RU2195959C1 - Inactivated vaccine against hens' hydropericarditis syndrome - Google Patents

Abstract

FIELD: veterinary virology and biotechnology. SUBSTANCE: the suggested vaccine contains an antigenic material as 10- 20%-virus-containing suspension from KR95N114 strain of HHPS virus, dimer ethylenimine inactivant (DEI) and adjuvants - aluminum oxide hydrate and saponin at an efficient ratio. The strain is depoted in the collection of microorganisms in VGNKI under registration number of KR95N114-DEP. It is reproduced on 5-50-d-aged chickens at the titer of infectious activity of 3.25-5.0 lgLD₅₀/02 cm³. Vaccine represents a rose-brown-colored liquid with a friable residue of a sorbent easily divided into a homogenic suspension at shaking. Vaccine is nonreactogenic, immunogenic and safe for vaccinated poultry. EFFECT: higher efficiency. 10 cl, 6 ex, 5 tbl

Description

 The invention relates to the field of veterinary virology and biotechnology and can be used in the manufacture of agents for the specific prophylaxis of chicken hydropericarditis syndrome (HHPC).

 SGPK (Angara disease, Lychee disease) is an acute infectious disease of hens of viral etiology, causing significant economic damage to the developed poultry industry in many countries of the world.

 The hepatitis C virus virus causes a disease of chickens mainly aged 10 to 50 days, which is characterized by sudden death of the bird, accumulation of excess fluid in the pericardial cavity, damage to the liver and kidneys, depression, diarrhea, and in some cases anemia.

 In August 1987, in Pakistan, a disease with signs of hepatitis with inclusion bodies and heart damage (hydropericarditis) was recorded at a broiler farm in Angara Goth near Karachi in Pakistan. The disease was noted, as a rule, in birds of 3-6 weeks of age, but sometimes it was found in younger or older birds, mortality reached 50-70%. Initially, the disease was given the name "Angara disease" in accordance with the geographical area where the outbreak was first recorded. Later, the disease was given the name "Hydropericarditis Syndrome" due to the fact that when autopsies of birds were observed accumulation of fluid (up to 20 ml) in the cardiac chemise caused by impaired circulation in the cardiovascular system. A similar disease has been reported in Iraq, India, Mexico, Kuwait, Australia, Japan, and Chile (1).

 Since the early 90's. outbreaks of high-mortality SGPKs are recorded in Russia (2). The lack of diagnostic tools and specific prophylaxis of SGPK in domestic veterinary practice has caused significant economic damage to poultry farms. The economic losses incurred by poultry farms from outbreaks of agro-agricultural complex are made up of the death of birds (from 10 to 60%), the death of developing chicken embryos during incubation, reduced productivity and payment of feed, as well as funds spent on unscheduled veterinary and sanitary measures etc. (3) Vaccine prophylaxis of CGD takes a leading place in the fight against this disease. Both live and inactivated vaccines are used to prevent CGD. In recent years, inactivated vaccines have been most commonly used (4). However, the problem of improving their quality continues to be very relevant and is the main direction of research to increase the immunogenic activity and harmlessness of the drug and address the issue of protecting the poultry industry of our country from this dangerous disease.

 The closest to the invention according to the totality of signs is an inactivated vaccine against CHPC containing antigenic material in the form of a purified virus-containing suspension from the liver of chickens infected with the causative agent of the disease and fallen with signs of CHP, and formaldehyde in effective amounts (5).

The vaccine prototype has significant disadvantages:
- to obtain antigen, the liver of chickens that died during an outbreak of CGD, which is often heterogeneous in content of antigen, can also be contaminated with pathogens of other infectious diseases of birds (Gamboro disease, Marek’s disease, infectious bronchitis, infectious chicken anemia, etc.). .);
- formaldehyde is used as an inactivant, causing destructive changes in viral proteins and reducing the immunogenic activity of the vaccine. The probability of incomplete inactivation of the virus during formaldehyde treatment of a viral suspension containing a significant amount of ballast protein is also high;
- this vaccine preparation does not include an adjuvant, the use of which improves storage and increases the antigenic activity of the vaccine;
- the prototype vaccine has low antigenic and immunogenic activity and does not provide satisfactory protection against epizootic HCG virus circulating in the Russian Federation.

 The objective of the creation of the present invention was to develop a highly immunogenic and harmless inactivated vaccine against the HCV virus circulating in the Russian Federation.

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

The specified technical result was achieved by the creation of an inactivated vaccine against SGPK, characterized by the following set of features:
1. Inactivated vaccine against SGPK.

 2. Antigenic material in the form of a purified 10-20% virus-containing suspension from the liver of chickens infected with the CGPC virus from strain KP95 114 and fallen with signs of CGC.

 3. Antigenic material from strain KP95 114 virus SGPK, reproduced in the body of 5-50-day-old chickens.

4. Antigenic material from strain KP95 114 of the SGPV virus with biological activity of 3.25-5.0 lg LD ₅₀ / 0.2 cm ³ .

 5. Antigenic material from strain KP95 114 of the SGPV virus in an effective amount.

6. Inactivant
7. Of the inactivants, ethyleneimine dimer (DEI).

 8. DEI in the form of a 10% aqueous solution.

 9. Optional adjuvant.

 10. Of the adjuvants, alumina hydrate (GOA).

 11. GOA in the form of a 3-4% colloidal solution.

 12. Of the adjuvants of saponin.

 13. Saponin in the form of a 10% aqueous solution.

14. The inactivated vaccine against SGPK contains antigenic material from strain KP95 114 virus SGPK, DEI, GOA and saponin in the ratio, wt.%:
Antigenic material - 50.0-80.0
DEI (10% aqueous solution) - 0.15-0.4
GOA (3-4% colloidal solution) - 17.6-49.35
Saponin (10% aqueous solution) - 0.5-2.0
The present invention includes the following set of essential features that provide a technical result in all cases for which legal protection is sought:
1. Inactivated vaccine against SGPK.

 2. Antigenic material from strain KP95 114 virus SGPK.

 3. Antigenic material from strain KP95 114 of the SGPV virus in an effective amount.

 4. Inactivant.

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 KP95 114 virus SGPK reproduced in the body of 5-50-day-old chickens.

2. Antigenic material from strain KP95 114 of the SGPV virus with biological activity of 3.25-5.0 lg LD ₅₀ / 0.2 cm ³ .

 3. Antigenic material from strain KP95 114 of the SGPV virus in an effective amount.

 4. Of the inactivators of DEI.

 5. DEI in the form of a 10% aqueous solution.

 6. Optional adjuvant.

 7. GOA.

 8. GOA in the form of a 3-4% colloidal solution.

 9. Of the adjuvants saponin.

 10. Saponin in the form of a 10% aqueous solution.

11. Antigenic material from strain KP95 114 virus SGPK, DEI, GOA and saponin in the ratio, wt.%:
Antigenic material - 50.0-80.0
DEI (10% aqueous solution) - 0.15-0.4
GOA (3-4% colloidal solution) - 17.6-49.35
Saponin (10% aqueous solution) - 0.5-2.0
The signs of the invention, characterizing the proposed vaccine and coinciding with the signs of the prototype, including the generic concept, reflecting the purpose, are:
1. Inactivated vaccine against SGPK.

 2. Antigenic material in the form of a purified virus-containing suspension from the liver of chickens infected with the causative agent of the disease and who died with signs of CGD.

 3. Inactivant.

 Compared with the prototype vaccine, the essential distinguishing features of the invention is that the proposed vaccine contains antigenic material from strain KP95 114 of the SGPV virus, taken in an effective amount.

The present invention is also characterized by other distinctive features that express specific forms of execution or special conditions for its use:
1. Antigenic material from strain KP95 114 virus SGPK reproduced in the body of 5-50-day-old chickens.

2. Antigenic material from strain KP95 114 of the SGPV virus with biological activity of 3.25-5.0 lg LD ₅₀ / 0.2 cm ³ .

 3. Antigenic material from strain KP95 114 of the SGPV virus in an effective amount.

 4. Of the inactivators of DEI.

 5. DEI in the form of a 10% aqueous solution.

 6. Optional adjuvant.

 7. Of the adjuvants of GOA.

 8. GOA in the form of a 3-4% colloidal solution.

 9. Of the adjuvants saponin.

 10. Saponin in the form of a 10% aqueous solution.

11. Antigenic material from strain KP95 114 virus SGPK, DEI, GOA and saponin in the ratio, wt.%:
Antigenic material - 50.0-80.0
DEI (10% aqueous solution) - 0.15-0.4
GOA (3-4% colloidal solution) - 17.6-49.35
Saponin (10% aqueous solution) - 0.5-2.0
The proposed vaccine has a higher antigenic and immunogenic activity and harmlessness compared to the prototype vaccine.

 The achievement of the technical result from the use of the invention is explained by the fact that antigenic material from the new strain KP95 114 of the SGPV virus was introduced into the composition of the inactivated vaccine against CHP.

 An additional technical result from the use of the present invention is achieved due to the fact that the vaccine contains DEI as an inactivant, which can significantly reduce labor and energy costs for the manufacture of the vaccine and improve the quality of antigenic material.

 An additional technical result from the use of the invention in terms of increasing antigenic and immunogenic activity is also achieved through the use of adjuvants GOA and saponin.

As a result of the studies, it was found that the GOA gel, introduced into the experimental series of vaccines against SGPK, increased their antigenic activity with an increase in the concentration of GOA in the prescription of the drug. So, in chickens vaccinated with a vaccine without GOA, the titer of antibodies to the CGPV virus according to the results of the RDP was at the level of 0.56 + 0.1 log ₂ , and in chickens immunized with drugs with GOA, from 1.23 ± 0.2 log ₂ up to 3.85 ± 0.2 log ₂ .

 As for saponin, it was established as a result of studies that the introduction of an aqueous solution of saponin in the amount of 0.1 to 2 mg per dose of the inactivated sorbed anti-SGP vaccine resulted in an increase in the antigenic activity of the drug compared to a vaccine without saponin. The highest indices of antigenic activity were recorded in vaccines containing saponin in their composition in a final concentration of 0.1 to 1.5 mg per vaccine dose.

 The original virus for obtaining strain KP95 114 was isolated from the liver of sick broilers at the Krasnoyarskaya poultry farm in the Krasnoyarsk Territory in 1995. Production strain KP95 114 was obtained by repeated successive passages on chickens. The strain KP95 114 was deposited in the State collection of microorganisms of the research institute for control, standardization and certification of veterinary drugs (VGNKI) 12/14/2000 under registration number KP95 114-DEP. The strain is virulent and has high antigenic and immunogenic activity after inactivation. The possibility of its use for the manufacture of an inactivated vaccine against SGPK has been experimentally confirmed.

 Strain KP95 114 of the HCVV virus is characterized by the following features and properties.

Morphological properties
The strain KP95 114 of the SGPV virus belongs to the family Adenoviridae, genus Aviadenovirus, group 1, serotype 4, has morphological characteristics characteristic of adenoviruses: like all other viruses of this family lacking a supercapsid coat, the diameter of the virion of strain KP95 114 and nucleocapsid are equal. The size of the virion is 75-80 nm. Virion has an icosahedral type of symmetry. The virion is represented by a nucleoid consisting of nucleic acid and core proteins and a capsid protein coat. The capsid contains 252 capsomeres, which are located in 20 facets having the shape of equilateral triangles. On the edges of equilateral triangular faces common to neighboring triangles, there are six capsomeres; the rib length is about 42 nm. Each of the 240 capsomeres of the capsid shell forming equilateral triangles is surrounded by six of the same capsomeres and is called hexon. Each of the 12 capsomeres located at the tops of the icosahedron is surrounded by only five neighboring capsomeres and is called peptone. Each capsomer (except for a complex peptone) is a prismatic particle with a diameter of 6-7 nm with a central cavity of 1.5-2 nm in size. The method of laying capsomeres in a capsid is unique for viruses with an icosahedral type of symmetry and differs in geometric ordering. Hexons are the main soluble component of the pathogen during its reproduction in sensitive cells.

 Peptones are associated with their ability to agglutinate red blood cells, due to the presence of complete hemagglutin in their composition.

 The core of the virion contains a double-stranded DNA molecule and two internal proteins. DNA contains about 60 genes that carry information that is sufficient to encode from 30 to 50 proteins.

Antigenic properties
By its antigenic properties, strain KP95 114 belongs to group 1 of avian adenoviruses. The virus is stably neutralized by homologous antiserum. The virus does not exhibit hemagglutinating activity. When vaccinated, the viral antigen induces the formation of specific antibodies detected in the RDP at a dilution of 1: 2-1: 64.

Biotechnological characteristics
Strain KP95 114 is virulent and has high biological activity in its native form, and also exhibits high antigenic and immunogenic properties after inactivation.

Strain KP95 114 is intended for use as a raw material for the manufacture of inactivated vaccines, as well as diagnostic biologics. Strain KP95 114 is reproduced in chickens 5-90 days old. Within 48-120 hours of incubation, the virus accumulates in the liver of chickens in the titer of 3.25-5.0 lg LD ₅₀ / 0.2 cm ³ . Strain KP95 114 is stable.

Chemo- and genotaxonomic characterization
The core of the virion contains a double-stranded DNA molecule and two internal proteins. The nucleic acid mass reaches 17.3% of the mass of the virion.

The bulk of the proteins of the HCV virus are concentrated in the capsid shell of the virion. Core proteins account for up to 20% of all virus proteins. Core protein "1" with a molecular weight of 46 • 10 ³ g / mol is easily separated from the internal nucleoprotein containing DNA in complex with arginine-rich core protein "2".

Hexon, peptone base and its filament (fiber) of the K95 114 virus are constructed from polypeptides with molecular weights of 120 • 10 ³ , 70 • 10 ³ , 62 • 10 ³ g / mol, respectively.

 The protein structures of the virus strain KP95 114 form several antigens. In the structure of the structural viral antigen, type-specific, group-specific, subgroup-specific antigens of hexon polypeptides, peptone bases and subgroup-specific and type-specific antigens of peptone filament were found in them.

The structure of the structural viral antigen revealed three main components, designated A, B and C. Antigen A is structurally associated with hexon, antigen B with peptone, antigen C with virion filaments (fiber). Hexon-linked component A contains a group-specific antigen (α) and a type-specific antigen (ε). The antigenic factor associated with peptone is called the antigen (β), and with the filament of the virion (fiber) - (γ).
Physical properties
The mass of the virion is 252x10 ³ g / mol. Floating density 1.32-1.35 g / ml in gradient C _S Cl. The sedimentation constant of the mature virion is 810 • 10 ^-13 - 910 • 10 ^-13 s.

Resistance to external factors
Strain KP95 114 is resistant to ether, chloroform, saponin and sodium deoxycholate, is sensitive to formaldehyde, β-propiolactone, hydroxylamine, ethyleneimine derivatives, UV radiation, and γ-radiation.

 Strain KP95 114 is quite resistant to heat and to a change in the pH of the medium over a wide range, resistant to 0.25% trypsin solution, 2% phenol solution and 50% ethyl alcohol solution. Absolute ethanol, like its iodine solution, causes complete inactivation of the virus.

 Strain KP95 114 is resistant to repeated cycles of freezing and thawing, does not lose infectious properties during freeze-drying.

Additional features and properties
Immunogenic activity - 100%
Pathogenicity is expressed
Virulence expressed
Contagiousness expressed
No oncogenicity
Based on the data obtained, it can be argued that strain KP95 114 in terms of antigenic and immunological spectra is an original, taxonomically new, previously unknown variant of the CGD virus. To reduce its epizootic danger, timely vaccine prophylaxis of newly emerging foci of the disease is necessary, for this a highly active and harmless vaccine is needed.

 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 sources characterized by features that are identical (identical) to all the essential features of the invention. The definition from the list of identified analogues of the prototype as the closest in the totality of the features of the analogue allowed us 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 solution to the patentability condition "inventive step", an additional search was carried out for known solutions to identify features included in the distinctive part of the independent claim. The search results showed that the proposed solution does not follow explicitly for the specialist from the prior art set forth in the corresponding section of the description (no solutions having features matching the distinguishing 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 implementation.

 Example 1

To obtain a vaccine from strain КР95, 5–50-day-old chickens obtained from farms that are successful in infectious diseases are infected intramuscularly in a volume of 1 cm ³ with 114 matrovirus. Chickens contain 120 hours after infection. Livers are taken from sick and dead chickens. The liver is subjected to grinding in a colloidal mill for 10-12 minutes, then the virus-containing mass is mixed with phosphate buffer solution, resulting in a 10-20% virus-containing suspension. The virus-containing suspension is purified from ballast impurities in a known manner. To do this, you can use low-speed centrifugation, processing the suspension with various detergents (Freon-113, chloroform) and a high molecular weight polyhexamethylene guanidine hydrochloride flocculant (HSV) in combination with low-speed centrifugation or separation. Purification of the virus-containing suspension from ballast proteins is carried out at 6-12 ^o C. the Purified suspension is served in a sterile reactor. Ethyleneimine dimer (DEI) is used to inactivate the virus. Pre-prepared 10% solution of DEI in demineralized water, pH 7.9-8.0. Then, in a virus-containing suspension warmed up to 28-34 ^° C, a 10% DEI solution is added to a concentration of 0.15-0.4%. The mixture is thoroughly mixed and maintained at 35-36 ^o C for 24 hours. Upon completion of inactivation, the antigenic material is cooled to 2-8 ^o C. As a adjuvant sorbent in the vaccine use a sterile 3-4% colloidal GOA solution. The pH of GOA is adjusted to between 7.5-7.6 and cooled to 2-8 ^° C. GOA is added to the cooled antigen with constant stirring to a concentration of 0.1%. Then, a 10% aqueous solution of saponin adjuvant is added to the mixture at the rate of 0.5 mg per dose of the vaccine, bringing the concentration of saponin in the vaccine to 0.5-2%. After mixing the mixture for one hour, determine the pH of the drug and, if necessary, bring its value to 7.4-7.6. The resulting vaccine is monitored for avirulence, safety and sterility, and then packaged in sterile vials.

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

 Avirulence and harmlessness of the drug is determined by introducing a 10-15-day-old chickens vaccine in a triple dose.

 The resulting vaccine is a pink-brown liquid with a loose precipitate of sorbent, which, when shaken, easily breaks into a homogeneous suspension.

 The resulting vaccine against CHP has the optimal component composition (formulation), presented in table A.

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

 Example 2

Studies have been carried out to determine the antigenic activity of an inactivated vaccine against SGPK, obtained as described in example 1, and containing, wt.%:
Purified antigenic material from strain KP95 114 of the SGPV virus reproduced in the body of 30-day-old chickens with an infectious activity titer of 3.3 lg LD ₅₀ / 0.2 cm ³ (10% suspension) - 50.0
DEI (10% aqueous solution) - 0.15
GOA (3-4% colloidal solution) - 49.35
Saponin (10% aqueous solution) - 0.5
The antigenic activity of the vaccine was determined on chickens of 15 days of age, free of antibodies to the virus CGPC. The vaccine was administered at a dose of 0.3 cm ³ into the thigh muscle. 21 days after vaccination, the blood was taken from the chickens, serum was obtained, and they were examined in the RDP with a specific serum hyperimmune to the CGPC virus. The research results are presented in table 1.

 The results presented in table 1 indicate a high antigenic activity of the inactivated vaccine against SGPK from strain KP95 114.

Example 3
Studies have been carried out to determine the antigenic activity of an inactivated vaccine against SGPK, obtained as described in example 1, and containing, wt.%:
Purified antigenic material from strain KP95 114 of the SGPV virus reproduced in the body of 30-day-old chickens with an infectious activity titer of 3.54 lg LD ₅₀ / 0.2 cm ³ (10% suspension) - 70.0
DEI (10% aqueous solution) - 0.2
GOA (3-4% colloidal solution) - 28.8
Saponin (10% aqueous solution) - 1.0
Determination of antigenic activity was carried out on chickens of 12 days of age, free of antibodies to the CGPV virus. The vaccine was administered at a dose of 0.3 cm ³ into the thigh muscle. 21 days after vaccination, blood was collected from birds, serum was obtained, and they were examined in the RDP for the presence of antibodies to the CGPV virus. The research results are presented in table 2.

 The results presented in table 2 indicate a high antigenic activity of the inactivated vaccine against SGPK from strain KP95 114.

 Example 4

Studies have been carried out to determine the antigenic activity of an inactivated vaccine against SGPK, obtained as described in example 1, and containing, wt.%:
Purified antigenic material from strain KP95 114 of the SGPV virus reproduced in the body of 30-day-old chickens with an infectious activity titer of 3.25 lg LD ₅₀ / 0.2 cm ³ (10% suspension) - 80.0
DEI (10% aqueous solution) - 0.4
GOA (3-4% colloidal solution) - 17.6
Saponin (10% aqueous solution) - 2.0
The antigenic activity of the vaccine was determined on chickens 18 days of age, free of antibodies to the virus CGPC. The vaccine was administered intramuscularly in a volume of 0.3 cm ³ in the thigh area. 21 days after vaccination, the blood was taken from the chickens, serum was obtained and they were examined in the RDP for the presence of antibodies to the CGPV virus. The research results are presented in table 3.

 The results presented in table 3 indicate a high antigenic activity of the inactivated vaccine against SGPK from strain KP95 114.

 Example 5

Studies are carried out to determine the immunogenic activity of an inactivated vaccine against SGPK, obtained as described in example 1, and containing, wt.%:
Purified antigenic material from strain KP95 114 of the SGPV virus reproduced in the body of 35-day-old chickens with an infection rate of 3.25 lg LD ₅₀ / 0.2 cm ³ (10% suspension) - 70.0
DEI (10% aqueous solution) - 0.2
GOA (3-4% colloidal solution) - 28.8
Saponin (10% aqueous solution) - 1.0
For this, a series of experiments was carried out on chickens of 10 days of age that did not have antibodies to the CGPV virus, which served as a biological model for testing the bird's immune response to vaccine administration. The experience was taken 350 chickens, which were divided into nine groups of 50 goals each. Eight groups were test vaccine subjects, and one group served as a control. The chickens of the experimental groups were vaccinated once intramuscularly in doses of 0.15, 0.3 and 0.5 cm ³ inactivated sorbed vaccine 2 and 5 series. On day 3, 5, 7, 10, and 14 after vaccination, 10 chickens were selected from each group, which were placed in a separate box and intramuscularly infected with SGPK virus from strain KP95 at a dose of 3.78 ± 0.23 lg LD ₅₀ / 0.2 cm ³ . After infection, the birds were monitored for 10 days. Fallen chickens were recorded and the percentage of protection was counted. The results of the studies are presented in table 4. As can be seen from the data given in table 4, a single vaccination of chickens at a dose of 0.5 cm ³ provided 100% protection against infection after 3 days after administration of the drug. In groups of chickens that were injected with an inactivated sorbed vaccine at doses of 0.15 and 0.3 cm ³ , 100% protection against control infection occurred 5-7 days after vaccination. Mortality in the control group of chickens was at the level of 80-100%.

 Example 6

Studies were carried out to determine the actual immunogenic activity (ImD ₅₀ / cm ³ ) and the protective activity of the vaccinated dose of the inactivated sorbed anti-CGP vaccine, manufactured as described in example 1, and containing, wt.%:
Purified antigenic material from strain KP95 114 of the SGPV virus reproduced in the body of 35-day-old chickens with an infection rate of 3.25 lg LD ₅₀ / 0.2 cm ³ (10% suspension) - 70.0
DEI (10% aqueous solution) - 0.2
GOA (3-4% colloidal solution) - 28.8
Saponin (10% aqueous solution) - 1.0
For this, experiments were carried out on 8-day-old egg-cross chickens that did not have antibodies to the CGPV virus. In the experiments used chickens with a total of 250 goals. The chickens were inoculated with an inactivated sorbed vaccine against SGPK 2, 3 and 5 series. The immunogenic activity of vaccine samples was determined by the volumetric method using calculations according to the Kerber-Ashmarin formula. The PD _{50 of the} vaccine was calculated by dividing the vaccine dose of the vaccine by the value of ImD ₅₀ . The results of the studies are shown in table 5. In the experiments it was found that the immunogenic activity of the inactivated vaccine against CPS is not significantly affected by the method of administration of the drug. So, the ImD ₅₀ when the vaccine was administered subcutaneously was 0.0107 cm ³ , and intramuscularly - 0.012 cm ³ . From the data of table 5 it is seen that one vaccine dose of the vaccine in the volume of 0.3 cm ³ with subcutaneous administration of the drug contains 28 ± 1.15 PD ₅₀ , and one vaccine dose of the vaccine in the volume of 0.3 cm ³ with intramuscular injection contains 25 ± 1 15 PD ₅₀ . A vaccine series is produced for practical use, if the maximum vaccination volume for chickens equal to 0.5 cm ³ contains at least 7 PD ₅₀ .

Thus, the above information indicates the fulfillment when using the invention of the following set of conditions:
- inactivated vaccine against SGPK, embodying the invention, is intended for use in agriculture, namely in veterinary virology and biotechnology;
- for the proposed invention in the form described in the independent claim, the possibility of its implementation using the means and methods given in the description or known prior to the priority date is confirmed;
- when using the proposed vaccine against SGPK achieved the technical result envisaged by the task of creating the invention.

 Therefore, the present invention meets the condition of patentability "industrial applicability".

Sources of information
1. Cowen BS, Lu H. et al. Characterization studies of fowl adenoviruces associated with inclusion body hepatitis / hydropericardium syndrome in chickens. Presented at the 68-th Northeastern Conference in Avian Diseases. June 10-12 1996, p.14.

 2. Lagutkin NA, Arsentiev A.R. et al. Massive hydropericarditis in young meat chickens. Questions of veterinary virology, microbiology and epizootology: Materials scientific. conf. VNIIVViM. - Pokrov, 1992,4.2,248-251.

3. Borisov VV, Borisov AV et.al. Hydropericardium syndrome in chickens in Russia // Proc. of the 11 ^th Int. Congress of the World Vet. Poult. Ass. August 18-22, Budapest, Hungary. - 1997. - P.258.

 4. Winterfield R.W., Fadly A.M. et al. Immunization of chicken against adenovirus infection. Poult. Sci., 1977, V. 56, No. 5, 1481-1486.

 5. Ahmad I., Malik M.I. et. al. Efficacy of formalinized level-organ-vaccine against Angara disease in broilers // Veterinarski Arhiv. - 1990. - V.60. - N 3. - P. 131-138 (prototype).

Claims (11)

 1. An inactivated vaccine against hydropericarditis syndrome of chickens containing antigenic material in the form of a purified virus-containing suspension from the liver of chickens infected with an infectious agent and fallen with signs of chicken hydropericarditis syndrome, and an inactivant, characterized in that it contains antigenic material from the chicken hydropericarditis virus strain of hens. Adenoviridae, genus Aviadenovirus, group 1, serotype 4, collection of VGNKI KR95 114-DEPT in an effective amount.  2. The inactivated vaccine according to claim 1, characterized in that it contains antigenic material from strain KP95 114-DEPT of the chicken hydropericarditis syndrome virus, reproduced in the body of 5-50 day old chickens. 3. Inactivated vaccine according to any one of paragraphs. 1 and 2, characterized in that it contains antigenic material from strain KP95 114-DEPT of the chicken hydropericarditis syndrome virus with biological activity of 3.25-5.0 1g LD ₅₀ / 0.2 cm ³ .  4. Inactivated vaccine according to claim 1, characterized in that the inactivant contains an ethyleneimine dimer.  5. The inactivated vaccine according to claim 4, characterized in that it contains an ethyleneimine dimer in the form of a 10% aqueous solution.  6. The inactivated vaccine according to claim 1, characterized in that it further comprises an adjuvant.  7. The inactivated vaccine according to claim 6, characterized in that the adjuvant contains alumina hydrate.  8. The inactivated vaccine according to claim 7, characterized in that it contains aluminum oxide hydrate in the form of a 3-4% colloidal solution.  9. The inactivated vaccine according to claim 6, characterized in that the adjuvant contains saponin.  10. The inactivated vaccine according to claim 9, characterized in that it contains saponin in the form of a 10% aqueous solution. 11. Inactivated vaccine according to any one of paragraphs. 1-10, characterized in that it contains antigenic material from strain KP95 114-DEPT of the chicken hydropericarditis syndrome virus, ethyleneimine dimer, aluminum oxide hydrate and saponin in a ratio, wt. %:
Antigenic material - 50.0-80.0
Ethyleneimine Dimer - 0.15-0.4
Alumina Hydrate - 17.6-49.35
Saponin - 0.5-2.0

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