Method to prepare live preparations of microscopic fungi of coccidioides genus for light microscopy
SUBSTANCE: method provides for growing of a culture of microscopic fungi on a dense nutrient medium. A standard suspension containing 104CFU/ml is prepared from the grown culture of microscopic fungi. The suspension in the amount of 0.5 ml is seeded into 4.5 ml of molten agar nutrient medium cooled down to 45°C. Mixed, poured into a Petri cup to produce a seeded plate of the agar nutrient medium. The seeded agar plate is cut into blocks with the specified size, which are placed onto the microscope slide and covered with the cover slop with formation by the agar chamber - production of the preparation. The produced preparation is incubated in the moist chamber at 28°C for 3 weeks.
EFFECT: invention makes it possible to simplify methodology of preparations making and to expand area of application of the produced preparations.
3 dwg, 2 ex
The invention relates to medicine and biotechnology, in particular to methods of preparation of living preparations of microscopic fungi of the genus Coccidioides for light microscopy, can be used to identify, establish the specifics of the structure and development of cells in different physiological States.
In the medical mycological practice in identifying the causative agents of mycotic nature is most frequently used methods aimed at studying the features of the morphology of the structures that vegetative propagation of micromycetes. This concept of diagnosis was named morphological or phenotypic, when to determine the unit type is the determination of the morphological characteristics of the studied strain and comparative analysis by establishing differences with closely related species. This methodological approach is implemented in the cultivation of microscopic fungi in conditions conducive to a culture with a distinctive morphology. The literature describes methods of preparation of live unstained microorganisms for study by light microscopy.
For determination of cell shape and motility of microorganisms examined in the drugs "crushed" or "hanging" drop. Drugs mushrooms for research, as the rule is, prepared from cultures grown on solid agar nutrient media. To do this on a glass slide put a drop of water or isotonic saline (0.15 M) solution of sodium chloride. Bacteriological loop is suspended a small amount of material taken from the colony suspected of cultural characteristics on the colony of the pathogen. Cover glass mounted on a rib at the edge of the drop and slow down, squeezing the air between the objective and the cover glass. Also use the method of preparation of microbial suspensions in vitro, after which a drop of the suspension is applied on the glass. In the case of the preparation of the drug "hanging" drop suspension is applied on the glass with a recess (hole) ["Manual of medical Microbiology" in 3 books edited by A.S. Labinsky, E.V. Valinol. - Moscow: BINOM-2008 - CN p.160-161]. A common shortcoming of such drugs is that they do not allow to evaluate the dynamics of culture. In addition, for their preparation is needed in all cases the preparation of the suspension of the studied culture, accompanied by mechanical impact on the culture, which inevitably leads to the destruction of characteristic structures of the fungus, which is an integral part of its morphological identification.
There is a method of study of living unstained microorganisms in which microcamera with agar nutrient media (agar chambers) ["Medical Microbiology" Ed. Weaponammo, Ochrosia. - Moscow: GEOTAR Medicine. - 1998. - p.114]. The described method of preparation of agar chambers of agar or gelatin-agar plates. A clean glass slides twice (or more times) is dipped in molten agar nutrient medium of the corresponding composition, which is located in the Petri dish. From the bottom of the glass agar nutrient medium wipe with a damp cloth. Using a binocular magnifying glass, the culture of make selected on the upper side of the glass pane with an extruded from a Pasteur pipette, glass fiber or glass rods. The tip of the fiber was soaked in an aqueous solution of peptone, lightly touch them to the colony, transferred adhering microorganisms in a drop of an aqueous solution of peptone on the fresh surface of the agar nutrient medium and distribute a drop on the surface. Cut agar environment around your area, put a cover glass and the edges pour the wax. [Methods of General bacteriology" in 3 volumes edited by F. Gerhardt and other Moscow: Mir. - 1983. 1. - p.46-47]. The disadvantages of this method include high possibility of contamination of the preparation by other organisms, and contamination of personnel when working with pathogenic agents, as in the formation of agarwood camera, namely when closing it on the smooth glass, perhaps squeezing liquid culture outside of the glass cover. In addition, the described method allows to prepare the camera, containing a thin layer of agar nutrient medium, which limits the use of the method when working with micromycetes different slow growth rate.
As a prototype of the selected preparation method of living preparations of fungi NM according to the method of pidoplichko. For this sterile glass slide calcined over the flame of the burner, quickly put the hot loop a small drop of molten agar nutrient medium and allow it to cool slightly. Immediately the burner needle put pre-grown on solid nutrient medium culture, trying to get to the center of the drop. Cover glass, as well as subject, calcined above the burner, a little away from the flames so that it is slightly cooled, and placed on a drop of nutrient agar medium seeded with a culture on a glass slide. For the formation of agarwood camera cover glass gently press down until until agar nutrient medium will not be distributed evenly tapering to a thin layer, and placed under an angle of 10-15° to the subject glass. Prevent drying and contamination of the drug coating glass, except for a raised hand poured paraffin. GE is prepared, the preparation is put into a Petri dish so that to the surface of glass slides on which the drug was directed upwards, and incubated in a humid chamber. ["Methods of experimental Mycology" Ed. by V.I. Bilai. - Kiev: Naukova Dumka. - 1982. - s]. A significant disadvantage of the prototype is seeding culture in negoziroma number that does not allow the growth of the fungus in the form of isolated colonies. In addition, the supply of nutrients in a thin layer of agar camera is limited, which makes it impossible for long-term cultivation of slow-growing fungi, for example, for 3 weeks in the case study mycelial forms of fungi of the genus Coccidioides, and some other pathogenic micromycetes. The disadvantages of the prototype is also the fact that the method involves a subjective assessment of the temperature of the agar nutrient medium in which you are sowing culture. In that case, if the environment is cooled insufficiently studied culture may die, or, on the contrary, when excessive cooling medium prematurely freezes on a slide that prevents preparation.
The task of the invention is to develop a method of preparation of living preparations of microscopic fungi of the genus Coccidioides for examination by light microscopy, which allows to simplify the preparation procedures and to expand on the region of their use.
The technical result is to simplify the method, obtaining predictable results, as well as the opportunity to study in the dynamics of the micro-morphology of fungal cultures.
The technical result is achieved in the method of preparation of living preparations of microscopic fungi of the genus Coccidioides for light microscopy, which consists in pre-grown culture on solid nutrient medium, the seed culture of suspended arthroconidia 104CFU/ml, in the amount of 0.5 ml 4.5 ml of melted and cooled to 45°C agar medium, which is stirred and poured into a Petri dish for education seeded agar plates medium, from which cut out the block for the formation of agarwood camera, and incubated received the product in a humid chamber at 28°C for 3 weeks.
Technical solution allows to predict the result at the expense of seeding culture with a known concentration of CFU (CFU - colony forming unit) and get its growth in the form of isolated colonies. Excludes subjective assessment of the temperature of the agar nutrient medium, in which is the seed culture as the set temperature environment is provided by the device, this simplifies the method and guarantees the suitable drugs. Agar Luggage in the drug has izvestneishim, contains a sufficient amount of nutrients, which eliminates the step of filling the chamber with paraffin wax to prevent drying. In addition, it becomes possible to grow a culture in preparation for a long time and to investigate the dynamics of the development of slow-growing fungi from the stage of germination of spores to the stage of formation of the organs of vegetative propagation. Depending on the purpose of the study is available preparation of a large number of drugs on the same sample.
Example 1 (optimal). Culture microscopic fungus, Coccidioides immitis C-5 grown on agar nutrient medium Saburo at 28°C for 3 weeks. Then the culture is washed off of 0.15 M sodium chloride, pH 6.8 and filtered through a fabric filter. From the filtrate using a sample turbidity gisk named after. Tarasevich (CCA) prepare standard suspension containing 104CFU/ml (CFU - colony forming unit) for Coccidioides immitis C-5. The suspension in the amount of 0.5 ml inoculated into a test tube with 4.5 ml of molten and cooled in a water bath to 45°C agar nutrient medium Saburo containing 2% agar-agar, mixed and poured into a sterile Petri dish. Square Petri dishes 10 cm in diameter makes 78.5 cm2. The thickness of the agar plates when applying 5 ml (cm3medium in a Petri dish - 0,064 mm After hardening seeded plate agaropectin environment is cut into blocks of size 1×1 cm, are in compliance with the rules of asepsis is placed on a glass slide and covered with a cover glass for the formation of agarwood camera. The number of CFU in agar block with an area of 1 cm2is 64 units. Finished products are placed in Petri dishes and incubated in a thermostat at a temperature of 28°C for 3 weeks.
Research drugs is conducted daily using light microscopy with a dry lens when magnification ×100, ×400, which allows to obtain data on the dynamics of cultural development. The size of the cells is determined using an ocular micrometer screw MOU-1-15xin a linear increase in βcf.=46,375.
After 24 h cultivation, arthrospira acquire a rounded shape. There are distinct tortuous growth tubes. After 2 days in growth tubes are formed septa (2-3), are viewed by the kernel. Occasionally observed the formation of secondary branches originating from the primary growth tubes. After 3 day begins education cept in the secondary branches. For 4-6 days formed the branches of the mycelium of the following orders of magnitude (figure 1). On the 7th day of cultivation begins the differentiation of cells of the mycelium. Clearly highlighted cells migrating in arthrospira. In their cytoplasm visible seal gray-blue color, reminiscent of the cell nucleus. Cells migrating in arthrospira, displaced who are stated by the cell-separators, having slightly smaller. To 14th days begin to form arthrospores typical organs of vegetative propagation of Coccidioides immitis (figure 2). Arthroconidia have a rectangular shape. For strains of Coccidioides immitis C-5 sizes arthroconidia are 9,0±0,34×3,6±0,64 mm. After 3 weeks of cultivation in the drug are observed filaments of mycelium in the form of "chains", educated Mature arthroconidia interspersed blank-delimited (figure 3).
Example 2 (optimal). Culture microscopic fungus, Coccidioides posadasii 442 grown on agar nutrient medium Saburo at 28°C for 3 weeks. Then the culture is washed off of 0.15 M sodium chloride, pH 6.8 and filtered through a fabric filter. From the filtrate using a sample turbidity gisk named after. Tarasevich prepare a suspension of arthroconidia 104CFU/ml Suspension in the amount of 0.5 ml inoculated into a test tube with 4.5 ml of molten and cooled in a water bath to 45°C agar nutrient medium Saburo containing 2% agar-agar, which is stirred and poured into a sterile Petri dish. Square Petri dishes 10 cm in diameter makes 78.5 cm2. The thickness of the agar plates when applying 5 ml (cm3medium in a Petri dish - 0,064 mm After hardening seeded plate agar nutrient medium is cut into blocks of size 1×1 cm are in compliance with the rules of asepsis placed on PR is Mednoe glass and covered with a cover glass for the formation of agarwood camera. The number of CFU in agar block with an area of 1 cm2is 64 units. Finished products are placed in Petri dishes and incubated in a thermostat at a temperature of 28°C for 3 weeks.
Research drugs is conducted daily using light microscopy with a dry lens when magnification ×100, ×400, which allows to obtain data on the dynamics of cultural development. The size of the cells is determined using an ocular micrometer screw MOU-1-15xin a linear increase in βcf.=46,375.
After 24 h cultivation, arthrospira acquire a rounded shape. There are distinct tortuous growth tubes. After 2 days in growth tubes are formed septa (2-3), are viewed by the kernel. Occasionally observed the formation of secondary branches originating from the primary growth tubes. After 3 day begins education cept in the secondary branches. For 4-6 days formed the branches of the mycelium of the following orders of magnitude (figure 4). On the 7th day of cultivation begins the differentiation of cells of the mycelium. Clearly highlighted cells migrating in arthrospira. In their cytoplasm visible seal gray-blue color, reminiscent of the cell nucleus. Cells migrating in arthrospira, interspersed cells separators having slightly smaller. To 14th days begin to form arthrospores typical bodies in getting breeding Coccidioides posadasii (figure 5). Arthroconidia have a rectangular shape. For strains of Coccidioides posadasii 442 sizes arthroconidia are 12,0±0,71×3,6±0.29 micrometers. After 3 weeks of cultivation in the drug are observed filaments of mycelium in the form of "chains", educated Mature arthroconidia interspersed blank-delimited (6).
The method of preparation of living preparations of microscopic fungi of the genus Coccidioides for light microscopy, which consists in pre-grown culture on solid nutrient medium, the seed culture of suspended arthroconidia 104CFU/ml, in the amount of 0.5 ml 4.5 ml of melted and cooled to 45°C agar medium, which is stirred and poured into a Petri dish for education seeded agar plates medium, from which cut out the block for the formation of agarwood camera, and incubated received the product in a humid chamber at 28°C for 3 weeks.
SUBSTANCE: connective or supporting tissue cells fit for transplantation are cultivated in bioreactor (1) which includes main case closed by sterile sealing cap (21) and forms at least one reactor space where transplant surface (11) and mini actuator (14) are placed. Bioreactor (1) is equipped with at least two coupling connections for hoses (19) of nutrient medium and gas feed and discharge.
EFFECT: obtaining cells with mechanical endurance to self-tissue transplantation for application as substitute tissue in treatment of connective and supporting tissues, direct injury of joints, rheumatism and degenerative diseases of joints.
22 cl, 27 dwg, 11 ex
SUBSTANCE: claimed medium contains 18-20 of bactoagar "Difko", pH 7.2-7.4 per 1 l of casein hydrolyzate. Casein hydrolyzate contains amine nitrogen in amount of 170-180 mg %. Medium effectiveness is suggested by presence of Paenibacillys larvae var. larvae in testing material of <10 CFU up to complete surface weediness. Medium of present invention is useful in diagnosis of bee American foulbrood disease.
EFFECT: express method for isolation of Paenibacillys larvae spores in honey and other bee products.
1 tbl, 3 ex
SUBSTANCE: claimed method includes providing of coccidial oocysts from fecal suspension, homogenization of fecal suspension, oocyst separation from fecal remainders of organic substances by salt flotation with sodium sulfate and other floating agents, oocyst sporulation using hydrogen peroxide and air barbotage, bleaching of sporulated oocysts, washing of bleached oocysts, concentration of sterile washed oocysts and blending of coccidial oocyst concentrates of various species to produce vaccines.
EFFECT: vaccines of high quality without toxic admixtures.
FIELD: food industry.
SUBSTANCE: additive is produced from wheat offal fermented with mould fungi. The additive agents are at least one ferment and a mixture of nutritional ingredients for yeast: ergosterol, N-acetylglucosamine, vitamins, nucleic acids and amino acids. The said additive is also used for activation of alcohol fermentation or pre-fermentation intended for preparation of yeast under aerobic conditions.
EFFECT: reduction of the simultaneous saccharification-fermentation stage time in the process of ethanol production, increase of yeast growth and efficiency.
21 cl, 2 dwg, 4 tbl, 4 ex
SUBSTANCE: composition contains an agent designed for biocontrol, for reduction of level of pollution of food and fodder with aflatoxin, a binding agent, an agent having osmoprotector and adhesive properties, a carrier and a source of nutrient elements for an agent, designed for biocontrol.
EFFECT: reduced level of food and fodder contamination with aflatoxin in raw materials.
29 cl, 4 tbl, 6 ex
FIELD: food industry.
SUBSTANCE: invention relates to a food product with extended storage life and to usage of probiotic spores and/or resting cells as a probiotic ingredient in the said product. The product is chosen from the group containing milk products, fruit juices and fruit beverages. The probiotic spores are chosen from the group containing bacterial, yeast and fungous spores. The resting cells are chosen from bacterial, fungous and/or yeast and/or their mixtures. The produced is stored under uncooled conditions during a long period of time by way of probiotic bacteria spores inclusion.
EFFECT: probiotic organisms contained in fruit juices and beverages remain viable during a longer period of time in comparison with live probiotics.
23 cl, 9 ex
SUBSTANCE: strain of fungus Trichoderma harzianum Rifai VKPM F-180 is used as a producent of an inhibitor of a stimulant of bacterial burn of fruit cultures.
EFFECT: invention makes it possible to reduce losses of decorative and fruit cultures caused by bacteria Erwinia amylovora.
SUBSTANCE: fermentation medium for production of recombinant proteins using methanol inducible fungi of species Pichia pastoris is characterised by maintained concentration of urea or its derivatives in the range from about 0.3 M to about 1 M. The medium contains per liter of water basic salts in the following amounts: orthophosphoric acid (85%) from 2.67 to 133.5 ml, calcium sulfate from 0.093 to 4.65 g, potassium sulfate from 1.82 to 91 g, magnesium sulfate-7H2O from 1.49 to 74.5 g, potassium hydroxide from 0.413 to 20.65 g, glycerol from 4 to 200 g, and microelements per liter of water in the following amounts: copper sulfate-5H2O from 0.6 to 30 g, sodium iodide from 0.008 to 0.4 g, manganese sulfate-H2O from 0.3 to 15 g, sodium molybdate-H2O from 0.02 to 1 g, boric acid from 0.002 to 0.1 g, cobalt chloride from 0.05 to 2.5 g, zinc chloride from 2 to 100 g, sulphate divalent iron-7H2O from 6.5 to 325 g, biotin from 0.02 to 1 g, sulfuric acid from 0.5 to 25 ml. The method for production of recombinant proteins comprises the stage of reproduction of methanol inducible fungi of species Pichia pastoris, and the stage of recombinant protein expression using the said fermentation medium in which feeding with methanol is carried out at a rate of about 6 g/l/hr to about 20 g/l/hr.
EFFECT: increased yield of target products.
11 cl, 27 dwg, 16 ex
SUBSTANCE: preparation with polyfunctional medical-biological activity that influences tissue exchange is produced as a result of liquid-phase depth cultivation of fungus Pleurotus ostreatus All-Russian collection of industrial microorganisms F-819 with further separation of mycelium and cultural liquid and extraction of biologically active substances in the form of a condensed extract from the mycelium. The produced extract, which represents a pharmacological substance of a preparation, is enriched with the agent of the derivative of sterol 4-hydroxy-17P-methylinecisterol with molecular weight of 332.2452 Dalton, polysaccharide β 1-3 glucan and dihydroquercetine with molecular weight of 304.26 Dalton. At the same time the specified components are taken at the following weight ratio: (1):(2):(3):(4) as (79.0-158.0):(1.0-2.0):(10.0-20.0):(10.0-20.0).
EFFECT: invention makes it possible to increase quality of a preparation as a result of increasing its antitumoral activity and reduced toxicity of preparations of antitumoral chemotherapy with simultaneous application with the suggested preparation.
2 cl, 21 dwg, 11 tbl, 2 ex
SUBSTANCE: entomopathogenic biopreparation for protecting plants from pests contains a dry mixture of a concentrate of a culture fluid based on infection units of entomopathogenic fungi of the type Verticillium lecanii or Beauveria bassiana, or Metarhizium anisopliae, aerosil and an additive which protects the infection units from loss of viability and biological efficiency when drying, with the following ratio of components, wt %: culture fluid concentrate 69.0-77.5, additive - 10.8-13.8, aerosil - 11.6-17.2. The additive is a polyatomic alcohol, carbohydrate, vegetable oil, sodium gluconate and water, with the following ratio of components, wt %: polyatomic alcohol 6.2-13.4, carbohydrate 38.0-41.9, vegetable oil 38.7-41.0, sodium gluconate 0.4-0.5, water 9.3-10.5. The biopreparation is obtained by mixing the culture fluid concentrate with the additive in ratio of 5-7:1, and then adding aerosil in ratio of 4.5-7.6:1 and drying the mixture. Efficiency of the biopreparation on white flies is 95-100% in a working concentration of 1·107 spores/ml; the titre of viable units is (0.35-5.5)·1010 CFU/g.
EFFECT: invention increases biological efficiency of the biopreparation, preserves and increases viability of infection units of fungi during drying and storage.
8 cl, 3 tbl, 15 ex
SUBSTANCE: method of qualitative assessment of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys in operation of spacecrafts and the suspension of spore materials fungi of implementation of the said method is proposed. The test and control samples of aluminium and magnesium alloys are prepared. The prepared samples are dried, sterilised. Fungal cultures - strains of microorganisms Paecilomyces variotii Bainier of All-Russian collection of microorganisms F-4039D, Ulocladium botrytis Preuss of All-Russian collection of microorganisms F-4032D, Penicillium chrysogenum Thorn of All-Russian collection of microorganisms F-4034D, Aspergillus sydowii (Bainier et Sartory) Thorn et Church of All-Russian collection of microorganisms F-4037D, Cladosporium sphaerospermum Penz. of All-Russian collection of microorganisms F-4041D are inoculated for growing spores into tubes with a sloped agar medium of Czapek-Dox. The tubes are thermostated at a temperature of (29+2)°C for 14-28 days until appearance of mature spores. Spore materials suspensions of individual cultures of the said fungi are prepared, which are then mixed in equal proportions. And the concentration of spores of each fungi species in the suspension should be in the range of 1-2 mln/cm3. The resulting suspension is applied to the sterilised test samples. Then they are dried and placed as one sample in each Petri dish on the surface of agar medium of Czapek-Dox. The control samples not treated by the spore material are also placed on the surface of the agar medium of Czapek-Dox in the Petri dishes. The Petri dishes with the test and control samples are placed in different desiccators at the bottom of which water is poured to maintain the humidity more than 90%. The desiccators are closed and incubated in the thermostat at a temperature of (29±2)°C. The exposition of test and control samples is carried out for 40, 120 and 180 days. Then the samples are removed from the Petri dishes, and the corrosion products and mycelium are removed from the surface of the samples washing them in running water. The samples are soaked in 70% ethyl alcohol for 30 minutes, then they are washed with detergent and dried. Then, using a scanning electron microscope a qualitative assessment of biocorrosion damages is carried out. According to the change in the appearance of the surface of the samples the initial stages of biocorrosion and its type is evaluated, as well as the distribution of corrosion damage on the surface of the samples, and the dependence of the biocorrosion process on time is determined.
EFFECT: inventions enable to carry out the qualitative assessment of the initial stages of biocorrosion damages of thin-walled sealed enclosures of aluminium-magnesium alloys with the thickness of more than 2 mm.
2 cl, 9 tbl
SUBSTANCE: invention relates to microbological industry, particularly production of biologically active substances and can be used in producing fusicoccins for different purposes, for both agricultural and medical purposes. The culturing medium contains molasses, raw sugar, potassium nitrate (KNO3), potassium phosphate (KH2PO4), magnesium sulphate (MgSO4 x 7H2O) and tap water in a given ratio.
EFFECT: invention increases the range of fusicoccins.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to a compound and a pharmaceutically acceptable salt thereof to be used as an antifungal agent, particularly, a therapeutic agent for deep fungal disease. The fungus Acremonium persicinum is collected, and a cyclic compound is recovered from its cultural fluid.
EFFECT: what is presented is the compound applicable as an antifungal agent.
10 cl, 16 tbl, 5 ex
SUBSTANCE: 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).
EFFECT: invention enables to improve the accuracy of determination of IPC.
SUBSTANCE: method includes the following: (a) production of a sequence of nucleic acid from one or more microorganisms extracted from a medium of a hydrocarbon-containing bed; (b) determination of presence of one or more gene products of the specified sequence of nucleic acid, where the specified gene product represents a ferment on the way of metabolism involved into transformation of hydrocarbon into methane selected from a group consisting of peroxidases, phenoloxidases, alcoholoxidases, laccases, hydrolases, glycosyl hydrolases, esterases, etherases, oxidases, nitrogenases, cellulases, amylases, glucanases, pullulanases, reductases, dismutases, oxygenases, monooxygenases, dioxygenases, catalases, hydrogenases, carboxylases and methyl reductases; or a ferment involved into homoacetogenesis, methanogenesis, acetoclastic methanogenesis or CO2-reducing methanogenesis; and (c) identification of substrate, reactant or cofactor of the specified ferment, which increases producing of methane, being available to one or more microorganisms in the specified hydrocarbon-containing bed, and (d) identification of a stimulant in this manner, which increases biogenic producing of methane in the hydrogen-containing bed in comparison with producing of methane in the hydrocarbon-containing bed in absence of the stimulant.
EFFECT: invention makes it possible to efficiently stimulate biogenic producing of methane in hydrocarbon-containing beds.
15 cl, 16 dwg, 2 tbl, 3 ex
FIELD: food industry.
SUBSTANCE: method involves at least the following stages: beet roots washing, heating, partial expression for production of, on the one side, a squeezed presscake containing more than 20% of sugar in terms of dry substances and, on the other side, of beet root juice with dry substances concentration more than 15%, separation of the squeezed presscake and beet root juice, beet root juice filtration by way of centrifugation for production of, on the one side, a filter-press cake and, on the other side, of filtered beet root juice, filter-press cake mixing with squeezed presscake and/or mixing a small quantity of flocculant (nearly 1%) with beet root juice or filtered beet root juice, the said mixture membrane filtration for production of a retentate and a filtrate where the said filtrate is represented by filtered beet root juice with purity no less than 93%, retentate mixing with squeezed presscake. The produced mixture of squeezed presscake with filter-press cake and/or retentate is subjected to ensilage without additional/mechanical compaction in a silo storage for storing; additionally, the said mixture drying is envisaged. Due to the beet roots processing method one may produce animals fodder products based on a mixture of squeezed presscake with filter-press cake and/or retentate, based on filtered beet root juice syrup with dry substances content equal to more than 60%. By the said method one may produce a nutritional fermentative medium based on beet root juice, filtered beet root juice with purity equal to nearly 90% and higher, filtered beet root juice syrup with dry substances content equal to more than 60% or a mixture of squeezed presscake with filter-press cake and/or retentate produced by the above method. Additionally the method allows to produce edible sugar produced by crystallisation of filtered beet root juice syrup with purity equal to 93%.
EFFECT: invention ensures beet root processing simplification and acceleration as well as the possibility of production of a nutritional fermentative medium and edible sugar from processing products, allows to simplify the processing products storing by way of their ensilage without forced compaction with simultaneous stabilisation of the silo pH within the range of 3,4-3,6, reduce the degree of the silage infection with oleaginous acid spore-forming bacteria, facilitate the processing product usage and dispensing to animals in a summer feeding period at farms.
17 cl, 7 dwg
SUBSTANCE: method includes inspection of an inoculator with process equipment for tightness, sterilisation of the inoculator with steam via an aeration device at the pressure of 0.20 - 0.25 MPa for 30…40 min., its filling with nutrient medium heated by steam to the temperature of 100°C. Then the temperature of the nutrient medium is increased to 121 - 123°C at steam pressure of 0.10 - 0.15 MPa, and the nutrient medium is maintained at these parameters for 15-60 min., afterwards the pressure in the inoculator is reduced down to 0.03…0.05 MPa. The nutrient medium is cooled down to cultivation temperature of 31…32°C with cold water with temperature of 7 - 10°C. After cooling of the nutrient medium, it is seeded with a seeding material with simultaneous mixing and aeration with sterile air. Cultivation of the produced liquid seeding culture is carried out at pH 4.2 - 4.5 and temperature of 31 - 32°C to achieve the phase of exponential growth for 12 - 14 hours. Then it is sent by means of displacement with sterile air from the inoculator into the prepared fermenter in the amount of 3…10% of the nutrient medium amount with its filling by 7/10 of its volume, and the microorganism culture is grown at fermentation temperature of 28 - 40°C for 96 - 120 hours with continuous aeration with sterile air, mechanical mixing and supply of warm water with temperature of 27 - 47°C into a heating jacket of the fermenter. After fermentation the cultural fluid with accumulated biomass is supplied into previously sterilised collectors of finished culture.
EFFECT: increased yield of cultural liquid with accumulated biomass of aerobic microorganisms, reduced specific power inputs and provision of environmental safety at all stages of production.
1 dwg, 2 ex