RU2279291C1 - Vaccine against coypu salmonelasis - Google Patents

Abstract

FIELD: chemical and pharmaceutical industry, in particular vaccine against coypu salmonelasis.

SUBSTANCE: claimed vaccine contains suspension of pure culture cells of Salmonella typhimurium causative agent, obtained by sampling of pathological material from fallen coypus of local epidemical locus, suspension preparation, sowing on differential diagnostic media, isolation of Salmonella typhimurium causative agent pure culture, and culturing in meat-peptone broth with titer of 4-5 bln of microbial cells per 2 cm³. Vaccine also contains glucose, formalin and aluminum hydroxide in the next mass ratio: abovementioned pure culture cells of Salmonella typhimurium causative agent 83.0-86.0; glucose 1.0-2.0; formalin 1.0-2.0; and balance: aluminum hydroxide.

EFFECT: non-toxic vaccine having high reactivity.

5 ex, 1 tbl

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.

A known vaccine against salmonellosis of chickens and a method for the prevention of salmonellosis of chickens containing a suspension of cells of the vaccine strain Salmonella enteritidis VGNKI 204 with a concentration of live microbial cells of 175-250 billion in 1 cm ³ in the following ratio, vol.%:

cell suspension of Salmonella enteritidis strain VGNKI 204 with a concentration of living microbial cells 175-250 billion in 1 cm ³ 42.0-55.0 sucrose 5.0-7.5 gelatin 1.5-2.0 distilled water rest

To prevent chicken salmonellosis, this vaccine is administered to chickens of 2-3 days of age with water by drinking 0.5-1.0 billion microbial cells per chicken, twice with an interval of 48 hours after an 18-24 hour fasting (RF patent No. 2030916 from 04/07/92, MKI A 61 K 39/125, 39/135).

Known inactivated vaccine against salmonellosis of sheep, containing an immunostimulant in the form of a suspension of cells of the strain Rodococcus equi VIEV No. 2 in physiological saline with a concentration of 150 billion microbial cells, adjuvant - oil-oil mixture in a ratio of 83-87: 17-13, serovar strains Salmonella abortusovis VI , serovar Salmonella typhimurium VIEV No. 159, serovar Salmonella dublin VIEV No. 1449, in the following ratio of components, ml / l:

formalinized cell suspension of strains Salmonella abortusovis VIEV No. 876 with concentration of 100 billion microbial cells in 1 ml 190-210 formalinized cell suspension of strains Salmonella typhimurium VIEV No. 159 with a concentration of 100 billion microbial cells in 1 ml 95-105 formalinized cell suspension of strains Salmonella dublin VIEV№1449 with a concentration of 100 billion microbial cells in 1 ml  95-100 suspension of cells of the strain Rodococcus equi VIEV No. 2 in physiological saline with concentration 150 billion microbial cells in 1 ml 95-105

oil-lanolin mixture in the ratio 83-87: 17-13 the rest (RF patent No. 2092187 from 09/30/94, MKI A 61 K 39/125, 39/135).

The technology of this vaccine is complex, and this vaccine is not used to prevent salmonellosis in nutrias.

The technical solution to the problem of the invention is to eliminate the above disadvantages, in particular increasing the specificity, immunogenicity and effectiveness of the vaccine against salmonellosis in nutria, by introducing a new culture of the pathogen, components, excluding negative and side effects.

The solution to this problem is achieved by the fact that the nutria salmonellosis vaccine, including the inactivated antigen, adjuvant, as an antigen contains: a suspension of cells of a pure culture of the pathogen Salmonella typhimurium obtained by selection of the affected organs from fallen nutrias from the local epizootic focus, preparation of the suspension, plating for differential diagnostic environment, isolation of a pure culture of the pathogen Salmonella typhimurium and cultivation is carried out in a meat-peptone broth with a titer of 4.5 billion microbial cells per 1 cm ^3, making glucose form in and aluminum hydroxide in the following ratio, wt.%:

cell suspension of a pure pathogen culture Salmonella typhimurium isolated from affected organs from fallen nutria from local epizootic focus in a nutrient medium with a titer 4-5 billion microbial cells in 1 cm ³ 83.0-86.0 glucose 1.0-2.0 formalin 1.0-2.0 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 a pure culture of the pathogen Salmonella typhimurium is isolated, then grown on nutrient media use a pure culture of the pathogen Salmonella typhimurium, which allows to increase the specificity and immunogenicity of the vaccine. When cultivating a pure culture of Salmonella typhimurium in a meat-peptone nutrient medium with the addition of glucose to 0.2% concentration, it is possible to obtain a concentration of microbial cells of at least 4-5 billion in 1 cm ³ for 12-14 hours of incubation. Inactivation of a pure culture with formalin in a 0.4-0.5% final concentration at a temperature of 37 ° C for 72-96 hours can suppress the pathogenicity of the pathogen and maintain its immunogenic properties for 12 months. The introduction of a 3% solution of aluminum hydroxide in an amount of 15% of the culture volume 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 injection of at least 9 months (observation period). The use of a pure culture of the pathogen Salmonella typhimurium, isolated from fallen nutria from the local epizootic focus during salmonellosis, can significantly increase the specificity and immunogenicity of the vaccine, after a single vaccination provides reliable protection of animals from salmonellosis (table 1).

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.

As for 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 (instructions for the manufacture and control of formol vaccines against paratyphoid piglets, approved by the GUV Ministry of Agriculture of the USSR on 10.23.1996; instructions for the manufacture and control of a concentrated formol-alum vaccine against salmonellosis (paratyphoid) of calves, approved by the Main Veterinary Administration of the Ministry of Agriculture of the USSR on March 28, 1980). The proposed vaccine against salmonellosis nutria is simple to implement, affordable and is industrially applicable.

The nutria salmonellosis vaccine contains a formalin-inactivated pure culture of the pathogen Salmonella typhimurium 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 nutria salmonellosis vaccine is intended for the prevention of a dangerous infectious disease of nutria with salmonellosis. It causes the formation of specific immunity against salmonellosis nutria. The vaccine is harmless and areactogenic. Immunity in nutria vaccinated with the salmonellosis vaccine is formed 12-15 days after vaccination and lasts at least 9 months. The vaccine is used in successful, dysfunctional and threatened salmonella nutria farms. In farms that are safe and threatened by salmonellosis, the vaccine is administered once intramuscularly into the thigh area of young animals from 2 months of age at 1.0 cm ³ and for adult animals 1.5 cm ³ . In dysfunctional farms, the vaccine is administered intramuscularly in the thigh area from 2 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 of containing components in various proportions per 100 cm ^{3 of} vaccine are given.

Example 1

The affected organs are taken from the fallen nutria from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Salmonella typhimurium is isolated, the meat-peptone culture medium is infected, the suspension is cultivated, inactivated and mixed (wt.%) Suspension cells of a pure culture of the pathogen Salmonella typhimurium 83.0 with a titer of 4-5 billion and a glucose content of 1.0, inactivate by adding formalin 1.0, then add aluminum hydroxide, the rest is thoroughly mixed and half ayut vaccine with the following composition in the following ratio, wt.%:

cell suspension of a pure pathogen culture Salmonella typhimurium isolated from affected organs from fallen nutria from the local epizootic focus in salmonellosis in a nutrient medium with a titer 4-5 billion microbial cells in 1 cm ³ 83.0 glucose 1,0 formalin 1,0 aluminum hydroxide rest

With this ratio of components, the salmonellosis vaccine has weak immunogenic activity.

Example 2

The affected organs are taken from the fallen nutria from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Salmonella typhimurium is isolated, the meat-peptone culture medium is infected, cultivated, obtained, inactivated and mixed (wt.% ) a suspension of cells of a pure culture of the pathogen Salmonella typhimurium 84.0 with a titer of 4-5 billion and a glucose content of 1.5, inactivate the introduction of formalin 1.5, then add aluminum hydroxide - the rest, mix thoroughly ayut give a vaccine with the following composition in the following ratio, wt.%:

cell suspension of a pure pathogen culture Salmonella typhimurium isolated from affected organs from fallen nutria from the local epizootic focus in a nutrient medium with a titer of 4-5 billion. microbial cells in 1 cm ³ 84.0 glucose 1,5 formalin 1,5 aluminum hydroxide rest

With this ratio of components, the nutria salmonellosis vaccine has little immunogenic efficacy.

Example 3

The affected organs are taken from fallen nutria from the local epizootic focus, a suspension is prepared from pathological material, inoculated on meat-peptone broth and differential diagnostic media, a pure culture of the pathogen Salmonella typhimurium is isolated, the meat-peptone culture medium is infected, cultured, obtained, inactivated and mix (wt.%) a suspension of cells of a pure culture of the pathogen Salmonella typhimurium 85.0 with a titer of 4-5 billion and a glucose content of 1.0, inactivate by adding formalin 1.0, then add aluminum hydroxide nal, thoroughly mixed and receive a vaccine of the following composition in the following ratio of components, wt.%:

cell suspension of a pure pathogen culture Salmonella typhimurium isolated from affected organs from fallen nutria from the local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 85.0 glucose 1,0 formalin 1,0 aluminum hydroxide rest

With this ratio of components, the nutria salmonellosis vaccine has weak immunogenic activity.

Example 4

The affected organs are taken from the fallen nutria from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Salmonella typhimurium is isolated, the meat-peptone culture medium is infected, cultivated, obtained, inactivated and mixed (wt.% ) a suspension of cells of a pure culture of the pathogen Salmonella typhimurium 85.5 with a titer of 4-5 billion and a glucose content of 1.0, inactivate the introduction of formalin 1.5, then add aluminum hydroxide - the rest, mix thoroughly ayut give a vaccine with the following composition in the following ratio, wt.%:

cell suspension of a pure pathogen culture Salmonella typhimurium isolated from affected organs from fallen nutria from the local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 85.5 glucose 1,0 formalin 1,5 aluminum hydroxide rest

With this ratio of components, the vaccine against salmonellosis nutria has a high immunogenic activity, forms a tense immunity.

Example 5

The affected organs are taken from the fallen nutria from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Salmonella typhimurium is isolated, the meat-peptone culture medium is infected, cultivated, obtained, inactivated and mixed (wt.% ) a suspension of cells of a pure culture of the pathogen Salmonella typhimurium 86.0 with a titer of 4-5 billion and a glucose content of 2.0, inactivate the introduction of formalin 2.0, then add aluminum hydroxide-the rest, thoroughly mixing they get the vaccine of the following composition in the following ratio of components, wt.%:

cell suspension of a pure pathogen culture Salmonella typhimurium isolated from affected organs from fallen nutria from the local epizootic focus in a nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 86.0 glucose 2.0 formalin 2.0 aluminum hydroxide rest

With this ratio of components, the nutria salmonellosis vaccine has immunogenic activity and causes minor post-vaccination complications.

Table 1
Comparative vaccine efficacy Vaccine The dose of the vaccine in cm ³ Method of administration Immunobiological properties The number of nutrias in the experience (goal) The presence of post-vaccination complications Duration of immunity (months) Protection in (%) by Inside muscular Oppression, prototype twice chromate failure 1st dose from feed to 1.0 cm ³ ; 500 2-3 6 60-70 2nd dose days 2.0 cm ³ by Inside muscular offer 1500 no 9 90-100 mom way 1.0 cm ³

Claims (1)

A nutria salmonellosis vaccine containing an inactivated antigen, adjuvant, characterized in that the antigen contains a suspension of cells of a pure culture of the pathogen Salmonella typhimurium obtained by selecting the affected organs from fallen nutrias from the local epizootic focus, preparing the suspension, plating on differential diagnostic media, Isolation of a pure culture of the pathogen Salmonella typhimurium and cultivation is carried out in meat-peptone broth with a titer of 4-5 billion microbial cells in 1 cm ³ , glucose, formalin and hydrocoins are added smiling aluminum in the following ratio of components, wt.%: Pure culture cell suspension pathogen Salmonella typhimurium, isolated from affected organs from fallen nutria from the local epizootic focus in the nutrient medium with a titer of 4-5 billion microbial cells in 1 cm ³ 83.0-86.0 Glucose 1.0-2.0 Formalin 1.0-2.0 Aluminum hydroxide Rest