Method of obtaining of brucellous l-antigen
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
The invention relates to veterinary Microbiology, immunology, biotechnology, in particular to a technology for production of antigen for the diagnosis of brucellosis.
Diagnostics and specific prophylaxis of brucellosis imperfect, due to the variety of forms of the organism and its ability to intracellular parasitic, which leads to the unavailability of disease existing diagnostic methods, which are mainly based on the use of antigens from the S-forms of Brucella (Counsel for the diagnosis of brucellosis, 2004) .
Each existing form (S - R - L-) of the microorganism when sown on nutrient medium has its own characteristics, which are clearly defined only for S-shape (Counsel for the diagnosis of brucellosis, 2004) .
A method of obtaining L-subcultures of B. abortus (Bazikov I. A. "L-transformation Brucella", 1989), including the use of the environment containing the basis of semi trypticase prewar bovine hearts, 10-20% sucrose, 0.05 to 5% of magnesium sulfate, 0.01 to 1% sodium chloride, 15-25% horse serum, 1% glycerol, 0.01 to 0.03% of nalidixic acid, as well as transforming Agusta bicillin-3 from 50 to 20,000 IU /ml and glycerol from 0.1%to 3%. However, obtained by the method of L-subculture Brucella have reduced and sometimes completely lost antigenic properties. Anticavity, poluchena is using data L-subcultures Brucella, was not sufficiently specific, as they agglutinable source of bacterial S-shape in a dilution of 1:320. Anticavity was proposed for bacteriological diagnosis of brucellosis .
A method of obtaining L-subcultures - "Method for L-subcultures strain brucella abortus And-206" (patent No. 2263142 from 2003.07.14). The method involves the passage of Brucella strains of Brucella abortus And 206 on a nutrient medium containing: agar-agar - 1,2; sodium chloride - 0,1; magnesium sulfate - 0,05; sucrose - 10; glycerol - 1,0; normal horse serum - 10; hepatic infusion - else, where bicillin-3 added to the culture medium in increasing concentrations from 10 to 9000 IU/ml of medium. This method allows you to get L-subculture Brucella at a final concentration of bicillin-3 2000 IU/ml of medium get L-subculture Brucella, which after a single intravenous administration to rabbits cause the production of specific agglutinins detected in agglutination reactions in titer of 1:1600 . Cultivation of Brucella L-form occurs with the use of bicillin-3 in increasing concentrations from 10 to 9000 IU/ml of medium is a long process. And getting even a small amount of bacterial mass was used to obtain hyperimmune serum, which was used in the bacteriological diagnosis of brucellosis.
The closest technical solution t is aetsa a method of obtaining diagnostic serum against Brucella in an L-shape, to obtain which used culture of the strain of C. abortus 19 in an L-shape by transforming the original typical culture vaccine strain Brucella abortus 19 (S-form) cultivation on solid nutrient medium prepared based on meat-peptone hepatic glucose-glycerol agar (1,3-1,5%) (MPGGE) with the addition of normal horse serum 10-15% and streptomycin at a dose of 2.5 to 5.0 IU/ml at 37-38°C for 4-5 days, the antigen is produced by inactivation of the obtained culture by heating at a temperature of 85-90°C for 60 min and three times intravenously administered to rabbits in increasing doses with an interval of 72 hours . The disadvantage of this method is the long process of preparation of the antigen, the purpose of this invention has been receiving a stable L-culture, and used the antibiotic streptomycin, which retards the growth of microorganisms and transition it into the original S-shape, the output of the bacterial mass is unimportant, since the introduction of culture into the body of an experimental animal antibodies (serum), which is used for bacteriological diagnosis of brucellosis.
The technical result of the invention is to reduce timing brucellosis L-antigen, increased output of finished products, the use of antigen in complex serological studies when diagnostics is brucellosis, namely agglutination reaction (RA) to identify additional infected animals with latent brucellosis.
The claimed technical result is achieved in that a method of obtaining brucellosis L-antigen includes the cultivation of Brucella abortus strain 19 in an L-shape on the solid nutrient medium prepared based on meat-peptone hepatic glucose-glycerol agar (1,3-1,5%) with the addition of 10-15% normal horse serum inactivation suspension at a temperature of 85-90°C for 60 minutes, the cultivation is carried out at 20-22°C for 2-3 days, received in an L-shape culture emuleret 0,5% fenrisians saline solution, centrifuged at 3000-5000 rpm for 15-20 minutes and set the concentration of Brucella 50-60 billion, M. K., the resulting antigen is titrated 1:5, 1:10, 1:20, 1:40, standardizes on specificity and activity, used for agglutination reactions in serological diagnosis of brucellosis. The antigen recognized suitable for diagnostic purposes in a dilution of 1:20.
The proposed method for the preparation of brucellosis L-antigen 2 times reduces the time of manufacture and increases the yield of bacterial mass to prepare antigen 3 times. The antigen used in the complex serological tests in the diagnosis of brucellosis, namely in PA to identify additional infected animals with l is a competent form of brucellosis.
The invention is characterized by the following examples.
Example 1. Culture of Brucella abortus strain 19 in an L-shape inoculated in tubes with nutrient dense environment, based on meat-peptone hepatic glucose-glycerol agar (1,3-1)5%) with the addition of 10-15% normal horse serum and streptomycin at a dose of 2.5 to 5.0 U/ml, at 37-38°C for 4-5 days. Microbial suspension of Brucella abortus 19 L wash saline solution of 0.85% NaCl), prepare one billionth mixture (according to the turbidity standard gisk named after. L. A. Tarasevich). Then by the method of successive dilutions choose a concentration to 1 ml was 100 m K. .
To study take 7 temperature conditions: 18°C, 20°C, 22°C, 25°C, 30°C, 35°C, 37°C. For each mode serves 4 Petri dishes with the environment, based on meat-peptone hepatic glucose-glycerol agar (1,3-1,5%) with the addition of 10-15% normal horse serum. In the first option in the environment add streptomycin, in the second variant without streptomycin. And in each Petri dish contribute 100 M. K. culture Brucella abortus 19 L, which stand in different temperature regimes, the growth of culture Brucella abortus 19 L check on Petri dishes visually daily for 5 days (table.1).
In different temperature regimes in environments with streptomycin and without culture C. abortus 19 L on Petri dishes grew up differently. When temperature is re 20°C, 22°C on media without antibiotics for 2-3 days grew colonies in 3 times more than with streptomycin. At 37°C for 5 days has grown colonies of the same number with streptomycin and without it, the number of colonies was less than in the first example.
In the proposed method of obtaining brucellosis L-antigen for 2-3 days at a temperature of 20-22°C was observed maximum growth of bacterial mass. Cultivation of B. abortus 19 L at a temperature of 20-22°C 2 times reduces the time of preparation of antigen and 3 times the output of the bacterial mass.
|The dependence of the growth culture of B. abortus 19 L from the temperature regime for 5 days|
|Temperature||Day (1-5)||Medium with streptomycin, Petri-dish||Medium without antibiotic Petri-dish|
Example 2. The specificity of L-antigen was tested in the relevant serological reactions with the use of S - and R-brucellosis sera from commercial diagnostic kits and negative (N-) of the blood serum of healthy animals. Antigen for agglutination reaction (RA) of Brucella in an L-shape gives a positive reaction only with L-Brucella serum (table.2, 3, 4).
|The results of the reaction of agglutination (PA) L-Brucella antigen with brucellosis (S,R) and negative (N-) sera|
|Breeding L-Brucella antigen||Names and cultivation of Brucella||sera|
The table shows that cross-reactions with heterologous (S-and R-) sera obtained negative, and with homologous (L-) sera - positive results.
From the above data it follows that the proposed brucellosis L - antigen, has a stringent specificity in relation to S, R and N sera and diagnostic activity. The working concentration of microbial cells L-antigen 50-60 billion
Example 3. Production testing of brucellosis L-antigen was performed on the population of deer in herds with different epidemic response of brucellosis total number 4400 goals, including affected with brucellosis -1005 head.
The specificity of L-antigen was determined on the reindeer head with a good brucellosis (table.3).
|The results of serological tests for brucellosis in reindeer in prosperous stud|
|Year research||The district of Yamal-Nenets Autonomous district||The studied animals (animals)||Of them respond positively||With antigens|
These data confirm that the antigen is derived from L-cultures, has specificity and shows diagnostic activity in the reaction of agglutination (PA) with sera, nesteriak S-, R-, L-antibodies.
In foci of brucellosis infection animals are native not only S-and L-forms of the pathogen that are not registered commercial S-antigen-based assays, and additionally identifies the proposed L-antigen (table.4).
|The results of serological tests for brucellosis in reindeer from disadvantaged stud|
|Year research||The district of Yamal-Nenets Autonomous district||The studied animals (animals)||Of them respond positively||With antigens|
|% of investigated||-||2,8||2,7|
The table shows that of the 1005 and the following goals identified positively reactive - 57 goals, which amounted to 5.5% of the total number of investigated animals, including the well-known S - antigen identified 2,8% of the animals (29 goals), and the proposed L-antigen allows to identify 2,7% (28 goals) animal brutselloosile with persistence of the pathogen in the L-form.
Thus, the proposed brucellosis L-antigen used in the reaction of agglutination (PA), has a strict specificity in relation to the S-, R - and N-sera, diagnostic activity, increases the efficiency and reliability of the diagnosis of brucellosis by 50.0%. Can be used for the diagnosis of brucellosis in animals-brutselloosile with persistence of the modified L-forms of the pathogen.
The invention allows to reduce in 2 times the time of preparation of antigen due to a change in temperature during the cultivation of Brucella abortus 19 in an L-shape and to increase the output of the bacterial mass in 3 times, the resulting antigen can be used in PA, in the complex serological tests for brucellosis to identify additional infected animals with latent infection that will improve the efficiency and reliability of the diagnosis of brucellosis by 50%.
1. Counsel for the diagnosis of brucellosis in animals. - M., 2004. - 62 S.
2. Bazikov I. A. L-transformation Brucella / Epidemiology, Microbiology and immunology bacterial and viral is Pecci: abstracts. regional scientific. proc. young. scientists, 17-20 October 1989, Rostov-on-don, 1989, pp. 5-8.
3. Patent RU No. 2263142 C2, 7C12N 1/20, C12N 1/20, C12R 1:00 "How to get L-subcultures strain Brucellaabortus And -- 206", published 2005.10.27.
4. Patent RU No. 2268748 C2, A61K 39/40, C12N 1/20, G01N 33/531, publ. 27.01.2006.
5. Alton J., Jones, L. M. Methods of laboratory studies on brucellosis / FAO / who, Geneva, 1968. - 86 C.
The method of obtaining brucellosis L-antigen, including the cultivation of Brucella abortus strain 19 in an L-shape on the solid nutrient medium prepared based on meat-peptone hepatic glucose-glycerol agar (1,3-1,5% agar) supplemented with 10-15% normal horse serum, inactivation of the suspension at a temperature of 85-90°C for 60 minutes, characterized in that the cultivation is carried out at 20-22°C for 2-3 days, received in an L-shape culture emuleret 0,5% fenrisians saline solution, centrifuged at 3000-5000 rpm for 15-20 minutes and set the concentration of Brucella 50-60 billion, M. K., the resulting antigen is titrated standardise on the specificity and activity and is used for agglutination reactions in serological diagnosis of brucellosis.
FIELD: machine building.
SUBSTANCE: vortex bioreactor includes cylindrical vessel1 with lid 2 provided with a device for medium mixing. Mixing device consists of bladed wheel 7, horizontally fixed on vertical shaft in upper part of vessel 1 and horizontal annular partition 9, installed in vessel 1 with a gap relative to its cylindrical walls, rod 10 vertically installed along axis. Horizontal annular partition 9 is located with possibility of rotation on vertical rod 10 and has retention mechanism of horizontal annular partition 9, installed on rod 10. Bioreactor is provided with pipe 19 or telescopic pipe 20, mated to axial hole 13 of horizontal annular partition 9, attached to the bottom of the last one and located around rod 10. On upper surface 11 of horizontal annular partition 9 there made are radial channels 12, located from axial hole 13 to the edge of annular partition 9 inclined downwards to the bottom of vessel 1. Horizontal annular partition 9 is designed floating, for example, from polypropylene. Inner diameter of pipe 19 or telescopic pipe 20 corresponds to diameter of axial hole 13 of horizontal annular partition 9. Area of axial hole 13 of horizontal annular partition 9 equals to cross sectional area at inlet 14 to radial channels 12 and total cross sectional area at outlet 15 from these radial channels 12.
EFFECT: improving efficiency of mixing and speeding up biochemical processes upon their carrying out using fluid mediums of various viscosity.
6 cl, 5 dwg
SUBSTANCE: banana fruit and skin are weighed, washed, crushed and mixed with water preheated to 45±1°C in the ratio of 1:3. It is alkalised to pH 8.2-8.3, the enzyme is added, which is used as the pancreas of cattle in the amount of 9-15% of total volume. The preservative is added, which is used as chloroform in an amount of 1% of total volume. The mixture is subjected to hydrolysis for 10 days at a temperature of 45-50°C. During the first 24 hours the mixture is stirred every 15 minutes for 5 minutes, and then every 2 hours for 5 minutes. The hydrolyzate is filtered through cheesecloth, then the filter towels (paper) and sterilised by ozonation. It is covered with cork and stored hermetically sealed at a temperature of from 2°C to 8°C.
EFFECT: invention provides obtaining of transparent hydrolyzate with increased sterility, with high biological activity, which leads to improvement of its properties when it is used as a growth stimulator of Leptospira.
2 tbl, 3 ex
SUBSTANCE: method provides culturing in the liquid culture medium of the virulent strain of bacteria Staphylococcus aureus No. 6 followed by separation of the culture medium from the bacteria by filtration through the filter PLASMAFILTER PLASMAFLUX PSu 2S with obtaining the filtrate. Ammonium sulphate to 80% saturation is added to the resulting filtrate to obtain the precipitate. The resulting precipitate is separated by centrifugation at 10000 g for 30 minutes and dissolved in phosphate buffer at pH 7.4 with subsequent microfiltration of the protein-containing fraction through the 0.22 mcm membrane and desalting on the column PD-10, purification and concentration by carrying out ion-exchange chromatography on the column of Q-sepharose, elution with 0.15 M NaCl and ultrafiltration on two filters of 100 and 30 kDa to obtain protein containing fraction with the molecular weight of 30-90 kDa and protein content of 0.5-1.0 mg/ml.
EFFECT: invention enables to obtain the products based on protective secreted protein-containing compound.
1 dwg, 5 tbl, 1 ex
SUBSTANCE: method to cultivate sublimated strains of microorganisms includes introduction of a growth stimulant and a source of carbon into a dense nutrient medium with subsequent seeding of microorganism cells, incubation of seeds and accounting of viable microbial cells. Sublimated strains of microorganisms include Yersinia pestis EV, Escherichia coli C600, Bacillus anthracis "СТИ", Vibrio cholerae NAG 504, Brucella abortus 19BA. The growth stimulant and carbon source is 96% ethyl alcohol in the amount of 1.0-2.0% of the medium volume. If necessary, they add gentian violet in concentration of 1:100000.
EFFECT: improved yield of viable strain cells on dense nutrient media.
1 tbl, 5 ex
SUBSTANCE: two suspensions are prepared. Clinical polyantibiotic-resistant strains Escherichia coli are added to an isotonic solution of NaCl to achieve the concentration of 30-40 thousand CFU/ml. Copper nanoparticles are added to the solution of NaCl to achieve the concentration of 0.01-0.05 mg/ml. The suspension of ethylenediaminetetraacetic acid - EDTA is prepared by its dilution in distilled water at the rate of 0.1-0.2:1, respectively. NaOH is added to the prepared suspension to obtain the solution of EDTA with pH=7.6-8. The prepared suspensions are connected with the solution of EDTA in the following ratios by wt %: suspension of copper nanoparticles - 70-85, suspension of microorganisms - 10-20, EDTA - 5.10. It is incubated in the shaker at 100-150 rev/min and a temperature of 36-38°C for 40-60 minutes. The resulting biomass is inoculated on the solid nutrient medium with the volume of 20-25 ml in the amount of 0.1-0.12 ml. It is incubated in the thermostat at a temperature of 36-38°C for 18-24 hours. The sensitivity of E.coli strains to antibiotics is determined.
EFFECT: invention enables to increase the sensitivity of the said bacterial strains to antibiotics gentamicin and ampicillin.
2 tbl, 2 cl, 1 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology, namely to leukolectins, and can be used in medicine. What is prepared is the polypeptide leukolectin characterised by SEQ ID NO:1-8. The recombinant preparation is ensured by using a nucleic acid coding it and integrated into an expression vector which is used to transform a host cell. Testing absence-presence or determining an amount of the polypeptide leukolectin are ensured by using an antibody or an antigen-binding fragment of a variable region of the above antibody which is specifically bound to the polypeptide leukolectin. The polypeptide leukolectin or the nucleic acid coding it are used as ingredients of a pharmaceutical composition in therapy of pathological disorders of skin and mucous membranes.
EFFECT: invention enables treating or preventing autoimmune disorders of skin, inflammatory diseases of skin or mucous membrane, or injured skin in an animal effectively.
16 cl, 19 dwg, 3 tbl, 12 ex
SUBSTANCE: soil contaminated with oil products is detoxified by adding a natural sorbent with a biopreparation until a given concentration of the contaminant in the soil is achieved. The sorbent used is glauconite and the biopreparation used is polyculture. Before adding the sorbent with the biopreparation, concentration of the contaminant is measured, the volume of sorbent is determined and a shielding intermediate layer of biohumus is placed at the boundary where the contaminant penetrates the soil in the lower part of the contaminated layer. Concentration of the contaminant is determined layer-by-layer. The number of layers of contaminated soil with concentration of the contaminant in the layer differing from each other by at least double is not less than two. The contaminated soil is moistened after distributing the volume of sorbent with the biopreparation, effective microorganisms and fermented compost on the surface of the contaminated soil while simultaneously mixing. A water-swelling polymer is also added to the contaminated soil. Temperature conditions for activating biological processes are maintained by adding organic materials to the soil, wherein the volume of said organic materials in the shielding intermediate layer should be at least double that in the contaminated soil.
EFFECT: high efficiency and faster detoxification.
SUBSTANCE: invention refers to genetic engineering and can be used for methane-producing cell permeability control. What is prepared is a polypeptide able to permeate into a methane-producing cell and to increase its permeability, characterised by an amino acid sequence SEQ ID NO:117, 118 or 119 or being at least 90% identical to the above sequence, or at least 15 sequential amino acids of the above sequence. What is also prepared is a polynucleotide coding the above polypeptide cloning and expressing vectors used for producing host cells producing the polypeptide or used for the vector replication. The polypeptide can contain a fluorescent tag on an N-terminal amino acid residue.
EFFECT: invention enables providing higher methane-producing cell permeability.
18 cl, 35 dwg, 3 ex
SUBSTANCE: invention relates to biotechnology, in particular to methods of breeding beneficial insects. The method comprises preparing the nutrient medium comprising soy flour, sucrose, milk powder, palm kernel oil, dry aphides, Wesson salt, dry brewer's yeast, tocopherol, ascorbic acid, agar-agar, vitamins B1, B6, B12, metaben, inositol, and distilled water at a predetermined ratio and growing coccinellidae Harmonia axyridis Hall. at a temperature of 23-26°C in the prepared medium, applied to pieces of sterile polyethylene film. At that the change of feed is carried out daily.
EFFECT: invention enables to simplify breeding coccinellidae Harmonia axyridis Hall.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to field of biotechnology, namely to muteins of human tear lipocalin, and can be used in medicine. Mutein of human tear lipocalin (hTLc) has identifiable affinity of binding with human receptor Met (c-Met) receptor tyrosine kinase, or its domain, or fragment of human c-Met. Mutein contains from 6 to 18 amino acid substitutions relative to amino acid sequence of mature lipocalin of human tear liquid (SWISSPROT DATABANK ENTRY P31025; SEQ ID NO:36), selected from group, consisting of Arg 26→Thr, Val, Pro, Ser, Gly; Glu 27→Gln, Gly, Val, Ser; Phe 28→Met, Asp; Pro 29→Leu, Ile, Ala, Trp; Glu 30→Leu, Gly, Arg, Phe; Met 31→Ser; Asn 32→Leu, Arg, Val, Gln; Leu 33→Tyr, Val, Ile, Thr, Phe; Glu 34→Val, Arg, Ala; Leu 56→Asn; Ile 57→Gln; Ser 58→Ile, Val; Asp 80→Tyr; Lys 83→Ala; Glu 104→Asp; Leu 105→Thr; His 106→Trp and Lys 108→Gly. Mutein can also additionally contain the following substitutions: Cys 61→Ser; Cys 101→Ser; Cys 153→Ser; Arg 111→Pro; Lys 114→Trp; Thr 37→Ser; Met 39→Ile, Leu; Asn 48→Ser; Lys 52→Thr, Met; Met 55→Leu; Lys 65→Arg, Leu; Ala 79→Leu, Ser; Ala 86→Thr; Ile 89→Ser, Gln, Thr, His; Thr 40→Cys; Glu 73→Cys; Arg 90→Cys; Asp 95→Cys; Lys 121→Cys; Asn 123→Cys and Glu 131→Cys.
EFFECT: invention makes it possible to efficiently treat pathological disorders, which involve pathway HGF/c-Met, as well as to perform identification of human c-Met in sample.
40 cl, 16 dwg, 9 tbl, 25 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: medicine, pharmaceutics.
SUBSTANCE: invention refers to veterinary microbiology, immunology, biotechnology, namely to technology of an antigen for diagnosing brucellosis. The method for producing brucellous L-antigen is implemented as follows. The strain Brucella abortus 19 is cultured in the L-form on a solid nutrient medium prepared of meat-peptone liver glucose-glycerol agar (1.3-1.5% agar) with added 10-15% normal horse serum and streptomycin in a dose of 2.5-5.0 Units/ml at 37-38°C for 4-5 days. Then the prepared L-form culture is emulsified by 0.5% phenolised physiological saline; the suspension is inactivated at temperature 85-90°C for 60 minutes, centrifuged at 3000-5000 rpm for 15-20 minutes; the brucella concentration is specified at 50-60 bln microbial cells; the prepared antigen is titred, standartised by specificity and activity. The latter is used for the purpose of an agglutination test in serological diagnosis of brucellosis. The invention enables higher effectiveness and reliability of diagnosing brucellosis by 20-25%.
EFFECT: invention may be used for diagnosing brucellosis in carrier animals with persistent changed L-forms of the agent.
4 tbl, 3 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