Associated vaccine against anthrax and plague in cattle

FIELD: biotechnology.

SUBSTANCE: the suggested vaccine contains suspension of viable spores of anthracic vaccinal strain "55-VNIIBB&M" at initial concentration of 500-700 mln. spores/cu.cm, cultural virus-containing raw material of vaccinal virus of cattle plague of "LT" strain at activity of not less than 7.0 lg TCD50/cu. cm, lactosopeptonic stabilizing agent and distilled water at the following content of components,%: spores of anthracic strain "55-VNIIVV&M" -6.2 - 10.0; cultural raw material of cattle plague virus of "LT" strain -25.0 - 30.0; lactosopeptonic stabilizing agent -48.0 - 50.0; distilled water - the rest. One vaccinal dosage contains about 20-25 mln. anthracic spores and about 4.5-5.5 lg TCD50 of cattle plague virus The suggested vaccine is of high immunogenicity, develops tense immunity in once vaccinated cattle that lasts for 12 mo, not less, moreover, it is areactogenous, safe and stable at storage.

EFFECT: higher efficiency.

3 ex, 3 tbl

 

The invention relates to the field of veterinary medicine, in particular to the use of biological agents for simultaneous specific prophylaxis of anthrax and rinderpest.

Currently known vaccine against anthrax, which is a suspension of live spores paskapallero avirulent vaccine strain "55-Vniivvim" [1].

Known also cultural virusvaktsinu against rinderpest, made of vaccine strain "LT" [2, 3].

Abroad for the prevention of these diseases shall be applied vaccine made from strains 34F2(Sterne) and Weybridge anthrax, as well as from strain TCRV and Plowright W. virus rinderpest [4, 5].

Existing vaccines are used independently from each other, and their application may not create immunity in animals against anthrax and plague of cattle. The use of these vaccines separately requires twice the amount of work and provides additional stress on the animal organism.

The literature describes the results of research on the development of associated vaccines based on antigens of bacterial and viral nature: against anthrax and smallpox of sheep using the anthrax vaccine strain "GNCI and vaccine strain "To" smallpox is vec [6]: associated vaccine against anthrax and smallpox of sheep using a vaccine strain "55-Vniivvim" anthrax and strain "Nishi" sheep pox [7]; the associated vaccine against anthrax and foot and mouth disease [8]; inactivated polyvalent vaccines against foot and mouth disease and cattle plague [9].

Materials to create the associated vaccine against anthrax and rinderpest in the available literature is not found.

The purpose of the invention is the creation of associated vaccine against anthrax and rinderpest, a highly immuno-heterogeneity distorts, reactogenicity, safety, storage stability and inducing the formation of equivalent stress immunity, protecting animals from infection cultures virulent strains of anthrax and rinderpest virus of cattle. Its use in animal husbandry will significantly reduce the labor costs of veterinary specialists, to reduce stress on the animal organism when vaccinations, in a shorter time to create lasting well-being in relation to infectious diseases of bacterial and viral etiology.

The objective is achieved by mixing suspensions of spores anthrax vaccine strain "55-Vniivvim" culture vaccinated raw vaccine virus rinderpest strains of "LT" and a stabilizer, followed by lyophilization with the following content of components in the vaccine (volume is s %): spores of the anthrax strain 6,2-10,0; cultural raw materials of rinderpest virus of cattle - 25,0-30,0; lactation stabilizer - 48,0-50,0; distilled water - the rest.

Biological activity of the thus prepared vaccine is: anthrax component 500-700 mlnister/cm3by viral component not lower than 7.0 lg TCD50/cm3that corresponds to 20-30 prilivnym doses. In one pravilnoy dose of vaccine contains 20-25 million anthrax spores and 4.5-5.5 lg TCD50virus rinderpest.

In the result of the experiments that the proposed associated vaccine against anthrax and rinderpest made from anthrax strain 55-Vniivvim and cultural strain "LT" virus rinderpest creates immunity once vaccinated animals after 10-12 days at a time against anthrax and plague of cattle a duration of at least 12 months (observation period). On immunogenicity she is not inferior to the monovaccines used in these diseases, and stable during storage. Examples of specific performance:

Example 1.

To 25,0 ml culture vaccinated raw material of the virus rinderpest strain "LT", with the activity of 7.0 lg TCD50/ml, added 6.2 ml of a suspension of spores of strain 55 Vniivvim" anthrax, 48,0 ml lactation is stabilizator (peptone - 20%, lactose - 4%) and 20.8 ml of distilled water. Initial concentration of anthrax component in the finished mixture is 500-700 mlnister/cm3. The mixture is stirred and freeze-dried until the content of the mass of moisture in the range of 1-4%.

The vaccine is a homogeneous porous tablet from light brown to brown.

The test results of the biological properties and physico-chemical parameters thus prepared 4-x series associated vaccines are given in table 1.

Table 1

Biological properties and physico-chemical characteristics of the associated vaccine
Characteristics vaccinesThe results of the biological and physico-chemical properties of the series
Series, No. 1Series, No. 2Series No. 3Series No. 4
The content of viable spores pieces 55-Vniivvim.750 million500 million676 million630 million
spores/cm3spores/cm3spores/cm3spores/cm3
The contents of the virus7,5 lg6,5 lg7,0 lg7,0 l
(caption) the plague of cattle.TCD50/cm3TCD50/cm3TCD50/cm3TCD50/cm3
Mass fraction of spores of strain 55-Vniivvim.98%98%98%98%
Mass fraction of moisture, %2,82,62,52,6
The number of doses in 1 cm3vaccine.30202825

Example 2.

In experiments on Guinea pigs, rabbits and calves it is shown that the associated vaccine is not inferior to the immunogenicity of the monovaccines against anthrax and rinderpest made from these strains.

The validation results of the immunogenicity of 3 series of associated vaccine are shown in table 2.

The percentage protection of Guinea pigs infected referencearray culture anthrax strain M-71 in 10-12 days after vaccination, was an associate of the drug 92,6%, and for anthrax single was 86.7%. The vaccine was protected from disease 100% of the vaccinated calves infected with a virulent strain No. 76 anthrax and virulent strain Of virus rinderpest. Similar results were obtained PR the infection control animals after 8 months after immunization.

Table 2

Comparison of indicators of immunogenicity of mono - and combined vaccines against anthrax and rinderpest
VaccineView animalQty

animals (animals)
Antibody titers after 21 days after vaccinationThe results of the control of infection
Rnga (B.)PH (virus CCRS)Goals% protection
SurvivedPalo
AssociatedGuinea pigs27--25292,6
calves11:3201:3:10100
rabbits6-1:32-1:64---
Anthrax a monovalentGuinea pigs30--26486,7
calves21:160-1320 -20100
The monovalent against cattle plaguecalves2-1:3220100
rabbits31:32-1:64
ControlGuinea pigs15--0150
calves2--020

In vaccinated animals was hematological analysis (blood picture and leucoformula) and received the serum for research on the availability of appropriate antibodies. Blood samples were collected before vaccination and after 5, 7, 14 and 21 days after it. After 21 days after administration of the vaccine in calves title protivoseborainey antibodies in rnga was 1:320, and the titers of antiviral antibodies in PH in the cell culture PS was 1:32; in rabbits the antiviral antibody titer was 1:32-1:64 (14-28 days).

In the result of the experiments that the proposed associated vaccine against anthrax and plague of cattle creates at once inoculated large R the Gato cattle tight immunity to these infections duration of at least 12 months and is currently released to monovaccines against these diseases.

Example 3.

We studied the persistence of dry associated vaccines at temperatures of 4±2°C (storage temperature), 18±2°37±2°C. the Results are shown in table 3.

As can be seen from the table, at the recommended storage temperature (4±2° (C) the vaccine components retain their original activity in the past 12 months (observation period). Activity associated drug in 18±2°With storage for 4 months will remain at the original level, which is important in the use of the vaccine in the southern regions of Russia.

Table 3

The persistence of associated vaccine at different

temperature modes.
A series of vaccinesComponentStorage temperature °The initial activityActivity during storage
1 month.2 months.to 4 months.8 months.12 months.14 months.
No. 3Anthrax4±2°676±25655±18674±22675±21672±17681±24638±18
18±2°676±25650±19637±18644±22574±18453±17323±14
37±2°676±25545±22390±14351±17310±14265±12210±12
Viral4±2°7,07,07,07,07,07,07,0
18±2°7,07,06,56,5the 5.254,0-
37±2°7,06,25the 5.254,0---
Note: the activity of anthrax component (St-Vniivvim) expressed in mlnister/cm3viral (STL) - lg TCD50/cm3.

Literature

1. Patent No. 2095409, 1997, Bakulev I.A., Gavrilov V., Seliverstov CENTURIES and other "Method of making a vaccine against anthrax animals".

2. THE 10-07-081-92 "Virusvaktsinu from strain "LT" against rinderpest liofilizirovannaya".

3. Mitin N, Petrov SCI freeze-Dried virusvaktsinu from the strain of the LITAS against rinderpest. Materials of scientific-practical Conf., Cover, 1992, p.146-147.

4. Gennady Onishchenko, etc. "anthrax: actual aspects of Microbiology, epidemiology, clinical presentation, diagnosis, treatment and prevention", 1999 - 448 S. veterinary 1989. No. 3. - P.71-72.

5. Plowright, W., Ferris R. Cytopathogeniutu of rinder pest virus in tissues culture. Nature, 1957, v.179, N7, p.708-712.

6. Likhachev, NV, Kolosov YEAR, B. .F., Presnov I.I. "Affiliated vaccination of sheep against anthrax and smallpox". Tr. GNCI veterinary preparations, 1967. - T. - P.35-44.

7. Patent No. 2181296, 2002, Bulychev V.M., Selyaninov CREATING, Bakulev IA and other Associated vaccine against anthrax and smallpox of sheep.

8. Patent No. 2056861, 1996, Bakulev I.A., Gavrilov, V.A., Sobakin A.S. and other Associated vaccine against anthrax and foot and mouth disease and the way to prevent anthrax and foot and mouth disease.

9. THE 9384-102-0049552-94 "Inactivated associated polyvalent vaccine against foot and mouth disease and cattle plague".

The associated vaccine against anthrax and rinderpest, containing a suspension of viable spores of the anthrax vaccine strain "55-Vniivvim" in the initial concentration 500-700 million spores/cm3cultural vaccinated raw vaccine virus rinderpest strain "LT" activity not less than 7.0 lg TCD50/cm3, lactation stabilizer and distilled water in the following content is the research Institute of the components in the vaccine, vol.%:

Spores of anthrax
strain 55-Vniivvim"6,2-10,0
Cultural raw materials of rinderpest virus of cattle
strain "LT"25,0-30,0
Lactation stabilizer48,0-50,0
Distilled waterRest



 

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