Method of identifying vibrio bacteria. RU patent 2506313.
 Consortium of probiotic strains of lactobacillus rhamnosus and lactobacillus plantarum for producing bacterial preparation and direct administration ferment for producing fermented milk and fermented beet juice / 2506308Invention relates to microbiology and biotechnology. Disclosed is a consortium of Lactobacillus rhamnosus VKM B-2726D and Lactobacillus plantarum VKM B-2725D strains. |
|
Starter material of pure growth of lactate microorganisms for preparation of cultured milk products / 2505601 Strain Lactococcus casei VKPM V-8730 is obtained at available growth-supporting media and is used as starter material in cultured milk products production. |
|
Starter material of pure growth of lactate microorganisms for preparation of cultured milk products / 2505600 Strain Enterococcus durans VKPM B-8731 is obtained at available growth-supporting medium and is used as starter material in cultured milk products production. |
 Probiotic strains lactobacillus and their consortium for prophylaxis and treatment of urogenital infectious diseases of females / 2504580Group of inventions refers to microbiology and biotechnology. Proposed strains Lactobacillus crispatus "ВКМ" B-2727D, Lactobacillus gasseri "ВКМ" B-2728D and Lactobacillus plantarum "ВКМ" B-2731D have antagonistic activity in relation to pathogenic and semi-pathogenic microorganisms. Besides, the invention proposes a consortium based on the above strains for production of a bacterial preparation, a biologically active additive, and direct inoculation for obtaining fermented milk product of functional nutrition. |
 Plasmid for expression in cells of bacterium belonging to escherichia class, non-active predecessor of dnase i of human or its muteins; bacterium belonging to escherichia class, - producer of non-active predecessor of recombinant dnase i of human or its mutein; predecessor of recombinant dnase i of human or its mutein; method for obtaining recombinant dnase i of human or its mutein; method for obtaining conjugates of polyethylene glycol and recombinant mutein of human dnase i, fermentative active conjugate of mutein of recombinant dnase i of human / 2502803Invention represents a method for obtaining recombinant DNAse I of a human or its mutein, as well as their conjugates with polyethylene glycol, using a bacterium belonging to Escherichia class, transformed with expression plasmid, containing a promoter functioning in a bacterial cell, DNA fragment coding a hexahistidine cluster, a fragment coding enterokinase recognition sequence amalgamated in frame with human DNAse I or its functionally active mutein containing replacements of asparagine with cysteine, transcription termination section, vector pET28a(+) fragment containing initiation section of replication of bacteriophage fl, sequence coding aminoglycoside-3'-phosphotransferase, area of beginning of plasmid pBR322 replication, gene RNA-organising protein Rop, sequence coding lactose operon repressor. |
 Preparation for cleaning of soil from oil and oil products / 2501852Preparation 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. |
|
Bacillus licheniformis strain having apparent antagonism in relation to salmonella typhi, staphyloccus aureus, listeria monocytogenes and resistance to streptomycin and nalidixic acid / 2501851 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). |
 Lactobacillus paracasei subspecies paracasei strain having antimicrobial and immunomodulating properties, and food product on its basis / 2501850Strain is deposited in CNCM and has number 1-3689. Strain has ability to inhibit the growth of pathogenic microorganisms in culture, and namely Escherichia coli, Salmonella enteritidis and Listeria monocytogenes. Strain has anti-inflammatory properties at the ratio of IL-10/IL-12 equal to 11.8. |
|
Bacillus licheniformis vkm b-2713d strain having apparent antagonism in relation to salmonella typhi, staphyloccus aureus, listeria monocytogenes and resistance to streptomycin and nalidixic acid / 2501849 Bacillus licheniformis VKM B-2713D strain has apparent antagonism in relation to Salmonella typhi, Staphylococcus aureus, Listeria monocytogenes. Strain has resistance to antibiotics at streptomycin concentration of 20 mcg/ml and at concentration of nalidixic acid of 20 mcg/ml. Strain has α-amylase activity of 0.015 U/ml of the medium at pH 6.0, protease activity of 2.34 U/ml of the medium at pH 7.0. |
 Method for obtaining recombinant core protein of hepatitis e virus and recombinant vaccine for prophylaxis of hepatitis e virus / 2501809Invention proposes a method for obtaining recombinant core protein of hepatitis E virus (rtHEV-ORF2) and recombinant vaccine for prophylaxis of hepatitis E virus. Core protein is obtained by cultivation of recombinant yeast strain Hansenula polymorpha "КБТ"-11/pHEV-001, which contains DNA sequence integrated into genom of yeast cell and coding the fragment of amino-acid sequence from position 86 to 607 of core protein of hepatitis E virus of genotype 3 (rtHEV-ORF2) under control of promoter of MOX gene. The method allows obtaining immunogenic antigene of hepatitis E virus, which has properties of natural protein. Based on the obtained antigene there created is recombinant vaccine for prophylaxis of hepatitis E virus. Vaccine includes effective amount of rtHEV-ORF2 protein, adjuvant and a physically acceptable diluter. |
 Method for quick growth, detection and identification or counting of microcolonies of microorganisms at early stage / 2505607Invention pertains to the method for quick growth, detection and identification or counting of microcolonies of microorganisms at early stage. The described method includes the following stages: obtaining the container with medium with porous element located above or under the top surface of the medium, note that the medium has nutritious substances for quick growth of microcolonies of microorganisms and porous element has pores from 1000 to 107 Da; pouring the liquid sample without serial dilution into container to the area above porous element; capturing the microorganisms in porous element or at the medium above porous element; incubation of container for the time sufficient for quick growth of microcolony at an early stage; transfer of porous element and any medium above it from container to the secondary medium for evaluation, detection and identification of microorganisms; and microcolonies research relatively the growth, detection, identification or counting of microorganisms. The said method for growth, detection and identification or counting of microorganisms takes not more than approximately six hours. |
|
Method for increasing biocidal and therapeutic action of suspension-cream with linco-spectin / 2505285 Invention represents a method for increasing biocidal and therapeutic action of a suspension-cream with linco-spectin consisting in detoxification and polymerisation of linco-spectin 100 g in water 300 ml by 0.15±0.05% glutaric aldehyde 0.15±0.05% alkyldimethyl benzylammonium chloride at 38-40°C for 2-3 days. |
 Method for individual selection of preparations containing probiotic lactic bacterial strains for effective intravaginal therapy / 2500411Invention 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. |
 Method of determining genus of bacteremia agents / 2495939Method 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. |
|
Method of determining minimal inhibitory concentration of antimicrobial preparation / 2491348 Microorganism-causative agent is isolated from the biological material of the patient. The standard suspension of the isolated microorganism-causative and two-fold dilutions of antibacterial preparations with known activity is prepared. Inoculation is carried out. The standard suspension of the isolated causative agent with the volume of 0.1 ml is mixed with 5.0 ml of liquid nutrient medium specific for this pathogen. It is incubated for 24 hours at +37°C. The test 1:100 and control 1:200 dilutions are prepared in the same nutrient medium. Each of the two-fold dilutions of antibacterial preparations in amount of 0.075 ml with an equal amount of the test dilutions is applied in the test cavities of the plate. In the control cavity 0.150 ml of control dilution is applied and incubated for 48 hours at + 37°C. The liquid fraction of the suspension is removed from the cavities. For staining 0.2 ml of 1% solution of crystal violet is added to each cavity. It is exposed at room temperature for 30 min. The contents of the cavities are washed three times with distilled water and 0.2 ml of 99% solution of dimexidum is added to each cavity. It is exposed at room temperature for 15 minutes. The optical density of the medium in the control (ODC) and the test cavities is measured in the optical units (pu) in a microplate reader at the wave-length of 540 nm. When fulfilment of the condition ODC> 0.2pu, the greatest two-fold dilution of antibacterial preparation in the test cavity with an optical density less than 0.2 pu is defined as the minimum inhibitory concentration (MIC). |
 Method of detecting mycobacterium tuberculosis in air environment / 2490328Invention relates to microbiology and is meant for detecting mycobacterium tuberculosis in the air environment of prevention and treatment facilities. The method involves collecting air samples through directed impact action of a jet on a foam plate in a Petri dish, said plate being soaked in 4-6 ml of 5% trisubstituted sodium phosphate solution followed by incubation of the Petri dish wit the foam plate for 18-24 hours; centrifuging the analysed material, followed by separation of supernatant fluid and neutralising the precipitate with 1% citric acid solution in ratio of 1:1, followed by inoculating the neutralised precipitate on a Novaya culture medium and culturing until growth. The diameter of the foam plate is equal to the diameter of the bottom of the Petri dish and its thickness is 4-6 mm. |
 Method of detecting and counting viable legionella pneumophila microorganisms / 2490327Present 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. |
 Method to assess biological activity of lactobacilli and bifidus bacterial relative to choleraic vibrios in vitro / 2487943Invention may be used to assess biological activity of lactobacilli and bifidus bacteria relative to Vibrio cholerae with the purpose to establish possibility to use probiotics for prevention and treatment of cholera. The method provides for testing of antagonistic and acid-forming activity of lactobacilli and bifidus bacteria strains. Antagonistic activity relative to choleraic vibrios is detected by the method of holes in dense nutrient media - CDM - agar and alkaline agar, and activity is accounted on the basis of quantitative data, namely: zone of growth inhibition (in mm) of a test strain V. cholerae from 6.0 to 15.0 - low antagonistic activity, from 15.1 to 29.0 - medium, ≥29.1 - high. Acid-forming activity of strains relative to choleraic vibrios expressed in Turner degrees (°T) is defined on the basis of quantitative data. So if acid formation ≤is 99.9 (°T), then activity of acid formation is assessed as low, if the values of this index are within (100-149.9) (°T) - as medium, and if the value is ≥150.0 (°T) - as high. At the same time the result of assessment of lactobacilli and bifidus bacteria biological activity relative to V. cholerae eltor, classica, O139, non O1/ non O139 is considered positive with 4 versions of combination of testing results according to the specified indices, namely: high antagonistic activity, high acid-forming activity, accordingly, high - medium, medium - high, medium - medium. |
|
Method of diagnostics of candidiasis of upper respiratory tract in workers of agroindustrial complex / 2485183 Method provides sampling of the material under study. Inoculation of the material under study on nutrient medium sabouraud followed by incubation on this medium at 37°C for 24 hours. A quantitative assessment of growth of yeast-like fungi of the genus Candida, and with a value exceeding 10*1 CFU/swab the candidiasis of upper respiratory tract is diagnosed. The obtained colonies are examined under the microscope and the isolated colonies of yeast-like fungi of the genus Candida are separated from these colonies. The isolated colonies of yeast-like fungi of the genus Candida are suspended in five test tubes containing liquid medium sabouraud with the phenol red indicator, where the discs with carbohydrates are added, and in the first test tube - a disc containing maltose, in the second - sucrose, the third - lactose, the fourth-galactose, the fifth - trehalose, and are incubated at 37°C for 24 hours, followed by assessment of colour change of the indicator. The colour change of the indicator to yellow is taken as one, lack of colour change is taken as zero, the sum of the values obtained in assessment of the indicator colour in five test tubes is calculated. If the value of the sum is 0 the candidiasis of upper respiratory tract caused by Candida kruzei is diagnosed, if it is 1 - the candidiasis of the upper respiratory tract caused by Candida glabrata is diagnosed, if it is 3 - the candidiasis of the upper respiratory tract caused by Candida albicans is diagnosed, if it is 4 - the candidiasis of the upper respiratory tract caused by Candida tropicalis is diagnosed, if it is 5 - the rest. |
|
Elective-differential nutrient medium for extraction of choleraic vibrios / 2484141 Invention is an elective nutrient medium, which may be used in laboratory practice for extraction of a cholera agent. The nutrient medium contains a nutrient base, microbiological agar, saccharose, sodium chloride, sodium carbonate, bismuth nitrate pentahydrate, sodium citrate, cattle bile, EDTA iron (III)-sodium salt, bromthymol blue, mesoinositol, activated charcoal, sodium sulfite and L-cysteine, potassium tellurite and distilled water. The nutrient base in the nutrient medium is a dry nutrient broth from a pancreatic overcook of Caspian sprat or a dry nutrient broth from hydrolysate of fish and bone flour or a complex of yeast extract and peptone at the specified ratio of components. |
 Vns-met-histones / 2498997Nucleic acid molecule codes a polypeptide consisting of two residues of methionine as the first and the second N-end amino-acid residues connected through a peptide link to a mature eucariotic histone. Polypeptide is obtained by cultivation of a host cell transformed by an expression vector including the above molecule of nucleic acid. Polypeptide is used as part of pharmaceutical composition for therapy of cancer, bacterial, virus or fusarium infections. Besides, polypeptide is used as part of composition for diagnostics of a patient in relation to response to pharmaceutical composition containing the above polypeptide, or in relation to curability using it. |
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
The invention relates to the field of Microbiology. Can be used by bacteriological studies on identification of the bacteria of the genus Vibrio and manufacturing of diagnostic products for these studies.
There is a method test using as substance of the O129 (2,4-diamino-6,7-diisopropyl-pteridine phosphate) in paper disks on 10 mcg 150 mcg. test applies the bacteriological study on the identification of species of Vibrio after determining the morphology of pure culture identified by gram stain (curved gram-negative rods) and the identification of bacteria oxidase. According to the results of test is determined by bacteria belonging to the genus Vibrio and they are divided into groups of species Vibrio: sensitive to 10 mcg 150 mcg O129 - V. cholerae, V. vulnificus, Vibrio spp.; resistant to 10 mcg 150 mcg O129 - bacteria of Aeromonas or V. cholerae O1 and V. cholerae 139; resistant to 10 mcg sensitive to 150 mcg O129 - V. parahaemolyticus and V. alginolyticus, and others. The final identification of species is carried out in addition commercial test-systems biochemical identification and serums to O1 and O139 antigens V.cholerae (Health Protection Agency. 2011. Identification of Vibrio species. UK Standards for Microbiology Investigations. ID 19. Issue 2.1.P.4). Note to method. The method of setting test substance 129 (2,4-diamino-6,7-diisopropylpteridine phosphate salt) CAS No 84176-65-8. Uses paper disks containing 10 mcg 150 mcg O129 to disk (production companies Oxoid, Dalynn Biologicals, Hi-Media). Fresh pure culture of the microorganism tested seeded with sterile tampon on a Cup with a non-selective nutritive medium (blood agar with 0.5% sodium chloride to obtain a continuous growth; placed on the surface of the environment disks with 10 mcg O129 and 150 mcg O129 on distance from each other, are incubated in aerobic conditions at 35 C for 24 h, then note the presence of a zone inhibit the growth of bacteria and interpret the results: sensitivity - a inhibit the growth of bacteria around both disks; partial sensitivity - no zones of growth suppression drive around 10 mcg, there is a zone suppress growth drive around 150 mcg; sustainability - there are no zones inhibit the growth of bacteria around both disks. Controls: Vibrio metschnikovii ATCC 7708 - sensitive; Vibrio parahaemolyticus ATCC 17802 - partial sensitivity; Aeromonas hydrophila ATCC 49140 - resistant (Dalynn Biologicals. Diagnostic Microbiology Catalogue. 2005. No DD 10.11.0129 Disks).
The prototype of the proposed method test us elected the above analogue of the Health Protection Agency. 2011. Identification of Vibrio species. UK Standards for Microbiology Investigations. ID 19. Issue 2.1. P.4 using substances 129 and methods of its application instructions Dalynn Biologicals. Diagnostic Microbiology Catalogue. 2005. No DD 10.11.0129 Disks. The prototype is the latest in a standardized fashion, it is most similar to the claimed method for the totality essential sign and objectively indicate defects test, which eliminated the claimed method.
The disadvantage of a prototype method is the low diagnostic sensitivity test, because of cross-resistance vibrios to substance O129 and antimicrobial drug trimethoprim, included into the composition of a medicinal product co-trimoxazole - 82.7% of strains V.cholerae O1 allocated in India from cholera patients resistant to O129 and co- (Rama-murthy So, A. Pal, Pal S.C., Nair G.B. Taxonomic implications of the emergence of high frequency of occurrence of 2,4-diamino-6,7-diisopropylpteridine - resistant strains of Vibrio cholerae from clinical of cholera in Calcuta, India.- J. Clin. Mi-crobiol. - 1992. - Vol.30. - No 3. - P. 742-743. Identified strains with cross-resistance to O129 and co- among V.cholerae not O1 (Abbott S.L., Cheung W.K., Portoni B.A., Janda J.M. Isolations of vibriostatic agent O129 - resistant Vibrio cholerae non - 1 from a patient with gastroenteritis. - J. Clin. Microbiol. - 1992. - Vol.30. No 6. - P. 1598-1599).
The aim of the invention is to increase diagnostic sensitivity test for identification of bacteria of the genus Vibrio.
In accordance with the invention of this goal is achieved by the fact that after the preliminary selection of the material subject to the research, is a pure culture, gram-negative, rod-shaped, glucose, or bacteria seeded daily culture of the studied bacteria on the surface of nonselective nutrient agar (GRM-agar) with 1% sodium chloride is placed on the sown surface of the paper disk containing 10 mcg or 16 mcg substances (2,5-dichloro-4-nitrosalicylanilide); incubated crops in aerobic conditions at 35 C or 28 degrees C for 24 h; identify the bacteria belonging to the genus Vibrio by the presence of drive around the zone inhibit the growth of bacteria.
Note1. Controls test: Vibrio metschnikovii 5H - sensitive (the presence of zones of inhibit the growth of bacteria around the disk), Aeromonas caviae 12H - stable (no zone inhibit the growth of bacteria around the disk).
2. GRM - agar (nutrient agar for cultivation of microorganisms dry, production of the State scientific center for applied Microbiology and biotechnology, ., Russia). The composition g/l: pancreatic digest of fish-flour - 12,0; peptone enzymatic - 12,0; sodium chloride - 6,0; agar microbiological to 12.0; pH 7,1 - 7,5. To bring the content of sodium chloride up to 1% should be added to 4 g/l sodium chloride. Preparation of the nutrient agar from a dry environment - for instructions to the drug.
3. Technique of preparation of paper disks . Use discs with a diameter of 6 mm from cardboard technical filtering (GOST 6722-75) production " Pasteur them. Pasteur, Saint-Petersburg; commercial product «Fenasal» (Niclosamide, CAS No 50-60-7) production (Mainland,China); dimethyl sulfoxide solvent (CAS No 67-68-5). It fenasala 10 mg or 16 mg placed in a sterile mortar, add 10 ml of dimethylsulfoxide and dissolve when grinding the drug. The resulting solution is clear, yellow color contains 1000 mcg/ml or 1600 mcg/ml . Contributed by micropipette 10 ml solution of on one disk the disk contains 10 mg or 16 mcg . The control drives contribute 10ul dimethyl sulfoxide solvent. Disks dried in a thermostat at 37 C for 24 h is then filled into bottles with silica gel, stored at 4 C-8 Degrees C. shelf life disks 6 months. Controls drives in the manufacture of: V. metschnikovii 5H - sensitive (the presence of zones of inhibit the growth of bacteria around the disk), A. caviae 12H - stable (no zone inhibit the growth of bacteria around the disk); both control test strain not have zones inhibit the growth of bacteria around controlling disk containing only dimethyl sulfoxide solvent.
A distinctive and essential feature of the method is the use of the test as substances (2,5-dichloro-4-nitrosalicylanilide) in the amount of 10-16 g disk.
A distinctive and essential feature was not used in the prototype and is not known from the prior art. (commercial drug, «Fenasal») was known as the drug that has been used in medical practice for treatment of intestinal - M.D. Mashkovsky Drugs (manual on pharmacotherapy for doctors). Part II. - M: Medicine, 1977. .332-333. Us for the first time revealed its selective antimicrobial action on gram-positive microorganisms on the basis of which the invention has been created on the use of this preparation into the nutrient media for the allocation of gram-negative enterobacteria ( H.E. Author's certificate of the USSR №1386660 on the invention «the Method of selection of bacteria of the family Entrobacteriaceae». Stated 24.12.1985. Publ. 07.04.1988. Bul. №13. P.124). Sensitivity vibrios to this drug has not been studied. Only recently we have found that selective sensitivity to this drug vibrios among other gram-negative bacteria. First reported in the application for an invention (table 1).
A distinctive and essential feature should not be obvious from the prior art. Were not known advantages of the new drug the famous O129, which determined the achievement of the technical result of the invention claimed. Was not known sensitivity to the most important in the pathology of the human species vibrios V. cholerae, V. parahaemolyticus, V. vulnificus. We were the first on the basis of study of many strains of V. metschnikovii (28 strains), allocated from the river Neva, were identified strains of V. metschnikovii resistant to O129 (3 strains). All strains resistant to O129 (Oxoid) 150 g, as well as all other strains of V. metschnikovii were sensitive to , which indicates higher diagnostic sensitivity test . We established suitability test for the study of vibrios V. cholerae non O1/non 139 V. parahaemolyticus, V. vulnificus (table 2). There was also a lack of positive results in tests with and O129 all 26 of tested strains 5 species of bacteria of the genus Aeromonas {A. caviae, A. popoffii, A. salmonicida, A. bestiarum, A. hydrophila).
A distinctive and essential feature of the method directly determines the achievement of this goal. We found that all three strains of V. metschnikovii resistant to O129 have cross-resistance to co- ( method with the disc trimethoprim/sulfamethoxazole 1,25/23,75), but were sensitive to . Other strains of V. metschnikovii were sensitive to both (table 2). This scientific fact testifies not only about the achievement in this study, the technical result of the goal - increase diagnostic sensitivity test, but pointed to the fundamental possibility of «rehabilitation» test in respect of other types of vibrios, due to the lack of cross-resistance to trimethoprim (co-).
Table 1Sensitivity levels to (2,5-dichloro-4-nitrosalicylanilide) on GRM - agar with 1% sodium chloride bacteria of the genus Vibrio and other genus of gram-negative, glucose, or bacteria
Genus, species of bacteria
Number of investigated strains
IPC (mcg/ml)
V. metschnikovii
28 1-6Vibrio vulnificus*
1 6Vibrio parahaemolyticus*
1 6Vibrio cholerae non-O1/non 139
2 4-8Aeromonas caviae
1764-more than 64
Aeromonas popoffii
4 32-64Aeromonas salmonicida
2 64Aeromonas hydrophila
1 32Aeromonas bestiarum
1 32Escherichia coli
3more than 64
Shigella sonnet
3more than 64
Salmonella Enteritidis
3more than 64
Klebsiella pneumoniae
3more than 64
Klebsiella mobilis*
3more than 64
Enterobacter cloacae
3more than 64
Serratia marcescens*
3more than 64
Hafnia alvei*
3more than 64
Citrobacter freundii
3more than 64
Proteus mirabilis
3more than 64
Morganella morganii
3more than 64
Yersinia enterocolitica
3more than 64
Designation: * including standard strain.
Table 2Comparative diagnostic sensitivity identification vibrios (2,5-dichloro-4-nitrosalicylanilide) and 129 on GRM-agar with 1% sodium chloride
Types of bacteria of the genus Vibrio
Number of investigated strains
O129 (150 mcg/disc)
(10 mcg/disc)
of them, sensitive
sensitivity P±m%
of them, sensitive
sensitivity P±m%
V. metschnikovii
28 25*89,2±5,8
28 100V.parahaemolyticus **
1 1 1V. vulnificus**
1 1 1V. cholerae not O1/not O139
2 2 2 Just 32 2990,6 ħ 5.1
32 100Designations. * 3 strains have cross-resistance to O129 and co-, ** typical strain.
Essential features of the proposed method are: characteristics of the strains to be investigated (gram-negative, rod-shaped, fermenting micro-glucose, or ), which limits the range of species of bacteria and includes all types of vibrios; use of nonselective nutrient agar (GRM-agar) with a final concentration of 1% of sodium chloride that permits the cultivation of and halophilic vibrios; incubation of crops in aerobic conditions at 35 degrees C.
Examples confirming the possibility of implementing the method.
Example 1. The investigated material - daily culture strain №19, allocated from the river Neva, containing gram-negative, rod-shaped, fermenting micro-glucose, bacteria, seeded with a sterile swab the surface GRM-agar with 1% sodium chloride, place on the sown surface of the paper disk with 10 mcg and paper disk with 150 mcg O129, incubated in aerobic conditions at 35 C for 24 hours, then take into account the result: drive around with 10 mcg area, inhibit the growth of bacteria, drive around with a 150 mcg O129 no zone inhibit the growth of bacteria. Controls carried out by the same method with the control strains: Vibrio metschnikovii 5H - are also areas of inhibit the growth of bacteria around disks with 10 mcg and 150 mcg O129; Aeromonas caviae 12H - no zones inhibit the growth of bacteria around disks with 10 mcg and 150 mcg O129. Conclusion: the strain sensitivity # 19 indicates that it belongs to the genus Vibrio, thus there is acquired resistance to O129. Subsequent investigation of strain additional tests species identification and antibiotic susceptibility established his membership of the mind V. metschnikovii and resistance to co-.
Example 2. The investigated material - daily culture of V. cholerae strain not O1/ not O139 ctx And - No. 9741 obtained from the North-Western anti-plague station, containing gram-negative, rod-shaped, fermenting micro-glucose, bacteria, seeded with a sterile swab the surface GRM-agar with 1% sodium chloride, place on the sown surface of the paper disk with 10 mcg , incubated at 28 C. within 24 hours, after which take into account the result: drive around with 10 mcg area, inhibit the growth of bacteria. Controls carried out by the same method with the control strains: V. metschnikovii 5H - there is a zone inhibit the growth of bacteria drive around with 10 mcg ; A. caviae 12H - no zone inhibit the growth of bacteria drive around with 10 mcg . Conclusion: the strain sensitivity №9741 to confirming his / her belonging to the genus Vibrio and the opportunity to study test vibrios type of V. cholerae.
Example 3. The investigated material - daily culture strain of V. parahaemolyticus 13580 (ATCC 17802)obtained from the Rostov-on-don research period of antiplague Institute, containing gram-negative, rod-shaped, fermenting micro-glucose, bacteria, seeded with a sterile swab the surface GRM-agar with 1% sodium chloride, place on the sown surface of the paper disk with 10 mcg , incubated at 35 C for 24 hours, then take into account the result: drive around with 10 mcg area, inhibit the growth of bacteria. Controls carried out by the same method, consistent with the results of study 2. Conclusion: the strain sensitivity V. parahaemolyticus 13580 (ATCC 17802) to confirming his / her belonging to the genus Vibrio and the opportunity to study test vibrios kind V. parphaemolyticus.
Example 4. The investigated material - daily culture strain V. vulnificus 13344 (ATCC 27562)obtained from the Rostov-on-don research period of antiplague Institute, containing gram-negative, rod-shaped, fermenting micro-glucose, bacteria, seeded with a sterile swab the surface GRM-agar with 1% sodium chloride, place on the sown surface of the paper disk with 16 mcg , incubated at 35 C for 24 hours, then take into account the result: drive around with a 16 mcg area, inhibit the growth of bacteria. Controls carried out by the same method, consistent with the results of study 2. Conclusion: the strain sensitivity V. vulnificus 13344 (ATCC 27562) to confirming his / her belonging to the genus Vibrio and the opportunity to study test vibrios type of V. vulnificus.
Thus, when all marginal quantities drives and variations of the temperature regime of the proposed method yielded significant results determine test facilities of the studied bacteria of the genus Vibrio species, including species V. metschnikovii, V. cholerae, V. parahaemolyticus, V. vulnificus. A method for the identification of bacteria of the genus Vibrio using allows for the identification of belonging to strains that have acquired cross-resistance to O129 and co-, and therefore increases the diagnostic sensitivity identification cholerae.
A method for the identification of bacteria of the genus Vibrio, including a preliminary selection of the material subject to the research, is a pure culture of rod-shaped, gram-negative, glucose, or bacteria, sowing investigated daily culture of bacteria on surface nonselective nutrient agar (GRM-agar) with 1% sodium chloride, placing on the sown surface of the paper disk containing substance, incubation crops in aerobic conditions at 35 C for 24 h, the definition of the membership of bacteria of the genus Vibrio by the presence of drive around the zone inhibit the growth of bacteria, wherein as substance use (2,5-dichloro-4-nitrosalicylanilide) in an amount of from 10 to 16 mcg mcg per disk.