RU2301680C1 - Mixed vaccine against streptococcosus and enterococcal infection in nutria - Google Patents

Abstract

FIELD: veterinary medicine.

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

EFFECT: valuable properties of vaccine.

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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 containing bacterial Escherichia cells, strains: 0138 K88.09, K99, 200; Salmonella, strains: GISC No. 195, GISC No. 196, GISC No. 197; Proteins, strains: GISK №198, GISK №199; Klebsiella, strains: GISK No. 201, GISK No. 202 (patent for the invention of the Russian Federation No. 2035916, dated June 17, 93, MKI A61K 39/125, 39/135). This vaccine is intended for the prevention of infectious diseases in calves caused by Escherichia, Salmonella, Protein and Klebsiella, not used for nutrias.

Known vaccine against streptococcosis and pasteurellosis nutria (RF patent for the invention No. 2099084 from 09/30/94, MKI A61K 39/125, 39/135). This vaccine contains as antigen a suspension of the strain of Streptococcus zooepidemicus VGNKI No. K-DEPT with a titer of 2.5-3.0 billion microbial cells in 1 ml in culture medium, a suspension of cells of the strains Pasteurella multosida VGNKI No. 6011, Pasteurella multosida VGNKI No. 2394 and Pasteurella multocida VGNKI No. 1015 in equal proportions with a total content of 2.5-3.0 billion microbial cells in 1 ml of nutrient medium glucose, formalin, aluminum hydroxide and saponin. This vaccine is used to prevent streptococcosis and pasteurellosis nutria. Raw materials are obtained by culturing strains of pathogens of streptococcosis and pasteurellosis nutria in a nutrient medium with low activity. The use of saponin in the vaccine can cause various post-vaccination complications in vaccinated animals (inflammatory processes, abscesses at the injection site, lameness, etc.)

The technical solution to the problem of the invention is to remedy these shortcomings, in particular, increase the specificity and effectiveness of the vaccine for the prevention of streptococcosis and enterococcal infections of nutria by introducing new cultures of pathogens isolated from the epizootic foci, components, excluding negative and side effects.

The solution to this problem is achieved by the fact that the vaccine associated against streptococcosis and enterococcal nutria infection, including inactivated antigens, an adjuvant, contains as antigens a suspension of cells of pure cultures of Streptococcus pneumoniae pathogens and enterococcal Streptococcus fecalis infection obtained by selection of the diseased organs from the fallen nutria foci, preparation of the suspension, plating on differential diagnostic media, isolation of pure cultures of pathogens, cultivation by t separately in meat and peptone broth with glucose to a concentration of microbial cells of 4-5 billion in 1 cm ³ , mixed in equal proportions, formalin and aluminum hydroxide are added in the following ratio of components, wt.%:

cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 cm ³ 38.0-41.5 cell suspension of a pure culture of the pathogen enterococcal Streptococcus fecalis infection isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 cm ³ 38.0-41.5 glucose 2.0-1.0 formalin 2.0-1.5 aluminum hydroxide rest

The novelty of the claimed technical solution is due to the fact that, in contrast to the known vaccines, raw materials are selected from the fallen nutria from the local epizootic focus, suspension is prepared, plated on differential diagnostic media, and pure cultures of Streptococcus pneumoniae and Enterococcal Streptococcus fecalis infections are isolated , the cultivation of pure cultures of pathogens is carried out separately on meat and peptone broth, which allows to increase the specificity and immunogenicity of the vaccine.

Separate cultivation of pure cultures of pathogens and adding glucose to a nutrient medium to a 0.2% concentration gives a concentration of microbial cells of at least 4-5 billion in 1 ml for 12-14 hours of incubation. Inactivation of bakerya with formalin in a 0.4-0.5% final concentration at a temperature of 37 ° C for 72-96 hours makes it possible to suppress the pathogenicity of pathogens and preserve their immunogenic properties for 12 months. Mixing inactivated pathogens of Streptococcus pneumoniae streptococcosis and Streptococcus fecalis enterococcal infection in equal proportions, the introduction of aluminum hydroxide and thorough mixing does not cause post-vaccination complications in the grafted nutria, it allows the formation of intense immunity 12-15 days after a single administration of at least 9 months (observation period ) The use of pure cultures of the causative agents of Streptococcus pneumoniae and enterococcal infection Streptococcus fecalis isolated from the local epizootic focus from patients and fallen nutria can significantly increase the specificity of the vaccine, after a single vaccination it protects the animals from two dangerous infectious diseases of streptococcosis and enterococcal nutria infection.

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: formolvac vaccine against paratyphoid (salmonellosis) calves TU 46-21-166-75, approved 10.05.75, instructions for the manufacture and control of this vaccine approved by the General Directorate of Veterinary of the Ministry of Agriculture of the USSR 03/28/80; vaccine against paratyphoid (salmonellosis) piglets TU 46-21-952-74, approved July 15, 74, instructions for the manufacture and control of this vaccine approved by the General Directorate of Veterinary of the Ministry of Agriculture of the USSR 10.23.96; associated vaccine against salmonellosis, pasteurellosis and enterococcal infection of piglets TU 10.09-62-90, approved 01.08.90, instructions for the manufacture and control of this vaccine approved by the General Directorate of Veterinary Medicine 16.07.90; vaccine against enterococcal infection of calves, lambs and piglets TU 10.09-91-90, approved 15.12.90. However, for vaccines, these vaccines are not used.

Methods of isolation and characterization of streptococci are presented in the Methodological Instructions 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 of the USSR Council of Ministers for Food and Procurement, and also in the reference book “Identifier of zoopathogenic microorganisms” edited by the professor Sidorova M.A. M .: Kolos, 1995, pp. 90-95.

The proposed vaccine associated against streptococcosis and enterococcal nutria infection is simple to implement, affordable and is industrially applicable.

The vaccine associated against streptococcosis and enterococcal nutria infection contains pure formalin-inactivated pure cultures of causative agents of streptococcosis Streptococcus pneumoniae and enterococcal infection Streptococcus fecalis adsorbed on alumina hydrate. In appearance, the vaccine is a light yellow liquid with a loose precipitate, which, when shaken, easily breaks up 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 of nutria against streptococcosis and enterococcal nutria infection. It causes the formation of specific immunity against streptococcosis and enterococcal nutria infection. The vaccine is harmless and areactogenic. Immunity in nutria vaccinated with a vaccine against streptococcosis and enterococcal nutria infection is formed 12-15 days after vaccination and lasts at least 9 months. The vaccine is used in safe, dysfunctional and threatened by streptococcosis and enterococcal infections of nutria farms. In the farms that are safe and endangered by streptococcosis and enterococcal infection of nutria, the vaccine is administered once, intramuscularly in the thigh area, young animals from 45 days of age are 1.0 cm ³ , and adult animals 1.5 cm ³ . In dysfunctional farms, the vaccine is administered intramuscularly in the thigh area from two months of age twice with an interval of 7-10 days: the first vaccination at a dose of 1.0 cm ³ , the second at a dose of 1.5 cm ³ . Slaughter products of vaccinated nutria are used without restrictions, 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

The affected organs are taken from the fallen nutria from the local epizootic focus, from which the suspension is prepared, inoculated on differential diagnostic media, pure cultures of Streptococcus pneumoniae and Enterococcal Streptococcus fecalis infections are isolated, cultured separately, obtained, inactivated and mixed (wt.%) a suspension of microbial cells 75.0, with a titer of 4-5 billion, and a glucose content of 0.5, inactivate by adding formalin 1.0, then add aluminum hydroxide - the rest, mix thoroughly and get a vaccine with eduyuschego composition in the following ratio, wt.%:

cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 37.5 cell suspension of a pure culture of the pathogen enterococcal Streptococcus fecalis infection isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 37.5 glucose 0.5 formalin 1,0 aluminum hydroxide rest

With this ratio of components, the vaccine associated against streptococcosis and enterococcal nutria infection has weak immunogenic activity.

Example 2

The affected organs are taken from the fallen nutria from the local epizootic focus, from which the suspension is prepared, inoculated on differential diagnostic media, pure cultures of Streptococcus pneumoniae and Enterococcal Streptococcus fecalis infections are isolated, cultured separately, obtained, inactivated and mixed (wt.%) a suspension of microbial cells of 76.0, with a titer of 4-5 billion, and a glucose content of 1.0 is inactivated by the addition of formalin 1.5, then aluminum hydroxide is added - the rest is thoroughly mixed and a vaccine is obtained traveling composition in the following ratio of components, wt.%:

cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 38,0 cell suspension of a pure culture of the pathogen enterococcal Streptococcus fecalis infection isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 38,0 glucose 1,0 formalin 1,0 aluminum hydroxide rest

With this ratio of components, the vaccine associated against streptococcosis and enterococcal infection of nutria has immunogenic activity.

Example 3

The affected organs are taken from the fallen nutria from the local epizootic focus, from which a suspension is prepared, inoculated on differential diagnostic media, pure cultures of Streptococcus pneumoniae streptococcosis pathogens and Streptococcus fecalis enterococcal infections isolated from the local epizootic focus are isolated, cultivated separately, and obtained, inactivated mix (wt.%) a suspension of microbial cells 80.0 with a titer of 4-5 billion, and a glucose content of 1.7, inactivate by adding formalin 1.3, then add aluminum hydroxide - remained Noe, carefully mixed to obtain a vaccine with the following composition in the following ratio, wt.%:

cell suspension of a pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 40,0 cell suspension of a pure culture of the pathogen enterococcal Streptococcus fecalis infection isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 40,0 glucose 1.7 formalin 1.3 aluminum hydroxide rest

With this ratio of components, the vaccine associated against streptococcosis of enterococcal nutria infection has little immunogenic efficacy.

Example 4

The affected organs are taken from the fallen nutria from the local epizootic focus, from which a suspension is prepared, inoculated on differential diagnostic media, pure cultures of Streptococcus pneumoniae streptococcosis pathogens and Streptococcus fecalis enterococcal infections isolated from the local epizootic focus are isolated, cultivated separately, and obtained, inactivated mix (wt.%) a suspension of microbial cells 83.0, with a titer of 4-5 billion, and a glucose content of 1.0, inactivate the addition of formalin 1.5, then add aluminum hydroxide - remained It is thoroughly mixed and a vaccine of the following composition is obtained in the following ratio of components, wt.%:

cell suspension of pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 41.5 cell suspension of a pure culture of the pathogen enterococcal Streptococcus fecalis infection isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 41.5 glucose 1,0 formalin 1,5 aluminum hydroxide rest

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

Example 5

The affected organs are taken from the fallen nutria from the local epizootic focus, from which the suspension is prepared, inoculated on differential diagnostic media, pure cultures of Streptococcus pneumoniae nutria streptococcosis and Streptococcus fecalis enterococcal infections isolated from the local epizootic focus are isolated, they are cultivated separately, they are inoculated, they are cultivated separately, they are and mix (wt.%) a suspension of microbial cells 85.0, with a titer of 4-5 billion, and a glucose content of 2.0, inactivate the introduction of formalin 1.5, then add aluminum hydroxide - the rest is thoroughly mixed and receive a vaccine of the following composition in the following ratio of components, wt.%:

cell suspension of pure culture of the causative agent of streptococcosis Streptococcus pneumoniae isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 42.5 cell suspension of a pure culture of the pathogen enterococcal Streptococcus fecalis infection isolated from local epizootic foci in a nutrient medium with a titer 4-5 billion microbial cells in 1 ml 42.5 glucose 2.0 formalin 1,5 aluminum hydroxide rest

At this ratio of components, the vaccine associated against streptococcosis and enterococcal infection nutria has a weak immunogenic activity, forms a tense immunity, in some animals the injection of the drug causes slight irritation.

Table
Comparative vaccine efficacy Vaccine The dose of the vaccine, cm ³ Method of administration Immunobiological properties The number of nutria in the experiment (goal) The presence of post-vaccination complications Duration of immunity (months) Protection (%) According to the prototype twice the first dose of 1.0 cm ³
2nd dose 2.0 cm ³ Intramuscularly 1500 Oppression, lameness, refusal of food for 2-3 days 6 60-70 According to the proposed method 1.0 cm ³ Intramuscularly 1500 no 9 90-100

The table shows that the proposed vaccine associated against streptococcosis and enterococcal nutria infection has significant advantages compared with the vaccine of the prototype.

Claims (1)

An vaccine associated against streptococcosis and enterococcal nutria infection, containing as antigens a suspension of cells of pure cultures of streptococcosis causative agents Streptococcus pneumoniae and enterococcal infection Streptococcus fecalis, isolated from a suspension prepared from diseased organs of fallen nutria from a local epizootic glucose isolated separately microbial cell concentration of 4-5 billion. in 1 cm ^3, and mixed in equal ratios, formalin and aluminum hydroxide with the following ratio com onentov, wt.%: cell suspension of a pure pathogen culture streptococcosis Streptococcus pneumoniae, isolated from local epizootic foci in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 38.0-41.5 cell suspension of a pure pathogen culture enterococcal infection of Streptococcus fecalis, isolated from the local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 38.0-41.5 glucose 2.0-1.0 formalin 2.0-1.5 aluminum hydroxide rest