RU2553556C1 - Vaccine associated against colibacillosis, streptococcosis and staphylococcosis of cattle - Google Patents

Abstract

FIELD: veterinary medicine.

SUBSTANCE: characterised vaccine contains the antigens as cell suspension of pure cultures of pathogens of colibacillosis E. coli, streptococcosis - Streptococcus pneumonia, and staphylococcosis - Staphylococcus aureus in equal proportions, obtained by selection of pathological material from sick and dead cattle from local epizootic locus, as well as formalin and aluminium hydroxide.

EFFECT: invention has high immunogenic activity, it forms after vaccination the expressed intense immunity and does not cause post-vaccination complications.

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Description

The invention relates to veterinary medicine, in particular to vaccines, and can be used in institutions involved in the design of biological products, as well as in enterprises of the biological industry.

Known associated vaccine against calf diarrhea (patent for the invention of the Russian Federation No. 2035916 from 06.17.93, MKI A61 39/125, 39/135) containing ethanol inactivated cells of the Proteus vulgaris strain GISK N198, Proteus mirabilis GISK N199, Klebsiella pneumoniae GISK N201, Klebsiella pneumoniae GISK N201, Klebsiella pneumoniae GISK N201, pneumoniae GISK N202, from Escherichia and Salmonella antigens, ethanol inactivated cells of strains Escherichia coli 09 K99, Escherichia coli 0138 K88, Escherichia coli GISK N200, Salmonella typhimurium GISK N195, Salmonella Dublin GISK N196, Salmonella salmonella enter and as an adjuvant - aluminum hydroxide. This vaccine is intended for the prevention of infectious diseases in calves caused by Escherichia, Salmonella, Protein and Klebsiella.

Known inactivated emulsin vaccine against calf salmonellosis (RF patent for the invention No. 2154495 from 10.15.97, MKI AK61K 39/112, C12N 1/20, AK61/39/39). This vaccine contains as antigens a mixture of formalin-inactivated virulent strains of the genus Salmonella (S. dublin No. 1449, S. typhimurium No. 159) as an immunomodulator — a suspension of Rodococcus equi No. 2 cells in the oil-lanolin mixture, and the components that make up the vaccine are taken in the following ratio per liter: antigens in physiological saline with the addition of 0.4-0.5% formalin and a concentration of microbial cells in 1 ml - 90 billion (S. dublin No. 1449) 285-315 ml, antigen S. typhimurium No. 159 95-105 ml, suspension of Rodococcus equi No. 2 cells in physiological saline containing 200 JPL microbial cells; adjuvant - maslanolin mixture in a ratio of 83-87: 17-13 the rest. We accept the specified vaccine for the prototype.

The manufacturing technology of this vaccine is complex. This vaccine is used to prevent calf salmonellosis. The use of the oil-lanolin mixture in the vaccine can cause various vaccine-related complications in vaccinated animals (inflammatory processes, abscesses at the injection site, lameness, etc.).

The technical result of the invention is to increase the specificity, safety, effectiveness of the vaccine for the prevention of Escherichiosis, streptococcosis and staphylococcosis in cattle by introducing new pure cultures of pathogens, components, eliminating the negative and side effects.

The technical result is achieved by the fact that in the vaccine associated against Escherichiosis, streptococcosis and staphylococcosis of cattle containing a suspension of formalin inactivated antigens, the adjuvant according to the invention contains a suspension of pure cultures of Escherichia coli, Escherichia coli, streptococcosis streptococcosis streptococcosis streptococcosis streptococcosis and streptococcosis streptococcosis equal proportions obtained by selecting pathological material from a sick and fallen cattle from a local epizootic one chamber, as well as formalin and aluminum hydroxide with the following ratio of components, wt. %:

cell suspension of a pure culture of Escherichia coli Escherichia causative agent isolated from pathological material from a sick and dead cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0-29.0 cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0-29.0 cell suspension of a pure culture of the causative agent of staphylococcosis Staphylococcus aureus isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0-29.0 glucose 2.0-1.0 formalin 1.0-1.5 aluminum hydroxide rest

The novelty of the claimed technical solution is due to the fact that, in contrast to the known vaccines for the production of raw materials, pathological material is taken from a sick and fallen cattle from a local epizootic focus, suspension is prepared, culture on differential diagnostic media, isolation of pure cultures of Escherichia coli causative agents of Escherichia coli, streptococcosis Streptococcus pneumoniae and staphylococcosis Staphylococcus aureus, the cultivation of pure cultures of pathogens is carried out separately on meat-peptone broth, which allows to increase etsifichnost and immunogenicity of the vaccine.

Separate cultivation of pure cultures of pathogens and the addition of glucose to a nutrient medium to 0.2% concentration allows to obtain a concentration of microbial cells of at least 4 billion per 1 cm for 14-24 hours of incubation. Inactivation of pathogen cultures with formalin in a 0.4-0.5% final concentration at a temperature of 37 ° C for 72-96 hours allows to suppress the pathogenicity of pathogens and preserve their immunogenic properties for 12 months. Mixing inactivated Escherichia, streptococci and staphylococci antigens in equal proportions, the addition of aluminum hydroxide and thorough mixing allows us to obtain a vaccine that does not cause post-vaccination complications in vaccinated animals, which allows the formation of intense immunity in vaccinated animals (12-15 days after intramuscularly or subcutaneously twice administration with an interval of 7-14 days in a dose to cows of 5.0 cm ³ , calves aged 1.0-1.5 months - at a dose of 1.5-2.0 cm ³ ). The use of pure culture cells of Escherichia coli Escherichia coli causative agents, Streptococcus pneumoniae streptococcosis and Staphylococcus aureus staphylococcosis isolated from the local epizootic focus from sick and dead animals can significantly increase the specificity of the vaccine and protect animals from three dangerous infectious diseases at once: escherichiosis and large streptococcus cattle lasting at least 9 months (observation period).

According to the patent and scientific and technical literature, no similar combination of features, components was found, which allows us to judge the inventive step of the claimed technical solution.

Regarding the criterion of "industrial applicability", the components that make up the vaccine are known and widely used in veterinary practice in the manufacture of inactivated vaccines: the associated vaccine against salmonellosis, pasteurellosis and enterococcal infection of piglets TU 10.09-62-90, approved 01.08.90 g ., instructions for the manufacture and control of this vaccine approved by the General Directorate of Veterinary Medicine 07/16/90; vaccine against enterococcal infection of calves, lambs and piglets TU 10.09-91-90, approved 15.12.90,

Methods for isolating a pure culture of the causative agent of colibacteriosis and characteristics are described in the “Methodological guidelines for the bacteriological diagnosis of colibacteriosis (Escherichiosis) of animals”, approved July 27, 2000, No. 13-7-2 / 2117, by the Department of Veterinary Medicine of the Ministry of Agriculture of the Russian Federation, describes the characteristics of Escherichia in the reference The determinant of zoopathogenic microorganisms ”edited by Professor Sidorov MA, M .: Kolos, 1995, p. 196-201, as well as in the“ Workshop on Veterinary Microbiology and Immunology ”authors: Kostenko TS, Rodionova VB , Skorodumov D.I., M.: Kolos, 2001, on pages 219-222.

Methods of isolation and characterization of streptococci and staphylococci are described in the reference book “Determinant of zoopathogenic microorganisms” edited by Professor Sidorov MA, M .: Kolos, 1995, pp. 90-95; in the "Workshop on Veterinary Microbiology and Immunology" authors: Kostenko TS, Rodionova VB, Skorodumov DI, M .: Kolos, 2001, on pages 165 - 171; in the reference book “Microbiological diagnosis of animal bacterial diseases” authors: Skorodumov DI, Subbotin VV, Sidorov MA, Kostenko TS, M .: “IzograF”, 2005, on pages 246-257 ; in the “Guidelines for the laboratory diagnosis of animal streptococcosis”, approved on September 25, 1990 by the General Directorate of Veterinary Medicine with the State Inspectorate under the State Commission for Food and Purchasing of the USSR.

The proposed vaccine associated against Escherichiosis, streptococcosis and staphylococcosis in cattle is simple to implement, affordable and is industrially applicable.

The vaccine associated against Escherichiosis, streptococcosis and staphylococcosis in cattle contains formalin inactivated pure cultures of pathogens E. coli, Str. pneumoniae and Staph, aureus adsorbed on alumina hydrate. In appearance, the vaccine is a light yellow liquid with a loose precipitate, which, when shaken, easily breaks into a homogeneous suspension. Shelf life in a dry, dark place at a temperature of 2 to 10 ° C for 1 year. The vaccine is intended for the prevention of dangerous infectious diseases in cattle against Escherichiosis, streptococcosis and staphylococcosis in cattle. It causes the formation of specific immunity against escherichiosis, streptococcosis and staphylococcosis in cattle. The vaccine is harmless and areactogenic. The vaccine is used in dysfunctional and threatened by Escherichiosis, streptococcosis and staphylococcosis in cattle farms. Slaughter products of vaccinated animals are used without restriction, regardless of the timing of vaccination.

The factors that determine the receipt of a vaccine of a given property include the sequence of manufacture and the percentage ratio between the components included in the preparation.

Five examples are given containing components in various proportions per 100 ml of vaccine.

Example 1

Take pathological material from a sick and fallen cattle from a local epizootic focus, from which a suspension is prepared, then inoculate on differential diagnostic media, then cells of pure cultures of Escherichia coli causative agents are isolated, streptococcus Streptococcus pneumoniae and Staphylococcus aureus are separately grown and Staphyloccus a meat-peptone broth supplemented with glucose at a titer of 4-5 billion per 1 cm ^3, after which the cells were separately inactivated pathogens making formalin to 0.4% strength course concentration at 37 ° C for 72 hours, and occasional agitation, and then the cells were mixed with pure cultures in equal proportions, then a mixture of inactivated cultures contribute adjuvant - aluminum hydroxide, thoroughly mixed and the vaccine prepared with the following component ratio, wt. %:

cell suspension of a pure culture of the causative agent of Escherichiosis E. coli isolated from pathological material from a sick and dead cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 22.0 cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 22.0 cell suspension of a pure culture of the causative agent of staphylococcosis Staphylococcus aureus isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 22.0 glucose 0.5 formalin 1,0 aluminum hydroxide rest

With this ratio of components, the vaccine associated against Escherichiosis, streptococcosis and staphylococcosis in cattle has weak immunogenic activity.

Example 2

Take pathological material from a sick and fallen cattle from a local epizootic focus, from which a suspension is prepared, and then make inoculation on differential diagnostic media, after which cells of pure cultures of Escherichia coli, Escherichia coli, Streptococcus pneumoniae and Staphylococcus staphylococcosis Staphylococcus aureus separately isolated grown in a meat-peptone broth supplemented with glucose at a titer of 4-5 billion microbial cells per 1 cm ^3, thereafter cells were separately inactivated pure culture of pathogens in by adding formalin to a 0.4% final concentration at a temperature of 37 ° C for 72 hours and periodically stirring, then cells of pure cultures are mixed in equal proportions, then an adjuvant - aluminum hydroxide is added to the mixture of inactivated cultures, mixed thoroughly and a vaccine is obtained when the following ratio of components, wt. %:

cell suspension of a pure culture of the causative agent of Escherichiosis E. coli isolated from pathological material from a sick and dead cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0 cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0 cell suspension of a pure culture of the causative agent of staphylococcosis Staphylococcus aureus isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0 glucose 1,0 formalin 1,5 aluminum hydroxide rest

With this ratio of components, the vaccine associated against escherichiosis, streptococcosis and staphylococcosis in cattle has sufficient immunogenic activity.

Example 3

Take pathological material from a sick and fallen cattle from a local epizootic focus, from which a suspension is prepared, and then make inoculation on differential diagnostic media, after which cells of pure cultures of Escherichia coli, Escherichia coli, Streptococcus pneumoniae and Staphylococcus staphylococcosis Staphylococcus aureus separately isolated grown in a meat-peptone broth supplemented with glucose at a titer of 4-5 billion microbial cells per 1 cm ^3, thereafter cells were separately inactivated pure culture of pathogens in by adding formalin to a 0.4% final concentration at a temperature of 37 ° C for 72 hours and periodically stirring, then cells of pure cultures are mixed in equal proportions, then an adjuvant - aluminum hydroxide is added to the mixture of inactivated cultures, mixed thoroughly and obtained the next time the ratio of components, wt. %:

cell suspension of a pure culture of E. coli Escherichiosis isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 28.0 cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 28.0 cell suspension of a pure culture of the causative agent of staphylococcosis Staphylococcus aureus isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 28.0 glucose 1.7 formalin 1.3 aluminum hydroxide rest

At this ratio of components, the vaccine associated against Escherichiosis, streptococcosis and staphylococcosis in cattle has little immunogenic efficacy.

Example 4

Take pathological material from a sick and fallen cattle from a local epizootic focus, from which a suspension is prepared, and then sow on differential diagnostic media, after which cells of pure cultures of Escherichia coli, Escherichia coli, Streptococcus pneumoniae and Staphylococcus aureus Staphylococcus staphylococcosis and Staphylococcus aureus are isolated separately grown in meat-peptone broth with the addition of glucose with a titer of 4-5 billion microbial cells in 1 cm ³ , then cells of pure cultures of pathogens are separately inactivated in carrying formalin to a 0.4% final concentration at a temperature of 37 ° C for 72 hours and periodically stirring, then mix the cells of pure cultures in equal proportions, then add the adjuvant - aluminum hydroxide to the mixture of inactivated cultures, mix thoroughly and get the next the ratio of components, wt. %:

cell suspension of a pure culture of the causative agent of colibacteriosis E. coli isolated from pathological material from a sick and dead cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 29.0 cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 29.0 cell suspension of a pure culture of the causative agent of staphylococcosis Staphylococcus aureus isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 29.0 glucose 1,0 formalin 1,5 aluminum hydroxide rest

At this ratio of components, the vaccine associated against escherichiosis, streptococcosis and staphylococcosis in cattle has high immunogenic activity, forms pronounced intense immunity after vaccination, does not cause post-vaccination complications, and is stable during storage for 12 months (table 1).

Example 5

Take pathological material from a sick and fallen cattle from a local epizootic focus, from which a suspension is prepared, and then make inoculation on differential diagnostic media, after which cells of pure cultures of Escherichia coli, Escherichia coli, Streptococcus pneumoniae and Staphylococcus staphylococcosis Staphylococcus aureus separately isolated grown in a meat-peptone broth supplemented with glucose at a titer of 4-5 billion microbial cells per 1 cm ^3, thereafter cells were separately inactivated pure culture of pathogens in by adding formalin to a 0.4% final concentration at a temperature of 37 ° C for 72 hours and periodically stirring, then cells of pure cultures are mixed in equal proportions, then an adjuvant - aluminum hydroxide is added to the mixture of inactivated cultures, mixed thoroughly and obtained the next time the ratio of components, wt. %:

cell suspension of a pure culture of the causative agent of Escherichiosis E. coli isolated from pathological material from a sick and dead cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 30,0 cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 30,0 cell suspension of a pure culture of the causative agent of staphylococcosis Staphylococcus aureus isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 30,0 glucose 2.0 formalin 1,5 aluminum hydroxide rest

With this ratio of components, the vaccine associated against escherichiosis, streptococcosis and staphylococcosis in cattle has immunogenic activity, in some animals the injection of the drug causes slight irritation.

From the data of table 1 it is seen that the proposed vaccine associated against Escherichiosis, streptococcosis and staphylococcosis in cattle has significant advantages compared with the vaccine of the prototype.

Claims (1)

Vaccine associated against Escherichiosis, streptococcosis and staphylococcosis of cattle, containing a suspension of formalin inactivated antigens, an adjuvant, characterized in that the antigens contain a suspension of cells of pure cultures of causative agents of Escherichiosis E. coli, Streptococcus pneumococcus pneumonia streptococcus, and Streptococcus pneumonia by selecting pathological material from a sick and fallen cattle from a local epizootic focus, as well as formalin and aluminum hydroxide When the following component ratio, wt. %:
cell suspension of a pure culture of the causative agent of Escherichiosis E. coli isolated from pathological material from a sick and dead cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0-29.0 cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0-29.0 cell suspension of a pure culture of the causative agent of staphylococcosis Staphylococcus aureus isolated from pathological material from a patient and fallen cattle from a local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 24.0-29.0 glucose 2.0-1.0 formalin 1.0-1.5 aluminum hydroxide rest