Method of differentiating atoxigenic strains of cholera vibrios 01 and 0139 serogroups from toxigenic strains on hydrolase activity
SUBSTANCE: atoxigenic strains of cholera vibrios 01 and 0139 serogroups are differentiated from toxigenic strains on presence of hydrolase activity. The substrate used is glycerin, which is introduced into molten Marten's agar at 2.5/100 ml of the medium with pH 7.8±0.1 and Nile blue sulphate indicator 1.5 mg/100 ml of medium, which is poured into petri dishes. Analysed broth cultures grown in a thermostat for 4 hours are deposited in volume of 20 mcl on a sector of developed agar and inoculations are incubated at 37°C for 36 to 48 hours. Strains are differentiated from their results of hydrolase activity: of the colonies grown on the medium are in form of soft semitransparent films, merging with the colour of the medium, then the analysed strain of cholera vibrio is toxigenic and epidermic. The colonies are a thick yellow or whitish deposit, then the strain is non-toxigenic haemolytic and is not epidemic.
EFFECT: method is disclosed for differentiating atoxigenic strains of cholera vibrios 01 an 0139 sergroups from toxigenic strains on hydrolase activity.
3 tbl, 3 ex
The present invention relates to Microbiology and can be used for differential diagnosis of hemolytic toxigenic and non-haemolytic toxigenic Vibrio cholerae.
Remains relevant today issue a comprehensive study of the properties of strains of Vibrio cholerae O1 and O serogroups.
The study of various enzymes causative agents of cholera, including: esterase, peptidases, triacylglycerols, amino acid sequences which are part of the Hly protein (Lip)-hly - the area responsible for the hemolytic properties of strains of Vibrio cholerae.
However, in this sequence contains amino acid motifs of α and β hydrolases, the phenotypic manifestation of which has not been studied. Many found short motifs gene hly To testify in favor of the fact that he is able to manifest itself in the phenotype depending on the nature and structure of the substrates in the environment.
A prototype of the selected method of differentiation atoxigenic strains of V. cholerae from toxigenic (see EN Pat. No. 2257415, C12Q 1/04, C12N 1/20, 27.07.2005, BI No. 21), namely, that as the nutrient medium used Martin agar with a pH of 9.0 with indicator - Nile blue sulfate in a concentration of 1.0-1.2 mg/100 ml of medium and substrate - emulsified chicken fat to the concentrations of 1.0-1.5 ml/100 ml, with further incubation for 48 h at 37°C, followed by staining micromason differentiate toxigenic strains of V. cholerae, having lipase activity from toxigenic without coloring micromason.
The disadvantage of this method is the use of non-standard substrate chicken fat, which helps to identify triacylglycerols activity of Vibrio cholerae, the latter include the true lipase, which is strictly substratespecific enzymes that require standard substrates, and chicken fat is not and requires biochemical characteristics, which complicates the process of differentiation.
In addition, the lipase activity in highly exposed to variability depending on the physiological state of the microorganism, which complicates its detection.
The task of the invention consisted in a simplified way differentiation atoxigenic strains of V. cholerae O1 and O serogroups from toxigenic on hydrolases activity.
This object is achieved in that in the method of differentiation atoxigenic strains of V. cholerae O1 and O serogroups from toxigenic on hydrolases activity, including the application of the studied culture on a nutrient medium with an indicator and a substrate with subsequent consideration of the MD results, as the substrate used glycerin, which is injected in molten agar Martin at the rate 2.5/100 ml of medium with a pH of 7.8±0.1 and indicator Nile blue sulphate 1.5 mg/100 ml of medium, pour in bowl Petri, then grown in an incubator for 4 hours studied broth culture is applied in a volume of 20 µl per sector designed agar and spend within 36-48 hours incubation crops at 37°C, then the results hydrolases activity strains spend their differentiation, if the environment has grown colonies are kind of delicate translucent films, merging with color environment, the analyzed strains of V. cholerae is toxigenic and epidemically significant, and if the colonies are dense yellowish or whitish plaque, we have nontoxigenic strain of hemolytic and not epidemically significant.
The method is as follows
Pre-training of the studied cultures in 1.0 ml of broth Martin pH 7.8±0.1 or 1% peptone water contribute strains of Vibrio cholerae, the minimum concentration of cells during seeding is 100 microbial cells. Crops incubated for four hours in a thermostat at 37°C.
Conduct design of differential nutrient medium: molten agar Martin pH 7.8 (±0,1) enter the glycerine - 2.5 ml/100 ml among the s and indicator Nile blue sulphate 1.5 mg/100 ml of medium. All manipulations should be carried out under aseptic conditions in order to avoid gains environment.
Agar is poured into glass Petri dishes. After that grown in thermostat broth culture in a volume of 20 µl automatic pipette dosing is applied on the sector designed agar. Incubation of crops is carried out in a thermostat at 37°C for 36-48 hours.
As a result, the glycerin, which is added to the nutrient medium, is subjected to hydrolysis, and various strains of V. cholerae with different speeds. While glycerin in neeterificirovannah state becomes a substrate, a directional hydrolysis which occurs in α and β positions.
Thus, gidrolizny activity of V. cholerae considered as a differential characteristic.
Records of the results carry through 36-48 hours. If the sectors of the agar culture was grown in the form of an opaque dense micromason whitish-yellow color, the color of the environment does not change, we conclude that the studied strain of hemolytic atoxigenic and has no epidemiological significance. If the environment has grown colonies have views of the blue semi-transparent films, blending with the color of the environment, the analyzed strains of V. cholerae is toxigenic, meaning epidemically significant.
It is noted that during incubation of cultures within 72 hours of the mi is rogatory as hemolytic atoxigenic strains (ctx -Hly+)toxigenic and non-haemolytic (ctx+Hly-) take the same view: micromason toxigenic variants compacted yellowish tint, similar to hemolytic strains. This suggests that gidroliznaja activity specific for all V. cholerae, but hemolytic strains possess more pronounced hydrolases activity, which manifests itself in them after 36-48 hours on medium with glycerol, which allows for the differentiation of Vibrio cholerae inside view on this topic.
As the studied strains used culture V. cholerae eltor No. 14863 (ctx-Hly+and 5879 (ctx+Hly-); V.cholerae O139 "Bengal" No. 16077, R-16131 (ctx+Hly-and 17918 (ctx-Hly+); V.cholerae classica No. 569 In and 251 (ctx+Hly-As the substrate used glycerin in a concentration of 0.5-1.0%. Accounting results after 48 hours of cultivation (see table 1).
From table 1 it is seen that when the concentration in the environment of 0.5-1.0% of glycerol impossible to identify gidrolizny microbial activity and conduct differentiation of toxigenic from non-haemolytic nontoxigenic hemolytic variants, since the substrate concentration in the environment is not sufficient for its development and visualization. This is evidenced by equal growth in all variants of V. cholerae with the formation of the translucent blue colonies without active hydrolysis of glycerol.
As the studied strains used culture V. cholerae eltor No. 14863 (ctx-Hly+and 5879 (ctx+Hly-); V.cholerae O139 "Bengal" No. 16077, P-16131 (ctx+Hly-and 17918 (ctx-Hly+); V.cholerae classica No. 569 In and 251 (ctx+Hly-). As the substrate used glycerin in a concentration of 2.5%. Accounting results after 48 hours of cultivation (see table 2).
Table 2 shows that when the concentration in the environment of 2.5% glycerol observe the differentiation of hemolytic nontoxigenic (ctx-Hly+) toxigenic and non-haemolytic (ctx+Hly-) variants of Vibrio cholerae on hydrolases activity. This hemolytic strains of El tor No. 14863 and O139 serogroup No. 17918 grow by sectors of the agar in the form of a dense opaque micromason whitish-yellow color, the color of the environment does not change. These strains are classified as hemolytic, no epidemiological significance. The obtained results confirm the methods for determining the epidemiological significance of Vibrio cholerae, is given in MU 4.21097-02 "Laboratory diagnosis of cholera". - M., 2002. - 95 C., namely, in the sample of Greig, there is complete hemolysis of sheep red blood cells, and the results of PCR diagnostics indicate the absence of ctx AB-gene.
The toxigenic strains (ctx+Hly-) cholerae O1 El tor circulation No. 5879, classical biovars the No. V, 251, A serogroup No. 16077, R-16131, growing form on agar translucent blue, blending with the color of the environment, microgate. Active hydrolysis was not observed, which indicates that their toxigenicity. In the sample of Greig and PCR these cultures are characterized as toxigenic non-haemolytic strains.
Thus, the nature of growth on medium containing 2.5% glycerol strains of V. cholerae can be concluded about demolitionist and oxygenate strains that determines their pijnacker.
As the studied strains used culture V.cholerae eltor No. 14863 (ctx-Hly+and 5879 (ctx+Hly-); V.cholerae O139 "Bengal" No. 16077, R-16131 (ctx+Hly-and 17918 (ctx-Hly+); V.cholerae classica No. 569 In and 251 (ctx+Hly-As the substrate used glycerin in a concentration of 2.5% (see table 3). Analysis of the reaction produced through 36, 48, 72 and 96 hours of cultivation on the environment.
From table 3 it can be seen that under cultivation within 36 hours hemolytic strains of V.cholerae eltor No. 14863 (ctx-Hly+) grows on agar in the form of a dense opaque micromason whitish-yellow color, the color of the environment does not change. However, hemolytic strain O139 serogroup No. 17918 (ctx-Hly+) did not show hydrolases activity and formed a blue of translucent colonies. Toxigenic in the ways cholerae was grown in a semi-transparent film blue color.
Under cultivation of strains within 48 hours hemolytic variants (ctx-Hly+) Ol and O139 serogroups form microgate whitish-yellow, while all toxigenic variants of Vibrio cholerae are also blue translucent film. From these data we can conclude that the differentiation of toxigenic non-haemolytic (ctx+Hly-and nontoxigenic hemolytic (ctx-Hly+) strains of V. cholerae possible at the stage of cultivation of strains on a nutrient medium with glycerol for 48 hours.
When growing strains within 72 hours on agar with 2.5% glycerol culture as nontoxigenic hemolytic and toxigenic non-haemolytic strains acquire dense whitish tint and become indistinguishable. Culture strains of classical biovars retain the transparency. After 96 hours of cultivation all strains formed on the sectors of agar dense whitish, with a waxy film colony. At this stage we see the manifestation of hydrolases activity of all strains of Vibrio cholerae, and differentiation in this case is impossible.
Thus, it is established that differentiation of Vibrio cholerae on hydrolases activity on solid nutrient medium with the addition of glycerol is most evident when substrate concentration of 2.5 ml/100 ml of medium is under cultivation of strains for 48 hours at 37°C.
From the experiments conducted by the staff of Rostov NPCI found that gidrolizny activity of V. cholerae can be identified by a bacteriological method on solid nutrient medium with the addition of glycerol as a standard substrate in an optimal concentration of 2.5 ml/100 ml of medium. At the same time manifests intraspecific differentiation of toxigenic and non-haemolytic nontoxigenic hemolytic strains of O1 serogroup of biovars of classical and El tor circulation and cholerae O139 serogroup "Bengal", allowing to determine the epidemiological significance of the studied strain.
The use of the present invention allows for the expense of glycerol as a substrate to identify hydrolases activity of V. cholerae O1 and O139 serogroup "Bengal" to perform a simple way differentiation atoxigenic strains O1 and O139 serogroups from toxigenic.
While glycerin is standard and available organic compound that does not require additional features, which are many domestic producers.
|Biovariant serovariants||no strain||Characterization by PCR, etc. of Greig ctx/Hl||Characteristic growth||The definition of epidemiological significance|
|14863||ctx-Hly+||Growth without active hydrolysis of glycerol (education translucent blue colonies)||Hydrolytic activity was not detected, all strains have worked equally, differentiation is impossible, pijnacker not installed|
|V.cholerae eltor||5879||ctx+Hly-||Growth without active hydrolysis of glycerol (education translucent blue colonies)|
|V.cholerae classica||W||ctx+Hly-||Growth without active hydrolysis of glycerol (education translucent blue colonies)|
|16077||Growth without active hydrolysis of glycerol (education translucent blue colonies)|
|V.cholerae A "Bengal"||R-16131||ctx+Hly-|
|17918||ctx-Hly+||Growth without active hydrolysis of glycerol (education translucent blue colonies)|
The method of differentiation atoxigenic strains of V. cholerae O1 and O139 serogroups from toxigenic on enzymatic activity, including the application of the studied culture on a nutrient medium with an indicator and a substrate, incubation of crops at 37°C and differentiation of strains by staining colonies, characterized in that the differentiation is carried out on hydrolases activity, as the substrate used glycerin, which is injected in molten agar Martin at the rate 2.5/100 ml of medium with a pH of 7.8±0.1 and indicator Nile blue sulphate 1.5 mg/100 ml of medium, the medium is poured into Petri dishes, then grown in thermostat for 4-h studied broth culture is applied in a volume of 20 µl per sector designed agar, incubation of crops is carried out within 36-48 h and when grown on medium colonies in the form of a blue semi-transparent films, blending with the color of the environment, differentiate the studied strain as toxigenic and epidemically significant, and in the presence of colonies in the form of dense yellowish or whitish nale is and differentiate the studied strain as nontoxigenic hemolytic and not epidemically significant.
SUBSTANCE: method involves concurrently taking Scotch pine needle samples growing on case and control test areas, determining chlorophyll concentration and catalase activity therein. Metabolism intensity coefficient is calculated next in case and control samples as ratio of chlorophyll concentration and catalase activity. The comparison results are interpreted in terms of anthropogenic influence degree.
EFFECT: high result data reliability; low costs and labor inputs level.
3 cl, 5 tbl
FIELD: medicine, oncology.
SUBSTANCE: one should study the activity of catalase in the tissue of malignant mammary tumor and its perifocal area, and at ratio coefficient of catalase activity in the tissue of malignant mammary tumor to that in the tissue of perifocal area being equal to 1.0 ± 0.2 one should predict the chance for the development of new foci of lesion before their clinical manifestation, that provides necessary treatment in due time.
EFFECT: higher efficiency of prediction.
1 ex, 1 tbl
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to microbiology. Day old agar bacterial cultures are inoculated in a single loop in a sodium chloride solution on a sector of culture medium, which contains barium ions of barium chloride, and the same culture medium without barium ions (control). Seeds are incubated at 28°C for 24 hours, after which Pseudomonas bacteria is identified from lack of growth of bacteria on the culture medium with barium ions and growth of bacteria on the same culture medium without barium ions. Presence of yellow-green fluorescence of bacterial lawn and the culture medium surrounding it indicates a group of fluorescent pseudomonade. Synthetic culture medium is used with the following content of ingredients, g/l: L-proline 1.0 to 3.0, L-sodium glutamate or L-glutamine 1.0 to 3.0, NaCl 5.0, MgCl2•6H2O 0.05, KH2PO4 0.05, K2HPO4 0.1, bacteriological agar 15.0, distilled water up to 1 litre, pH - 7.2+0.2, barium chloride supplement (BaCl2•2H2O) 6.0 to 16.0. The ingredients, except barium chloride, are dissolved while heating. The medium is boiled for 5 minutes. Part of the ready medium is poured into sterile Petri dishes (culture medium without barium ions). An amount of barium chloride calculated using a given formula is added to the remaining part of hot medium. The mixture is stirred and poured into sterile dishes (culture medium with barium ions).
EFFECT: invention increases standardness and simplifies identification of Pseudomonas bacteria.
2 tbl, 4 ex
SUBSTANCE: method involves deactivation of definite VGC2 DNA sequence of Salmonella typhimurium positioned between ydhE and pykF genes or its part containing at least 50 nucleotides, or the DNA version of at least 85% identity, representing VGC2 DNA of any microbe out of Salmonella aberdeen, Salmonella gallinarum, Salmonella cubana and Salmonella typhi.
EFFECT: obtainment of microbe with reduced adaptability to specific environmental conditions.
6 cl, 12 dwg, 8 ex
SUBSTANCE: invention can be used for extracting bioluminescent bacteria from sea water. Growth medium consists of sea water taken in the extraction area of bioluminescent bacteria and dry nutrient agar. Medium pH is 7.2-7.4.
EFFECT: increasing bioluminescent bacteria output.
1 tbl, 5 ex
SUBSTANCE: nutrient medium contains pancreatic digest of fresh-water fish, pancreatic digest of infusion broth waste, agar, sodium chloride, sodium carbonate, lithium chloride, vitamin B1, esculin, ammoniacal ferrum citrate, polymyxin B sulphate, ceftazidime, acriflavine hydrochloride and distilled water.
EFFECT: improved inhibiting and differentiating effect of the medium, reduced time for Listeria recovery.
3 tbl, 2 ex
SUBSTANCE: nutrient medium contains pumpkin water, ammonium phosphate twice-substituted by (NH4)2HPO4, sodium chloride (NaCl), microbiological agar and boiled tap water.
EFFECT: extended range of nutrient mediums.
1 tbl, 4 ex
FIELD: medicine; microbiology.
SUBSTANCE: liquid nutrient medium containing L-arginine, L-cysteine, L-histidine, L-isoleucine, DL-methionine, DL-leucine, L-lysine, the L-proline, DL-threonine, L-tyrosine, L-valine, L-asparaginic acid, L-serine, glucose, sodium chloride, magnesium sulphate, thiamine, calcium pantothenate, twice-substituted sodium phosphate, monosubstituted potassium phosphate, distilled water at a parity of phases 1:1, is layered on sterile perfluorodecalin, representing the lower phase. Inoculation of F. fcularensis is performed on border of the phases. Propagation of F. tularensis in a liquid diphasic nutrient medium is carried out in static conditions during 120 h at 37°C temperature.
EFFECT: increase of an output of cells of a microorganism.
1 tbl, 8 ex
SUBSTANCE: invention refers to medicine, namely to otorhinolaryngology, and can be applied in diagnostics of upper airways diseases. Said method includes examination of palatine tonsil lacuna contents. It is combined with evaluating the immunologic activity of palatine tonsils with calculating the coefficient K determined as a relation of lymphocytes in one great square of Gorjaev's count chamber to total colonies of microorganisms in thousand units cultivated after inoculation of crypt contents 0.05 ml. If K is <1, decompensation of immunologic functions of palatine tonsils is observed and requires tonsillectomy, while K>1 shows satisfactory immunologic functions of palatine tonsils with conservative therapy recommended. Application of the method allows evaluating decompensation of functions of palatine tonsils referring to immunological activity of these organs within clinical examination of the patient.
EFFECT: possibility to evaluate decompensation of functions of palatine tonsils referring to immunological activity of these organs within clinical examination of the patient.
3 ex, 2 tbl
SUBSTANCE: method for preparing fine-dispersed mycobacteria cell cultures sensitive to mycobacteriophage infection involves primary cultivation of mycobacteria in a nutrient medium with nonionic detergent being either Tween 80, or Tween 20, or Span 85 in amount 0.25-0.5 wt %, and/or ionogenic detergent being sodium lauroyl sarcosinate within 0.01-0.03 wt %. After one or more reinoculation, cultivation is carried out without detergent. Herewith throughout the entire cycle, mycobacteria cell cultivation is enabled in the nutrient medium containing, g/l: trypton - 15, yeastrel - 5, glucose - 40, sodium chloride - 2.5, potassium hydrophosphate - 5, calcium chloride - 0.111.
EFFECT: higher dose accuracy of fine-dispersed cell mass combined with bacteriophage infection of large quantity of sampled cell mass with reduced work content of the process.
FIELD: medicine; biotechnology.
SUBSTANCE: invention can be applied as microbiological control medium for bacterisation assessment of production means, raw materials, and finished products. Nutrient medium includes enzymatic hydrolysate of milk proteins, yeast autolysate, disubstituted potassium phosphate, lactose and agar.
EFFECT: enhanced commonality level, improved cultivation properties of nutrient medium.
1 tbl, 1 ex
FIELD: medicine; biotechnology.
SUBSTANCE: sea water salinity is defined in the area of bioluminescent bacteria separation. Nutrient medium based on distilled water is prepared with addition of sodium chloride in amount corresponding to defined salinity value, and dry nutrient agar in amount of 34.0-36.0 g per litre of distilled water. Bioluminescent bacteria are separated from sea water by membrane filter method, inoculated to nutrient medium and cultivated at room temperature for 22-24 hours. Presence of bioluminescent bacteria is defined by bioluminescence displayed by bacteria.
EFFECT: enhanced output of bioluminescent bacteria, reduced cost of nutrient medium.
1 tbl, 6 ex
FIELD: biotechnology, microbiology.
SUBSTANCE: method for preparing liquid lactobacterin involves regeneration, culturing passages of lyophilized culture and culturing ferment of lactobacilli in liquid lyophilized nutrient medium containing dry defatted milk enzymatic hydrolyzate with the content of amine nitrogen 1 485 mg%, 30.0 ± 3.0 g/l; yeast concentrated autolyzate, 110.0 ± 10 g/l; food agar, 0.8 g/l, and distilled water, up to 1 l. Culturing ferment is carried out up to accumulation of biomass of lactobacilli 109-1010 CFU/ml. Then 10-30% of supernatant liquid is removed from the ready product and replaced it with equal volume of fresh nutrient medium. Invention provides simplifying technology in preparing liquid lactobacterin and to elevate the storage period of viable lactobacilli. Invention can be used in producing probiotic preparations.
EFFECT: improved preparing method.
1 tbl, 3 ex
FIELD: biotechnology, microbiology.
SUBSTANCE: invention relates, in particular, to preparing nutrient media used for culturing the plague microorganism vaccine strain and can be used in medicinal microbiology. The nutrient medium for culturing the plague microorganism vaccine strain comprises additionally as a stimulating additive sodium sulfite and as a nutrient base - soybean fruits enzymatic hydrolyzate in the following ratio of components, g/l: microbiological agar, 11.0-13.0; soybean fruits enzymatic hydrolyzate, 250.0-350.0; sodium chloride, 4.5-5.5; sodium hydrogen phosphate, 3.5-4.5; sodium sulfite, 0.0003-0.0005; distilled water, the balance. Invention provides enhancing the growth property of nutrient medium.
EFFECT: valuable properties of medium.
FIELD: biotechnology, microbiology.
SUBSTANCE: method involves sampling of specimen from environment and addition of bacteriophage to sample that is able to multiplicate in analyzed microorganism. Inoculate is kept for a definite time and then bacteriophage titer is determined. The presence of microorganism is proved by increase the bacteriophage titer value measuring by degree of light diffusion in sample. Except for, in increase of light diffusion the sample is contacted with either immunosorbent and radioactivity of immunosorbent is determined or with piezoelectric resonator with electrodes covered by an antiphage Ig and the presence of piezoeffect is assayed. Invention can be used in carrying out analysis for the presence of microorganisms in air and liquid medium, monitoring of environment in periodic regime being in closed compartments mainly and in places with human mass.
EFFECT: improved method for indication.
FIELD: medicine, medicinal microbiology.
SUBSTANCE: method involves growing microorganism culture to be studied in solid nutrient medium followed by preparing microbial suspension and its incubation in the presence of lactoferrin. Control sample is prepared in parallel series. Control and experimental samples are incubated, supernatant is removed from bacterial cells and lactoferrin concentration is determined in supernatant of experimental and control sample by immunoenzyme analysis. Then anti-lactoferrin activity is calculated by difference of concentrations of residual lactoferrin in experimental and control samples. This method provides enhancing the sensitivity and precision in carrying out the quantitative evaluation of anti-lactoferrin activity in broad spectrum of microorganisms that is urgent in diagnosis and prognosis of diseases with bacterial etiology. Invention can be used in determination of persistent indices of microorganisms for assay of their etiological significance in pathological processes.
EFFECT: improved assay method.
3 tbl, 3 ex
FIELD: medicinal microbiology, medicine, pharmacy.
SUBSTANCE: invention relates to agents used for enhancing sensitivity of microorganisms to antibacterial preparations. Invention involves applying hydrocarbonate chloride-sodium drinking mineral water "Obukhovskaya" with the total mineralization 1.8-2.2 g/l as an agent for enhancing sensitivity of microorganisms to antibacterial preparations that is used early for treatment of atopic dermatitis and as an agent for treatment of iodine-deficient states. It has been found by method using disks that mineral water "Obukhovskaya" provides enhancing sensitivity of microorganisms to such antibacterial preparations as antibiotics and specific preparations against yeast-like fungi.
EFFECT: improved and valuable agent.
6 tbl, 6 ex
FIELD: biotechnology, microbiology.
SUBSTANCE: invention relates to a method for preparing nutrient medium for collection, culturing and transportation of patient blood with suspicion for the brucellosis disease. Nutrient medium contains beef meat hydrolyzate as a nutrient base, the preparation SAG-1 as a stimulating agent, and also sodium chloride, glucose, sodium citrate, glycerol, sodium hydroxide and distilled water. Invention provides enhancing accumulating properties of nutrient medium and alleviating the nutrient medium transportation.
EFFECT: improved and valuable properties of nutrient medium.
FIELD: medicine, in particular clinical microbiology.
SUBSTANCE: Claimed broth contains potassium dihydrophosphate, anhydrous disodium phosphate, magnesium sulfate, iron ammoniumcitrate, sodium citrate, glycerol, sodium hydro-L-glutaminate, thiamine bromide, manganese (II) sulfate, distillated water, normal horse serum and at least one antituberculosis preparation: rifampicin, streptomycin, isoniazide, ethambutol. Method of present invention is useful in accelerated determination of tuberculosis mycobacterium drug resistance.
EFFECT: broth with increased growth properties, simplified visual profitability analysis.
2 tbl, 3 ex
SUBSTANCE: method involves determining general microbial tympanic membrane dissemination index in middle ear within first 4-6 weeks of remission period with conditionally pathogenic microflora and staphylococci. Coagulase-negative staphylococcus strains lacking in beta-hemolytic activity and showing anti-lyzozyme activity below 6 mcg/ml within microbial dissemination of 1.0x106 conditional units/tampon being the case, miryngoplasty is considered to be indicated.
EFFECT: high accuracy in determining objective conditions for administering miryngoplasty.