RU2283137C2 - Inactivated emulsified vaccine against bird infectious bursal disease - Google Patents

Abstract

FIELD: veterinary virology, biotechnology.

SUBSTANCE: claimed vaccine contains antigenic material from virus strain of bird infectious bursal disease (IBD), ethylene imine as inactivation agent; and oily adjuvant in effective ratio. Claimed strain is reproduced on 9-11-days embryos of SPF-hens with infective activity of 5.25-7.0 lg EID_{50/0.2 cm} ³. Vaccine represents homogenous white emulsion containing inactivated antigen of IBD virus emulsified in oily adjuvant and is administered to healthy bird being 110-120 days old in volume of 0.5 cm², in single dose. Vaccine of present invention induces high levels of antibodies against IBD of parent bevy and provides anchored passive immunity in 15-25-day young birds.

EFFECT: safe vaccine against bird infectious bursal disease.

8 cl, 5 tbl, 6 ex

Description

The invention relates to the field of veterinary virology and biotechnology and can be used in the development and manufacture of means for the specific prophylaxis of infectious bursal disease (IBD) in birds.

IBD of birds (infectious bursitis, Gumboro disease) is a widespread highly contagious viral disease of chickens, mainly aged 15-80 days, characterized by damage to the factory bag, kidneys, intramuscular hemorrhages and diarrhea. IBD affects mainly (3-lymphocytes of the factory bag, causing atrophy of its lymphoid follicles.

IBD is acute, with a characteristic clinic of the disease, and subclinically. The subclinical form is widespread.

In the acute form of the disease, clinical and pathological signs characteristic of secondary infections come to the fore, which creates difficulty in making a diagnosis. Therefore, laboratory studies are crucial in diagnosing the disease.

IBD birds registered in all countries of the world. In recent years, IBB has become widespread in most large poultry farms in Russia. The use of domestic and imported biological products in foci of infection did not provide the expected effect. This is because their use was not preceded by a study of the antigenic properties of circulating virus isolates, specific antibodies of the bird before and after vaccination, and the level of passive maternal immunity in chickens were not determined. The well-known feature of the IBD pathogen was not taken into account - its antigenic variation within the same serotype, occurring at different time intervals and in different territories and depending on the species and species composition of the bird population, its immune status, and many other factors.

In this regard, the choice of the vaccine strain of the IBD virus of birds should be carried out taking into account the antigenic structure of the epizootic isolate and for the preparation of vaccine preparations it is necessary to use production strains that have the highest degree of homology in the field of antigenic determinants with epizootic isolates of the IBD virus.

The economic damage from IBD caused by the death of up to 20-70% of the number of chickens in the acute course of the disease, significant culling of poultry, decreased productivity, and also as a result of secondary infections, is very large.

Currently, it is generally recognized that the only right and reliable direction in the fight against infection is vaccination. For specific prophylaxis of IBD, live and inactivated vaccines are used from homologous strains of the pathogen grown in various sensitive biological systems.

In most poultry farms, chickens are inoculated with 2-fold live vaccines, and revaccinated at 16–20 weeks of age with an inactivated vaccine. This is due to the fact that with IBD, the introduction of live vaccines to an adult bird does not form intense immunity in it, which is associated with its age-related features. For this reason, inactivated vaccines, both sorbed and emulsion, are widely used for specific prevention of IBD in an adult bird (1).

In the manufacture of an inactivated vaccine, virulent strains of the IBD virus are used.

Known inactivated emulsion vaccine against IBD birds containing antigenic material homologous to the pathogen infection strain BIA 52/70, obtained in a sensitive biological system and subjected to inactivation, inactivant and oil adjuvant in an effective ratio. As a sensitive biological system, fabric of fabric bags of 30-40 day old chickens was used (2).

Known inactivated emulsion vaccine against IBD birds containing antigenic material homologous to the pathogen infection strain BIA 52/70, obtained in a sensitive biological system and subjected to inactivation, inactivant and oil adjuvant in an effective ratio. The allantoic membranes of chicken embryos were used as a sensitive biological system (3).

Known inactivated emulsion vaccine against IBD birds containing antigenic material homologous to the causative agent of infection of strain D78, ATCC No. VR-2041, obtained in a sensitive biological system and subjected to inactivation, inactant and oil adjuvant in an effective ratio.

As a sensitive biological system, 9–11-day-old SPF-chicken embryos were used (4).

Known inactivated emulsion vaccine against IBD birds containing antigenic material homologous to the pathogen infection strain GP / 82, NCAIM No. V (P) -001033, obtained in a sensitive biological system and subjected to inactivation, inactivant and oil adjuvant in an effective ratio. As a sensitive biological system, 10-day-old embryos of SPF hens were used (5).

Known inactivated emulsion vaccine against IBD birds containing antigenic material homologous to the pathogen infection strain BIA 52/70, obtained in a sensitive biological system and subjected to inactivation, inactivant and oil adjuvant in an effective ratio. As a sensitive biological system, fabric of fabric bags of SPF chickens of 30-40 days old was used (6).

The main disadvantage of the known vaccines is that they are imported drugs and do not provide satisfactory protection against the IBD virus circulating in the poultry stock in the Russian Federation.

Also known is an inactivated sorbed vaccine against IBD in birds, containing antigenic material homologous to the causative agent of infection of the strain "BG" No. 102-DEPT ", obtained in a sensitive biological system and subjected to inactivation, inactivate and adjuvants aluminum hydroxide and saponin in an effective ratio.

As a sensitive biological system, 10-day-old embryos of SPF chickens were used (7).

The disadvantage of this vaccine is its low antigenic and immunogenic activity.

The closest to the invention according to the set of essential features is an inactivated emulsion vaccine against IBD birds, containing antigenic material from the D78 strain homologous to the pathogen, ATCC No. VR-2041, obtained in a sensitive biological system and subjected to inactivation, an inactant and an oil adjuvant in an effective ratio. As a sensitive biological system, 9–11-day-old SPF-chicken embryos were used (4).

The disadvantage of the prototype vaccine is that it has low antigenic and immunogenic activity

The objective of the present invention was to develop an inactivated emulsion vaccine based on a new strain of the IBD virus of birds, which has higher antigenic and immunogenic activity and is harmless in its native form and after inactivation.

The technical result from the use of the invention is to increase the antigenic and immunogenic activity and harmlessness of the proposed vaccine.

The specified technical result was achieved by the creation of an inactivated emulsion vaccine against IBD birds, characterized by the following set of features.

1. Inactivated emulsion vaccine against IBD birds.

2. Antigenic material from a strain homologous to the pathogen, obtained in a sensitive biological system and subjected to inactivation, in an effective amount.

3. Antigenic material from the strain "K-58 No. 122-DEPT" virus IBD birds.

4. As a sensitive biological system using 9-11-day-old embryos of SPF-chickens.

5. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 Ig EID _{50 / 0.2cm} ³ and subjected to inactivation.

6. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 Ig EID ₅₀ / _0.2cm ³ and subjected to inactivation, in the amount of 29.9-49.7 wt.%.

7. Inactivant.

7.1. Of the activants, ethyleneimine dimer (DEI).

7.2. DEI in the form of a 15% aqueous solution.

7.3. DEI in an amount of 0.1-0.3 wt.%.

8. Oil adjuvant.

8.1. From oil adjuvants oil adjuvant ARRIAH.

8.2. Oil adjuvant ARRIAH in the amount of 50.0-70.0 wt.%.

8.3. Of the oil adjuvants, the Montanide ISA-70 oil adjuvant.

8.4. Montanide ISA-70 oil adjuvant in an amount of 50.0-70.0 wt.%.

8.5. Of the oil adjuvants, the Montanide ISA-206 oil adjuvant.

8.6. Montanide ISA-206 oil adjuvant in an amount of 50.0-70.0 wt.%.

9. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 Ig EID _{50 / 0.2cm} ³ and subjected to inactivation, inactivating DEI and oil adjuvant in the ratio, wt.%:

Antigenic material 29.9-49.7 DEI 0.1-0.3 Oil adjuvant 50.0-70.0

The present invention contains the following set of essential features that provide a technical result in all cases for which legal protection is sought.

1. Inactivated emulsion vaccine against IBD birds.

2. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in a sensitive biological system and subjected to inactivation in an effective amount.

3. Inactivant.

4. Oil adjuvant.

The present invention is also characterized by other features expressing specific forms of execution or special conditions for its use.

1. Antigenic material from strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.2 Ig EID _{50 / 0.2 cm} ³ and subjected to inactivation.

2. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 Ig EID _{50 / 0.2cm} ³ and subjected to inactivation, in the amount of 29.9-49.7 wt.%.

3. Of the inactivants of DEI.

4. DEI in the form of a 15% aqueous solution.

5. DEI in an amount of 0.1-0.3 wt.%.

6. From oil adjuvants oil adjuvant ARRIAH.

7. Oil adjuvant ARRIAH in the amount of 50.0-70.0 wt.%.

8. Of the oil adjuvants, the Montanide ISA-70 oil adjuvant.

9. Oil adjuvant Montanide ISA-70 in an amount of 50.0-70.0 wt.%.

10. Of the oil adjuvants, the Montanide ISA-206 oil adjuvant.

11. Oil adjuvant Montanide-206 in an amount of 50.0-70.0 wt.%.

12. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 Ig EID _{50 / 0.2cm} ³ and subjected to inactivation, inactivator DEI and oil adjuvant in the ratio, wt.%:

Antigenic material 29.9-49.7 DEI 0.1-0.3 Oil adjuvant 50.0-70.0

Signs of the invention characterizing the proposed vaccine and coinciding with the signs of the prototype, including the generic concept, reflecting the purpose:

1. Inactivated emulsion vaccine against IBD birds.

2. Antigenic material from a strain homologous to the pathogen, obtained in a sensitive biological system and subjected to inactivation.

3. Inactivant.

4. Oil adjuvant.

Compared with the prototype vaccine, an essential distinguishing feature of the proposed vaccine is that it contains an effective amount of antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds.

The present invention is also characterized by other distinctive features expressing specific forms of execution or special conditions for its use.

1. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 Ig EID _{50 / 0.2cm} ³ and subjected to inactivation.

2. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 lg EID ₅₀ / _0.2cm ³ and subjected to inactivation, in the amount of 29.9-49.7 wt.%.

3. Of the inactivants of DEI.

4. DEI in the form of a 15% aqueous solution.

5. DEI in an amount of 0.1-0.3 wt.%.

6. Oil adjuvant in an amount of 50.0-70.0 wt.%.

7. Antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least 5.25 Ig EID _{50 / 0.2cm} ³ and subjected to inactivation, inactivator DEI and oil adjuvant in the ratio, wt.%:

Antigenic material 29.9-49.7 DEI 0.1-0.3 Oil adjuvant 50.0-70.0

Compared with the prototype vaccine, an inactivated emulsion vaccine based on the strain K-58 No. 122-DEPT has a higher antigenic and immunogenic activity and is harmless.

The achievement of the technical result from the use of the proposed vaccine is explained by the fact that viral material from the new attenuated strain "K-58 No. 122-DEPT", homologous to the pathogen and not causing immunosuppression in the vaccinated bird, was introduced into its composition.

The original virus (the copyright name of the isolate "K-58") for obtaining the strain "K-58 No. 122-DEPT" was isolated from the factory bags of sick broilers at the Vasilyevskaya poultry farm in the Penza region in 1994. The vaccine strain "K-58 No. 122-DEPT" was obtained by repeated consecutive passages of the initial isolate on 9-11-day-old embryos of SPF-hens and SPF-chickens.

The strain "K-58" was deposited in the All-Russian state collection of strains of microorganisms used in veterinary medicine and animal husbandry, the Federal State Institution "All-Russian State Center for Quality and Standardization of Medicines for Animals and Feed" (FGU VGNKI) Ministry of Agriculture of the Russian Federation on June 1, 2004 under registration number - strain "K-58 No. 122-DEPT" virus IBD birds.

The strain "K-58 No. 122-DEPT" does not cause immunosuppression and has high biological, antigenic and immunogenic activity in its native form and after inactivation. The possibility of its use for the preparation of an inactivated emulsion vaccine against IBD birds has been experimentally confirmed.

The strain "K-58 No. 122-DEPT" virus IBD birds characterized by the following features and properties.

Morphological properties

The strain "K-58 No. 122-DEPT" of the IBD virus of birds belongs to the Birnavindae family, the genus Avibirnavirus, serotype I and has morphological characteristics characteristic of the IBD pathogen: the form of the virion is icosahedral, the size of the virion is 58-60 nm. Capsid T13, no shell.

Antigenic properties

According to its antigenic properties, the strain "K-58 No. 122-DEPT" refers to serovariant I. The virus is stably neutralized by a homologous antiserum. The virus does not exhibit hemagglutinating activity. During vaccination, the virus induces the formation of specific antibodies detected by RDP at a dilution of 1: 4-1: 64, in ELISA - 1: 400-1: 10000 and more, with hyperimmunization in RDP to 1: 4096 and above, in ELISA> 1: 10000 . To determine the genotaxonomic homology of the vaccine strain "K-58 No. 122-DEPT" and field isolates of the IBD virus, the variable regions of VP2 field isolates (37 samples isolated in 1993-1996 in the Russian Federation and the CIS countries) and vaccine strains (7 samples) were compared commercial live vaccines). For this, PCR was amplified using specific oligonucleotide primers and thermostable Tag DNA polymerase, the variable region of the VP ₂ gene encoding the main conformational epitope. Using the direct sequencing method, the nucleotide sequence of this most important in the manifestation of antigenic properties of the genome of the field isolates and vaccine strain IBD was determined. It has been established that most of the field isolates of IBD virus isolated at Russian poultry farms are antigenically different from imported vaccine strains of Ron-Merier (France), Intervet (Holland), Solvey-Dufar (USA), Safit ( Israel) by 5-8% and have a structure close to the vaccine strain "K-58 No. 122-DEPT".

Biotechnological characteristics

The strain "K-58 No. 122-DEPT" is intended for the manufacture of live and inactivated vaccines and diagnostic biologics. The strain "K-58 No. 122-DEPT" is reproduced in 9-11-day-old embryos of SPF-hens at a temperature of 37.7 ° C and a humidity of 55-64%. During 72-96 hours of incubation, the virus accumulates up to 5.25-7.0 lg EID ₅₀ / _2.0cm ³ .

Chemo- and genotaxonomic characterization

The strain "K-58 No. 122-DEPT" is an RNA-containing virus with a pier. RNA mass 2.16 × 10 ⁶ Da. Nucleotide acid is 2-strand, 2-segment (A and B). The virus has a protein coat formed by 5 types of proteins (VP ₁ , VP ₂ , VP ₃ , VP _4, and VP ₅ ):

VP ₁ - RNA-dependent RNA polymerase;

VP ₂ , VP ₃ - structural proteins;

VP ₄ - protease;

VP ₅ - not identified.

Lipids are absent.

The strain "K-58 No. 122-DEPT" shows genetic stability. The bulk of mutations are concentrated in the RNA region encoding VP ₂ . Mutations leading to amino acid substitution determine VP ₂ antigenic variability.

The strain "K-58 No. 122-DEPT" shows a high degree of homology (98%) compared with the large group of isolates (30) isolated on the territory of the Russian Federation in 1993-1996.

Physical properties

The virion mass is 55.0 × 10 ⁶ Da. Sedimentation coefficient 460S in sucrose gradient. The floating density of 1.33-1.34 g / cm ³ in the gradient of CsCl. Resistance to external factors

The strain "K-58 No. 122-DEPT" is resistant to solvents (ether, chloroform) and detergents, sensitive to formaldehyde, UV radiation, γ-radiation and drying.

Additional features and properties

Immunogenic activity -100%.

Reactogenicity is absent.

Pathogenicity is absent.

Virulence is absent.

Oncogenicity is absent.

Contagiousness is absent.

The strain "K-58 No. 122-DEPT" is stable and does not cause immunosuppression.

For safety and immunogenicity, the strain "K-58 No. 122-DEPT" fully meets the requirements of O.I.E. (OIE).

Commission tests confirmed the full compliance of strain K-58 No. 122-DEP with the requirements and were the basis for its deposit in the All-Russian State Collection of Microorganisms Used in Veterinary and Animal Husbandry, All-Russian State Center for the Quality and Standardization of Medicines for Animals and Feed (FGU VGNKI) Ministry of Agriculture of the Russian Federation on June 1, 2004 under the registration number of the strain "K-58 No. 122-DEPT" of the IBD virus of birds.

Based on the above information, it can be argued that the strain "K-58 No. 122-DEPT" of the IBD virus in birds according to antigenic and immunogenic spectra is an original, taxonomically new, previously unknown variant of the bird virus.

The analysis of the prior art by the applicant, including a search by patent and scientific and technical sources of information and identification of sources containing information about analogues of the invention, allowed to establish that the applicant did not find a source characterized by features that are identical (identical) to all the features of the claimed invention. The definition from the list of identified analogues of the prototype, as the closest in the set of essential features of the analogue, made it possible to establish a set of essential distinguishing features of the proposed vaccine relative to the applicant’s technical result, as set forth in the independent claim.

Therefore, the claimed invention meets the condition of patentability "novelty."

To verify the conformity of the proposed vaccine with the patentability condition “inventive step”, an additional search for known solutions was carried out to identify features included in the distinctive part of the independent claim. The search results showed that the proposed vaccine does not follow for the specialist explicitly from the prior art set forth in the corresponding section of the description (no solutions having features matching the distinctive features of the present invention have been identified), and the effect of the transformations provided for by the essential features of the proposed invention has not been identified to achieve a technical result.

Therefore, the proposed vaccine meets the condition of patentability "inventive step".

The essence of the invention is illustrated by examples of its execution, which do not limit the scope of the invention.

Example 1

The matrices of the virus strain "K-58 No. 122-DEPT" in the embryos of SPF hens are obtained as follows. At the border of the membrana shell and chorioallantoic membrane (CAO), a notch is made without damaging the CAO, and then using a syringe, virus-containing material is introduced in an amount of 0.2 cm ³ per CAO. The hole in the shell is sealed with adhesive tape. Infected embryos are incubated at a temperature of 37.7 ° C for 48-120 hours. Fallen embryos are cut, frozen at a temperature of minus 40 ° C. Virus-containing material is thawed, crushed, extracted at 4 ° C for 45 minutes, frozen twice - thawed, phosphate buffer pH 7.5 is added, centrifuged for 20 minutes at 3000 rpm. The resulting virus-containing liquid with an infectious activity of 5.25-7.0 Ig EID ₅₀ / _0.2cm ^{3 is} used as a matra seed.

Example 2

At the first stage, matte material is prepared from the strain “K-58 No. 122-DEPT” of the IBD virus of birds as described in Example 1. The resulting virus-containing liquid is used for mass infection of 9-11-day-old SPF chicken embryos. Viral material in an amount of 0.2 cm ³ is introduced on HAO and incubated at a temperature of 37.7 ° C for 48-120 hours. Fallen embryos are cut, frozen at a temperature of minus 40 ° C and the obtained raw materials are used for the manufacture of a vaccine. The virus-containing material is thawed, crushed in a colloid mill, extracted for 45 minutes at 4 ° C, frozen twice - thawed, phosphate buffer pH 7.5 is added and centrifuged for 20 minutes at 3000 rpm, thus cleaning it from ballast impurities. The purified suspension is fed into a sterile reactor to inactivate the virus. Of the inactivants, DEI is used in the form of a 15% aqueous solution, pH 7.9-8.0. In a virus-containing suspension heated to 27 ° C with an infectious activity of 5.25-7.0 Ig EID ₅₀ / _0.3cm ^3, DEI is added in an amount corresponding to its 0.1-0.3, wt.% Content in the finished product. The mixture is thoroughly mixed and kept at 27 ° C for 24 hours. At the end of inactivation, the antigenic material is cooled to 2-8 ° C. The resulting antigen is mixed with an oil adjuvant, which is taken in an amount corresponding to its content of 50.0-70.0 wt.% In the finished product. From oil adjuvants it is possible to use the oil adjuvant VNIIZZH (US Pat. RF No. 2108111; A 61 K 39/39, A 61 K 39/00, A 61 K 39/102, A 61 K 39/12, A 61 K 39/135; 04/10/98), as well as oil adjuvants of the brands Montanide ISA-70 and Montanide ISA-206 manufactured by Seppic (France).

The resulting vaccine is a homogeneous white emulsion. The vaccine is monitored for avirulence and sterility and packaged in sterile vials.

The virulence of the drug is determined by the method of triple "blind" passages of the vaccine in 10-day-old embryos of SPF-hens.

Sterility of the vaccine is determined in accordance with GOST 28085-89.

The safety of the drug is determined by introducing a 10-fold immunizing dose (5.0 cm ³ ) of the obtained vaccine to 100-110-day-old chickens.

The immunogenic activity of the drug is determined by intramuscular administration of 0.5 cm ^{3 of the} obtained vaccine to 30-40-day-old chickens.

The stability of the vaccine is determined by keeping the vaccine samples in an incubator at a temperature of 37 ° C for a month and compared with the original samples. The vaccine is used in clinically healthy birds to create a high level of immunity. Birds are vaccinated at the age of 110-120 days once. The drug is administered intramuscularly in a volume of 0.5 cm ³ .

Immunity in a vaccinated bird occurs on 21 days after vaccination and lasts up to 12 months. The vaccine induces a high level of antibodies against IBD in birds of the parent flock and provides reliable passive immunity in young animals up to 15-25 days of age, which helps protect it from the virulent field virus of IBD in early life.

The resulting vaccine against IBD birds has the optimal component composition (wt.%), Presented in table 1.

The antigen content in the proposed vaccine in the above range is its effective amount in the drug, ensuring the achievement of a technical result.

Example 3

The immunogenic activity of the inactivated emulsion vaccine against IBD of birds, made as described in example 2, and containing, wt.%:

Antigenic material from the strain "K-58 No. 122-DEPT" virus IBD birds obtained in 10-day embryos SPF chickens with an infectious activity of 5.25 Ig EID ₅₀ / _0.2cm ³ and inactivated 29.9 DEI 0.1 Oil adjuvant 70.0

Immunogenic activity was determined on chickens of 30 days of age, free of antibodies to the IBD virus of birds. The vaccine was administered intramuscularly in the thigh area at 0.5 cm ³ . After 21 days, blood was collected from birds, serum was obtained and examined in RDP and ELISA. The research results are presented in table 2.

The results presented in table 2 indicate the high immunogenic activity of the inactivated emulsion vaccine against IBD birds from the strain "K-58 No. 122-DEPT" virus IBD birds.

Example 4

The immunogenic activity of an inactivated emulsion vaccine against IBD of birds, made as described in example 2, and containing, wt.%:

Antigenic material from the strain "K-58 No. 122-DEPT" virus IBD birds obtained in 10-day embryos SPF chickens with infectious activity 6.0 Ig EID ₅₀ / _0.2cm ³ and inactivated 39.8 DEI 0.2 Oil adjuvant 60.0

The determination of immunogenic activity was carried out on chickens of 30 days of age, free of antibodies to the IBD virus of birds. The vaccine was administered at 0.5 cm ³ into the thigh muscle. 21 days after vaccination, blood was collected from birds, serum was obtained and examined in RDP and ELISA for the presence of antibodies against IBD. The research results are presented in table 3.

The results presented in table 3 indicate the high immunogenic activity of the inactivated vaccine against IBD birds from strain "K-58№122-DEPT".

Example 5

The immunogenic activity of the inactivated emulsion vaccine against IBD of birds, made as described in example 2, and containing, wt.%:

Antigenic material from the strain "K-58 No. 122-DEPT" virus IBD birds obtained in 10-day embryos SPF chickens with infectious activity 6.0 Ig EID ₅₀ / _0.2cm ³ and inactivated 49.7 DEI 0.3 Oil adjuvant 50,0

Immunogenic activity was determined on chickens of 30 days of age, free of antibodies to the IBD virus of birds. The vaccine was administered intramuscularly in the thigh area at 0.5 cm ³ . After 21 days, blood was collected from birds, serum was obtained and examined in RDP and ELISA. The research results are presented in table 4.

The results presented in table 4 indicate the high immunogenic activity of the inactivated emulsion vaccine against IBD birds from strain "K-58 No. 122-DEPT".

Example 6

A comparative assessment of the immunogenic activity of the prototype vaccine and inactivated emulsion vaccine against IBD birds, made as described in example 2, containing, wt.%:

Antigenic material from the strain "K-58 No. 122-DEPT" virus IBD birds obtained in 10-day embryos SPF chickens with infectious activity of 6.5 Ig EID ₅₀ / _0.2cm ³ and inactivated 39.8 DEI 0.2 Oil adjuvant 60.0

The immunogenic activity of the vaccines was determined on chickens of 30 days of age, free of antibodies to the IBD virus of birds. Vaccines were administered intramuscularly in the thigh area at 0.5 cm ³ . After 21 days, blood was collected from birds, serum was obtained and examined in RDP and ELISA for the presence of antibodies.

The research results are presented in table 5.

The data in table 5 indicate that the inactivated emulsion vaccine against IBD, made from the attenuated strain K-58 No. 122-DEPT, significantly exceeds the vaccine against IBD (antibody titer is 1.88 times higher) made from strain D78, ATCC No. VR-2041, and provides reliable passive maternal immunity in young animals up to 15-25 days of age, which helps protect it from IBD disease in early life.

Thus, the above information indicates the fulfillment when using the invention of the following set of conditions:

- inactivated emulsion vaccine against IBD birds, embodying the invention, is intended for use in agriculture, namely in veterinary virology and biotechnology;

- for the proposed invention in the form as described in the independent claim, the possibility of its implementation using the means and methods described in the application or known prior to the priority date is confirmed;

- inactivated emulsion vaccine against IBD birds, prepared from strain "K-58 No. 122-DEPT" in accordance with the invention, has a high antigenic and immunogenic activity, is harmless and stable during storage. Therefore, the present invention meets the condition of patentability "industrial applicability".

Sources of information taken into account

1. Syurin V.N. and other viral diseases of animals. - M., VNITIBP, 1998, 64-84.

2. Wyeth P.J. The veterinary Record, 1979, 104, 188-193.

3. Lucia B.e.a. Avian Diseases. 1979, 23, 2, 466-478.

4. Pat. US No. 4530831, 425-89, 07.23.85.

5. Pat. Hungary No. 203781; C 12 N 7/00 A 61 K 39/12; 08.28.90 g.

6. Pat. Romania No. 100319, A 61 K 39/12, 10.29.90

7. Patent of the Russian Federation No. 2127603; A 61 K 39/12, C 12 N 7/00; 03/20/99 g.

8. Pat. USA No. 4530831, 425-89, 07.23.85, (prototype).

Table 1
The optimal component composition (wt.%) Inactivated emulsion vaccine against IBD birds, made on the basis of strain "K-58 No. 122-DEPT" Ingredients Minimum Average Maximum Antigenic material 29.9 39.8 49.7 DEI 0.1 0.2 0.3 Oil adjuvant 70.0 60.0 50,0 table 2
Determination of the immunogenic activity of inactivated emulsion vaccine against IBD birds from strain "K-58 No. 122-DEPT" №№ Name of material Antibody titer RDP IFA one Vaccine Serum 1:16 1: 4850 2 - "- 1:32 1: 5200 3 - "- 1:64 1: 5500 four - "- 1:64 1: 5850 5 - "- 1:64 1: 7200 6 - "- 1:64 1: 7370 7 - "- 1:64 1: 7600 8 - "- 1:64 1: 8100 9 - "- 1:64 1: 8500 10 - "- 1: 128 1: 9220 Table 3
Determination of the immunogenic activity of inactivated emulsion vaccine against IBD birds from strain "K-58 No. 122-DEPT" №№ Name of material Antibody titer RDP IFA one Vaccine Serum 1:32 1: 4500 2 - "- 1:32 1: 4800 3 - "- 1:64 1: 5000 four - "- 1:64 1: 6200 5 - "- 1:64 1: 6800 6 - "- 1:64 1: 7200 7 - "- 1:64 1: 7550 8 - "- 1: 128 1: 8000 9 - "- 1: 128 1: 8500 10 - "- 1: 128 1: 9800

Table 4
Determination of the immunogenic activity of "Inactivated emulsion vaccine against IBD birds from strain" K-58 No. 122-DEPT " №№ Name of material Antibody titer RDP IFA one Vaccine Serum 1 32 1: 5500 2 - "- 1:32 1: 5100 3 - "- 1:32 1: 5650 four - "- 1:64 1: 6800 5 - "- 1:64 1: 7200 6 - "- 1:64 1: 7420 7 - "- 1:64 1: 8550 8 - "- 1: 256 1: 9100 9 - "- 1: 256 1: 9210 10 - "- 1: 256 1: 10520 Table 5
Comparative evaluation of the immunogenic activity of inactivated emulsion vaccines from strain D 78 and strain "K-58 No. 122-DEPT" No. p / p The name of the strains D78 "K-58 No. 122-DEP" Antibody titer Antibody titer one 1: 1600 1: 2700 2 1: 1750 1: 3320 3 1: 2050 1: 4000 four 1: 3500 1: 4360 5 1: 3200 1: 4800 6 1: 2500 1: 5850 7 1: 1800 1: 5950 8 1: 4200 1: 5500 9 1: 2000 1: 6480 10 1: 2880 1: 5020 Average 1: 2548 1: 4798

Claims (8)

1. Inactivated emulsion vaccine against infectious bursal disease (IBD) of birds, containing antigenic material from a strain homologous to the pathogen, obtained in a sensitive biological system and subjected to inactivation, an inactivant and an oil adjuvant, characterized in that it contains antigenic material from antigenic material strain "K-58 No. 122-DEPT" of the IBD virus of birds (Bursitis infectiosa gallinae), family Birnaviridae, genus Avibirnavirus, serotype 1, collection of the Federal State Institution of Higher Vocational Research Institute "K-58 No. 122-DEPT", in an effective amount. 2. The inactivated emulsion vaccine according to claim 1, characterized in that it contains antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least at least 5.25 lg EID _{50 / 0.2cm} ³ and subjected to inactivation. 3. The inactivated emulsion vaccine according to claim 2, characterized in that it contains antigenic material from the K-58 strain No. 122-DEP strain of the IBD virus of birds obtained in 9-11-day-old embryos of SPF-chickens with infectious activity of at least at least 5.25 lg EID _{50 / 2.0cm} ³ and subjected to inactivation, in the amount of 29.9-49.7 wt.%. 4. Inactivated emulsion vaccine according to claim 1, characterized in that among the inactivants it contains an ethyleneimine dimer (DEI). 5. The inactivated emulsion vaccine according to claim 4, characterized in that it contains a DEI in the form of a 15% aqueous solution. 6. Inactivated emulsion vaccine according to claim 5, characterized in that it contains a DEI in an amount of 0.1-0.3 wt.%. 7. The inactivated emulsion vaccine according to claim 1, characterized in that it contains an oil adjuvant in an amount of 50.0-70.0 wt.%. 8. Inactivated emulsion vaccine according to any one of claims 1 to 7, characterized in that it contains antigenic material from the strain "K-58 No. 122-DEPT" of the IBD virus of birds, obtained in 9-11-day-old embryos of SPF-chickens with infectious activity at least 5.25 lg EID _{50 / 0.2cm} ³ and subjected to inactivation, inactivator DEI and oil adjuvant in the ratio, wt.%: Antigenic material 29.4-47.7 DEI 0.1-0.3 Oil adjuvant 50.0-70.0