Method for manufacturing associated vaccione against colibacteriosis, salmonellosis, streptococcosis and enterococcal infection in nutrias

FIELD: veterinary medicine.

SUBSTANCE: it is necessary to select the affected organs from dead nutrias out of local epizootic focus, prepare the suspension out of pathological material to inoculate onto differential-diagnostic media, isolate pure cultures of Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae and Streptococcus fecalis, grow separately the isolated cultured in beef-extract nutritive medium at addition of 0.2%-glucose to achieve the concentration of microbial cells of about 4-5 bln/cu/ cm, inactivate with formalin up to 0.4-0.6%-final concentration, keep at 37°C for about 72-96 h, mix the cultures at equal ratios, add 3%-aluminum hydroxide solution at the quantity of 20% against the volume of the culture to be thoroughly mixed. Then the vaccine obtained should be packed and sealed. The innovation is simple in usage, moreover, it enables to increase specificity and immunogenicity of the obtained vaccine.

EFFECT: higher efficiency.

1 tbl

 

The invention relates to veterinary medicine, in particular for vaccines and can be used in research and industrial establishments engaged in the construction of biological products, as well as enterprises of biological industry.

A method of obtaining an associate's vaccine for the prevention of infections caused by opportunistic bacteria (RF patent for the invention №2035189 from 02.01.86, MKI AC 39/125, 39/135). This method involves the separate allocation of protective antigens from Staphylococcus aureus No. B - 243 distinguish cytoplasmic antigen of toxinproducing strain of Pseudomonas aeruginosa of epidemiology and Microbiology No. PA - 7 receive toxoid Pseudomonas aeruginosa from clinical strains of Proteus mirabilis№6, 8, 40, 508, 1115, 4641 secrete soluble antigens by controlled proteolysis derived antigens are mixed in a saline solution from the calculation of their content in 1 ml of vaccine 0.15-0.2 mg protein, 15-30 µg protein, 10-20 μg of protein, respectively, and the mixture additionally add commercial staphylococcal toxoid and aluminium hydroxide in the amount of 5-10 The EU and 1-2 mg respectively. The technology of obtaining the vaccine is very difficult for the prevention of infectious diseases nutria is not applied.

A method of obtaining multicomponent vaccine for immunization of diseases caused by conditionally pathogenic micro is rganizati (RF patent for the invention №2083223 from 27.05.94, MCI AC 39/125, 39/135). This method includes separate cultivation of strains of Klebsiella peumoiae risk No. 204, Proteus vulgaris risk No. 177, Escherichia coli K-100 No. 147, Staphylococcus aureus risk No. 1991, №5, №9, while the extraction of antigens from cells of Klebsiella, Proteus and Escherichia coli spend hydroxylamine, and from cells of Staphylococcus - water extraction and the resulting antigens are mixed in a ratio of 0.6, and 0.6, 0.6 and 0.8 mg, respectively, per 1 ml of distilled water. This technology is complicated, for the prevention of infectious diseases nutria is not applied.

A method of obtaining the vaccine Streptococcus and pasteurellosis nutria (RF patent for the invention №2099083 from 30.09.94, MKI AC 39/125, 39/135). This method provides for separate growing in liquid culture medium with added glucose strains of Streptococcus zooepidemicus VGNKI # K-DEPT, Pasteurella multosida VGNKI No. 2394 and Pasteurella multocida VGNKI No. 1015 for 18-24 hours, the resulting culture medium is subjected to freeze-thawing, the separated liquid fraction, inactivate 0,3-0,4%formalin solution, combine them in equal proportions by volume and consistently contribute aluminium hydroxide. This method allows you to get the vaccine Streptococcus and pasteurellosis nutria. After the introduction of this vaccine in animals occur post-vaccination complications (inflammation at the injection site, abscesses, lameness). D. the fair vaccine does not protect nutria from colibacillosis, salmonellosis and Enterococcus infection.

The technical solution of the invention is to remedy these shortcomings, in particular higher specificity, efficiency of the method of making a vaccine against colibacillosis, salmonellosis, streptococcosis and Enterococcus infection nutria, through the introduction of new crops pathogens, components, excluding the adverse effect.

The solution is achieved in that in the method of manufacturing a vaccine associated against colibacillosis, salmonellosis, streptococcosis and Enterococcus infection nutria, including the production of antigens, inactivation, making adjuvant, shall conduct sampling of the affected organs from dead nutria from local epizootic focus, prepare a suspension of the pathological material, do the planting on differential diagnostic environment, isolated pure cultures of pathogens Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae and Streptococcus fecalis and separately grown cultures in mesopatania nutrient medium with addition of 0.2% glucose to achieve a concentration of microbial cells - 4-5 billion in 1 cm3that inactivate formalin to 0.4 to 0.6%final concentration, incubated at 37 ° °C for 72-96 hours, mixed culture in equal proportions and make a 3%solution of aluminium hydroxide in Koli is estwe 20% by volume of the culture, thoroughly mixed, Packed and sealed.

The novelty of the claimed proposal due to the fact that in contrast to known methods shall conduct sampling of the affected organs from dead nutria from local epizootic focus, prepare a suspension of the pathological material, do the planting on differential diagnostic environment will excrete pure cultures of pathogens Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae, Streptococcus fecalis during separate cultivation of pure cultures of Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae. Streptococcus fecalis in mesopatania nutrient medium with addition of 0.2% glucose to achieve a concentration of microbial cells - 4-5 billion in 1 cm3that inactivate formalin to 0.4 to 0.6%final concentration, incubated at 37 ° °C for 72-96 hours, mixed pure culture in equal proportions, which allows you to completely suppress the pathogenicity and to maintain a high immunogenicity against colibacillosis, salmonellosis, streptococcosis and Enterococcus infection. The addition of aluminium hydroxide in the amount of 20% by volume of the culture allows through 12-15 days after a single injection at a dose of 1 cm3the nutria to provide the grafted formation of a busy immunity against the three diseases. The method provides receiving harmless, highly immunogenic, more specific vaccines to protect the inside is from colibacillosis, salmonellosis, streptococcosis and Enterococcus infection.

According to the patent and scientific literature is not detected is similar to the claimed combination of features that allows us to judge of inventive step of the claimed technical solution.

As for the criterion "industrial applicability", the components included in the vaccine, well-known and widely used in veterinary practice in the manufacture of inactivated vaccines: formalchemy vaccine against paratyphoid (salmonellosis) calves THAT 46-21-166-75, approved 10.05.75, the instructions for the manufacture and control of this vaccine approved by the Main veterinary Department Ministry of agriculture of the USSR 28.03.80 year; vaccine against paratyphoid (salmonellosis) piglets THAT 46-21-952-74, approved 15.07.74, the instructions for the manufacture and control of this vaccine approved by the Main veterinary Department Ministry of agriculture of the USSR 23.10.96 year; the associated vaccine against salmonellosis, pasteurellosis and Enterococcus infection of piglets THAT 10.09-62-90, approved 01.08.90, the instructions for the manufacture and control of this vaccine approved by the Main veterinary Department 16.07.90 year; vaccine Enterococcus infection of calves, lambs and piglets THAT 10.09-91-90, approved 15.12.90,; PI and control formulatin polyvalent gidrookisaljuminievuju against colibacillosis (escherichiosis)piglets calves and lambs, approved BS Ministry of agriculture of the USSR 26.04.1984, But for nutria these vaccines are not applied.

Methods of isolation and characterization of Salmonella described in the guidance "Laboratory diagnosis of Salmonella infections of man and animals, detection of Salmonella in feed, food products and objects of the external environment". Moscow, VO "Agropromizdat", 1990, as well as in the directory "Determinant zoopathogenic microorganisms under the editorship of Professor Sidorov M.A., Moscow, Kolos, 1995, str-204. In "Workshop on veterinary Microbiology and immunology" authors: Kostenko T.S., Rodionova V.B. have been, Skorodumov DI, M.: Kolos, 2001, str-231.

Methods of isolation of pure culture of the pathogen colibacillosis and characterization are described in the "guidelines for bacteriological diagnosis of colibacillosis (escherichiosis) animals"approved 27.07.2000, No. 13-7-2/2117 by the veterinary Department of the Ministry of agriculture of the Russian Federation, described the characteristics of Escherichia reference "Determinant zoopathogenic microorganisms under the editorship of Professor Sidorov M.A., Moscow, Kolos, 1995, str-201, as well as in the "Workshop on veterinary Microbiology and immunology" authors: Kostenko T.S., Rodionova V.B. have been, Skorodumov DI, M.: Kolos, 2001, str-222.

Methods of isolation and characterization of Streptococcus presented in "guidelines for laboratory diag is astice streptococcosis animals", approved 25.09.1990, Main Directorate of veterinary state Inspectorate under the State Commission of the USSR food and procurement, as well as in the directory "Determinant zoopathogenic microorganisms under the editorship of Professor Sidorov M.A., Moscow. - Spike, 1995, p.90-95.

The essence of the proposed method lies in the fact that direct the selection of the affected organs from dead nutria from local epizootic focus, prepare a suspension of the pathological material, do the planting on differential diagnostic environment, isolated pure cultures of infectious diseases colibacillosis, salmonellosis, streptococcosis and Enterococcus infection, separately grown pure cultures of pathogens Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae and Streptococcus fecalis in mesopatamia broth with the addition of glucose to 0.2%concentration with a title 4-5 billion microbial cells in 1 cm3that inactivate by adding formalin to 0.4 to 0.6%final concentration, incubated at 37 ° °C for 72-96 hours, mixed culture in equal proportions, then make a solution of aluminium hydroxide in the amount of 20% by volume of the culture, thoroughly mixed, Packed and sealed.

An example of a specific implementation of the method of manufacture of the vaccine.

For the manufacture of a vaccine associated is against colibacillosis, salmonellosis, streptococcosis and Enterococcus infection nutria shall conduct sampling of the affected organs from dead nutria during their disease colibacteriosis, salmonellosis and Enterococcus infection, of which prepare the suspension and conduct bacteriological research in order to obtain pure cultures of pathogens.

To determine the morphology and tinctorial properties of the pathogen colibacillosis prepare brushstrokes-prints of the affected organs of dead nutria by touching skim glass to fresh incision of the affected tissue and subsequent fixation over the flame of a spirit lamp, paint them using gram. When using the microscope preparations under oil immersion microscope system see thick sticks with rounded ends in red, located a single that is characteristic of E. coli.

To identify cultural properties of the pathogen colibacillosis nutria E.coli make the crops selected culture of pathological material in mesopotania broth (BCH), on Wednesday Endo, mastopathy agar (MPA) and incubated in a thermostat at 37°C for 16-24 hours. Petri-dish with MPA vyrostayut moist colonies with smooth edges and a smooth gray color. In test tubes with BCH watched uniform turbidity nutrient medium with small sediment crashing PR is shaking. In Petri dishes, on the environment Endo see the growth of colonies crimson red with rosavin the rim with a diameter of 2-3 mm, typical for E. coli.

When determining the biochemical activity of pure BackColor conduct crops on differential diagnostic environment with a different set of components. For E. coli is characterized by the breakdown of glucose and lactose with production of acid and gas, separation of indole, the absence of urease.

The pathogenicity of pure selected crops is determined by the intraperitoneal infection of white mice weighing 14-18, After 2-4 days of observation all infected laboratory animals die at 100% safety control. To confirm the specificity of the death of mice in the parenchymatous organs of dead mice prepare brushstrokes-prints, paint them using gram. When using the microscope under oil immersion microscope system in preparations visible thick sticks with rounded ends in red, located single characteristic of E. coli. Serological typing of Escherichia coli is carried out in agglutination reactions with a set typespecification sera according to the standard technique in test tubes.

In the agglutination reaction of the studied culture of Escherichia coli reacts with caliciviridae and visible flocculent precipitate (positive) negative control.

For definition wide-angle the morphology and tinctorial properties of the causative agent of salmonellosis prepare brushstrokes-prints of the affected organs of dead nutria, paint them using gram. When using the microscope preparations under oil immersion microscope system see thin sticks red, located a single, typical for the genus Salmonella.

To identify cultural properties of the causative agent of salmonellosis nutria make the crops selected culture of pathological material in mesopotania broth (BCH), on differential diagnostic environment (Endo, Levin, bismuth sulfite agar), mastopathy agar (MPA) and incubated in a thermostat at 37°C for 16-24 hours. Petri-dish with MPA vyrostayut smooth, clear, colorless colonies with smooth edges. In test tubes with BCH see diffuse clouding of the medium. In Petri dishes, on the environment Endo see clear, colorless colonies on the environment Levin - blue, bismuth sulfite agar - black colonies with a characteristic metallic sheen that is typical for the genus Salmonella.

When determining the biochemical activity of pure cultures of conduct crops on differential diagnostic environment with a different set of carbs and components.

Selected pure culture of the causative agent of salmonellosis characteristic of the genus Salmonella (ferments glucose, mannitol, doesn't remove lactose, sucrose, no splits urea, does not form indole, not thins gelatine)./p>

Pathogenicity and specificity of pure cultures is determined by the intraperitoneal infection of white mice weighing 14-18, After 2-4 days of observation all infected laboratory animals die at 100% safety control. To confirm the specificity of the death of white mice from parenchymatous organs of dead mice prepared brushstrokes-prints, paint them using gram. When using the microscope under oil immersion microscope system see thin sticks red, located a single, typical for the genus Salmonella.

To establish Serafimovo facilities selected pure cultures of Salmonella put the agglutination reaction (RA) on the glass according to the standard technique. Selected pure culture check with comprehensive and monoresistance O - and N-agglutinating sera in agglutination reactions. For selected pure culture grown on stubble MPA for 18-24 h at 37 ° °C. RA on the glass put with each O-comprehensive sera consistently until you obtain a positive result with two sera. A positive result see RA as difficulty falling flakes with a dedicated culture of Salmonella typhimurium.

To determine the morphology and tinctorial properties of the pathogen Enterococcus infections prepare brushstrokes-prints of paragen the x bodies of dead nutria, paint them using gram. Microscopic examination of preparations under oil immersion microscope system see small cocci in chains, painted in purple color characteristic of the genus Streptococcus. To determine the culture of the pathogen properties make the crops selected culture of material in mesopotania broth with the addition of glucose to 0.2%final concentration and 10% inactivated horse serum (BCH), in Petri dishes with mesopartner agar (MPA) with the addition of adding glucose to 0.2%final concentration) and 5% defibrinating blood of the rabbit. After 18-20 hours of incubation in a thermostat at a temperature of 37°see in BCH growth in the form of a diffuse cloud environment. In Petri dishes with blood MPA see the growth of small rosinate, slightly hazy with smooth edges colonies, surrounded by a greenish zone of hemolysis, which is characteristic of Streptococcus fecalis. On Enterococcus differential diagnostic environment through 14-18 hours incubation see the growth of colonies cherry-red color, which is characteristic of the pathogen Enterococcus infection Str. fecalis.

For differentiation of streptococci from staphylococci using the catalase test. When setting the catalase sample on a glass slide put a drop of freshly prepared 3%hydrogen peroxide solution. Bacteriological loop is removed a colony of the studied culture and grind it in a drop of hydrogen peroxide. Staphylococci, unlike streptococci form catalase and cause foaming. For preliminary identification of streptococci and enterococci bakultala sown in test tubes with 10%, 40% bile, with 6.5% sodium chloride, carbohydrates (raffinose, sorbitol, mannitol), take into account the growth in liquid medium and hemolysis on MPA with blood.

As a result, in test tubes with bile see diffuse growth backcountry note fermentation of mannitol. On MPA with blood see incomplete hemolysis of erythrocytes with the formation around the colonies of the green zone, which is typical for streptococcal group D. To establish serogrouping facilities streptococcal use streptococcal group precipitating serum. Serogrouping belonging streptococcal determine the reaction of precipitation (RP) in the capillaries. The confirm pathogenicity bioassays on young white mice.

Selected so pure cultures of pathogens Escherichia coli colibacillosis, salmonellosis Salmonella typhimurium, Streptococcus Streptococcus pneumoniae and Enterococcus infection Streptococcus fecalis used for bacteria. The cultivation of pure cultures of pathogens colibacillosis, salmonellosis, streptococcosis and Enterococcus the infection is carried separately in mesopatamia broth.

When obtaining a pure culture of the pathogen colibacillosis the La intrusion take 1-2 cm 3fresh culture of E. coli and sow 80-100 cm3liquid nutrient medium and grown at 37°C for 12-14 hours. The concentration of microbial cells in the bacterial mass is determined from the turbidity standard and diluted with sterile distilled water to a concentration of 5-6 billion in 1 cm3. Inactivation obtained bacteria spend making formalin 0.5 to 0.6%final concentration, incubated at 37 ° °C for 72-96 hours with occasional stirring.

When obtaining a pure culture of the pathogen Salmonella to infect take 5 cm3fresh pure culture of the causative agent of salmonellosis Salmonella typhimurium at 1000 cm3mesopatania nutrient medium with the addition of glucose to 0.2%concentration and grown at 37°C for 12-14 hours. The concentration of microbial cells in the bacterial mass is determined from the turbidity standard and diluted with sterile distilled water to a concentration of 4-5 billion in 1 cm3. Inactivation obtained bacteria spend making formalin of 0.4-0.6%final concentration, incubated at 37 ° °C for 72-96 hours with occasional stirring.

When obtaining a pure culture of the pathogen Streptococcus for infection take 5 cm3pure cultures of the pathogen Streptococcus Streptococcus pneumoniae 1000 cm3liquid Pete is positive environment with the addition of 0.2% glucose and grown at 37° C for 12-14 hours. The concentration of microbial cells in the bacterial mass is determined by the standard and diluted with sterile distilled water to a concentration of 4-5 billion in 1 cm3. Inactivation bacteria carried out separately by the introduction of formalin of 0.4-0.6%final concentration, incubated at 37 ° °C for 72-96 hours with occasional stirring.

When obtaining a pure culture of the pathogen Enterococcus for infection to take 5 cm3pure cultures of the pathogen Enterococcus infection Streptococcus fecalis 1000 cm3liquid nutrient medium with the addition of 0.2% glucose and grown at 37°C for 12-14 hours. The concentration of microbial cells in the bacterial mass is determined by the standard and diluted with sterile distilled water to a concentration of 4-5 billion in 1 cm3. Inactivation bacteria carried out separately by the introduction of formalin of 0.4-0.6%final concentration, incubated at 37 ° °C for 72-96 hours with occasional stirring, mix inactivated backcountry in equal proportions. Then to the mixture of BackColor add a solution of aluminium hydroxide in the amount of 20% by volume and thoroughly mixed, Packed and sealed.

Thus, the use for the manufacture of a vaccine associated against colibacillosis, salmonellosis is streptococcosis and Enterococcus infection nutria pure cultures of pathogens Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae and Streptococcus fecalis, isolated from local epizootic focus, allows you to get more specific, highly immunogenic associated vaccine to protect against these three infectious diseases: colibacillosis, salmonellosis, streptococcosis and Enterococcus infection nutria. Separate cultivation of pathogens with a content of 0.2% glucose allows the concentration of bacterial cells of at least 4 billion in 1 cm3within 12-14 hours of incubation. The use of formalin to inactivate 0.4 to 0.6%final concentration allows for 72-96 hours at a temperature of 37°to suppress the pathogenicity and to maintain a high immunogenicity within 12 months of storage. The adsorption is carried three antigens on aluminium hydroxide allows through 12-15 days after a single injection at a dose of 1 cm3the nutria is to provide in vaccinated nutria formation of a busy immunity against colibacillosis, salmonellosis, streptococcosis and Enterococcus infection duration of at least 9 months (observation period). After a single injection of the nutria intramuscularly causes no vaccine-related complications (table).

The proposed method is simple, affordable, and is industrially applicable.

td align="center"> 6 months
Table
Comparative characteristic of making a vaccine of the proposed method and the prototype
№ p/pDistinctive featuresThe placeholderThe proposed method
1PathogensStrains of Streptococcus zooepidemicus VGNKI # K-DEPT, Pasteurella multosida VGNKI No. 2394 and Pasteurella multocida VGNKI No. 1015Clean backcountry pathogens Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae and Streptococcus fecalis allocated from the local epizootic focus
2put zamorian-defrosting, separating the liquid fractionIt is necessarilyNot done
3Inactivation of bacterial mass0,3-0,4%solution of formalin, incubated for 44-50 h0,4-0,6%concentration of formalin within 72-96 hours at a temperature of 37°
4Adjuvant3%solution of aluminium hydroxide in the amount of 10% by volumealuminium hydroxide 20% by volume
5Postvaccinal complicationsThe nutria after the vaccine chromate, inflammatory processesNo
6The duration of immunity9 months

Thus, these tables show the advantages of the proposed method of making a vaccine associated against colibacillosis, salmonellosis, streptococcosis and Enterococcus infection nutria compared with the existing prototype.

A method of manufacturing a vaccine associated against colibacillosis, salmonellosis, streptococcosis and Enterococcus infection nutria, which consists in the fact that undertaking the selection of the affected organs from dead nutria from local epizootic hearth, preparing a suspension of pathological material, do the planting on differential diagnostic environment, isolated pure culture of Escherichia coli, Salmonella typhimurium, Streptococcus pneumoniae and Streptococcus fecalis, separately grown cultures in meat-peptone culture medium with addition of 0.2% glucose to achieve a concentration of microbial cells - 4-5 billion in 1 cm3that inactivate formalin to 0.4 to 0.6%final concentration, incubated at 37 ° °C for 72-96 h, mixed culture in equal proportions, make a 3%solution of aluminium hydroxide in the amount of 20% by volume of the culture and thoroughly mixed, after which the vaccine is Packed and sealed.



 

Same patents:

FIELD: veterinary medicine.

SUBSTANCE: the vaccine suggested contains as antigens: the cell suspension of pure cultures of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis obtained due to selecting the affected organs from dead nutrias out of local epizootic focus, preparing the suspension, inoculation onto differential-diagnostic media, isolating pure cultures of the agents of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis. The obtained pure cultures of microorganisms should be separately grown in beef-extract broth with glucose up to the concentration of microbial cells being 4-5 bln/cu. cm to be then mixed at equal ratios. The vaccine, also, contains formalin, glucose and aluminum hydroxide at the following ratio of the components, weight%: cell suspension of Escherichia coli pure culture isolated from the affected organs out in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0; cell suspension of Salmonella typhimurium pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, cell suspension of Streptococcus pneumoniae pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu/ cm 18.0-21.0 cell suspension of Streptococcus fecalis pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, glucose 2.0-1.0; formalin 2.0-1.5; aluminum hydroxide - the rest. The vaccine in question is of high specificity, safety and immunogenicity.

EFFECT: higher efficiency.

5 ex, 1 tbl

FIELD: microbiology, in particular production of associated pertusis, diphtheria, and tetanum vaccine.

SUBSTANCE: claimed method includes blending of pertusis component, diphtheria and tetanum anatoxins; mixture sorption on aluminum hydroxide; mixture dosage into container, mixture lyophilizing and container sealing. As pertusis component cell-free pertusis anatoxin in concentration of 40-50 mug/ml is used. Mixture of cell-free pertusis anatoxin, diphtheria and tetanum anatoxins is sorbed on aluminum hydroxide. Then polyoxydonium as immunomodulator in amount of 0,5-0,52 mug/g in introduced into abovementioned mixture, mixture is held for 18-20 h at 4-10°C, sucrose is added up to finish concentration and mixture is held for 1 h at 4-10°C.

EFFECT: vaccine of decreased reactogenicity.

2 ex, 1 tbl

FIELD: microbiology.

SUBSTANCE: the resent innovation deals with mixing pertussis component, diphtheric and tetanus anatoxins and sorption of the mixture upon aluminum hydroxide followed by dosing the mixture into the tank, freeze drying the mixture and hermetic sealing of this tank, moreover, as pertussis component one should apply cell-free pertussis anatoxin; as for mixing cell-free pertussis anatoxin, diththeric and tetanus anatoxins it should be carried out simultaneously at availability of phosphate-buffered physiological solution; the mixture obtained should be kept for 1 h at about 4-10°C with subsequent sorption upon aluminum hydroxide; just before freeze drying the mixture should be supplemented with saccharose up to final concentration being 9-10.5%, then the mixture should be mixed and kept at about 4-10C for 1 h. The innovation enables to achieve decreased reactogeneity.

EFFECT: higher efficiency.

2 ex, 4 tbl

FIELD: veterinary microbiology, biotechnology.

SUBSTANCE: vaccine comprises inactivated purified chlamydium antigens from strains of species Chlamydia psittaci: MZ-89 - a pathogen of chlamydiosis meningoencephalitis in calves, 250 - a pathogen of chlamydiosis abortion in cows and SK-89 - a pathogen of chlamydiosis conjunctivitis in calves, These strains are grown in chicken embryo yolk and allantois envelopes simultaneously and taken in the ratio 0.25:0.25:0.25, respectively, and in the concentration of chlamydia 16-18 mg/ml adsorbed on potash alum and emulsified in oily-lanoline adjuvant with the content of 0.2% glutaraldehyde as a preserving agent. Proposed vaccine possesses broad immunogenic activity and broad spectrum of antigenicity in different clinical forms of diseases in cattle and sheep and goats.

EFFECT: enhanced and valuable properties of vaccine.

9 tbl, 4 ex

FIELD: biotechnology.

SUBSTANCE: vaccine comprises as antigens the cell suspension of pure cultures of pathogens of coli-bacteriosis Escherichia coli, salmonellosis Salmonella typhimurium and streptococcosus Streptococcus pneumoniae obtained by selection of damaged organs from dead nutrias and inoculation of cultures on differential-diagnostic media. Isolated pure cultures are grown separately in beef-extract broth with glucose to the concentration of microbial cells 4-5 billions in 1 cm3 followed by their mixing in equal ratio. Vaccine comprises formalin and aluminum hydroxide in the following ratio of components, wt.-%: cell suspension of pure culture of coli-bacteriosis pathogen Escherichia coli isolated from a local epizootic focus in nutrient medium with a titer 4-5 billions of microbial cells in 1 cm3, 24.0-29.0; cell suspension of pure culture of salmonellosis pathogen Salmonella typhimurium isolated from a local epizootic focus in nutrient medium with a titer 4-5 billions of microbial cells in 1 cm3, 24.0-29.0; cell suspension of pure culture of streptococcosus pathogen Streptococcus pneumonia isolated from a local epizootic focus in nutrient medium with a titer 4-5 billions of microbial cells in 1 cm3, 24.0-29.0; glucose, 1.0-2.0; formalin, 1.5-2.0, and aluminum hydroxide, the balance. Proposed vaccine shows high immunogenicity, high specificity and harmless.

EFFECT: valuable properties of vaccine.

1 tbl, 5 ex

FIELD: veterinary medicine.

SUBSTANCE: vaccine comprises as antigens cell suspension of pure cultures of a streptococcosus pathogen Streptococcus pneumoniae and an enterococcal pathogen Streptococcus fecalis isolated from suspension and prepared from damages organs of dead nutrias from the local epizootic focus wherein these cultures are grown separately in beef-extract broth with glucose with the concentration of microbial cells 4-5 billions in 1 cm3 and mixed in equal ratio. Also, vaccine comprises formalin and aluminum hydroxide in the following ratio of components, wt.-%: cell suspension of pure culture of streptococcosus pathogen Streptococcus pneumoniae isolated from a local epizootic focus in the nutrient medium with a titer 4-5 microbial cells in 1 cm3, 38.0-41.5; cell suspension of pure culture of enterococcal infection pathogen Streptococcus fecalis isolated from a local epizootic focus in the nutrient medium with a titer 4-5 billions of microbial cells in 1 cm3, 38.0-41.5; glucose, 1.0-2.0; formalin, 1.5-2.0, and aluminum hydroxide, the balance. Vaccine is harmful and possesses high specificity and effectiveness.

EFFECT: valuable properties of vaccine.

1 tbl, 5 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: vaccine comprises cell suspension of pure cultures of Escherichia coli as coli-bacteriosis pathogen, Salmonella typhimurium as salmonellosis pathogen and Streptococcus faecalis as enterococcus pathogen as antigens and prepared by selection of damaged organs from dead nutrias from local epizootic focus, preparing suspension, inoculation in differential diagnostic media, isolation of pure cultures of pathogens. Their culturing is carrying out separately in beef-extract broth with glucose up to the concentration 4-5 billions microbial cells/1 cm3 and mixed in equal ratios. Vaccine comprises formalin and aluminum hydroxide in the following ratio of component, wt.-%: cell suspension of pure culture of Escherichia coli as pathogen of coli-bacteriosis isolated from local epizootic focus in nutrient medium with titer value 4-5 billions microbial cells/1 cm3, 24.0-29.0; cell suspension of pure culture of Salmonella typhimurium as pathogen of salmonellosis isolated from local epizootic focus in nutrient medium with titer value 4-5 billions microbial cells/1 cm3, 24.0-29.0; cell suspension of pure culture of Streptococcus faecalis as pathogen of enterococcus infection isolated from local epizootic focus in nutrient medium with titer value4-5 billions microbial cells/1 cm3, 24.0-29.0; glucose, 1.0-2.0; formalin, 1.5-2.0, and aluminum hydroxide, the balance. Vaccine shows high immunogenicity, high specificity and safety.

EFFECT: valuable properties of vaccine.

1 tbl, 5 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: invention relates to a method for preparing harmless, specific, highly immune, inactivated, mixed vaccine against coli-bacteriosis, salmonellosis and streptococcosis in nutria wherein method involves using antigens, inactivation and addition of adjuvant. Method involves selection of damaged organs from dead nutrias in local epizootic focus and preparing suspension from these organs followed by inoculation in differential diagnostic media. Pure cultures of coli-bacteriosis, salmonellosis and streptococcosis pathogens are isolated. Cultures Escherichia coli, Salmonella typhimurium and Streptococcus pneumoniae are grown separately in beef-extract broth with addition of 0.2% of glucose and with titer value 4-5 billions microbial cells/1 cm3. Inactivation is carried out by addition of formalin in the final concentration 0.4-0.6%. Then cultures are kept at temperature 37°C for 72-96 h and mixed their in equal ratio. Then aluminum hydroxide solution is added in the amount 20% to volume, stirred thoroughly, packaged and sealed. Method provides preparing harmless, immunogenic and highly specific vaccine.

EFFECT: improved preparing method.

1 tbl, 1 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: method involves selection of damaged organs from dead nutrias from local epizootic focus and preparing suspension from these organs. Then method involves inoculation in differential diagnostic media and isolation of pure cultures of pathogens. Cultures Streptococcus pneumoniae and Streptococcus faecalis are grown in beef-extract broth with addition glucose to final 0.2% concentration and with titer value 4-5 billions microbial cells/1 cm3 and inactivated by addition of formalin to final concentration 0.4-0.6% and kept at temperature 37°C for 72-96 h. Cultures are mixed in equal ratios, aluminum hydroxide solution is added in the amount 15% to volume, stirred thoroughly and sealed. Method provides preparing harmless, highly immune and highly specific vaccine.

EFFECT: improved preparing method.

1 tbl, 1 ex

FIELD: veterinary medicine.

SUBSTANCE: for manufacturing an associated vaccine it is necessary to select affected organs in dead nutrias from local epizootic focus for the purpose to prepare the suspension and fulfill inoculation onto differentially-diagnostic media. One should isolate pure cultures and separately grow the cultures of salmonellosis agents Salmonella typhimurium and Escherichia coli in beef-extract broth at addition of 0.2%-glucose at the titer being about 4-5 bln. microbial cells/cu. cm followed by inactivation due to applying formalin up to 0.4-0.6%-final concentration to be kept at 37°C for about 72-96 h. Cultures should be mixed at equal ratios, then aluminum hydroxide solution should be applied at the quantity of 20% against the culture volume to be then thoroughly mixed, packed and sealed. The innovation enables to obtain safe, specific and high-immunogenic vaccine.

EFFECT: higher efficiency.

1 ex, 1 tbl

FIELD: veterinary medicine.

SUBSTANCE: the vaccine suggested contains as antigens: the cell suspension of pure cultures of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis obtained due to selecting the affected organs from dead nutrias out of local epizootic focus, preparing the suspension, inoculation onto differential-diagnostic media, isolating pure cultures of the agents of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis. The obtained pure cultures of microorganisms should be separately grown in beef-extract broth with glucose up to the concentration of microbial cells being 4-5 bln/cu. cm to be then mixed at equal ratios. The vaccine, also, contains formalin, glucose and aluminum hydroxide at the following ratio of the components, weight%: cell suspension of Escherichia coli pure culture isolated from the affected organs out in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0; cell suspension of Salmonella typhimurium pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, cell suspension of Streptococcus pneumoniae pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu/ cm 18.0-21.0 cell suspension of Streptococcus fecalis pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, glucose 2.0-1.0; formalin 2.0-1.5; aluminum hydroxide - the rest. The vaccine in question is of high specificity, safety and immunogenicity.

EFFECT: higher efficiency.

5 ex, 1 tbl

FIELD: veterinary, vaccines.

SUBSTANCE: claimed vaccine contains as antigens pure culture cell suspension of causal organisms of salmonellosus Salmonella typhimurium and enterococcus infection Streptococcus fecalis, obtained by sampling of damaged organs of fallen nutrias from local epizootic focus, suspension preparation, seeding on differential diagnostic media, isolation of causal organism pure cultures. Culturing is carried out separately in meat-and-peptone broth with glucose at microbial cell concentration of 4-5 bln per 1 cm3, cells are mixed in ratio of 1:2, formalin and aluminum hydroxide are introduced in the next component ratio (mass %): pure culture cell suspension of causal organisms of salmonellosus Salmonella typhimurium isolated from local epizootic focus in broth with titer of 4-5 bln per 1 cm3 22.5-27.5; pure culture cell suspension of causal organisms of enterococcus infection Streptococcus fecalis isolated from local epizootic focus in broth with titer of 4-5 bln per 1 cm3 50.5-55.0-27.5; glucose 2.0-1.0; formalin 2.0-1.5; and balance: aluminum hydroxide.

EFFECT: associated vaccine with increased specificity and effectiveness without negative side effects.

1 tbl, 5 ex

FIELD: veterinary science.

SUBSTANCE: down-calving cows should be immunized simultaneously with vaccine "Coli-Vac C99" and formulated alum vaccine against salmonellosis in calves at 10-d-long interval 1.5 mo before calving. The first dosage corresponds to 10 cu. cm, the second one - 15 cu. cm. Simultaneously with vaccination it is necessary to inject Ligavirin for cows at the dosage of 5 ml/cow. As for calves born in these cows they should be once injected intramuscularly with Ligavirin for 0.5-1 h after their birth at the dosage of 1 ml/calf. The innovation enables to create more tense specific immunity in cows before calving and colostral immunity in calves born in these cows.

EFFECT: higher efficiency of prophylaxis.

3 ex, 5 tbl

FIELD: veterinary medicine.

SUBSTANCE: claimed method includes sampling of pathological material from fallen coypu from local epizootic focus and preparation of suspension therefrom. Suspension is seeded on differential diagnostic media, and pure culture of salmonella typhimurium is isolated. Pure culture is cultivated in liquid broth with addition of 0.2 % of glucose to produce microbial cell concentration of 4-5x109 per 1 cm3. Culture is inactivated with formalin up to finish concentration of 0.4-0.6 % and conditioned at 37°C for 82-96 hours. Then 3 % solution of aluminum hydroxide is introduced in amount of 15 % based on culture volume, mixture is thoroughly agitated, pre-packed and sealed.

EFFECT: specific vaccine of high immunogenic properties.

1 tbl, 1 ex

FIELD: chemical and pharmaceutical industry, in particular vaccine against coypu salmonelasis.

SUBSTANCE: claimed vaccine contains suspension of pure culture cells of Salmonella typhimurium causative agent, obtained by sampling of pathological material from fallen coypus of local epidemical locus, suspension preparation, sowing on differential diagnostic media, isolation of Salmonella typhimurium causative agent pure culture, and culturing in meat-peptone broth with titer of 4-5 bln of microbial cells per 2 cm3. Vaccine also contains glucose, formalin and aluminum hydroxide in the next mass ratio: abovementioned pure culture cells of Salmonella typhimurium causative agent 83.0-86.0; glucose 1.0-2.0; formalin 1.0-2.0; and balance: aluminum hydroxide.

EFFECT: non-toxic vaccine having high reactivity.

5 ex, 1 tbl

FIELD: biotechnology.

SUBSTANCE: the suggested method deals with growing Salmonella typhimurium culture, washing with sterile 0.9%-NaCl solution (pH 7.0), treating with 10%-aqueous ammonia solution up to final concentration of 1% and incubating the mixture obtained for 5 h, then centrifuging takes place of salmonella suspension, then one should pour out supernatant to resuspend the residue with sterile 0.9%-NaCl solution (pH 7.0). The innovation suggested enables to obtain inactivated Salmonella typhimurium culture that causes no lethality at subcutaneous injection in mice.

EFFECT: higher efficiency of inactivation.

FIELD: biotechnology, microbiology, medicine, veterinary science.

SUBSTANCE: for preparing vaccine toxigenic strains of S. dysenteriae R-forms are grown, cells are subjected for lysis by treatment with chloroform, mixture is centrifuged and prepared supernatant is treated with saturated monobasic carboxylic acid or their derivatives and pH value is brought about to 3.0-5.0. Mixture if centrifuged and precipitate containing corpuscular antigens and shigellosis exotoxin are obtained. Precipitate is dissolved in buffer and pH value is brought about to 7.5-9.0. Then formalin is added in the amount 0.4-0.8% of the solution volume, or benzoic acid or benzoic acid salts are added in the amount 0.07-4.0% of the solution volume, or a mixture consisting of formalin and benzoic acid or benzoic acid salt solutions in the amount 0.10.3% and 0.03-2.5% of the solution volume, respectively. The solution is kept at temperature 30-60°C for 2 h - 60 days to provide the conversion of exotoxin to anatoxin and vaccine is prepared. Another variant of the claimed invention involves additional treatment with formalin or benzoic acid or benzoic acid salts to provide conversion of exotoxin to anatoxin, vaccine is prepared followed by its bagging and corking. For preparing immunoglobulin preparation animals are immunized with vaccine prepared by abovementioned methods followed by taking off blood, milk and/or colostrums, and/or blood, immunoglobulin fraction is prepared, sterilized, bagged and corked. This preparation is a component of the immunobiological preparation. The immunobiological preparation comprises the immunoglobulin preparation and at least one component taken among the following row: human and/or animal immunoglobulin preparations, lactoferrin, enzymes, inhibitors of proteolytic enzymes, human and/or animal normoflora preparations, yeast, vitamins, vitamin-like substances, human and/or animal proteins of acute phase, human and/or animal cytokines, higher plants components, lower plants components, components of natural origin products, apiculture products, enterosorbents, antibiotics, antibacterial chemopreparations, sulfanilamide drugs, antibacterial, anti-tuberculosis, antiviral preparations, antifungal antibiotics, synthetic antifungal preparations, stimulators of metabolic processes, antioxidants, mineral supplements, carbohydrates, lipids, replaceable and/or essential amino acids, organic acids, alkaloids, glycosides, taste supplements, aromatic supplements, base for suppositories, base for ointment formulations, technological additives for tableting, or their mixture. Invention provides preparing preparations eliciting antigenic activity with respect to broad species of pathogenic and opportunistic gram-negative microorganisms of intestine group and their exotoxins and therefore eliciting with prophylactic and curative effect with respect to diseases causing with these microorganisms.

EFFECT: improved preparing method, valuable properties of vaccine.

13 cl, 112 ex

FIELD: veterinary science.

SUBSTANCE: vaccine against porcine salmonellosis contains cell suspension of attenuated vaccine strain Salmonella choleraesuis VGNKI SC 230-DEP in the following ratio of components, vol.-%: suspension of strain Salmonella choleraesuis VGNKI SC 230-DEP with concentration 100-120 billion cells in cm3, 42.0-55.0; sucrose, 5.0-7.5; gelatin, 1.5-2.0, and distilled water, the balance. Morbidity of piglets immunized with vaccine is reduced 2-fold and their safety is by 8.6% higher as compared with the same indices for piglets immunized with the known vaccine from the strain Salmonella choleraesuis VGNKI № 9. Vaccine shows enhanced economy in producing and exhibits stable genetic markers.

EFFECT: improved, enhanced and valuable properties of vaccine.

5 tbl, 8 ex

FIELD: biotechnology, genetic engineering, molecular biology.

SUBSTANCE: invention proposes recombinant DNA pcDNA-TCI that has molecular mass 4.3 x 103 kDa and size 6 657 nucleotide pairs. The proposed recombinant plasmid DNA provides expression of artificial gene TCI comprising cytotoxic T-cellular epitopes of HIV-1 in eucaryotic cells. Also, invention proposes recombinant attenuated strain of bacterium Salmonella enteritidis E-23 pcDNA-TCI. The proposed strain is obtained by genetic transformation of the strain Salmonella enteritidis E-23 with plasmid pcDNA-TCI. The strain provides delivery DNA-vaccine in eucaryotic cells as the recombinant plasmid DNA pcDNA-TCI. Proposed groups of inventions provide inducing the expressed specific humoral and cellular immune response to HIV-1 in body. Proposed group of inventions can be used in medicine and virology for construction of live DNA-vaccine against HIV-1.

EFFECT: valuable biological and medicinal properties of strain and vaccine.

2 cl, 3 dwg, 3 tbl, 5 ex

FIELD: poultry science.

SUBSTANCE: one should irradiate eggs before incubation with the light of He-Ne laser LHN-104, wave length being 632.8 nm, power density at eggs surface being 50 mW/sq.cm/sec, with red light of fluorescent lamp DNESG-500, wave length being 630-650 nm, power at eggs surface being 23.1 erg and with ultraviolet from DRT-400 lamp at wave length being 185-400 nm, average dosage at eggs surface being 20 merg in 3-min-long exposures.

EFFECT: higher resistance to pullorosis in poultry.

4 dwg, 4 tbl

FIELD: veterinary medicine.

SUBSTANCE: the vaccine suggested contains as antigens: the cell suspension of pure cultures of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis obtained due to selecting the affected organs from dead nutrias out of local epizootic focus, preparing the suspension, inoculation onto differential-diagnostic media, isolating pure cultures of the agents of Escherichia coli, Salmonella typhimurium and Streptococcus fecalis. The obtained pure cultures of microorganisms should be separately grown in beef-extract broth with glucose up to the concentration of microbial cells being 4-5 bln/cu. cm to be then mixed at equal ratios. The vaccine, also, contains formalin, glucose and aluminum hydroxide at the following ratio of the components, weight%: cell suspension of Escherichia coli pure culture isolated from the affected organs out in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0; cell suspension of Salmonella typhimurium pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, cell suspension of Streptococcus pneumoniae pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu/ cm 18.0-21.0 cell suspension of Streptococcus fecalis pure culture isolated from the affected organs in dead nutrias out of local epizootic focus in nutritive medium at the titer being 4-5 bln microbial cells/cu. cm 18.0-21.0, glucose 2.0-1.0; formalin 2.0-1.5; aluminum hydroxide - the rest. The vaccine in question is of high specificity, safety and immunogenicity.

EFFECT: higher efficiency.

5 ex, 1 tbl

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