Method of accelerated growth of staphylococcus aureus for diagnostics of infections associated with delivery of health care
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
The invention relates to medicine, namely to clinical Microbiology, and relates to a method of fast growing Staphylococcus aureus (Staphilococcus aureus) for use in the diagnosis of infections associated with health care. Known standard method of growing Staphylococcus for the diagnosis of infections caused by these microorganisms, which consists in using as a nutrient medium yolk-salt agar. Duration of cultivation by the standard method [2,6,7] up to 48 hours.
Irrational use of antimicrobial agents has led to the development of resistance in many pathogenic and conditionally pathogenic microorganisms (including Staphylococcus) and wide dissemination of socially relevant staphylococcal infections, receiving threatening and causing the development of nosocomial purulent-inflammatory diseases, especially in surgical wards, maternity hospitals, pediatric hospitals (ulcerative colitis, enteritis, peritonitis, endocarditis, pneumonia, lung abscesses, staphylococcal sepsis, and others) [1,8,9]. The duration of the existing production bacteriology negative impact on the provision of timely medical aid to the patient, complicating quick apply adequate terap is I. Therefore, the development of methods for rapid cultivation of staphylococci for the diagnosis of infections associated with health care.
The aim of the invention is to increase the growth rate of Staphylococcus aureus for the diagnosis of infections associated with health care, and reducing the time of issuance of the analysis for the timely appointment of adequate antibiotic therapy.
This objective is achieved in that when grown Staphylococcus aureus for diagnostic purposes in preparation for analysis in the nutrient medium on the basis of the yolk-salt agar as a growth promoter microorganisms injected chemical compound: Tris(2-hydroxyethyl)ammonium 4-chlorophenylsulfonyl (1) or Tris(2-hydroxyethyl)ammonium 4-chlorophenylsulfonyl (2) or Tris(2-hydroxy-ethyl)ammonium 2-chlorophenoxyacetate (3) or Tris(2-hydroxyethyl)ammonium 2-methyl-4 - chlorophenoxyacetate (4) or Tris(2-hydroxyethyl)ammonium 1-benzyliden-3-yl-sulfonylurea (5) in the form of aqueous solutions at a concentration of 10-4-10-6wt.% (table 1). When using growth stimulants 1-5 growth of Staphylococcus reduced from 48 (without stimulants) up to 9 hours.
Table 1 presents the chemical structure of stimulants 1-5.
Table 2 presents the effect is biostimulators 1-5 on the growth of Staphylococcus aureus.
As growth stimulants 24 strains St. aureus isolated from patients with purulent-septic complications from the Department of surgery, first investigated biologically active compounds are representatives of a new class of biologically active proton (2-hydroxyethyl)ammonium ionic liquids [3-5].
Methods of preparation and properties of stimulants 1-5 are given in examples 1-5. The composition and structure of compounds 1-5 were confirmed by IR, NMR spectroscopy and elemental analysis. IR spectra of (v, cm-1) were recorded on spectrophotometer Varian 3100 FT-IR75 in CVG. NMR spectra (δ, ppm) was shot in the D2O spectrometer DPX 400 (400,13 MHz to1N and 101,62 MHz to13C).
The following examples illustrate the invention:
Example 1. Tris(2-hydroxyethyl)ammonium 4-chlorophenylsulfonyl (1)
To a solution of 2.03 g (0.01 mol) 4-chlorophenylsulfonyl acid in 10 ml of ethanol was bury ethanol solution of 1.49 g (0.01 mol) of Tris-(2-hydroxyethyl)amine (10 ml). The mixture was stirred at 25°C. for 3 hours. The solvent is kept in vacuum. The solid residue was repeatedly washed with ether and dried in a vacuum desiccator over P2About5. Received a colorless powder (yield 3.44 g, 98%) with TPL 91°C, soluble in water and organic solvents (acetone, THF, alcohols). NMR1H: 7.11-6.32 (m, 4H, C6H4S), 3.75 (t,6N, co2), 3.44 (who, 2H, SCH2COO), 3.28 (t, 6N, NCH2). NMR,13From: 177.19 (C=O), 155.81-111.55 (C6H4S), 60.79(SCH2COO), 55.09(och2), 54.83(NCH2). IR: 1560 (C=O), 2550-2700 (H+N), 3300 (OH). Found, %: C, 48.11; H, 6.01; N, 3.93. C14H22ClNO5S. Calculated, %: C, 47.79; H, 6.30; N, 3.98.
Example 2. Tris(2-hydroxyethyl)ammonium 4-chlorophenylsulfonyl (2)
Was obtained from 4-chlorophenylsulfonyl acid and Tris(2-hydroxyethyl)amine same as 1. Colourless powder with TPL 93°C. the Yield is 97%. NMR1H: 7.19-6.22 (m, 4H, C6H4SO2), 4.04 (s, 2H, SO2CH2COO), 3.71 (t,6N, co2), 3.22 (t, 6N, NCH2). IR: 1566 (C=O), 2500-2710 (H+N), 3310(HE). Found, %: C, 44.10; H, 5.50; N, 3.63. C14H22ClNO7S. Calculated, %: C, at 43.80; H, 5.77; N, 3.65.
Example 3. Tris(2-hydroxyethyl)ammonium 2-chlorophenoxyacetate (3)
Was obtained from 2-chlorophenoxyacetic acid and Tris(2-hydroxyethyl)amine same as 1. Colourless powder with TPL 81-83°C. a Yield of 99%. NMR1N: 7.26-6.02 (m, 4H, C6H4), 4.24 (s, 2H, och2Soo), 3.70 (t,6N, co2), 3.22 (t, 6N, NCH2). NMR13From: 178.09 (C=O), 157.01-111.05 (C6H4O), 60.00(och2MEO)57.19(och2), 55.93(NCH2). IR: 1558 (C=O), 2520-2700 (H+N), 3330(HE). Found, %: C, 50.23; H, 6.50; N, 4.18. C14H22ClNO6. Calculated, %: C, 50.07; H, 6.60; N, 4.17.
Example 4. Tris(2-hydroxyethyl)ammonium 2-methyl-4-chloro-PHENOXYACETIC (4)
Was obtained from 2-methyl-4-the EOS-phenyloxirane acid and Tris(2-hydroxyethyl)amine same as 1. Colourless powder with TPL 88°C, the Yield of 98%. NMR1N: 7.72-6.22 (m, 3H, C6H3), 4.44 (s, 2H, och2Soo), 3.78 (t,6N, co2), 3.38 (t, 6N, NCH2), 2.25 (CH3). NMR13From: 175.19 (C=O), 156.21-111.11 (C6H3O), 62.23(och2MEO)58.29(och2), 57.93(NCH2), 14.71 (CH3). IR: 1563 (C=O), 2500-2720 (H+N), 3310(HE). Found, %: C, 51.72; H, 6.62; N, 4.04. C15H24ClNO6. Calculated, %: C, 51.50; H, 6.91; N, 4.00.
Example 5. Tris(2-hydroxyethyl)ammonium 1-benzyliden-3-ylsulphonyl (5) was Obtained from 1-benzyliden-3-resultdisplay acid and Tris(2-hydroxyethyl)-amine analogously to 1. Colourless powder with TPL 109-110°C. a Yield of 96%. NMR1N (D3C)2SO, (δ, ppm, J/Hz), 3.05 (t, 6N, 3CH2N, J=5.3); 3.63 (t, 6N, SN2HE, J=5.3). IR: 1560, 1600 (C=O), 2500-2720 (H+N), 3290(HE). Found, %: C, 58.05; H, 6.23; N, 5.87. C23H30N2O7S. Calculated, %: C, 57.72; H, 6.31; N,5.85.
Example 6. Preparation of dilutions of growth stimulants for the study. During the dissolution tests were carried out for each potential growth stimulator according to the following procedure:
Was dissolved 0.1 g (100 mg) of the substance in 100 ml of distilled water, getting a 0.1% solution, which was further of the view matrix.
The first breeding: 0.1 ml of the matrix 0.1% solution was added to 100 ml of the yolk-salt agar (ISA). The concentration of the drug in the environment with the Tawil 10 -4%.
Second breeding: 0.1 ml matrix 0.1% solution is added to 1000 ml ISA. The drug concentration in the medium was 10-5%.
The third breeding: 0.01 ml matrix 0.1% solution is added to 1000 ml ISA. The drug concentration in the medium was 10-6%.
In the experiment using each drug dilution in culture media and pure culture of pathogenic bacteria associated with health care.
Example 7. The experiment on the cultivation of St. aureus.
The capture and culture of infected material and the light of the results produced in accordance with regulatory orders [2,6,7].
Culture St. aureus was obtained by culturing in an incubator at 37°C with the use of the yolk-salt agar, which was prepared in accordance with the instructions on the vial label.
For the preparation of nutrient medium required quantity of dry matter stirred in 1 l of distilled water, boiled for 3 min until complete melting of agar, filtered through a cotton-gauze filter, poured into sterile vials and sterilized by autoclaving at 121°C for 15 min, the Medium was cooled to a temperature of 45-50°C, poured into sterile Petri dishes with a layer of 3-4 mm After solidification of the medium, following the rules of asepsis, cups were dried at the temperature of the round (37±1)°C for 40-60 minutes
Sowing on Petri dishes was performed by standard method. Identification of isolated microorganisms was evaluated using a commercial test systems company LACHEMA (Czech Republic): "STAPHYtest"
Of the identified culture St. aureus was preparing a suspension 104 in physiological solution according to the turbidity standard.
Then took three dilution in GSA, poured agar cups (three cups of each dilution in culture medium). The cups were sown 0.1 ml of the prepared suspension of Staphylococcus aureus and incubated at 37°C, viewing the cups were made every 2-3 hours, noting the growth of the colonies. As a control, were sown on JSA without growth promoter.
Counting colonies grown on the surface of the agar, produced with the help of a magnifying glass (X5) or of the device for counting colonies. To do this, put the Cup upside down on a dark sheet of paper (thus providing better visibility of the colonies). For convenience, each counted colony was noted from the bottom of the Cup with a pencil or marker. 1 When using stimulants 1-5 in the environment IS for accelerated growth of crops St. aureus has established the viability of the microorganisms. The results of the growth-promoting activity displayed in table 2.
From the data presented in table 2, you can see significant differences in germination strains St. aureus on the media with growth factors at different concentrations, and n is the control environment, ISA. So, when you use any stimulants growth at concentrations of 10-4-10-5weight. %of stimulator No. 5 at concentrations of 10-4-10-6weight. % after 3 hours was pulverized growth of microorganisms after 6 hours there were single colony, 9 hours after seeding, growth was observed St. aureus expressed lecithinase activity. When added to the growth medium growth stimulants 1-4 at a concentration of 10-6weight. % after 3 and 6 hours after seeding was observed powdery growth of microorganisms, 9 hours - growth of single colonies.
When sowing the studied cultures in the yolk-salt agar no growth stimulants after 3 and 6 hours of growth was not observed, and only 9 hours appeared powdery growth of microorganisms. Over 9 hours of cultivation St. aureus in the control were observed no growth of single colonies of microorganisms or the growth of the studied cultures with a strong lecithinase activity.
The method of growing microorganisms tested on the 24 strains St. aureus isolated from patients with purulent-septic complications from the Department of surgery with positive results in the implementation of microbiological monitoring
Studies made it possible to establish that the investigated compounds 1-5 stimulate the growth of culture St. aureus, substantially reducing Vremya growth compared to the control on the standard nutrient medium (GSA). These results can be used to develop an improved methodology for rapid diagnosis of nosocomial infections caused by S. aureus, which will greatly reduce the time of issuance of the analysis and timely prescribe an adequate treatment of the patient.
1. Briskin BS, N. Nachtman. Nosocomial infections and their prevention: a view of the surgeon /Surgery: - 2005. No. 2. - 26-30.
2. Aserinsky, Lpina, Asesino, Private medical Microbiology techniques microbiological studies // M, JSC "Publishing house "Medicine", 2005. - 600 C.
3. Mirkova A.N., Mirkov RG, Adamovich, S., Voronkov M.G., Synthesis and pharmacological activity of 2-hydroxyethylammonium salts organosulfur(sulfonyl) acetic acid - new pharmacologically active compounds. / / Chemistry for sustainable development. - 2011. - So 19. No. 5. - S-478.
4. Mirkova A.N., Levkovskaya GG, Stupina A.G., Chkhenkeli, VA, Voronkov M.G. Influence of Tris(2-hydroxyethyl)ammonium, araxi, aaltio and arylsulfonate on the activity of bifidobacteria // Dokl. An. - 2003. - 390 so. No. 2. - S-282.
5. Mirkova A.N., Levkovskaya GG, Mirkov RG, Voronkov M.G. Alkanolammonium salt organosulfur (sulfonyl) acetic acid - new stimulants biological processes // Zhur.org.chem.. - 2008. - So 44. - VIP. - S-1508.
6. About invites the medical care of patients with purulent surgical diseases and strengthening of measures to combat hospital-acquired infections: order No. 720 the USSR Ministry of health. M., 1978.
7. About the unification of microbiologic methods used in clinical diagnostic laboratories medical and diagnostic institutions: order No. 535 the USSR Ministry of health. M., 1985.
8. Onishchenko GG national concept of prevention of infections associated with health care. - 2011. p.21.
9. Rock L., Sidorenko SV, Nekhoroshev A.G. Practical aspects of modern clinical Microbiology. - M.: Labelform, 2004. P.64-69.
|1. Tris(2-hydroxyethyl)ammonium 4-chlorophenyl-sulfonylated|
|2. Tris(2-hydroxyethyl)ammonium 4-chlorophenyl-sulfonylated|
|3. Tris(2-hydroxyethyl)ammonium 2-chlorophenyl-oxyacetic|
|4. Tris(2-hydroxyethyl)ammonium 2-methyl-4-chloro-PHENOXYACETIC|
|no growth stimulator||Time, h||Concentration %|
|Control (no growth factor)||3||d||"-"||"-"|
|Notes: - powdered microbial growth, b - isolated colonies of microorganisms, microbial growth with a strong lecithinase activity, Dr. no growth of microorganisms, "-" - studies have not been conducted.|
Way fast growing Staphylococcus aureus Staphylococcus aureus for the diagnosis of infections, which consists in the cultivation of Staphylococcus aureus in nutrient medium containing yolk-salt agar with the addition of the biostimulator, characterized in that as biostimulant use the following compound: Tris(2-hydroxyethyl)ammonium 4-chlorophenyl-sulfonylated or Tris(2-hydroxyethyl)ammonium 2-chlorophenoxyacetate or Tris(2-hydroxyethyl)ammonium 2-methyl-4-chlorophenoxyacetate or Tris(2-hydroxyethyl)ammonium 1-benzyliden-3-yl-sulfonylurea that bring nutrients the environment in concentrations of 10-4-10-6wt.% in aqueous solution.
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
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
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.
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.
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
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.
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
SUBSTANCE: bacillus subtilis subsp.subtilis BKM B-2711D strain has apparent antagonism in relation to Escherichia coli, Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and resistance to streptomycin and tetracycline antibiotics. It is deposited in the All-Russia Collection of Microorganisms of the Institute of Biochemistry and Physiology of Microorganisms Named after G.K. Skryabin of the Russian Academy of Science (IBFM RAN) and has the following registration number: BKM B-2711D, and can be used at production of probiotic bacterial preparations that can be used in veterinary medicine.
EFFECT: invention allows increasing protease and xylanase activities.
1 tbl, 3 ex
SUBSTANCE: bacillus amyloliquefaciens BKM B-2714D strain has apparent antagonism in relation to Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes, and resistance to tetracycline and trimethoprim antibiotics. It is deposited in the All-Russia Collection of Microorganisms of the Institute of Biochemistry and Physiology of Microorganisms Named after G.K. Skryabin of the Russian Academy of Science (IBFM RAN) and has the following registration number: BKM B-2714D, and can be used at production of probiotic bacterial preparations that can be used in veterinary medicine.
EFFECT: invention allows increasing protease and xylanase activities.
1 tbl, 3 ex
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
SUBSTANCE: what is being studied is the pure S. areus culture in the concentration of 1 bln/ml. There are at least 25 erythrocytes examined to determine an average microbial count adhered onto one erythrocyte. An adhesion substrate is presented by sheep erythrocytes in the concentration of 100 mln/ml pre-washed in 50% formaline.
EFFECT: qualitative and quantitative instant diagnosing of an intensity of the S areus adhesion properties.
SUBSTANCE: invention relates to microbiology. Disclosed is a method for quantitative determination of adhesion of Staphylococcus spp. to fibronectin and fibrinogen, during which the analysed staphylococcus culture is incubated in dimples of a polystyrene tray with separately protein-immobilised fibronectin, fibrinogen and in dimples in a liquid medium 199 for 60±15; further, bacteria that are not adhered are removed by washing three times with the medium 199 with tween 20 and the number of adhered staphylococci is determined, either by counting grown cells in the culture medium from optical density of the medium, or by counting cells grown on solid medium from a series of tenfold dilutions, or by determining activity of bacterial enzymes in adhered strains.
EFFECT: owing to good reproducibility of results, high sensitivity, easier conduction of the reaction directly in dimples of the tray and the possibility of determining the factor of specific adhesion of strains to fibronectin and fibrinogen, the invention can be used in medical bacteriology for quantitative estimation of adhesive properties of staphylococci to human fibronectin (Fn) and fibrinogen (Fb) proteins, for characteristic of virulent properties in vitro based on the level of specific adhesion to said proteins.
3 tbl, 3 ex
SUBSTANCE: synthetic oligonucleotide primers are proposed, as well as the method to detect Lactobacillus acidophilus in starter cultures used in production of cultured milk foods. The proposed method includes performance of PCR. In case a DNA ferment is detected with size of 412 base pairs. A conclusion is made on availability of Lactobacillus acidophilus in the investigated material. The method may be used in milk processing industry for detection and identification of strains and cultures Lactobacillus acidophilus in starter cultures used in production of cultured milk foods.
EFFECT: improved efficiency of strains application.
2 cl, 1 tbl, 3 ex
FIELD: food industry.
SUBSTANCE: one proposes synthetic oligonucleotide primers and a method for revealing Bifidobacterium longum subspecies longum in starter cultures used during cultured milk products manufacture. The proposed method involves PCR implementation. In case of revealing a DNA paragraph having size equal to 447 base pair one concludes about presence of Bifidobacterium longum subspecies longum in the biomaterial studied.
EFFECT: method may be used in milk-processing industry for revealing and identification of Bifidobacterium longum subspecies longum strains and cultures in starter cultures used during cultured products manufacture.
2 cl, 1 tbl
SUBSTANCE: there are offered synthetic oligonucleotide primers and method for detecting Streptococcus thermophilus in starter cultures in hard cheese production. The presented method involves conducting the PCR. If having found the DNA fragment of size of 655 b.p., the presence of Streptococcus thermophilus in the analysed biomaterial is stated.
EFFECT: method may be used in diary production industry for detecting and identifying the Streptococcus thermophilus strains and cultures in the starter cultures in hard cheese production.
2 cl, 1 tbl, 3 ex
SUBSTANCE: invention is intended for a quantitative estimation of adhesive properties of staphylococci recovered from a human and environment objects, with its bacteriological identification and for characterisation of its virulent properties in vitro by the adhesion to biopolymeric molecules. The staphylococcus culture being investigated is incubated in wells of a polystyrene plate with immobilised haemoproteins (muscle haemoglobin, haemoglobin) in a nutrient medium 199 for 1 hour; thereafter, nonadherent bacteria are removed by triple washing in the medium 199 with Twin-20, and a number of adhesive staphylococci is estimated by harvest cells and shown by optical density of the medium and by counting the cells sown on the dense nutrient medium from a series of tenfold cultivations, or by analysing the adhesive strains for the activity of bacterial enzymes.
EFFECT: good results reproducibility, high sensitivity of the method, ease of the reaction directly in wells of the polystyrene plate and possible express estimation of adhesive capacity of strains.
3 tbl, 3 ex
SUBSTANCE: nutrient medium contains agar-agar, lactose; mannitol, peptone, NaCl, neutral red, bromthymol blue and 0.5% extract of birch wood ash maintained for 3 days.
EFFECT: wider assortment of nutrient mediums.
2 tbl, 3 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: present invention pertains to biotechnology and can be used for identifying types of Streptococcus on appearance of three different colour shades of detected microorganisms and fluorescence emission, in addition to changes in the colour of the medium. The selective medium contains specific ratios of mixtures of nutrient bases, rich in natural protein compounds (polypeptides, peptides, protease, amino acids) and vitamins, which allow for heavy growth of the type of interest. The medium also contains a group of inhibitors, a mixture of degradable chromogenic and fluorogenic substances and desaminase activity indictors.
EFFECT: identification with high degree of sensitivity and specificity.
7 cl, 3 dwg, 6 tbl, 10 ex
SUBSTANCE: invention can be applied in laboratory microbiological diagnostics of infection diseases. The method involves introduction of 12-hour staphylococci culture into plasma diluted by isotonic sodium chloride solution. The microbe suspension obtained is incubated at the temperature of 37°C for 5 hours. Electric impedance of the microbe suspension is registered through 3rd to 5th hour of incubation. And if the solution impedance reaches 64-84 kOhm, it allows making conclusion of staphylococci pathogenicity due to its plasma coagulating activity.
EFFECT: reduced analysis time and improved analysis accuracy.
SUBSTANCE: invention refers to medicine, specifically to microbiology and epidemiology. Method implies differential testing of biological properties of staphylococci isolated from clinical material and from environment using classical nutrient mediums and standard incubation conditions, and provides detection of guiding signs: coagulation factor, plasmacoagulase and urease enzymes, novobiocin sensitivity. It enables to group staphylococci using combined signs selected from group: evaluation of hemolytic activity, colonies rate, β-galactosidase, chromogenesis, culture growth in incubation conditions at temperature 45°C in anaerobic environment, decomposition of saccharose, galactose, mannitol, mannose, xylose and arabinose. Specific property of agent is defined on basis of signs mentioned above. Invention provides increased reliability of specific differential staphylococci diagnostics and reduced analysis time.
EFFECT: increased reliability of specific differential staphylococci diagnostics and reduced analysis time.
8 tbl, 6 ex, 1 dwg
SUBSTANCE: method for obtaining spore material of bacteria of Clostridium type provides for production of inoculum of bacteria in a full synthetic growth medium, seeding of inoculum and cultivation under the corresponding conditions in growth medium including potato, glucose, ammonium sulphate and chalk. During the main fermentation process for 18-28 parts of bacterial culture growth depending on time of occurrence in one field of view of at least 80-100 thickened forms of cells, n-butanol in the amount of 0.16-0.81 wt % is added to growth medium. Number of formed spores is 1.08-2.86·108 in one millilitre of the medium.
EFFECT: increased content of spores in spore material.
1 tbl, 15 ex