Two-phase nutritional medium for thin layer cultivation of helicobacter pylori and method of its realisation. RU patent 2518304.
 Test-system for differentiating species and biotypes of bacteria of genus yersinia / 2518297Invention relates to field of microbiology and deals with test-system for differentiation of species and biotypes of bacteria of genus Yersinia. Claimed invention consists of a set of nutritional media, which contain substrates and reagents for determination of presence in microorganism of lysine decarboxylase, ornithine decarboxylase, tryptophan deaminase, urease, acetoin production, indole production, meliobiose fermentation, rhamnose fermentation, saccharose fermentation, sorbite fermentation, lipase, xylose fermentation, maltose fermentation, α-methyl-D-glucopyranoside fermentation, salicin fermentation, sorbose fermentation and raffinose fermentation. |
 Anti-obesity preparation, anti-obesity food product or drink, preparation improving tolerance to glucose and food product or drink for improving tolerance to glucose / 2518293Group of inventions relates to field of biotechnology. Strain Bifidobacterium breve MCC 1274 FERM BP-11175 demonstrates low coefficient of conversion of linoleic acid into conjugated linoleic acid. Strain demonstrates coefficient of conversion of linoleic acid into conjugated linoleic acid not higher than 10%. To reduce or prevent obesity or to improve tolerance to glucose said strain is introduced in efficient quantity to subject requiring it. Also claimed are food product and drink, which contain preparation based on said strain. |
| Nutrient medium for submerged cultivation of tularemia microbe / 2518282 Nutrient medium for submerged cultivation of tularemia microbe contains as a base, that provides the content of amino nitrogen in the medium of not less than 0.32%, dry enzymatic hydrolyzate of fibrin obtained from waste of whey-vaccine production, and the components - glucose and calcium pantothenate at a predetermined ratio. |
 Nutrient medium for extraction of bacteria of shigella kind from water objects / 2517750Nutrient medium is proposed for extraction of Shigella kind bacteria from water objects. The nutrient medium contains the following components: 0.1% alcohol solution of bromcresol green - 9.0 ml; 0.2% alcohol solution of methyl red - 3.0 ml; fermentative peptone - 0.5 g; extract of fodder yeast - 4.5 g; potassium dihydrophosphate - 8.7 g; caustic soda - 1.4 g; sodium chloride - 5.0 g; distilled water - up to 1000 ml; medium pH is 6.70-6.85. |
 Method to prepare medicated product from live strains and microorganisms of lactobacilli and bifidobacteria "lb-complex plus" / 2517734Method provides for cultivation at 37±1°C of strains of lactobacilli and bifidobacteria in the medium and packing of liquid product with account of daily dose necessary for patients. The medium contains components in the following quantities: caseine hydrolysate dissolved by distilled water, 0.33-0.4 g/l, sodium chloride 5 g/l, fructose 10 g/l, peptone 2 g/l, agar-agar 0.75 g/l for lactobacilli, for bifidobacteria - 1.0 g/l, ascorbic acid 0.25 g/l, distilled water 0.67-0.6 g/l. Strains-producers are Lactobacillus plantarum 8 RA-3, Lactobacillus fermentum 39, Lactobacillus fermentum 90 TC-4, Bifidobacterium bifidum 791, Bifidobacterium longum 379, Bifidobacterium bifidum 1. For lactobacilli and bifidobacteria they prepare accordingly media with different content of agar-agar, amine nitrogen in caseine hydrolysate 160-170 and 180-200 mg% and medium pH 7.8-8.0 and 8.5-8.6. Starting from the first generation the bifidobacteria and lactobacilli are cultivated for 24±1 hours, two strains together, one separately. The produced biomass is mixed with the fresh nutrient medium at the ratio of 1:1000, and strains of lactobacilli are cultivated 24±1 hours, strains of bifidobacteria - 48±1 hours. Upon completion of cultivation of biomass of strains they are mixed at the ratio of 2:1:2:1. |
 Nourishing composition, including probiotics and improving sleep pattern / 2517616Invention relates to application of probiotic bacterial strain for production of a probiotic composition for reducing sleep disorders and/or improvement of sleep quality in people and animals. As a bacterial strain used is Lactobacillus reuteri DSM 17938 or Bifidobacterium longum NCC 3001 (ATCC BAA-999). |
| Strain of fungus stagonospora cirsii davis having herbicidal activity against canada thistle / 2515899 Strain of the fungus Stagonospora cirsii Davis of All-Union Research Institute of Plant Protection 1.42 has herbicidal activity against canada thistle and species closely related to it - S. Cirsii. It was deposited in the State Collection of Microorganisms of All-Union Research Institute of Plant Protection under the collection number of Stagonospora cirsii Davis of All-Union Research Institute of Plant Protection 1.42 and can be used for biological control of canada thistle. |
 Medication (versions), composition (versions) and application of medication (versions) for reduction of halitosis / 2515113Invention relates to versions of medication for reduction of halitosis, versions of compositions based on said medications and versions of application of said medications. Device for reduction of halitosis represents strains of microorganisms Lactobacillus acidophilus, selected from group Lactobacillus acidophilus DSM 19825, Lactobacillus acidophilus DSM 19826 and Lactobacillus acidophilus DSM 19827. Version of said medication represents culture supernatant of said strains. Also claimed are compositions, which contain said microorganisms, and their application for reduction of halitosis. |
| Method of culturing bifidobacteria in milk / 2515048 Invention relates to biotechnology and can be used in the dairy industry. Pasteurised milk is cooled to a given temperature and MnSO4, ZnSO4, KJ, CuSO4, FeSO4 and selexen are then added in a given ratio, followed by addition of bifidobacteria. The mixture is stirred and soured until a clot of given titratable acidity is obtained. |
| Method of obtaining toxin actinobacillus pleuropneumoniae apxi, applying culture medium, containing calcium-borogluconate complex / 2514667 Claimed invention relates to field of microbiology and deals with method of obtaining RTX-toxin ApxI. Claimed method is realised by cultivation of bacteria Actinobacillus pleuropneumoniae in culture medium, which provides growth of bacteria, and said culture medium contains borogluconate in concentration lower than 60 mmol/l in order to form in medium calcium-bologluconate complex. |
 Strain of lactobacilli lactobacillus paracasei cncm i-2116 (ncc 2461) eliciting ability to prevent intestine colonization with pathogenic microorganisms causing diarrhea, supernatant of its culture and oral agent for prophylaxis and/or treatment of diarrhea-associated disorders / 2243779Invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea. |
| Strain of microorganism lactobacillus buchneri 66 for ensilage of maize green mass and preserving flattened grain / 2243999 The strain Lactobacillus buchneri 600 is isolated from barley grains at milkwax stage of ripeness. The strain Lactobacillus buchneri 600 is registered in 05. 08. 2002 year in the All-Russian State collection of microorganism strains used in veterinary science and animal husbandry at number Lactobacillus buchneri VGNKI 02.08.04-DEP and deposited in the collection 000 "Biotrof". Invention provides the more effective multiplication of the strain in maize ensilaging green mass and preserving flattened grains with enhanced formation of lactic acid, inhibition of putrid microflora that allows preparing fodder from vegetable raw with enhanced quality. Invention can be used in fodder production for ensilage maize green mass and preserving flattened grains. |
| Strain lactobacillus acidophilus as producer of fodder protein / 2244000 Invention relates to a new isolated strain of Lactobacillus acidophilus 1660/08 as a producer of the protein fodder. The strain Lactobacillus acidophilus 1660/08 is obtained by the selection method and selected by its ability to form significant amount of crude protein and to accumulate the biomass. The strain is deposited in the VGNKI collection at number VGNKI-03.04.10.-DEP. Invention provides eliminating the environment pollution in producing the protein fodder, to enhance the specific protein yield, to reduce energy consumptions in preparing protein fodder, to simplify and to accelerate the process in its preparing, to simplify equipment fitting out and to utilize waste in manufactures using natural raw. |
| Strain lactobacillus plantarum as producer of fodder protein / 2244001 The strain Lactobacillus plantarum 578/25 is obtained by method of step-by-step selection and selected by its ability to produce significant amount of crude protein and to accumulate the biomass. The strain is deposited in the VGNKI collection at number VGNKI-03.04.09.-DEP. Invention provides eliminating the pollution of environment in producing the protein fodder, to elevate the protein specific yield, to reduce energy consumptions in preparing protein fodder, to simplify and to accelerate the process of its preparing, to simplify apparatus equipment, to utilize waste in manufacturing using the natural raw. |
 Strain of microorganism lactobacillus lactis for preparing curd from milk / 2244002The strain of microorganism Lactobacillus lactis VKPM B-8354 is prepared without using mutagens and genetic methods and shows resistance against broad spectrum of lactophages. The strain ferments effectively milk from different trading sorts, with broad range of fatness and different methods of thermal treatment. Individual specific properties of the strain allows its applying as a monostrain ferment. Curd obtained with applying the strain L. lactis VKPM B-8354 shows good organoleptic qualities, nonacid taste and homogenous consistence. The strain is suitable especially for plants with small volume of manufacture but with varied assortment. |
| Method for isolation and selection of microorganisms as producers of cyclodextrin glucanotrasnferase, strain of microorganism bacillus circulans b-65 ncaim (p) 001277 (b-65) as producer of extracellular cyclodextrin transferase, cyclodextrin glucanotransferase obtained from its and its applying for preparing cyclodextrin / 2244742 The strain Bacillus circulans B-65 a producer of cyclodextrin glucanotransferase is isolated and selected form the soil sample by culturing in nutrient medium with amylolytic activity 12.17 U/ml and cyclodextrinogenic activity up to 0.530 U/ml. Cyclodextrin glucanotransferase isolated from B. circulans B-65 shows the high degree for conversion of starch to cyclodextrins and this enzyme is specific for formation of β-cyclodextrin. Invention can be used in food industry for preparing cyclodextrins and cyclodextrin glucanotransferase used in different branches of industry. |
| Bacterial preparation, method for its producing, nutrient medium for culturing cells escherichia coli vkm cr-322d and method for prophylaxis and treatment of gastroenteric disease in agricultural and domestic animal and poultry / 2244743 For preparing a preparation cells of microorganism Escherichia coli VKM CR-322D is cultured in nutrient medium containing Hottinger's broth, glucose, yeast extract, manganese sulfate, potassium hydrophosphate, sodium chloride and tap water in the content of amine nitrogen 125-155 mg%. Glucose is added by batch portions in the process of culturing cells that is carried out at temperature 30-31oC at stirring and aeration for 10-12 h. Prepared cultural fluid containing 3 x 109 bacterial cells/ml is mixed with protective sucrose-gelatin medium and subjected for lyophilic drying. Dried mass is stored under nitrogen that enhances safety of viable cells in the preparation. Applying the preparation for prophylaxis and treatment of agricultural and domestic animals and poultries with gastroenteric diseases provides its effectiveness. |
| Method for preparing liquid lactobacterin / 2244744 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. |
| Strain bifidobacterium longum 379-in used for preparing bacterial preparations, biologically active supplements for food, ferments, fermented-dairy and nonfermented dairy foodstuffs, hygienic and cosmetic agents / 2244745 The strain Bifidobacterium longum 379-IN is obtained by selection without using methods of genetic modification of the strain Bifidobacterium longum B379M and distinct by ability to utilize insulin. The strain is deposited in GKNM GU "MNIIEM named for G. N. Gabrichevskiy Russia Ministry of Public Health" at № 172. The strain shows high technological effectiveness, accumulates biomass with substrates of vegetable origin and artificial nutrient media for short periods with concentration of bifidobacteria, it elicits acid-forming and antagonistic properties with respect to pathogenic and putrid microflora. This allows its using in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, ferments, hygienic and cosmetic agents providing probiotic effect and normalization of microbiocenosis in human body, among them in gastroenteric and urogenital tracts, cutaneous and mucosa integuments. Invention can be used in manufacturing bacterial preparations, biologically active supplements for food, fermented-dairy and nonfermented-dairy foodstuffs, hygienic and cosmetic agents. |
| Nutrient medium for culturing plague microorganism vaccine strain / 2245362 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. |
FIELD: chemistry.
SUBSTANCE: invention relates to field of microbiology and can be applied in medicine. Method includes preliminary inoculation of analysed material on Columbia agar with addition of 5% of sheep blood. Thermostatting is performed under microaerophilic conditions in CO2 incubator at 37°C for 3-5 days at higher humidity. Sampling colonies and their re-inoculation on Petri dishes with chocolate agar are performed and thermostatting is carried out under microaerophilic conditions in CO2 incubator at 37°C for 3-5 days at higher humidity. When thermostatting stage is completed Petri dishes with chocolate agar are poured on the top with liquid nutritional medium based on Schaedler broth, enriched with 10% cattle serum with their further thermostatting in CO2 incubator at 37°C for 72 hours.
EFFECT: obtaining nutritional medium.
4 cl
The invention relates to Microbiology and may be used in medicine.
The discovery of H. pylori in 1982, was the starting point in the formation of the new concept etiology of gastroduodenal diseases. Currently accepted fact is the role N. pylori in the pathogenesis of chronic gastritis. In 1990, these bacteria are officially included in the international classification as H. pylori gastritis or gastritis associated with Helicobacter pylori infection, gastritis type Century With H. pylori infection is associated with about 70-80% of cases of gastritis, more than 70% of cases of gastric ulcer, 90% of peptic ulcer duodenal ulcer, increased risk of gastric cancer and B-cell lymphoma. Scientific reports of the last years actively discussed the issue of communication with H.pylori wieloletnie manifestations (bowel disease, hepatobiliary system and pancreas), as well as lesions of the cardiovascular system, musculoskeletal system, skin, etc.
For correct and timely diagnosis helikobakterioza developed many methods, grouped using several parameters for classification. Depending on the purpose, material, technique they are divided into screening and confirmatory; invasive and non-invasive; direct and indirect. One of the methods of laboratory diagnosis of H. pylori infection is bacteriological investigation - isolation, identification, study of biological properties of the pathogen. The complexity and the complexity of the allocation of culture of H. pylori limit the application of this method in the practice of practical health care, but it remains only for phenotypic susceptibility strains to antibiotics.
For the cultivation of H.pylori proposed different media, mandatory components of which are the basic agar, growth supplements, and for selective inhibitors of growth associated microflora. The most basic agarov meet the requirements for the cultivation of H.pylori, for example Columbia agar, ritrit agar, cardio-cerebral agar. H.pylori is very whimsical microorganism, requiring additional growth factors (vitamins, microelements) [1]. A required component of the environment should be additive 5-10% of blood or serum of animals (horses, sheep). The use of human blood is limited by the presence of the majority of the adult population of protective antibodies, can inhibit the growth of H. pylori [2]. While the red blood cells may be lysed to growth substances could be used faster, which is achieved using chocolate agar. Developed a nutrient medium, containing serum and tissues of animals-SATFM (serum - animal tissue - free medium), which can be used to obtain pure cultures (dense)and for transportation (semi) with a difference in the percentage of agar [3]. Navratalova and Abierunt (1995) proposed the environment prepared on the basis of ritrit-agar, in which ex tempore make the blood, geminus and selective additives [4]. Did and colleagues (2000) developed Helicobacter-agar containing selective additive consisting of antibacterial and antifungal agents, and the blood made ex tempore [5]. Abiero with the group of authors (2002) recommend to use as the basis of the nutrient medium Columbia agar or cardio-cerebral agar with the addition of 5-7% of horse serum or 5-7% defibrinirovannaya horse blood and 1% solution isoetales [6]. Closest to the claimed is a way of cultivation H.pylori on chocolate agar, prepared on the basis of environment of Colombia agar with the addition of 2.5% of donor blood. Then cooled to 50 C nutrient medium authors suggest to add 2.5 percent emulsionnoj donor blood. The method provides more rapid cultivation on cheaper environment, but not without some disadvantages [7]. The disadvantage of the prototype in the opinion of the authors is the use of donor blood that may reduce the frequency of vyciveme because of the presence in the blood of antibodies to H. pylori, has inhibiting effect. Particularly acute problem in obtaining pure cultures, when after reseeding isolated colonies to obtain pure cultures grow a single colony, and biological material is not sufficient for identification and formulation of the antibiogram.
Lack of methods of cultivation on firm nutrient mediums is:
- low effectiveness of vyciveme of Helicobacter in primary isolation,
- further transfers for obtaining pure cultures of strains lost, germination is weakened and lost,
- poorness of growth of pure cultures leads to the impossibility of determining the sensitivity of selected strains to antibiotics and chemotherapeutic drugs.
The problem of the accumulation of culture can be solved through the use of liquid media. Similar to that of solid environments composition (the Foundation, growth and selective additives) is used in liquid environments, although the cultivation process more time consuming and in practical laboratories have not found wide application [8]. A variety of bases of liquid media: Brucella - broth, cardio-cerebral broth, soy broth with growth additives (serum, yeast extract, cyclodextrin and others), and antimicrobial therapy (vancomycin, nalidixic acid, amputee[9; 10]. The proposed environment that has a constant composition of vitamins and minerals without adding whey - serum - free Ham's F-12 [11]. With the growth in liquid nutrient mediums bacteria are typical of convoluted form, mobile and biochemically active, full antigenically [12]. H.pylori is growing in microaerophiles atmosphere, consisting of 5% Of 2 , 5-10% FROM 2 , 85-90% N 2 . The different systems can be used to create microaerophilic conditions. Among them - microaerobic Boxing or incubator, in which an adequate atmosphere is maintained automatically. Such an atmosphere can be created in microbiostatic type GasPac 100 or GasPac 150 Anaerobic Systems of the company BBL using a gas generator package type Saturas company BBL or Oxoid, which begin to produce the gas mixture with the addition of water, which creates necessary for the growth of H. pylori humidity. But under cultivation in liquid medium complexity can be the creation and maintenance of microaerophilic conditions. This problem can be solved constant monitoring microaerobic atmosphere in test tubes by mixing platforms angle of 20 degrees at the rotation speed of 120 rpm [13] or cultivation in the bottle with the holes for access gases, which developed a mini-bioreactors for forcing in the atmosphere cultivation of carbon dioxide through the tubes, immersed in vials of liquid nutrient medium [14].
Disadvantages of the methods of cultivation on liquid nutrient mediums:
- the complexity of creating and maintaining microaerophilic conditions in liquid nutrient mediums,
- the necessity to use additional equipment (shakers, rotational platforms, bioreactors and so on) to force the introduction of carbon dioxide.
The objective of the invention is development of a method of thin-layer cultivation of Helicobacter pylori with the use of two-phase medium, available for bacteriological laboratories are not equipped with special equipment. Technical result achieved is the absence of necessity to use additional equipment to maintain microaerophilic conditions in the liquid environment, using standard nutrient media (Colombia agar, and broth of Schedler) and that there is an accumulation of pure culture of H. pylori in sufficient quantity to morphological, biochemical identification and formulation of the antibiogram.
Some microorganisms hard to grow on a solid or liquid nutrient mediums, but they are much more actively grow and multiply in two-phase nutrient mediums consisting of a combination of solid and liquid phases. A possible explanation may be that such a nourishing environment create conditions for growth and reproduction, close to natural, when a part of the life cycle of a microbe requires solid culture media and other stages of the life cycle are in liquid nutrient medium. This is particularly characteristic of epithelial pathogens, which include H.pylori, partially breeding to the cells of the mucous membranes, and partly on the surface of mucous in the lumen of the body. The composition components liquid and solid phases of a nutrient medium is determined by the biology of a particular microorganism.
So, RF patent №2272834 (15) protected nutritional two-phase environment for the allocation of single-celled microorganisms trihomonad. The RF patent №2412991(16) protected two-phase nutrient medium for cultivation of cryptosporidia and patent RF №2346052 (17) offers a two-phase medium for isolation of Brucella.
The authors of the claimed invention offer the following composition of the nutrient medium. Two-phase medium consists of two phases: solid and liquid. The solid phase is a chocolate agar, prepared on the basis of Columbia agar with 5% of the blood of a RAM.
The composition of Columbia agar:
Ingredients
grams/lit P
Biopath 20,00Tripticase prewar beef heart
3,00Corn starch
1,00Sodium chloride
5,00Agar-agar
15,00End pH value (25 C) 7,3±0,2
Preparation:
Colombia agar diversity included in the formulation of the protection of sources of protein nutrition. The combination peptone it provides quick and abundant growth is very fastidious organisms. The result is the preparation of chocolate agar from red blood cells extracted geminus, ABOVE, and growth factors Haemophilus influenzae and fastidious bacteria including H. pylori, are faster.
The liquid phase is a broth of Schedler, enriched 10% serum of cattle.
The composition of the broth of Schedler:
Ingredients
grams/lit P
Casein hydrolysate
5,67Protectorate
5,00Papirovy prewar soy flour
1,00Yeast extract
5,00 Glucose of 5.83Sodium chloride
1,67Potassium hydrogen phosphate
0,83Tris (gidroksietilimino)
3,00L-Cystine
0,40 Gemin 0,01End pH value (25 C) 7,6±0,2
Preparation:
Stir 28,41 g powder in 1000 ml of distilled water. Boil with frequent stirring to dissolve the particles. Sterilized by autoclaving at 1,1 bar (121 C) for 15 minutes Cool and aseptico add up to 10% of the total volume of sterile serum of cattle. Before filling Wednesday thoroughly. To avoid overheating of the environment or its oxidation in the light, as it leads to stunted growth of bacteria.
Advantages used liquid phase:
This environment is a great base in which to highlight whimsical anaerobic, including coprofilia bacteria, you can add the blood or other processing additives. The combination of hydrolyzed casein, protectoriphone, papinogo of prevara soy flour, yeast extract and L-cystine provides presence of nitrogen nutrients, vitamins and other factors necessary for the growth of microorganisms. Glucose is the source of energy. Geminus and vitamin To stimulate the growth fastidious organisms.
The authors suggest the following method of thin-layer cultivation H.pylori on two-phase medium.
1 stage. Initial seeding of the studied material (biopsy of the mucous membrane of the stomach) produced by the Colombian agar with 5% of the blood of the lamb. Temperature control in microaerophilic conditions (2-incubator) at 37 OC for 3-5 days with high humidity.
stage 2. Evaluation of the results of sowing. Suspicious colonies (small, transparent 1-2 mm in diameter, similar to the "dew drops"). Reseeding selected colonies on chocolate agar for obtaining pure cultures. The incubation in microaerophilic conditions at 37 C in 3-5 days.
stage 3. The accumulation of pure culture. In case sparse growth of single colony dissipate the loop on the surface of dense medium and pour a liquid nutrient medium prepared on the basis of broth of Schedler, enriched 10% serum of cattle. The optimal amount of liquid poured into a Petri dish with a diameter of 90 mm 3,5 ml (the ratio of the solid and liquid phases 4:1). The incubation in microaerophilic conditions 37 C for 3 days.
stage 4. Identification cultures grown in the liquid phase.
The study of morphological properties. From cultural liquid prepare fixed smears stained by gram stain and the drug is "squashed" or "hanging drop" to determine mobility.
The study of biochemical properties. Staging tests catalase, oxidase, urease.
Statement of the antibiogram.
Thus, the claimed by the authors of the invention method of thin-layer cultivation of Helicobacter pylori involves the use of the advantages of the liquid phase compared to the solid phase for the accumulation of culture, and its thin-layer distribution supports microaerophilic conditions in the bulk liquid without additional equipment.
Literature
1. X. Jiang, M.P. Doyle Growth supplements for Helicobacter pylori. // J Clin Environ. - 2000. - Vol.38. - P.1984-1987.
2. Westblom T.U., Madan E., Riff B.R. Improved growth of Helicobacter pylori using a liquid medium supplemented with human serum. // Ital. J. Gastroenterol. - 1991. - Vol.29(Suppl. 2). - P.48.
3. Dierikx C.M., Martodihardjo J. Kuipers E.J. et al. Serum and animal tissue free medium for transport and growth of Helicobacter pylori. // Immunol. Med. Environ. - 2007. - Vol.50. - P.239-243.
4. Safonova N.V., gebrun A.B. Gastritis, peptic ulcer and Helicobacter pylori infection. - REC. for doctors. - SPb. - 1995.
5. Medzhidov MM, temirkhanova SO, Aliyev oil on canvas, etc. Assessment of nutrient medium for isolation and cultivation of Helicobacter pylori. , Zh. microbiol., Epidemiol., immunol. - 2000. - №2. - S-29.
6. Gebrun A.B., Alexandrov V., Goncharova LB other Diagnostics, prevention and treatment of diseases associated with Helicobacter pylori infection. (Manual for doctors). SPb., 2002.
7. Chervinets V.M., Chervinets L. F. invention Patent of the Russian Federation №2145975 "the Method of cultivation of Helicobacter pylori". - 2000.
8. Shahamat M., Mai U.E., Paszko-Kolva S. et al. Evaluation of liquid media for growth of Helicobacter pylori. // J. Clin. Environ. - 1991. - Vol.29. - P.2835-2837.
9. Morshed M.G., Karita M., Konishi H. et al. Growth medium containing cyclodextrin and low concentration of horse serum for cultivation of Helicobacter pylori. // Environ. Immunol. - 1994. - Vol.38(11). - P.897-900.
10. Murano A., M. Miyake, J. Kato et al. Enhancement of the growth of Helicobacter pylori in Brucella broth by hydrogen peroxide. // Environ. Immunol. - 1999. - 43(11). - P.1009-15.
11. Testerman T.L., McGee D.J., Mobley H.L.T. Helicobacter pylori growth and urease detection in the chemically defined medium Ham's F-12 nutrient mixture. // J. Clin. Environ. - 2001. - Vol.39. - P.3842-3850.
12. Y.L. Lin, N. Lee, Chan E.C. Determination of optimal liquid medium for enzyme expression by Helicobacter pylori. // J. Clin. Pathol. - 1996. - Vol.49. - P.818-820.
13. Xia H.X, L. English, Keane C.T. et al. Enhanced cultivation of Helicobacter pylori in liquid media. // J. Clin. Pathol. - 1993. - Vol.46(8). - P.750-753.
1. Two-phase medium for thin-layer cultivation of Helicobacter pylori containing solid phase, presents chocolate agar, made on the basis of Columbia agar with 5% of the blood of the lamb, and the liquid phase, representing a broth of Schedler, enriched 10% serum of cattle, characterized in that the ratio of the solid phase to the liquid is 4:1 (14 ml agar and 3.5 ml of broth).
2. Two-phase medium for thin-layer cultivation of Helicobacter pylori according to claim 1, characterized in that Colombia agar contains the following ingredients in g/l:
Ingredients
grams/liter
Biopath 20,00Tripticase prewar beef heart
3,00Corn starch
1,00Sodium chloride
5,00Agar-agar
15,00End pH value (25 C) 7,3±0,2
3. Two-phase medium for thin-layer cultivation of Helicobacter pylori according to claim 2, characterized in that the broth of Schedler contains the following ingredients in g/l:
Ingredients
grams/liter
Casein hydrolysate
5,67Protectorate
5,00Papirovy prewar soy flour
1,00Yeast extract
5,00 Glucose of 5.83Sodium chloride
1,67Potassium hydrogen phosphate
0,83Tris (gidroksietilimino)
3,00L-Cystine
0,40 Gemin 0,01End pH value (25 C) 7,6±0,2
4. Method of thin-layer cultivation of Helicobacter pylori with the use of two-phase of the nutrient medium, according to claim 1, providing initial seeding of the studied material on Colombia agar with 5% of the blood of the lamb, temperature control in microaerophilic conditions in 2 incubator at 37 OC for 3-5 days, with high humidity, reseeding selected colonies Petri dishes with chocolate agar, subsequent temperature control in microaerophilic conditions in 2 incubator at 37 OC for 3-5 days, with high humidity, wherein after a temperature Petri dishes with chocolate agar poured over liquid nutrient medium on the basis of broth of Schedler, enriched 10% serum large cattle according to claim 1, and finally termostatarea them in 2 incubator at 37 C during 72 hours.