RU2288004C1 - Vaccine against colibacteriosis in nutrias - Google Patents

Abstract

FIELD: veterinary medicine.

SUBSTANCE: the suggested vaccine as an antigen contains the suspension of cells of pure culture of Escherichia coli agent obtained due to selecting affected organs in dead nutrias from local epizootic focus to prepare the suspension followed by inoculation onto differentially-diagnostic media; then one should isolate pure culture of Escherichia coli agent to be grown in beef-infusion-peptone broth at addition of glucose at the titer being about 5-6 bln. microbial cells/cu. cm followed by inactivation due to introducing formalin and aluminum hydroxide at the following ratio of components, weight%: suspension of cells of pure culture of Escherichia coli agent 83.0-85.5; glucose 1.0-2.0; formalin 1.0-2.0; aluminum hydroxide - the rest. The vaccine is safe, highly immunogenic and stable at storage.

EFFECT: higher efficiency.

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 animal Escherichiosis (RF patent for the invention No. 2043771, 02.25.93, MKI A 61 K 39/125, 39/135). The vaccine includes various Escherichia antigens: somatic - 078, 0141, adhesive proteins - K88, K99, 987P, F41, capsular polysaccharide - K80, K87, TL, TS - toxoids. The authors believe that the components of the vaccine provide the synthesis of specific antibodies in the blood of immunized animals that inhibit the main processes of coli infection - colonization of the pathogen in the intestines of animals and the neutralization of endo- and enterotoxins. For the prevention of neonatal escherichiosis, the vaccine is administered to pregnant animals twice with an interval of 12-14 days for 1.5-2 months before birth, as well as to piglets and lambs before weaning. However, the technology of such a vaccine is very complex. It is necessary to grow individual Escherichia strains, isolate the necessary components from them, process them, then mix in a certain ratio. For nutria, this vaccine is not used.

A known vaccine against calves Escherichiosis (RF patent for the invention No. 2070819, 04.20.93, MKI A 61 K 39/125, 39/135). This vaccine against calves Escherichiosis involves the cultivation of toxigenic strains of Escherichia coli in Hottinger medium for 5-7 days, the microbial mass is separated from the toxin on filters with sterilizing plates, then the toxin is inactivated by adding formalin to 0.3-0.4% at a temperature of 37 ° C for 10-12 days, the resulting toxoid is precipitated with a 10% solution of alum-potassium alum at a rate of 1% for the entire volume, then after settling for 2-3 days, the supernatant is removed, the resulting complex toxoid is mixed in equal amounts fractions with antigen obtained from Escherichia coli strains after growing daily broth cultures at 37 ° C for two days and washing the microbial mass with saline containing 0.3-0.4% formalin, bringing to a concentration of 50-60 billion microbial cells in 1 ml and adding hydrogen peroxide to the flush to its content up to 0.15-0.25%, moreover, the mixture of toxoid and antigen is diluted with sterile distilled water to a concentration of 5 billion microbial cells in 1 ml.

This vaccine against calves Escherichiosis is used only for the prevention of calves Escherichiosis, raw materials are obtained by culturing toxigenic Escherichia strains, a long period of inactivation of antigens can reduce the immunogenic activity of the drug.

The use of potassium alum, hydrogen peroxide in the vaccine can cause vaccine-related complications in vaccinated animals. Numerous technological methods used in obtaining the vaccine complicate the manufacturing technology and cost of the final product.

The technical solution to the problem of the invention is to remedy these shortcomings, in particular, increase the specificity and immunogenicity of the vaccine against colibacteriosis of nutria, which is stable during storage, by introducing a new culture of the pathogen and effective components.

The solution is achieved by the fact that the vaccine against colibacteriosis of nutria, including the inactivated antigen, adjuvant, as an antigen contains a suspension of cells of a pure culture of the pathogen Escherichia coli obtained by selecting the affected organs from fallen nutria from the local epizootic focus, preparing the suspension, plating for differential diagnostic medium, isolation of the pure culture of the pathogen and cultivation, which is carried out in a meat and peptone broth with glucose in the titer of 5-6 billion microbial cells in 1 cm ³ make formalin and aluminum hydroxide in the following ratio, wt.%:

suspension of cells of a pure culture of the pathogen Escherichia coli in the meat-peptone broth with a titer of 5-6 billion microbial cells in 1 cm ³ 83.0-85.5 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, a pure culture of the pathogen Escherichia coli is isolated and grown in meat and peptone broth , which allows to increase the specificity and immunogenicity of the vaccine. Adding glucose to a nutrient medium up to 0.2% concentration allows to obtain a concentration of microbial cells of at least 5-6 billion in 1 cm ³ . Inactivation of bakerya with formalin in a 0.5-0.6% final concentration at a temperature of 37 ° C for 72-96 hours with periodic stirring allows to suppress the pathogenicity of the pathogen and maintain immunogenic properties for 12 months. The use of a 3% solution of aluminum hydroxide in an amount of 15% by 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 intramuscular or subcutaneous injection of at least 9 months (observation period) .

According to the scientific, technical and patent literature, a similar set of features, components was not 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 (instructions for the manufacture and control of polyvalent aluminum hydroxide formol vaccine against colibacteriosis (Escherichiosis) of piglets, calves and lambs, approved by the GUV of the Ministry of Agriculture USSR 04/26/1984; instructions for the manufacture and control of a vaccine against bradzot, infectious enterotoxemia, malignant edema of sheep and dysentery of lambs, multivalent concentrate ƈ This gidrookisalyuminievaya approved by the Main Department of Veterinary Medicine with the State Veterinary Inspectorate of 10.10.1991). The proposed vaccine against colibacteriosis of nutria is simple to implement, affordable and is industrially applicable.

The nutria colibacteriosis vaccine contains a formalin-inactivated pure culture of the pathogen Escherichia coli 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 colibacteriosis vaccine is intended to prevent the infectious disease of nutria. It causes the formation of specific immunity against colibacteriosis of nutria. The vaccine is harmless and areactogenic. Immunity in nutria vaccinated with a colibacteriosis vaccine is formed 12-15 days after vaccination and lasts at least 9 months. The vaccine is used in safe, dysfunctional and colibacteriosis-threatened nutria farms. In safe and threatened colibacteriosis farms, 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 at 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 determining 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 cm ^{3 of} vaccine.

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 Escherichia coli is isolated, the meat and peptone broth is infected, cultured, the cell suspension is obtained, inactivated and mixed (wt.%) Cell suspension . a pure culture with a titer of 82.0 6.5 billion microbial cells per 1 cm ³ and 0.9 glucose, 0.9 formalin inactivated introduction, is then added aluminum hydroxide - the rest, mixed thoroughly to give aktsinu the following component ratio, wt.%:

cell suspension of a pure culture of the pathogen Escherichia coli isolated from diseased organs from fallen nutria from the local epizootic focus for colibacteriosis in meat and peptone broth with a titer of 5-6 billion in 1 cm ³ 82.0 glucose 0.9 formalin 0.9 aluminum hydroxide rest

With this ratio of components, the nutria colibaceriosis vaccine is not very effective; it may cause slight irritation in some animals at the injection site.

Example 2

The affected organs are taken from fallen nutrias from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Escherichia coli is isolated, the meat and peptone broth is infected, cultured, the microbial suspension is obtained, inactivated and mixed (wt.%) pure culture cells 83.0 with a titer of 5-6 billion microbial cells in 1 cm ³ and a glucose content of 1.0, inactivate by adding formalin 1.0, then add aluminum hydroxide - the rest, mix thoroughly and receive the vaccine in the following ratio of components, wt.%:

cell suspension of a pure culture of the pathogen Escherichia coli isolated from the local epizootic focus in the meat-peptone broth with a titer of 5-6 billion microbial cells in 1 cm ³ 83.0 glucose 1,0 formalin 1,0 aluminum hydroxide rest

With this ratio of components, the nutria colibacteriosis vaccine has good immunogenic activity and does not cause post-vaccination complications in animals.

Example 3

The affected organs are taken from fallen nutrias from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Escherichia coli is isolated, the meat and peptone broth is infected, cultured, the microbial suspension is obtained, inactivated and mixed (wt.%) pure culture cells 84.5 with a titer of 5-6 billion microbial cells in 1 cm ³ and a glucose content of 1.0, inactivate by adding formalin 1.5, then add aluminum hydroxide - the rest, mix thoroughly and receive the vaccine in the following ratio of components, wt.%:

cell suspension of a pure culture of the pathogen Escherichia coli isolated from the local epizootic focus in the meat-peptone broth with a titer of 5-6 billion microbial cells in 1 cm ³ 84.5 glucose 1,0 formalin 1,5 aluminum hydroxide rest

At this ratio of components, the vaccine against colibacteriosis of nutria is not very effective; it may cause mild irritation in some animals at the injection site.

Example 4

The affected organs are taken from fallen nutrias from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Escherichia coli is isolated, the meat and peptone broth is infected, cultured, the microbial suspension is obtained, inactivated and mixed (wt.%) cells of pure culture 85.5 with a titer of 5-6 billion microbial cells in 1 cm ³ and a glucose content of 2.0, inactivate by adding formalin 2.0, then add aluminum hydroxide - the rest, mix thoroughly and receive the vaccine in the following ratio of components, wt.%:

cell suspension of a pure culture of the pathogen Escherichia coli isolated from the local epizootic focus in the meat-peptone broth with a titer of 5-6 billion microbial cells in 1 cm ³ 85.5 glucose 2.0 formalin 2.0 aluminum hydroxide rest

With this ratio of components, the vaccine against colibacteriosis nutria has a pronounced efficacy, does not cause inflammatory processes in animals at the injection site (table).

Example 5

The affected organs are taken from fallen nutrias from the local epizootic focus, a suspension is prepared from pathological material, inoculated on differential diagnostic media, a pure culture of the pathogen Escherichia coli is isolated, the meat and peptone broth is infected, cultured, the microbial suspension is obtained, inactivated and mixed (wt.%) 86.5 cells with a titer of 5-6 billion microbial cells in 1 cm ³ and a glucose content of 2.5, inactivate by adding formalin 2.5, then add aluminum hydroxide - the rest, mix thoroughly and receive vaccines in the following ratio of components, wt.%:

cell suspension of a pure culture of the pathogen Escherichia coli isolated from the local epizootic focus in the meat-peptone broth with a titer of 5-6 billion microbial cells in 1 cm ³ 86.5 glucose 2,5 formalin 2,5 aluminum hydroxide rest

With this ratio of components, the nutria colibacteriosis vaccine has weak immunogenic activity and does not cause post-vaccination complications in animals.

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 Immunity duration Protection,% proposed Intramuscularly 1000 no 9 months one hundred 1,0 prototype Inside- 500 Oppression, Twice: the first dose is 1.0; 2nd dose -2.0 muscularly lameness, refusal of food within 1 -2 days 6 months 60-70

Thus, the proposed vaccine against colibacteriosis of nutria is more specific and more effective than the prototype.

Claims (1)

A nutria colibacteriosis vaccine containing an inactivated antigen, adjuvant, characterized in that the antigen contains a suspension of cells of a pure culture of the pathogen Escherichia coli obtained by selecting the affected organs from fallen nutria from the local epizootic focus, preparing the suspension, plating on differential diagnostic media, isolation of a pure culture of the pathogen and cultivation, which is carried out in a meat-peptone broth with glucose in the titer of 5-6 billion microbial cells in 1 cm ³ , then formalin and hydroc aluminum in the following ratio, wt.%: Suspension of pure pathogen cell culture Escherichia coli in meat and peptone broth with titer of 5-6 billion microbial cells in 1 cm ³ 83.0-85.5 Glucose 1.0-2.0 Formalin 1.0-2.0 Aluminum hydroxide Rest