Method of production of bacteriologically pure cultures of marine blue-green microalgae
SUBSTANCE: method of production of bacteriologically pure cultures of marine blue-green algae provides chemical sterilisation of microalgae cultures by treatment them in the solution of sterile sea water containing 0.1% phenol and 1.0% ethyl alcohol. Microalgae are kept in sterile sea water containing phenol and ethyl alcohol in a predetermined ratio for 4-5 hrs. Irradiation with UV light is carried out for 9-10 minutes. The treatment of microalgae with antibiotics and fungicide is carried out. At that the antibiotics are used as chloromycetin and ampicillin in the amount of 50 µg/ml each, and fungicide is used as nystatin in the amount of 25 µg/ml.
EFFECT: invention enables to improve the efficiency of extraction of bacteriologically pure culture of blue-green microalgae.
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The invention relates to Algologie and Microbiology, and in particular to methods of allocation bacteriologically pure cultures of marine blue-green microalgae.
Currently, much attention is paid to the development of test systems for the evaluation of toxicity of the components of the marine ecosystems of water bodies, which requires the isolation of pure cultures of aquatic organisms, in particular microalgae. The standard procedure consists of isolating and growing algological pure cultures of algae, which then selects the most promising for biological testing. However, cultures of microalgae always be contaminated by bacterial microflora, spores and mycelium of fungi, which prevents further research, distorts the results. Purification from bacteria is a very time consuming process, because between algae and bacteria are dense biocenotic relationships. The mucous membranes of the algae serve as food source and shelter for microorganisms. Such a close relationship determines the difficulty of separating these communities for the isolated culture: algae and bacteria. Getting bacteriologically pure cultures of microalgae is a prerequisite for successful research, development, test systems for the evaluation of toxicity of the components of the ecosystem the marine waters and other scientific works.
There is a method of allocating axenically cultures of microalgae method agar plates (Wasser S. P., Kondratieff N. In., Masuk N. P. and other Algae. The Handbook. - Kiev: Naukova Dumka, 1989. - 608 C.) (1), in which microstrokes the suspension is introduced into the Petri dish and the microbial loop are distributed uniformly on the surface mycopathologia agar (MPA) followed by incubation in a thermostat. Then separate micro-colonies grown on the surface mycopathologia agar, remove sterile instruments together with pieces of dense nutrient medium and bring in a test tube with the appropriate sterile elective nutrient media for the accumulation of algae biomass. Exericise culture of algae get through repeated re-sieving on the surface of MPA in sterile conditions.
The known method has a limited range of application. For example, when selecting axenically cultures of the marine blue-green algae is not able to achieve their full freedom from bacteria, since the latter are strongly associated with strongly developed mucous membranes of these microalgae. The process of separating them tedious, time-consuming and sometimes inefficient, requires multiple re-seed.
In another method of purification of cultures of bacteria and other microorganisms carry out the irradiation of cultures of microalgae ultra olejowym light (K. Kvitko Century Obtaining cultures of individual cells of Chlorella // Research on genetics. - L.: Publishing house of Leningrad University, 1961. - Vol.1. - S. 50-54) (2). However, this method is not effective enough, because you cannot achieve full freedom of algae on bacteria and fungi. While the duration of UV exposure should not exceed 10 minutes, because the longer UV exposure to skin and eye irritant and can cause irreversible genetic changes irradiated culture.
Closest to the proposed method is chosen as the prototype of the method of purification of cultures of bacteria and other microorganisms (especially fungi) using chemical sterilization by treatment with 0.1% phenol and 1.0% ethyl alcohol. (Ecological physiology of marine planktonic algae. Kyiv: Publishing house "Naukova Dumka", 1971, S. 5-22) (3).
However, the chemical sterilization of microalgae also allows you to get a fully bacteriologically pure cultures of algae.
The inventive method solves the problem of increasing the efficiency of the allocation of bacteriologically pure cultures of marine blue-green microalgae.
This object is achieved in that a method of obtaining a bacteriologically pure cultures of marine blue-green algae consists of chemical sterilization Kul is ur microalgae by treatment in a solution of sterile sea water, containing 0.1% phenol and 1.0% ethanol, and additional irradiation with ultraviolet light for 9-10 minutes, and then treatment with antibiotics and fungicides, and the use of antibiotics chloramphenicol and ampicillin in the amount of 50 µg/ml each, and as a fungicide use nystatin in the amount of 25 μg/ml
Additional processing of microalgae after chemical sterilization first irradiation with ultraviolet light, and then antibiotics and fungicide, allows to obtain bacteriologically pure cultures of algae.
While the use of the antibiotic chloramphenicol in the amount of 50 μg/ml and ampicillin at 50 μg/ml, and as a fungicide - nystatin in the amount of 25 μg/ml allows to obtain, on the one hand, the almost complete absence of bacteria in the culture of marine blue-green microalgae, on the other hand, to avoid a decrease in growth rate and inhibition division of microalgae.
As a result of the analysis of the prior art is not detected similar, characterized by signs, identical with all the essential features of the claimed invention, and the definition of the prototype of the identified analogues revealed a set of essential towards the technical result of the distinctive features.Therefore, the claimed invention meets the condition of "novelty."
With additional search other solutions related to the proposed method, these distinctive characteristics are not detected. Thus, the claimed invention meets the condition of "inventive step".
The method is as follows.
Cleaning algological pure cultures of marine blue-green microalgae from bacteria is carried out in 3 stages. stage 1 - this chemical sterilization. For this purpose apply antiseptics such as phenol and ethyl alcohol at a concentration of 0.1 and 1.0%, respectively. Algae incubated in sterile seawater with the above antiseptics for 4-5 hours On the 2nd stage, hold 9-10-minute exposure of the cultures of algae with UV light (bactericidal irradiator OBN - 150,2 lamp 30 W) in Petri dishes at a distance from the radiation source 50, see the 3rd stage spend processing algae antibiotics and fungicide in the following optimal combinations of chloramphenicol at a concentration of 50 μg/ml ampicillin at a concentration of 50 μg/ml and nystatin at a concentration of 25 μg/ml
The presence of bacterial microflora controlled by reseeding from liquid cultures of the marine blue-green microalgae on agar or liquid culture of Abijah. Controlled growth IU filnik aerobic and facultative anaerobic microorganisms, fungi and yeast.
In the determination of mesophilic aerobic and facultative anaerobic microorganisms in liquid culture of microalgae the study was performed according to GOST 10444.15-94.
In the determination of mesophilic aerobic and facultative anaerobic microorganisms on agar nutrient medium, and 1 CC of culture of blue-green algae were sown in two parallel Petri dishes. Crops were flooded by GOST 26670 one of the nutrient agar media and incubation at 30°C.
In the determination of mesophilic aerobic and facultative anaerobic microorganisms in a liquid nutrient medium and 1 CC of culture of blue-green algae were sown in two flasks or test tubes with one of the liquid nutrient media according to GOST 26670. The ratio between the number of planted crops of algae and nutrient medium from 1:5 to 1:7. Crops were incubated at a temperature of 30±1°C for 72±3 h under aerobic conditions.
After incubation on agar nutrient media in Petri dishes take into account the presence or absence of colonies of microorganisms. In liquid nutrient media noted the presence or absence of visible signs of growth (flatulence, the appearance of turbidity, sediment).
If the apparent bacterial growth after three days of incubation was not considered unlikely that the sample of algae, dobavlenny into the culture medium, contains bacteria. However, the culture continued to watch for 7-10 days, as it could be slow-growing species. Conclusions about the purity of the cultures was done in the absence of bacterial colonies or visible signs of their growth.
When determining fungi and yeast in liquid culture of microalgae the study was performed according to GOST 10.444.88.
1 CC of culture of algae were sown in two Petri dishes with medium Saburo with antibiotics. Crops thermostatically at a temperature of 24±10C for 5 days. After incubation noted the presence or absence of visible signs of growth. Colonies of yeast and fungi were separated visually (yeast form large shiny convex colonies with smooth surface and straight edge, fungi form a mycelium of different colors).
In the absence of mesophilic aerobic and facultative anaerobic microorganisms, fungi and yeasts considered liquid culture of microalgae bacteriologically sterile.
The method is illustrated by the following examples.
Example 1 (control). Water samples and scrapings of microscopic algae from coastal rocks for the subsequent allocation of these cultures of algae were collected in the period of active vegetation period (may-June) of the main species of microalgae in the Azov and Black seas were First grown cumulative mixed culture of algae, for which sampling was added to the environment of Bristol in the modification of Hollerbach, a mixture of trace elements and soil extract in certain proportions. The cumulative culture of microalgae were obtained by culture mixed culture in flasks with liquid nutrient media intended for laboratory cultivation of different algae (green, nitrogen-fixing and pattricia blue-green). Time of cultivation of algae were averaged 3-4 weeks.
Of cumulative culture, using microbiological methods of re-seeding and dilution, got algological pure culture. Cumulative sterile culture were inoculated in a Petri dish with agar nutrient medium. The Cup was placed on the light for algal growth. From colonies grown algae loop carried part of the culture again in liquid medium or in a test tube with a beveled agar medium. Cultivation of algae was carried out on nutrient media Tamiya, Aguinaga, Guseva and Walna. Environment are universal for all types of algae and allow you to get a shared culture of microalgae physiologically active. For the separation of microalgae used a glass spatula and sieving by rubbing the surface of the medium. This allows you to split the mixture MICR is Oracle on a single cell, that preserves their integrity and, therefore, greatly facilitates and accelerates the process to obtain pure cultures.
The works were selected algological pure cultures of the following blue-green microalgae: Oscillatoria laetevirens, Spirulina tenussima and Snowella rosea. Next conducted clearing each of the obtained cultures of algae and bacteria by chemical sterilization (1). With this purpose used antiseptics such as phenol and ethyl alcohol at a concentration of 0.1 and 1.0%, respectively. The algae were kept in sterile seawater with the above antiseptics for 4-5 h (table 1, experiment 1).
Check the purity of the obtained cultures of algae had a control crops their liquid cultures to that of the above-mentioned agar nutrient media that has seen the best growth of this culture. When this was recorded the growth of microorganisms. Thus, chemical sterilization is not possible to obtain absolutely pure cultures of algae. The data are shown in table 1, experiment 1.
|Growth of microorganisms in cultures of the marine blue-green microalgae in experiments with different methods of sterilization|
local and fungal growth
Example 2. Culture dedicated marine blue-green algae were subjected to a first chemical sterilization (procedure described above), and then spent ten minutes exposure to the cultures of algae with UV light (bactericidal irradiator OBN - 150) in Petri dishes at a distance from the radiation source 50 cm (experiment 2). Check purity obtained after chemical sterilization and UV exposure of the cultures of microalgae had a control cultures liquid cultures on the above agar nutrient medium. When this was recorded the growth of microorganisms. The data are shown in table 1, experiment 2.
Thus, chemical sterilization, and radiation cultures of microalgae UV light is not allowed to get bacteriologically pure cultures of algae.
Example 3. Culture dedicated marine blue-green algae were subjected to a first chemical sterilization, then there was a ten-minute exposure of the cultures of algae ultraviolet with the etoy, as described in examples 1-2, and then subjected to treatment with the antibiotic chloramphenicol at a concentration of 50 μg/ml in an appropriate nutrient liquid medium. The results are shown in table 1, experiment 3.
Examples 4-20. Experiments were performed similar to that described above, i.e., the culture of selected microalgae were subjected to a first chemical sterilization, then there was a ten-minute exposure of the cultures of algae to ultraviolet light, and then subjected to treatment by different concentrations of antibiotics and fungicide in an appropriate nutrient liquid medium. The results are shown in table 1, experiments 4-20.
Were tested in different concentrations and different types of antibiotics and fungicides (table 1). After each treatment have been checking the purity of the obtained cultures of algae control crops from liquid cultures on agar nutrient medium. Thus, were selected antibiotics and fungicides, as well as their concentration, after processing which, when the control crops on agar nutrient medium was not recorded growth of microflora. Used the antibiotic chloramphenicol (50 and 90 μg/ml), tetracycline (50 and 90 μg/ml), erythromycin (50 mg/ml), ampicillin (50 and 90 μg/ml), and the fungicide - nystatin in the amount of 25 μg/ml were also conducted experiments with a series of samples of the marine blue-green MICR is Oracle, treated with antibiotics and fungicide, but not subjected to a preliminary chemical sterilization and/or treatment with ultraviolet light (experiments 8-10). Only treatment with chloramphenicol at a concentration of 50 μg/ml in combination with ampicillin 50 μg/ml and nystatin 25 μg/ml, performed after chemical sterilization, and radiation cultures UV light, have resulted in the almost complete absence of bacterial growth in several subcultures.
Were tested and the higher concentrations used antibiotics, which allowed to achieve 100% absence of bacterial microflora, however, such concentrations reduced the growth rate and inhibited division of microalgae. The research results are summarized in table 1.
The table shows that the effect associated with the full release of the marine blue-green microalgae from concomitant microflora preserving life mikrovodoroslevykh cells, is achieved by performing chemical sterilization, irradiation of cultures UV light, and then the processing of microalgae antibiotics and fungicide at the following minimum ratio of components: chloramphenicol and 50 μg/ml, ampicillin 50 μg/ml, and the fungicide, nystatin in the amount of 25 μg/ml
Thus, a stepwise method of chemical sterilization al is logicheskie pure cultures of marine blue-green microalgae with ultraviolet irradiation and further addition of certain concentrations and compositions of antibiotics and fungicides, allowed to get bacteriologically pure cultures of microalgae that can be used when conducting physiological-biochemical, cytological and genetic studies of algae and other scientific research. In particular, bacteriologically pure cultures of marine blue-green microalgae can be used to develop test systems based on fluorescence of algae to assess the toxicity of the components of the marine ecosystems of water bodies.
Using the proposed method improves the efficiency of the allocation of bacteriologically pure cultures of microalgae. This method is reliable, because it allows one to get bacteriologically pure cultures of marine blue-green microalgae regardless of the source separation and the degree of bacterial contamination.
The method of obtaining bacteriologically pure cultures of marine blue-green microalgae, including chemical sterilization cultures of microalgae by treatment in a solution of sterile seawater containing 0.1% phenol and 1.0% ethanol, characterized in that it further carry out the irradiation with ultraviolet light for 9-10 minutes, and then treatment with antibiotics and fungicides, and the use of antibiotics chloramphenicol and ampicillin in the amount of 50 µg/ml each, and in which the quality of fungicide use nystatin in the amount of 25 μg/ml
SUBSTANCE: method involves smear seeding from the nasal mucosa on meat infusion agar, incubation at 37°C for 24 hours, measurement of a total amount of microorganisms by counting grown colonies, determination of total microbial contamination of the analysed material expressed in CFU/ml. If the total microbial contamination is more than 10*3 CFU/ml, the exposure of an industrial microbiological factor on the nurse is considered to be damaging, while the derived value of 10*3 CFU/ml and less shows the admissibility of the exposure. The invention can be used for diagnosing the exposure of the industrial microbiological factor of the nurses working in large multi-field children's hospitals, not involved in a non-infectious material or a material suspicious for the microbial contamination of the 3-4th pathogenicity group, and fungi.
EFFECT: higher diagnostic accuracy.
FIELD: veterinary medicine.
SUBSTANCE: strip of filter paper with the width of 20 mm is attached on the filter, mounted in the ventilation system of the test livestock building, for a period of 4 hours. 1% sterile glucose solution is prepared on physiological solution, which is used as a nutrient medium. The strip of filter paper is placed in 10 ml solution of glucose and thermostated at the temperature 37±1°C for 2 hours. Then the electric conductivity of the solution is measured. The conclusion about the necessity in sanitary-hygienic processing of livestock buildings is made when the electric conductivity value is 287.3 mcS/cm, which corresponds to maximum allowable concentration of microorganisms in the air of the working area or below.
EFFECT: invention enables to determine reliably the microbiological environment in the buildings and to make the prompt conclusion about the necessity of sanitary-hygienic processing of livestock buildings.
SUBSTANCE: nutrient medium comprises nutrient agar, dry, of Black Sea sprat, para-aminobenzoic acid, tris-(hydroxymethyl) aminomethane (tris buffer), salicin, neutral red, L-tryptophan, 5-brom-4-chloro-3-indolyl β-D-glucuronide of cyclohexylammonium salt, 2-nitrophenyl β-D-galactopyranoside and distilled water in a predetermined ratio of components.
EFFECT: invention enables to improve the differentiating properties of the nutrient medium and to reduce the time of isolation of causative agents of urological infections.
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SUBSTANCE: method involves assessing radioreaction of the vaginal mucosa by visual examination, as well as assessing the vaginal epithelial condition by lactic acid bacilli testing and measuring by bacterioscopical and bacteriological analyses, performing the cytological examination of the surface and intermediate squamose epitheliocytes of the vaginal discharge followed by measuring and finding atrophic and dyskeratotic damages of the above cells. The laboratory examinations precede the radiation exposure and are repeated with the total basic dose (TBD) 20 Gy, 40 Gy and 60 Gy and 1 month after the completion of treatment.
EFFECT: invention is characterised by accessibility and low cost of production, and can be used in clinical practice for the purpose of the objective assessment of the vaginal mucosa health in radiation therapy of malignant new growths, particularly for the assessment of the early radiation reactions and injuries of the vaginal mucosa formed in the course of the therapy or in 3 near months thereafter, for the purpose of prediction of the developing radiation complications in the vagina, and if needed, the correction of the radiation exposure regimen, for the purpose of the radiation therapy compliance.
SUBSTANCE: invention relates to the field of biochemistry. Claimed is a method of evaluating cell viability in a microbioreactor by means of an optical light guide. The method includes the placement of cells into a membrane compartment of a replaceable cell unit of the microbioreactor, application of a working solution of a vital dye, introduction of the dye into the microbioreactor compartment. After the introduction incubation of the cells in the vital dye solution and removal of the vital dye solution, which has not bound with the cells, are performed. Removal is performed by the replacement of the incubation solution with a growth medium, which does not contain dye. The optical light guide, connected to a spectrometer, is brought into contact with an optically transparent material with the replaceable cell unit under the membrane compartment of the microbioreactor. After that, the support spectrum of a fluorescent signal is measured as an integral of the intensity of fluorescence on the membrane compartment of the microbioreactor, in which the cells to be analysed are absent. Also measured is the spectrum of the fluorescent signal as the integral of the intensity of fluorescence on the membrane compartment of the microbioreactor with the analysed cells. After that, the support spectrum of the fluorescent signal for the membrane compartment of the microbioreactor without the cells to be analysed is subtracted from the obtained spectrum of the fluorescent signal for the membrane compartment with the analysed cells. The quantity of the viable cells in the membrane compartment of the microbioreactor is calculated on the basis of the obtained value of the fluorescence signal intensity.
EFFECT: invention makes it possible to quickly determine viability of the cells under an impact of influencing factors in a real time mode in the microbioreactor.
6 cl, 3 dwg, 5 tbl, 5 ex
SUBSTANCE: method for estimating the bifidus bacteria and lactic acid bacilli in experimental animal's gastrointestinal tract involves preparing bacterial mutants on dense nutrient media with increasing rifampicin concentrations from 10 mcg·ml-1. The grown spontaneous mutants are selected, and the dense nutrient rifampicin medium is re-inoculated in the concentration of up to 160 mcg·ml-1. The mutants characterised by hereditarily stable rifampicin resistance are selected for administration into laboratory animals in the concentration of 150-160 mcg·ml-1 in the nutrient medium. The amount of the administered mutants is calculated. After keeping the animals, the selected faeces are suspended in isotonic sodium chloride. A supernatant is selected, and the dense nutrient medium containing rifampicin 110 mcg·ml-1 is inoculated in a Petri dish. It is incubated for 72 h in the microaerophilic environment at 37°C. The plate count is measured, and the growth-positive bacteria count is re-calculated per 1 g of faeces. A portion of the growth-positive microorganisms of the administered count is determined and used to state the survival.
EFFECT: invention enables effectively determining the portion of the positive-growth bifidus bacteria and lactic acid bacilli passed through the animal's gastrointestinal tract.
2 dwg, 3 tbl, 3 ex
SUBSTANCE: granule of biocatalyst with the cells contained in it of one or more species of Rhodococcus is placed in a well of a 96-well plate, a colorant iodine-nitro-tetrazolium (INT) is added. After the appearance of specific purple staining formazan the optical density of granule is measured using a tablet photometer. In the case of a stained substrate or preliminary presence of the immobilised biocatalyst in the soil the extraction of formazan is carried out with 96% ethanol and the optical density of the extract is measured. The amount of viable Rhodococcus in the granule of the gel is determined using the calibration dependency diagram of the optical density on the number of living cells in suspension as determined by traditional method of plating on solid growth medium. Then, from the granule stained by the colorant of the biocatalyst or from the granule after extraction of the colorant the DNA is isolated and PCR is performed using sets of species-specific primers. The specific position of the immobilised Rhodococcus is determined.
EFFECT: invention enables to reduce the time differentiation of Rhodococcus and to improve the accuracy of the method.
2 cl, 3 tbl, 4 dwg, 4 ex
SUBSTANCE: invention relates to field of biotechnology and deals with method of quantitative determination of fixed rabies virus strain "Moskva 3253". Method includes decontamination and separation of RNA from virus-containing material, carrying out reaction of reverse transcription and polymerase chain reaction with hybridization-fluorescence account of results in "real time" mode with application of specific primers RV5-5'-GTTGGGCACTGAAACTGCTA-3', RV6-5'-GAATCTCCGGGTTCAAGAGT-3' and probe RV7-5'-ROX-AATCCTCCTTGAACTCCATGCGACAGA-BHQ2. Quantitative assessment of virus is determined on the basis of registration of signal of analysed sample fluorescence and its comparison with signal of fluorescence of PCR-standards, which contain different quantities of DNA-targets. Claimed method makes it possible to determine quantitative content of virus in rabies antigen of organ-tissue and culture origin.
EFFECT: application of invention contributes to standardisation of stage of rabies antigen preparation in production of heterological anti-rabies immunoglobulin.
2 tbl, 3 dwg, 2 ex
SUBSTANCE: invention relates to biochemistry and molecular biology. Conservation of cells of Escherichia coli in presence of buffered 80-90% glycerol is performed. Cell envelopes are removed with 3% triton X-100. Cell supramolecular structures are successively extracted with increasing concentrations of salts: 0.14 M (bacterioplasm), 0.35 M (loosely linked with cell residue), 2 M NaCl (strongly linked with cell residue), and 6 M guanidine hydrochloride with 0.1% β-mercaptoethanol (cell residue). Acid hydrolysis is carried out in said fractions. Anthrone method is carried out, with preliminary purification of anthrone preparation. Calibration graph is built and quantity of hexoses is determined by means of calculation formula.
EFFECT: invention makes it possible to determine quantity of hexoses in supramolecular structures of bacterial cell of Escherichia coli.
3 dwg, 3 tbl, 1 ex
SUBSTANCE: invention can be used for detection of coliform bacteria and E.coli in specimens of food products and water at performance of bacteriological tests. Feed medium includes a nitrogen source represented by meat peptone or pancreatic hydrolysate of fish flour, sodium chloride, dibasic sodium phosphate, potassium monophosphate, sodium pyruvate, L-tryptophane, sodium dodecyl sulphate, 6-chloro-3-indolyl-β-D-galactopyranoside (Salmon - GAL), 5-bromine-4-chloro-3-indolyl-β-D-glucoronide-(X-GLUC), isopropyl- β-D1-tiogalactopyranoside (IPTG) and microbiological agar in the specified ratio.
EFFECT: invention allows reducing identification time, improving difernetiation accuracy of coliform bacteria and Ecoli, and simplifying a feed medium preparation method.
2 cl, 4 dwg, 1 tbl, 3 ex
SUBSTANCE: group of inventions relates to microbiology. The method for chemical modification of microalgae lipids includes culturing Prototheca microalgae to obtain a biomass containing at least 10% microalgae lipids with respect to the dry weight of cells and not more than 500 mcg/g dye inclusions and carrying out a chemical reaction, resulting in covalent modification of lipids. Culturing is carried out under heterotrophic conditions until the cells enter the lipid accumulation phase. The chemical reaction is selected from saponification, reesterification, interesterification, hydroxylation, cyclopropanation, epoxidation, hydroprocessing, deoxygenation, isomerisation, hydrogenation and hydrolysis. Also disclosed is a method of producing soap, which includes using Prototheca microalagae biomass, containing at least 5% microalgae lipids with respect to the dry weight of cells and not more than 500 mcg/g dye inclusions, and saponification of microalgae lipids. Also disclosed is soap which contains fatty acid salts.
EFFECT: group of inventions enables to obtain microalgae biomass with high content of lipids.
17 cl, 39 dwg, 12 tbl, 31 ex
SUBSTANCE: invention refers to a method for culturing unicellular green algae Haematococcus pluvialis for astaxanthin production involving inducing astaxanthin synthesis in monade vegetative cells with the culture grown on the nutrient medium MONM-1 in substrate cell saturation with biogenic elements (inoculate) is introduced in an amount of 0.3-0.35107cell.l-1 into the nutrient medium MONM-2 different from the medium MONM-1 by 30-fold reduced content of nitrogen (0.2 mM.l-1) and phosphorus (0.12 mm.l-1); sodium acetate 15 mM is once introduced that is followed by 20-day semi-flow growth (0.1-0.3 day"1) with maintaining the target levels of nitrogen and phosphorus and exposing to luminescent day-light lamp at a light intensity of 120 mcE.g-2·s-1, with continuous air blow (0.3 l min"1) and temperature 22-26 °C. Developed is a cost-effective and efficient method, which can provide a basis for industrial culture of the unicellular green algae Haematococcus pluvialis as raw material for producing pure natural astaxanthin and a biologically active additive with high carotinoid bioavailability.
EFFECT: method prevents the vegetative cell transformation into aplanospores, provides the advantage in the monade cell cultures (>80%) over at least 20 days and the astaxanthin content in the biomass of at least 2% of dry matter.
SUBSTANCE: invention "Using bottom sea water of hydrogen sulphide pool of the Black Sea as sea algae culture medium" refers to marine culture. Technical substance of the invention consists in using bottom seawater of the Black sea both containing hydrogen sulphide, and oxidised as a sea algae culture medium. Studying the biogenic properties of the aqueous medium from a reduction zone of the Black sea implemented by the invention has shown that the bottom seawater has no bad influence on the Black Sea plankton algae in the presence of high initial concentrations of hydrogen sulphide. After complete oxidation of this xenobiotic, the Black Sea bottom water can be used as a nutrient fertile culture medium of unicellular plankton and multi-cellular benthoalgae in laboratory or industrial environment, and in marine culture farms.
EFFECT: sea algae culture in laboratory or industrial environment.
SUBSTANCE: culture grown on the modified nutrient Trenkenshu medium by enrichment culture procedure to the density of 1.5-3 g OR·l-1 is transformed into a quasi-continuous culture process. The following growth procedure is performed at a specific medium flow rate of approximately 0.3 days-1 under continuous light at a superficial lighting of 80 Wt·m-2, with continuous blow of air-gas mixture at 1 l of the mixture·min-1·l-1 of the culture, which contains 3 % CO2, and a temperature of 26-28°C on the modified nutrient Trenkenshu medium. The biomass production rate has been shown at approximately 0.5 g of O.M. with 1 l of the culture a day at the relative carotinoid content in the biomass of not less than 0.9 % O.M., chlorophyll A - 2.8% O.M. and protein - 55 % O.M.
EFFECT: producing the biomass with using the quasi-continuous culture process.
SUBSTANCE: invention relates to biotechnology. The Arthrospira platensis (Nordst.) Geitl. rsemsu T/05-117 strain has high content of neutral lipids. The strain is stored in the collection of the research laboratory of renewable energy sources of the Faculty of Geography of Moscow Lomonosov State University.
EFFECT: invention increases output of neutral lipids.
SUBSTANCE: method of production of biosilicated nanotubes is proposed. The method comprises culturing cyanobacteria in sodium silicate solution, neutralised with hydrochloric acid and mixed with a nutrient medium Z-8, incubation with the replacement and removal of the nutrient medium. The subsequent treatment with the solution of hydrogen peroxide, heating and washing with distilled water is carried out. The culturing cyanobacteria is carried out in nutrient medium Z-8, additionally comprising 0.05 g/l ammonium chloride, with a ratio of sodium silicate solution neutralised with 2M HCl, and the medium Z-8 5:1. The culturing is carried out in the bioreactor at a temperature 25°C under constant illumination and stirring for 10 days with replacement of the nutrient medium once per 2 days.
EFFECT: invention enables to obtain biosilicated nanotubes in a shorter time and with less thickness.
1 dwg, 1 tbl, 2 ex
SUBSTANCE: group of inventions, including a strain of unicellular green algae Parachlorella nurekis and its application for destruction of cyanobacteria relates to biotechnology. The strain Parachlorella nurekis 1904 KIEG is deposited in the Culture Collection of Algae and Protozoa (Culture Collection of Algae and Protozoa, CCAP), Scottish Marine Institute, Dunbeg, OBAN, Argyll, PA37 1QA, Scotland, UK under the registration number CCAP No.259/1 and can be used for destruction of cyanobacteria.
EFFECT: inventions enable to reduce the amount of cyanobacteria in water reservoirs.
2 cl, 6 dwg, 1 tbl, 2 ex
SUBSTANCE: invention propose a strain of Chlorella vulgaris IPPAS C-616 microalgae for obtaining lipids as raw material for production of motor fuel.
EFFECT: strain of microalgae has an ability to produce lipids with high content of saturated and mono non-saturated fatty acids.
1 dwg, 1 tbl, 3 ex
SUBSTANCE: photobioreactor includes elastic working capacity (2) with the first and the second external side surfaces (20, 20'). Capacity (2) is made from elastic transparent material non-permeable for fluid medium and is installed in frame (3). Frame (3) has elongated and essentially vertical support components (32). Components (32) are located at least in one horizontal row; besides, they are installed so that they are in series adjacent to the first and the second external side surfaces (20, 20') of working capacity (2) with the possibility of their support.
EFFECT: improvement of efficiency and quality of a cultivation process of microalgae at simultaneous reduction of costs.
15 cl, 5 dwg
SUBSTANCE: inoculum of microalgae Desmodesmus sp. strain 2C166E is introduced into mineral medium BG-11 till final concentration of chlorophyll in mixture of 4-6 mcg/ml. Incubation is performed under constant lighting and environment barbotage by atmospheric air during 12-16 days under temperature of 25-27°C with consequent separation of microalgae biomass from nutritional medium with obtainment of microalgae biomass containing 33-35% of fatty acids from dry weight of cells.
EFFECT: invention allows increasing content of fatty acids in microalgae biomass.
1 dwg, 4 ex
SUBSTANCE: invention can be used for processing sewage waters of the production of nitroaromatic or nitrohydroxyaromatic compounds, for instance, nitrobenzene or dinitrotoluene. To realise the method two-stage processing, including a stage of preliminary reduction and a stage of wet oxidation, is carried out. At the first stage alkaline sewage water is mixed with an organic reducing agent, which does not form salts in the sewage water, selected from peat, brown coal and/or hard coal. Processing in reducing conditions is carried out with heating to a temperature from 80 to 200°C and exposure at the said temperature for the time period from 5 min to 5 hours. At the second stage the sewage water obtained at the first stage is acidified and subjected to oxidation with oxygen-containing gas, for instance oxygen.
EFFECT: method suggests a technically safe, simple and economic technology of processing and purification of sewage waters, providing the reduction of harmful admixture to the level, acceptable for the supply of processed sewage waters to biological purification.
4 cl, 5 tbl, 6 ex