Nutrient culture medium for swine erysipelas strain erysipelothrix rhuisipathie
SUBSTANCE: nutrient culture medium for swine erysipelas strain Erysipelothrix rhuisipathie refers to general biotechnology and veterinary microbiology. As a nitrogenous nutrition source, the nutrient medium contains a mixture of fish autolysate and alkaline mussel hydrolysate in the following proportions: alkaline mussel hydrolysate 20-50%; fermentation peptone 1%; potassium phosphate 0.3%; sodium phosphate 1.8%; fish autolysate - the rest.
EFFECT: method can be used for preparing the microbiological nutrient media for erysipelas strain upstream.
The invention relates to a total of biotechnology and veterinary Microbiology and can be used for preparing microbiological culture media for growing the biomass of the strain of the causative agent of swine erysipelas.
The causative agent of swine erysipelas Erysipelothrix rhuisipathie is culture, demanding in terms of power. When the activation strain after long-term storage is of great importance, the rate of increase of biomass. As a rule, for scientific and commercial purposes using traditional nutrient medium on the basis of raw meat. Mastopathy broth BCH and ΜΠΑ from parivara of Hottinger provide good growth of microorganisms without changing their cultural and biological properties. Hydrolysates of sardines and fish meal are also widely used in the manufacture of dry nutrient media for cultivation of microorganisms, because fish proteins according to their structure and amino acid composition is most similar to proteins of meat and can meet the needs of microorganisms in organic nitrogen compounds. However, finding high quality and inexpensive culture media is an important task.
Known Nutrient medium for cultivation of microorganisms (see patent No. 2089609, C1, C12N1/20, EN), contains a source of nitrogen nutrition, sodium chloride, agar. As the nitrogen nutrition use pancr eticheski hydrolyzed fishmeal content of amino nitrogen 4,4±0,2%, total nitrogen 12,±0,5%. A significant drawback is that the hydrolysate has a high cost, because of the need to import from abroad. The closest set of features of the claimed invention is "Poivre seredova "Aquamed" for kultivovana microorganisms (divas. Pat. No. 34711 U, C12N1/20, UA), where nutrient medium is a mixture of the alkaline hydrolysate of shellfish with acid hydrolysate from fish raw materials in a ratio of 3:1 to 1:3. The final concentration of amino nitrogen was 60-120 mg %. In the composition of the medium also includes glucose or maltose, peptone, and agar. A disadvantage of the known environment is insufficient for the cultivation of the strain of the causative agent of swine erysipelas end concentrada content of amino nitrogen.
Characteristics in common with the essential features of the invention, the Nutrient medium for culturing the strain of the agent of swine erysipelas Erysipelothrix rhuisipathie is the fact that the environment consists of peptone is an enzymatic and alkaline hydrolysate of marine raw materials.
The basis of the invention the culture medium for culturing the strain of the agent of swine erysipelas Erysipelothrix rhuisipathie the task through the use of products of marine origin to construct a nutrient medium, which will ensure optimal growth of strain of the causative agent of swine erysipelas Erysipelothrix rhuisipathie for the cooking the vaccine.
This object is achieved in that for the cultivation of vaccine strain of the causative agent of swine erysipelas Erysipelothrix rhuisipathie prepared nutrient medium, consisting of fish autolysate, which impose additional 20-50% alkaline mussels of the hydrolyzate.
Mussel hydrolysate and autolysate sprat get on with well-known technology (see Kruczenski, Century, Kulyasova Century, I.e. To the question of obtaining protein hydrolysis tov from mollusk Leda//Izvestiya TINRO.- 1976.- 99.- C. 102-104); Lichvar N. A. Dry nutrient medium of fresh and frozen fish waste industry. Scientific notes of Dinevi production of culture media. Makhachkala, 1956, 2.).
The authors investigated the level of biomass formation of an industrial strain of the causative agent of swine erysipelas Erysipelothrix rhuisipathie No. 286 on different environments (table. 1).
As can be seen from the table. 1 culture was expressed in all environments except the reference, low growth rate, which indicates the demand for the nutrition conditions. In some environments also noted polymorphism sticks causative agent of swine erysipelas: elongated chain E. rhuisipathie, which is a sign of unfavorable culture conditions, in particular due to unbalanced nutrition.
Next explored the environment, which in addition of hydrolysates sprat sod the whinnying from About to 50% of mussels hydrolysate (table. 2). Cultivated strain at standard temperature conditions within 48 hours. The optical density was measured at the photo-electrochlorinator at a wavelength of 545 nm and a thickness of 5 mm cuvette Control sample was UN-cropped medium of the same composition.
As can be seen from the table. 2, adding in a nutrient medium mussels of the hydrolyzate in the amount of 20% by volume provided the doubling of biomass as in the reference environment (0,163 against of 0.081), and experience (0,130 against 0,063), and making 50% of mussel hydrolysate of Wednesday tripled growth. The difference between the experienced and the reference environment, despite the absence in the test environment of glucose, with increase in the content of mussels pyrolysate gradually decreased.
Due to the high content of amino nitrogen (an indicator of nutrient media for cultivation of microorganisms) hydrolysates seafood are valuable raw materials for biotechnological production. In the autolysate fish contains 4-10 times more amino nitrogen than is required for the productive growth of the commercial strains. Mussel hydrolysate, when its content in the environment no more than 20%, dual bacterial mass. Thus the Association of fish autolysate and alkaline mussel hydrolysate of allows you to obtain a balanced nutrient medium for cultivation of the causative agent R is LM of pigs.
An example implementation of the method. Cooked experienced a series of vaccine against swine erysipelas Erysipelothrix rhuisipathie No. 286 in volume 3 DM3. Has preliminarily determined that the pH of fish hydrolysate rate of 6.0, a bmict asnago nitrogen -740 mg%. In mussel hydrolysate pH of 7.7, and the content of amine nitrogen mg%. Of these components was prepared liquid and solid nutrient medium according to the standard formulations for the BCH. In a medium composition for culturing industrial strain on the proposed nutrient medium consisted of a mixture of fish autolysate and 20% by volume of the alkaline mussels of the hydrolysate, peptone is an enzymatic - 1% of the total, potassium phosphate and 0.3%, sodium phosphate - 1, 8%, pH was 7.3 to 7.4. The strains were planted in test tubes with liquid medium, then perseval vials with broths, and after culturing under appropriate conditions, perseval vials with the same medium and cultured in appropriate conditions. At the end of the cultivation was determined representative and the purity of the culture microscopy smears of environments and established the concentration of microbial cells.
The industrial culture of the strain of the causative agent of swine erysipelas, received on Wednesday from hydrolysates, compared with environments, obtained with raw meat did not differ on cultural and morphological properties and meet the requirements of the passport data on the strain. The concentration of the microbial cells in 1 cm shall be not less than 50 million The authors have obtained the following data: on nutrient mediums seafood concentration of microbial cells was 50 million, and the media, made from a decoction of Hottinger - 70 million
Inactivation of microbial cultures was performed using formaldehyde in a concentration of 0.4%. To check the completeness of inactivation was performed by crops received the vaccine in the nutrient medium, the microscopy of Gram-stained smears. Control emissions vaccine was conducted in 5 white mice weighing 18-20 g, injecting them intraperitoneally at 0,sm. The results of observation of the animals during the 16 days showed that the vaccine is harmless: experimental animals remained alive, eat healthy, changes in behavior are noted.
Control immunogenic activity conducted on the same white mice vaccinated with vaccine. The mice were infected with control strain of the agent faces in a lethal dose. All control mice (10 animals), which were not inoculated, died within 4-6 days, all vaccinated survived for 10 days. Based on the results of these studies, concluded that manufactured the vaccine is immunogene-active.
Thus, the claimed nutrient Sereda using products of marine origin, and ensures the growth of the strain of the causative agent of swine erysipelas Erysipelothrix rhuisipathie suitable for making effective the nd vaccine.
Nutrient media for cultivation of strains of the causative agent of swine erysipelas Erysipelothrix rhuisipathie, composed of peptone is an enzymatic, potassium phosphate, sodium phosphate, and the hydrolyzate of marine raw materials, characterized in that as the source of nitrogen nutrition use a mixture of fish autolysate and alkaline mussel hydrolysate of the following ratio of components:
|alkaline mussel hydrolysate||20-50%|
|fish autolysate||the rest of it.|
SUBSTANCE: invention relates to biotechnology and medical industry and can be used in production of bacterial concentrates, biologically active food additives, fermented food products. Method of obtaining bacterial concentrate of bifidobacteria in liquid form includes preparation of nutritional medium with addition of growth components based on clarified cottage cheese whey or on water base with addition of up to 1.5% of glucose, or on soybean whey with addition of lactose in amount 1%. Strain of bifidobacteria B. bifidum 83, activated with β-halactosidase, is introduced into prepared nutritional medium in amount 3-5%, biomass is grown, cooled, poured in containers.
EFFECT: invention makes it possible to increase manufacturability and intensity of the process, increase biochemical activity of bifidobacteria and consumer properties of obtained product.
7 tbl, 5 dwg, 4 ex
FIELD: food industry.
SUBSTANCE: invention relates to a starter preparation method. The method envisages preparation of the substrate of a mixture of barley flour and dry milk, mixed with water at a ratio of 1:3-1:4 and a temperature of 75-80°C, the substrate cooling, introduction of a mixture of amylase and xylanase enzyme preparations, maintenance at a temperature of 48-50°C during 90-120 min, the barley-and-milk hydrolysate cooling to 32-35°C, introduction of Linex combined preparation (1 capsule per 100 g of the barley-and-milk hydrolysate) and pressed bakery yeast (0.1% of the barley-and-milk hydrolysate total weight), the barley-and-milk starter incubation during 18-20 h at a temperature of 32-35°C, until titratable acidity is 10-12 degrees.
EFFECT: method allows to optimise amino acid and vitamin composition and intensity the process of barley-and-milk starter preparation.
1 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and concerns a method for preparing a pertussis component of complex vaccines. The presented method involves the B. pertussis culture growth on dense Bordet-Gengou and/or CCA nutrient media, the grown colonies selection according to the morphological characteristics, the selected colonies passage, the bacterial mass growth, the culture washout, the extinction reduction of the prepared pertussis suspensions to 10 international optical units, the agglutination reaction to measure agglutinogens 1, 2, 3 and the selection of pertussis suspensions, wherein the agglutinogen content is determined by type-specific serum dilution 1:3200 and more. The detected B. pertussis culture expressing agglutinogens 1, 2, 3 actively after the lyophilisation is used to prepare the pertussis component of complex vaccines.
EFFECT: characterised invention enables preparing the potent pertussis component of complex vaccines.
1 dwg, 5 tbl, 1 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions relates to biotechnology. Claimed is strain of Lactobacillus delbrueckii subspecies lactis CNCM I-3741, reducing content of cholesterol in blood. Strain is applied for obtaining fermented dairy products.
EFFECT: inventions provide efficient reduction of cholesterol content in blood.
3 cl, 3 dwg, 2 ex
SUBSTANCE: group of inventions relates to biotechnology. Disclosed is a Bacillus subtilis VKPM V-11353 strain, capable of splitting a wide range of mono- and di-sugars and a wide range antagonistic effect on pathogenic and opportunistic pathogenic bacteria and fungi, which cause diseases in plants and farm animals. Also disclosed are versions of using the Bacillus subtilis VKPM V-11353 strain as a bacterial preservative for silos, for producing agents for normalising intestinal microflora of animal farms and for producing agents for protecting plants from diseases.
EFFECT: group of inventions improves the quality of silos, improves safety of animal farms and increases effectiveness against pathogenic and opportunistic pathogenic microorganisms which cause wheat diseases.
4 cl, 8 tbl, 4 ex
SUBSTANCE: strain of bacteria Paenibacillus sp. IB-1 has antagonistic activity against phytopathogenic fungi. The strain is deposited in the Russian National Collection of Microorganisms under the registration number VKM B-2823D and can be used to produce the biological product for protection of plants against diseases caused by phytopathogenic fungi.
EFFECT: invention enables to reduce the duration of cultivation and increase the yield of microbial biomass.
4 tbl, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology, namely to protective devices for humans and farm animals against blood-sucking mosquitoes. The strain Bacillus thuringiensis var. israelensis No. 7-1/23A possesses larvicidal properties. The strain Bacillus thuringiensis var. israelensis is deposited in the collection of State Scientific Institution All-Russian Research Institute of Agricultural Microbiology of Russian Agricultural Academy under No. RCAM00626 in the group of spore microorganisms. It can be used in creating a larvicidal biopreparation against blood-sucking mosquitoes.
EFFECT: invention enables providing higher death rate of blood-sucking mosquitoes.
5 tbl, 1 ex
SUBSTANCE: method comprises application into soil of suspension of microbial preparation on the basis of suspension of psychrotolerant bacterial strain Pseudomonas sp. IB - 1.1 with a titre of not less than 2.0·108 CFU/ml.
EFFECT: invention enables to carry out for 1 month the biodegradation of oil with a high degree of hydrocarbon decomposition.
SUBSTANCE: invention relates to recombinant bacterium strain Rhodococcus rhodochrous RNCIM Ac1960, which has a constitutive acylating activity and is obtained by replacement in a chromosome of strain Rhodococcus rhodochrous RCM Ac-1515D of a gene coding nitril hydrase with a gene coding acylamidase from strain Rhodococcus erythropolis 37 RNCIM Ac-1793, as well as to a synthesis method of N-replaced acrylamides from acrylamide and amines using it as a biocatalyst.
EFFECT: invention allows efficient production of concentrated solutions of N-replaced acrylamides.
2 cl, 1 dwg, 1 tbl, 5 ex
SUBSTANCE: agent for microbiological protection of plants comprises a mixture of culture liquids Trichoderma viride, Azotobacter chroococcum, Bacillus megaterium, Beauveria bassiana, Metarhizium anisopliae with the necessary amount of water. The ratio of cultures in the mixture is 1:1:1:1:1, with a titre of each culture is not less than 1×107 CFU/ml. Using the said agent the method of microbiological protection of plants is implemented, comprising 2-3-fold spraying of plants during the growing season at a dosage of 12.5 l of the agent per 1 ha of crops. Spraying is carried out when the average temperature of air is not less than 15°C with an interval of 2-3 weeks.
EFFECT: stimulation of plant growth, increase in plant yield, improvement of soil fertility.
2 cl, 5 tbl, 4 ex
SUBSTANCE: method of obtaining of brucellous L-antigen is performed as follows. The strain Brucella abortus 19 is cultivated in the L-form in the dense nutrient medium prepared on the basis of meat-peptonic hepatic glucose-glyceric agar (1.3-1.5%) with adding of 10-15% normal horse Serum at the temperature 20-22°C within 2-3 days. The culture, obtained in the L-form, is emulsified with 0.5% phenolised normal saline solution, inactivated at the temperature 85-90°C within 60 minutes. The suspension is centrifuged at 3000-5000 rpm within 15-20 minutes and the concentration of brucellas is established at the level 50-60 billion m.k. The obtained antigen is titrated, standardised by specificity and activity. It is used for agglutination test in serum diagnostics of brucellosis.
EFFECT: invention allows to minimise the time of antigen preparation and to increase the yield of bacterial mass.
4 tbl, 3 ex
SUBSTANCE: invention describes a composition for induction of immune response against P. gingivalis, which contains an effective amount of one of the above chimeric or hybrid proteins, a prophylaxis method of a state or a disease related to P. Gingivalis, and a method for reduction of incidence or severity of the state or disease related to P. gingivalis with their application. Besides, the invention describes use of the above chimeric or hybrid proteins for determination of antibodies to P. Gingivalis in a biological specimen.
EFFECT: invention allows effective induction of immune response against the specified etiologic agent.
16 cl, 7 dwg, 4 tbl, 22 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biochemistry, particularly to a recovered polypeptide which is a biological target for methane-producing cell inhibition, as well as to a recovered polynucleotide which codes this polypeptide. There are disclosed expression vector and cloning vector containing this polynucleotide, and host cells containing the above expression vector. There are described conjugated molecules or fused molecule for methane-producing cell inhibition, as well as antibody or its functional fragment which binds to the above polypeptide. The invention also covers a pharmaceutical composition and methods for inhibiting and identifying the methane-producing cell with the use of the above conjugated molecule or fused molecule and the antibody or its fragment.
EFFECT: invention enables inhibiting the methane-producing cell effectively.
19 cl, 9 dwg, 6 ex
SUBSTANCE: wound-healing agent is a concentrate of the culture liquid of strain Trichoderma harzianum Rifai deposited in the Russian National Collection of Industrial Microorganisms under the number of RNCIM: F-180, as a producer of L-lysine of alpha-oxidase, and can be applied as a wound-healing agent for skin lesions.
EFFECT: invention enables to expand the range of means that provide wound healing of skin lesions.
1 tbl, 2 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine, namely to veterinary science, and can be applicable for using a composition for protection against an infection caused by Lawsonia intracellularis. That is ensured by using a non-living composition containing carbohydrate which is also found in living cells of Lawsonia intracellularis in association with an external cell membrane of the above cells. The vaccine is presented in the form applicable for intramuscular introduction, and contains an oil-in-water adjuvant containing oil drops with an average size of 400 nm.
EFFECT: using the given non-living composition leads to effective immunisation in intramuscular introduction with using small drops of the oil-in-water adjuvant which provides protection of animals against Lawsonia intracellularis.
7 cl, 9 tbl, 4 ex
FIELD: veterinary medicine.
SUBSTANCE: method of prevention of infectious conjunctivitis-keratitis of cattle comprises vaccination with vaccine associated against infectious conjunctivitis-keratitis of cattle based on antigens of bacteria Moraxella bovis and herpesvirus type I, while 29-31 days prior to vaccination the immunostimulatory agent "Kerokonvitin" is injected to the animals subcutaneously in the upper third part of the neck at a dose of 0.045-0.055 ml/kg body weight, obtained on the basis of cytotoxic serum from the blood of donor horses by their hyperimmunisation with antigen prepared from tissues of eyes - the conjunctiva and cornea of cattle which had an infectious disease of conjunctivitis-keratitis. Eye tissues of cattle are obtained during slaughtering animals.
EFFECT: improving the efficiency of prevention of infectious conjunctivitis-keratitis of cattle, higher immune status of body of animals.
3 tbl, 1 ex, 2 dwg
SUBSTANCE: invention relates to biotechnology and can be used to produce thermolabile enterotoxin (LT-enterotoxin) and Hafnia alvei anatoxin when producing a vaccine. The strain is deposited in the State Collection of Pathogenic Microorganisms of FBSI Scientific Centre for Evaluation of Medical Products of the Ministry of Public Health and Social Development of Russia under number 294.
EFFECT: strain has high capacity for producing thermolabile LT-enterotoxin.
1 tbl, 2 ex
SUBSTANCE: invention relates to field of biotechnology and deals with method of obtaining preparation based on vaccine strain of plague microbe. Claimed invention includes preparing inoculation native culture of plague microbe, concentration of microbe suspension, preparing vaccine suspension and obtaining dry form of preparation, with process of preparing inoculation culture including cultivation of microbes in liquid nutritional medium in flasks for 48 h at temperature 26…28°C and contibuous aeration with not less than 10 l min-1. with passaged stabilised starting culture, obtained as a result of three successive passages through organism of guinea pigs and mixed with glycerol-lactose-polyglucinum liquid in ratio 2:1; for preparation of vaccine suspension used is optimised in component composition protective drying medium, lyophilisation being carried out with observance of the specified regimen.
EFFECT: claimed solution makes it possible to obtain product with higher activity with reduced duration of process of its manufacturing.
3 dwg, 6 tbl
SUBSTANCE: method includes cultivation of previously prepared culture of the recombinant strain B. anthracis 55ΔTPA-1Spo-. The cell mass is separated using the filtration module with a membrane having a pore diameter of 0.2 mcm. Protein EA1 is extracted from the washed cell mass using a buffer with 1% sodium dodecyl sulfate, and purified by diafiltration using membrane filters and two-stage ion-exchange chromatography on hydroxyapatite. The protective antigen is isolated from the culture filtrate and purified by successive steps of concentration and diafiltration.
EFFECT: use of the invention enables to obtain in one processing chain the highly purified antigens of anthrax microbe - protective antigen and protein EA1 needed to create chemical vaccines.
3 dwg, 5 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: presented group of inventions refers to medicine. There are presented a composition containing an immunologically effective amount of the live avirulent strain Mycoplasma hyopneumoniae, and a method of inducing the immune response on Mycoplasma hyopneumoniae, involving the stage of administering the above composition to an animal. What is presented is a method of preventing or relieving an attack of Mycoplasma hyopneumoniae. There are also presented methods of enhancing the immune response on Mycoplasma hyopneumoniae, involving the stages of administering single or double doses of the above composition to the animal.
EFFECT: group of inventions is effective to provide the immunity in the animal and to protect from the infection with the virulent strain Mycoplasma hyopneumoniae, thereby reducing a severity and/or preventing the diseases caused by one or more virulent strains Mycoplasma hyopneumoniae.
18 cl, 8 tbl, 4 ex
FIELD: biotechnology, vaccines.
SUBSTANCE: vaccine comprises bacterial mass of Pasteurella multocida of serovariants A, B and D, Haemophilus pleuropneumonia of serogroups 1 and 2, and streptococcus of serogroups C and R, and also lysate-anigens of salmonellae Salmonella cholerae - suis, strain № 370 and Salmonella typhimurium № 415 mixed in the definite concentration. Vaccine elicits the high immunogenicity and provides the protection of pigs against infectious pneumonia of bacterial etiology and salmonellosis.
EFFECT: valuable veterinary properties of vaccine.
3 cl, 1 tbl, 5 ex