Virusvaktsinu against rabies for oral immunization of carnivores

 

(57) Abstract:

The invention is intended for the specific prophylaxis of rabies carnivorous. The vaccine contains a freeze-dried suspension of virus strain TC-80 and stabilizer. As a stabilizer that protects the virus inactivating effect of gastric juice and environmental factors, it contains peptone, lactose and pectin. After twice with an interval of 12 to 14 days of vaccination, the vaccine stimulates the vaccinated animals the production of neutralizing antibodies in the range of 1 : 16 to 1 : 32. The invention allows to obtain sublimation vaccine against rabies with high immunological efficacy of oral immunization of carnivores.

The invention relates to the field of veterinary Virology, namely the production and application of biological preparations for oral immunization of carnivores against rabies.

Known dry virusvaktsinu for oral immunization of wild carnivores from strain RB-71, which before use rehydration and syringe injected into the bait in the form of pieces of fish, meat or chicken heads (abstracts of the international Symposium "Problems of pathology and protection of the Islands, 1992).

However, the procedure for preparation of baits vaccine immediately before inoculation complicates the process of vaccination, and short shelf life of the vaccine in such baits reduces the effectiveness of immunization.

Have oral application of a vaccine against rabies, which consists of a liquid suspension of rabies virus, strain "GARDEN", placed in the internal cavity of the lure in the form of briquettes (Temporary instruction for use of the drug "Livolin for immunization of wild animals (in order of production tests in 1998-2000,), approved by the veterinary Department of the Ministry of agriculture of the Russian Federation 26.03.98,, N-13-3-2/1197). The disadvantage of this vaccine is that it is unstable at positive temperatures, which makes inefficient use it in the summer-autumn period for immunization of young carnivorous.

Also known vaccine from strain TC-80 (Ru 2077338 Cl, A 61 K 39/208). Vaccine strain TC-80 antigen, stable, has no variability in potency against different species of animals, deposited in VGNKI 17.02.88, (Case N 10V). The vaccine is produced by growing the virus in human cell cultures kidney saiga in serum-free nutrient is. The disadvantage of this vaccine is that oral immunization she has no immunological effectiveness, since the antigen is destroyed by the action of gastric juice.

The aim of the present invention is a virus vaccine against rabies in the form of dry polydisperse powder, dried by sublimation, immunological effective oral immunization.

This goal is achieved by the fact that the suspension of rabies virus strain TC-80, subjected to freeze-drying, pre-mixing with components that protect the virus inactivating effect of gastric juice and environmental factors, grind in a ball mill and fill cake-bait. Protective components are peptone, lactose and pectin.

A positive effect is achieved in the following ratio, wt. %:

suspension of rabies virus strain TC-80 - 5-7

peptone - 65 - 67

lactose - 11 - 13

pectin - rest

Examples of specific performance

Example No. 1.

100010 cm suspension of rabies virus strain TC-80, grown in human cell cultures kidney saiga, mixed with an equal volume of a stabilizing environment, steriade subjected to freeze-drying. Dry vaccine grind in a ball mill, determine biological activity, sterility and safety, then fill briquettes-bait, which prior application stored at a temperature of 6-8oC.

Serological analysis of blood serum samples carnivorous immunized twice with an interval of 12 days, revealed titers of neutralizing antibodies in foxes and raccoon dogs from 1:16 to 1:32 2 months after vaccination. Titles preimmune serum samples did not exceed 1:2.

Example No. 2.

5005 cm suspension of rabies virus strain TC-80, grown in human cell cultures kidney saiga, mixed with two volumes 4,00,5% pectin and one volume of a stabilizing medium containing 35,00,5% peptone and 8,01,0% lactose. Received technical liquid poured into 150 cm vials and subjected to freeze-drying. Dry vaccine crushed ice rink, define the characteristics and fill briquettes-bait. Bait vaccine stored at a temperature of 6-8oC until use.

Serological analysis of blood serum samples carnivorous immunized twice with an interval of 14 days, showed the presence of neutralizing anti-Christ. x serum samples did not exceed 1:2.

The data presented indicate a high immunological effectiveness of dry VirusWall from strain TC-80 oral immunization of carnivores.

Virusvaktsinu against rabies for oral immunization of carnivores, dried by sublimation, containing a suspension of rabies virus strain TC-80, characterized in that it further contains peptone, lactose and pectin in the following ratios, wt%:

Suspension of rabies virus strain TC-80 - 5 - 7

Peptone - 65 - 67

Lactose - 11 - 13

Pectin - Rest

 

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FIELD: biotechnology.

SUBSTANCE: the present innovation deals with biotechnology of viral preparations. One should apply RB-71 strain of rabies virus which should be introduced into culture reservoir (0.1-0.01 MLD50/cell) simultaneously with VNK-21 at initial cell concentration being 0.5-0.5 mln cells/ml to be grown in suspension. Cultivation should be performed in suspension medium at 37 C for 5-6 d at constant mixing and maintaining pH value of 7.2-7.4. The obtained viral raw material at infectious 6.5-6.8 lg MLD50/ml and antigenic activity being 1:90-270 should be inactivated with 1,8,36-diendomethylene-1,3,6,8-tetriasecyclodecane 0.01% at 37 C for 3 d. Ready-to-use vaccine preparation meets the requirements of the standard for veterinary preparations being of 1.5-2.0 IU activity.

EFFECT: higher efficiency.

1 ex, 1 tbl

FIELD: biotechnology, veterinary science.

SUBSTANCE: he present innovation deals with manufacturing biopreparations for oral immunization in animals. One should grow vaccinia rabic virus, TC-80 strain in a certain passage cell culture to mix it with protective components (peptone, lactose, pectin). Mixing should be carried out at 14 : 5 : 1 ratio (viral suspension : stabilizing medium, consisting of 40% peptone solution with 8% lactose solution : 4% pectin solution, correspondingly), a semifinished product obtained after mixing should be applied onto a bait (a porous briquette) to be frozen and freeze dried for 24-30 h. The method includes decreased number of stages for manufacturing necessary vaccine being more efficient and economical. The vaccine keeps immunogenic properties for 5 d, not less at environmental temperature being up to 20 C and during storage period for 6 mo at 6-8 C.

EFFECT: higher efficiency of manufacturing.

2 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: method involves setting direct liquid phase immunofluorescence reaction with antigen under study. Antigen material suspension is prepared from the sample treated with formalin, washing out the antigen material from formalin, treating the antigen material with tripsin and washing out the antigen material from tripsin. Then, specific antigen-antibody complex preparation is obtained in liquid phase by binding the antigen material suspension with fluorescein-isothiocyanate globulin, washing the specific complex preparation from not reacted components and covering solid substrate with the specific complex preparation. Then, the specific complex preparation is studied using luminescent microscope and reaction result is studied. Specific fluorescence is recorded as bright greenish inclusions in neurons and their processes and outside of neuron boundaries. Uniformly luminescent greenish background without bright inclusions is taken to be negative result.

EFFECT: high sensitivity; accelerated toxicity drop; low cost.

5 tbl

FIELD: biotechnology, vaccines.

SUBSTANCE: method involves infection of transplantable cells VNK-21 with virus as measured for 0.01-0.001 MMLD 50/cell, multiplied for 36-70 h in suspended state and inoculated on microcarriers in the dose (1.2-2.5) x 105 cells in 0.3-0.6 ml of nutrient medium per each 1 cm2 of microcarriers. Culturing cells on microcarriers is carried out in bioreactor for 5-6 days and virus-containing cultural liquid is collected every day. Prepared vaccine possesses the high immunogenic activity and method for its preparing shows the high output.

EFFECT: improved preparing method of vaccine.

1 tbl

FIELD: veterinary virology.

SUBSTANCE: the suggested vaccine is being a bait-tablet with flavor and taste being attractive for carnivores. It contains lyophilized vaccinic strain TC-80 of rabies virus grown in passed culture of saiga's kidney, protective components, food and forming components. It has been, also, suggested the method for its obtaining due to growing vaccine strain TC-80 of rabies virus, lyophilic drying with stabilizing additives, grinding the lyophilized vaccine strain, mixing its with food and forming additives followed by pressing the mixture.

EFFECT: higher efficiency.

2 cl, 2 ex

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