinia. Consequently, the proposed nutrient medium can be considered as a simple as well as their biochemical properties.

The composition of the two dyes - differential diagnostic environment with high resolution for bacteria of the genus Yersinia

Introduction to the culture medium of calcium chloride, which is not a component of the power Arseni, provides its isotonicity, protecting bacterial cells from osmotic shock.

While the essential features of the proposed suggestions should be considered the establishments the th and quantitative composition of the nutrient medium, with high resolution, since the magnitude of the contents of the components in the claimed nutrient medium are optimal and determine the relative balance of the cost of its growth characteristics. Under the concept of high resolution nutrient medium, the applicant implies the possibility of separating bacteria of the genus Yersinia during the one sowing to the claimed nutrient medium without the use of additional tests, resulting in the time frame of the study is reduced to 18-24 hours.

A nutrient medium is prepared as follows.

The dry ingredients of the environment dissolved in distilled water in any sequence. After dissolution of the agar mixture is heated to boiling and add the indicator (phenol red and methylene blue). Establish a pH of around 7.0-7.2 and poured into test tubes 5-6 ml, sterilized by autoclaving for 15 min at a pressure of 0.5 ATM.

In a hot environment has a dirty green colour, and immediately after autoclaving acquires a yellow-orange coloration in the recovery of methylene blue.

The tubes with the medium gently shaken and set in an inclined position at an angle of 45° (for cutting, leaving a vertical column height of 1.5-2.0 cm and an oblique surface 4-5 cm). When the cooling medium it again when breaet dirty-green color due to the oxidation of the indicator atmospheric oxygen.

Solidified nutrient medium within 2-3 days to become Mature is to acquire a solid uniform color, indicating that the oxidation of the indicator throughout the depth of the environment.

Planting material spend across the beveled surface (jamb) and subsequent injection into the column environment.

Under cultivation on the environment through the day, various members of the family Enterobacteriaceae cause characteristic changes in the color of the proposed environment (table 2).

Table 2
Changes proposed nutrient medium during the cultivation of bacteria of the family Enterobacteriaceae
BacteriaColumnThe beveled partCharacter growth on the jambThe presence of gas
ShigellaGreenPurpleGentle, with a blue tintNo
SalmonellaYellowPurpleLush, with whitish tint
E. 0124 YellowPurpleLush, with whitish tint
EscherichiaYellowGreenLush, with whitish or green
shade
ProteusCrimsonPurpleDrain, with a purple tintThere*
YersiniaCrimsonPurpleDrain, with a purple tintNo

Note: *) in the course of injection.

Yersinia ferment glucose with the formation of acid (obligate characteristic for all members of the family Enterobacteriaceae) decompose urea and restore methylene blue indicator. However, the final chemical and redox changes in the column and on the beveled surface environment (jamb) significantly differ in color. In the column occurs HC is the increase in pH due to the alkalinity of the medium (the result of the decomposition of urea), resulting in the recovered methylene blue indicator light will fade. When this column is colored phenol red indicator in crimson (Fig.3).

Consequently, the proposed nutrient medium can be considered as a differential diagnosis. On the beveled portion of the column also occurs cleavage (hydrolysis) of urea and, therefore, the alkalization of the environment. Phenol red indicator is colored in Magenta color, which, mingling with methylene blue, gives an intense violet coloration environment.

Thus, various representatives of bacteria of the family Enterobacterioceae with different biochemical properties in relation to the lactose, glucose, urea and methylene blue, change the color of the medium at different pH values. Yersinia give a clear picture, as a rule, after 18-24 hours, and other members of the family cause characteristic changes in the environment and in a shorter time.

The claimed nutrient medium meets the criteria of the invention:

"novelty" in the claimed nutrient medium simultaneously use two indicator phenol red and methylene blue, which amplify the differential diagnostic properties of the environment and when superimposed on one another give crimson and purple color, which allows to differentiate the bacteria Yersinia from the hypoxia representatives of bacteria of the family Enterobacterioceae);

"inventive level" of a particular applicant's prior art is not revealed fame simultaneous use of two dyes for diagnosing bacterial-Yersinia.

"industrial applicability" - this nutrient medium is simple in execution and can be used both for research purposes and for diagnosis of diseases in hospitals.

The differential diagnosis of nutrient medium for the identification of bacteria of the genus Yersinia, containing microbiological agar (dry), lactose, glucose, urea, dye phenol red distilled water, wherein the nutrient medium further comprises a dye methylene blue and calcium chloride in the following ratio of components, g/100 ml

Dry agar3,0
Lactose1,0
Glucose0,15
Urea0,3
Calcium chloride0,2
Phenol red (1% alcohol solution)0.2 ml
Metileno the th blue (1% alcohol solution) 0.1 ml
Distilled waterto 100 ml

a pH of 7.0 to 7.2.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. Claimed is strain of Fusarium sambucinum, deposited in VKPM collection under number F-1161. Claimed strain is producent of protein food biomass.

EFFECT: invention makes it possible to accumulate biomass with high protein content with higher quantity of valuable unsaturated fatty acids, complete in composition.

2 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. Claimed is method of obtaining food fungal biomass with high protein content. Multicycle deep cultivation of Fusarium sambucinum All-Russian collection of industrial microorganisms F-1161 on liquid nutritional medium, containing sources of carbon, nitrogen, mineral salts, separation and drying of wet fungus biomass are carried out. Cultivation is performed at pH from 3.5 to 7.0 under conditions of air aeration from 0.5 to 2.0 l/l/min. Temperature mode in each cycle of fermentation is supported from the beginning of the cycle to the point of switch at the level from 26 to 30°C, and further to the end of the cycle at the level from 22 to 25°C. Point of switch is determined by accumulation of biomass to concentration from 45 to 60% from maximally achievable in fermentation apparatus, or point of switch is determined by concentration of dissolved oxygen by its reduction to the value from 20 to 40% of saturation ( calculated per atmospheric air pressure).

EFFECT: invention makes it possible to obtain the largest accumulation of biomass with high protein content with specified quantity of nucleic acids.

4 ex

FIELD: biotechnology.

SUBSTANCE: growing of Staphylococcus aureus is carried out in a nutrient medium containing yolk-salt agar. At a stage of preparation for analysis the growth stimulators of Staphylococcus aureus are introduced into the nutrient medium in the form of aqueous solutions at concentrations of 10-4-10-6 wt %. The following compounds are used as growth stimulators: tris(2-hydroxyethyl)ammonium 4-chlorophenyl-sulfanylacetate or tris(2-hydroxyethyl)ammonium 2-chlorophenyloxyacetate or tris(2-hydroxyethyl)ammonium 2-methyl-4-chlorophenyloxyacetate or tris(2-hydroxyethyl)ammonium 1-benzylindol-3-yl-sulfanylacetate.

EFFECT: invention enables to accelerate growing of Staphylococcus aureus for diagnostics of infections, by reducing the time of growing from 48 to 6-9 hours in comparison with the control in a standard nutrient medium.

2 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and deals with recombinant strain of E. coli TG1(pRVMoscow3253G-L) for obtaining PCR-standards for quantitative determination of cDNA of rabies virus of strain "Moskva 3253".Recombinant strain is created on the basis of strain of E. coli TG1 by transformation with plasmid pRVMoscow3253G-L. Plasmid is obtained by ligation of fragment G-L of the region of genome of fixed rabies virus of strain "Moskva 3253", which has sequence SEQ ID NO1, into plasmid pGem-T. Also claimed is set of PCR-standards for quantitative determination of cDNA of rabies virus of strain "Moskva 3253" in rabies antigen. Set contains solutions of plasmid pRVMoscow3253G-L DNA in concentrations 108, 107, 105, 103 GE/ml. Concentration is determined by method of polymerase chain reaction, with hybridisation-fluorescence account of results.

EFFECT: invention contributes to standardisation of stage of preparation of rabies antigen in production of heterological anti-rabies immunoglobulin.

2 cl, 3 dwg, 2 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry and molecular biology. Conservation of cells of Escherichia coli in presence of buffered 80-90% glycerol is performed. Cell envelopes are removed with 3% triton X-100. Cell supramolecular structures are successively extracted with increasing concentrations of salts: 0.14 M (bacterioplasm), 0.35 M (loosely linked with cell residue), 2 M NaCl (strongly linked with cell residue), and 6 M guanidine hydrochloride with 0.1% β-mercaptoethanol (cell residue). Acid hydrolysis is carried out in said fractions. Anthrone method is carried out, with preliminary purification of anthrone preparation. Calibration graph is built and quantity of hexoses is determined by means of calculation formula.

EFFECT: invention makes it possible to determine quantity of hexoses in supramolecular structures of bacterial cell of Escherichia coli.

3 dwg, 3 tbl, 1 ex

FIELD: biotechnology.

SUBSTANCE: invention is production of the nutrient medium, which creates optimal conditions for growing legionella, comprising: enzymatic hydrolyzate of pig lung, enzymatic hydrolyzate of chicken egg yolk, potassium monophosphate, trihydrate disubstituted potassium phosphate, L-cysteine hydrochloride, activated carbon, microbiological agar and distilled water at a predetermined ratio of ingredients.

EFFECT: invention enables to produce the high-quality, easy-to-prepare nutrient medium, to reduce the time of growing legionella.

FIELD: biotechnologies.

SUBSTANCE: nutritive medium includes lactoserum, yeast autolysate, acetic acid 70%, agar and cabbage brew at the specified component ratio.

EFFECT: invention allows increasing selectivity of nutritive medium and simplifying its production.

8 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and can be used in biological treatment of waste water from electroplating plants from heavy metal salts. The method involves adding yeast biomass to waste water, said biomass being in form of brewery wastes containing a combination of yeasts of different strains of Saccharomyces cerevisiae with viability of 90-95% in a given amount. The biomass is mixed with the waste water to obtain a suspension. The obtained suspension is held for 8 hours at temperature of 10°C-29°C and solution pH of 5.5-8.0, followed by recycling spent yeast containing heavy metals by treating with lime Ca(OH)2, with the ratio of yeast biomass to lime of 1:5-8, to obtain a mixture. The obtained mixture is subjected to wet treatment at temperature of 90°C for 1 hour, followed by isolation of the obtained mixture, which contains heavy metals, in concrete paste.

EFFECT: invention increases efficiency of purifying waste water from heavy metal ions.

3 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: invention presents a diagnostics method of sensitivity of M. tuberculosis (MBT) to injection antituberculous preparations of a reserve row. The method involves a DNA extraction stage, amplification of the investigated DNA sections by means of a polymerase chain reaction method and analysis of conformational polymorphism of single-chain fragments (SSCP). Gene section rrs is amplified in 50 mcl of a reaction mixture with addition of 5 mcl of the specimen. Promoter gene section eis is amplified in 30 mcl of the reaction mixture containing direct 5'CGGAGCCGTCGGGGTATGC and reverse 5'GCCGCGGCCAGTAGGAACA primers and 3 mcl of the specimen as per the amplification programme: 1-st stage - 95°- 4 min; 2-nd stage - 95° - 20 sec, 59° - 30 sec, 72° - 20 sec (30 cycles); 3-rd stage: 72° - 4 min; 10° - storage. Separation of amplification products in the ratio of 4 mcl of the specimen and 6 mcl of a denaturing dye is performed by electrophoresis in 8% polyacrylamide gel with 5% glycerin at voltage of 400 Volts during 5 hours at 8°C. Gel painting is performed by means of caustic silver.

EFFECT: invention allows diagnosing sensitivity of M tuberculosis to injection antituberculous preparations of a reserve row with high accuracy.

2 dwg

FIELD: biotechnologies.

SUBSTANCE: invention proposes an association of strains of bacteria-oil decomposers, which have been extracted from oil-contaminated soil, Acinetobacter species B-1037, Pseudomonas species B-989, Bacillus species B-1040, deposited at The State Research Centre of Virology and Biotechnology VECTOR. Besides, at least 30% bacteria of each strain is contained in the association. Remediation of oil-contaminated soils includes water suspension of lyophilic dried biomass of the strain association based on 109 cells per square metre. Strains of the association can utilise a wide range of oil components at the temperature of 10-15°C.

EFFECT: improving cleaning efficiency of oil-contaminated soils.

2 cl, 5 dwg, 2 tbl, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: 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.

EFFECT: method enables preparing the immunosensor used for the purpose of the detection of infectious agents with the sensitivity of 1×103-1×104 mln. cells/ml and preserving the specific activity for.

3 cl, 3 ex.

FIELD: medicine.

SUBSTANCE: 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.

EFFECT: method enables assessing the bactericidal activity of the disinfectants.

3 dwg, 3 tbl, 1 ex

FIELD: biotechnologies.

SUBSTANCE: 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.

EFFECT: proposed solution can be used for identification of kinds of lactobacilli in different specimens of human and animal organisms, in pharmaceutical preparations and in food products.

2 cl, 1 dwg, 3 tbl

FIELD: biotechnologies.

SUBSTANCE: 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.

EFFECT: invention allows reducing identification time, improving difernetiation accuracy of coliform bacteria and Ecoli, and simplifying a feed medium preparation method.

2 cl, 4 dwg, 1 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: 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.

EFFECT: invention allows preserving sterility of feed medium during 10 days, increasing accuracy of differentiation of coliform bacteria and Ecoli and simplifying a feed medium preparation method.

2 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: 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.

EFFECT: invention increases diagnostic sensitivity of the method of identifying vibrio bacteria.

2 tbl, 4 ex

FIELD: biotechnologies.

SUBSTANCE: 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.

EFFECT: invention allows more quickly, efficiently and less expensively indentifying the total number of revivable microcolonies of microorganisms, their identifying and distinguishing.

7 cl, 10 dwg

FIELD: medicine.

SUBSTANCE: 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.

EFFECT: linco-spectin resistance to the enzymatic degradation of a microbial cell, lower toxicity, faster tissue regeneration and manifested biocidal action on the pathogenic bacteria, viruses, and moulds for 3 years.

5 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely gynaecology and may be used for individual selection of the preparations containing the probiotic lactic bacterial strains for effective intravaginal therapy. For this purpose, vaginal epithelial cells are recovered from the patient, released from the accompanying microflora. That is followed by preparing an epithelial cell suspension in a culture medium, and mixed with a suspension of thermally-activated probiotic lactic bacterial strains. Then, the suspension is incubated; the epithelial cell culture fluid filtrate is prepared and added to the suspension of the tested probiotic lactic bacterial strains in ratio 1:7. Concurrently, a reference of the mixture of the epithelial cell culture medium and the suspension of the tested probiotic lactic bacterial strains in ratio 1:7 is prepared. The test and reference samples are incubated, measured for optical density; and a degree of biomass increase in the test sample is related to that in the reference. The preparation containing the probiotic lactic bacterial strains the biomass increase of which under the influence of patient's vaginal epithelial cells is stimulated most is selected form the effective intravaginal therapy.

EFFECT: invention enables the individual selection of the preparations containing the probiotic lactic bacterial strains for the effective intravaginal therapy.

3 ex, 1 tbl, 2 dwg

FIELD: biotechnology.

SUBSTANCE: method comprises incubation of the blood sample in the nutritional medium, followed by detection of microbial growth in the primary hemoculture. Sample preparation of the sample under study is carried out by centrifugation. The quantitative chromatography-mass-spectrometer analysis of the sample under study is carried out with the definition of marker molecules, which are used as molecules of free and substituted higher fatty acids of cellular lipids of microorganisms. The free and substituted higher fatty acids are identified by comparing the data obtained with the database NIST of chromatography-mass-spectrometer. The genus of bacteremia agents is determined using the Table 1.

EFFECT: invention enables to provide for early diagnosis of bacteremia.

5 dwg, 5 ex

FIELD: biotechnology.

SUBSTANCE: growing of Staphylococcus aureus is carried out in a nutrient medium containing yolk-salt agar. At a stage of preparation for analysis the growth stimulators of Staphylococcus aureus are introduced into the nutrient medium in the form of aqueous solutions at concentrations of 10-4-10-6 wt %. The following compounds are used as growth stimulators: tris(2-hydroxyethyl)ammonium 4-chlorophenyl-sulfanylacetate or tris(2-hydroxyethyl)ammonium 2-chlorophenyloxyacetate or tris(2-hydroxyethyl)ammonium 2-methyl-4-chlorophenyloxyacetate or tris(2-hydroxyethyl)ammonium 1-benzylindol-3-yl-sulfanylacetate.

EFFECT: invention enables to accelerate growing of Staphylococcus aureus for diagnostics of infections, by reducing the time of growing from 48 to 6-9 hours in comparison with the control in a standard nutrient medium.

2 tbl, 7 ex

Up!