SK292792A3 - The composition for protecting hydrophobia virus - Google Patents

Abstract

The composition for protecting hydrophobia virus in a vaccine, applied for oral immunisation of foxes, contains crystalline collagen, potassium 2,4,6-tris (dimethylaminomethyl)phenol glutamate, ethylene diamino tetra acetic acid together with animal's inactive serum without antitoxins against the hydrophobia virus soluble in distilled water. It provides protection against a rapid lowering of titer closed to the virus in vaccine, after having inserted it to the free nature.

Description

Composition for protection of rabies virus

Technical field

The invention relates to a composition for the protection of rabies virus for the production of fox vaccine. Orally applied rabies control in the rabies population for oral immunization of the vaccine are successfully used in wild animals. The immunity of rabies foxes limits the spread of rabies virus in the fox population, thereby preventing the spread of infection to other animals. Foxes contribute 95% to all cases of rabies occurrence in animals. Improving the health situation of foxes reduces the risk of human infection. Reducing the number of human vaccinations required results in a reduction in the cost and number of post-vaccination responses in humans known after the use of rabies vaccines.

BACKGROUND OF THE INVENTION

To ensure effective oral immunization of foxes, the rabies virus should be prevented from dropping in the wild-type vaccine. The vaccine is a plastic capsule filled with a vaccine virus with a protective medium, covered with aluminum foil. The virus capsule is coated with a suitable bait substance that is attractive to the fox. Upon ingestion, the plastic capsule is lost and the vaccine virus flows into the bait bite, which contaminates the oral cavity with a virus that penetrates the bloodstream and leads to the development of immunity. The vaccine produced is stored frozen at -20 to -30 ° C. Vaccination is carried out in such a way that Ba vaccine is given to nature in the amount of 16 doses of vaccine per km ² . The aim is to achieve the highest possible number of immune foxes, which is achieved by double vaccination during the calendar year. The first term of vaccination is late April, the second

- 1 'in early October. In these seasons, temperatures generally range from 0 to 20 ° C. Under these climatic conditions, it is necessary to ensure a minimum decrease in titer in the vaccine, thereby maximizing the efficacy of the vaccine and vaccination. The vaccine dose must contain at least 1CP'OPFU (coating units) per ml. The virus in the vaccine must be suitably stabilized to achieve these parameters. Various substances can be used for stabilization.

Purified casein hydrolyzate (Baer g.m., Winkler e.g., U.S. Pat. No. 4,014,991, issued Mar. 29, 1977, Int. Ciz., A61K 39/28) is desirable to stabilize rabies virus.

The use of hydrolyzed gelatin and sorbitol at a concentration of 3.57X leads to the protection of viral antigens (Abitz W., Mori D., DE 28 16 349 A1 of 14.4.1978, Int. Ciz, A61K 39/16).

A mixture of hydrolysed gelatin, disaccharides,

L-glutamic acid and L-arginine protect viral suspensions (Mc Aleer W.J., Markus H.Z., US 30.7.1980, Int. C13, A 61K 39/12), H.Z., EP 0 028 563 of 28.10.1980, Int.

Tris buffer at a concentration of 0.03 virus suspension (Biades J.E., 2.10.1982, Int. C13, A 61K 39/205).

U.S. Patent 4,338,335 of U.S. Pat

Mc Aller W.J., Markus C13, A 61K 39/121).

up to 0.08 M stabilizes EP 0 049 296 A1

Rabies virus in a vaccine for oral immunization USiek can be stabilized by adding 5 to 10% egg yolk (Steck F., EP 0 100 752 A1 of 2.8.1983 Int. C13, A61K 39/205) or a mixture of 10% egg yolk with bovine serum albumin (Schneider LG, DE 35 26 889 A1 of 26.7.1985, Int

C13, 39/205).

The aim of the solution was to prepare a suitable composition for the protection of rabies virus for oral vaccination of drugs, which ensures its protection during long-term storage at temperatures of -20 to -30 ° C and after unloading the fox vaccine at temperatures of -5 to 20 ° C for 14 days .

SUMMARY OF THE INVENTION

SUMMARY OF THE INVENTION Compositions comprising crystalline collagen, 2,4,6-Tris (dimethylaminomethyl) phenol, glutamate potassium, ethylenediaminotetraacetic acid, animal inactivated serum free of rabies virus antibodies in distilled water upon contact with rabies vaccine virus prevent loss of its rabies virus. biological activity. Short term at -5 ° C to + 20 ° C or long term at -20 ° C to -60 ° C. The biological activity of rabies virus is crucial to ensure a vaccine effect in wild foxes. The vaccine stored in the bait is placed in the wild in late April and early October, when the temperature can vary from -10 to + 25 ° C. Under these conditions, the unprotected virus is easily activated, thereby jeopardizing the efficacy of the vaccine. For this reason, various agents are used which limit the decrease in the biological activity of rabies virus in a vaccine for oral immunization of foxes. After the bait has been eaten in which the capsule containing the vaccine virus with the composition is stored, the capsule is broken by the teeth, the virus enters the bait substance. The gamma of the bait mass contaminates the oral cavity with USky rabies virus, which penetrates the bloodstream. In the fox, antibodies to the rabies virus are produced which protect it from the disease. 0.5 to 10% by weight are used to prepare the composition. % crystalline collagen, 0.7 to 1.5 96 wt. % 2,4,6, -Tris (dimethylaminomethyl) phenol, 0.1 to 0.8 wt. potassium glutamate, 0.1 to 1.8 wt. % of ethylenediaminotetraacetic acid, 2-109 wt. animal inactivated serum free of rabies virus antibodies in distilled water. The composition is mixed with the vaccine virus in a ratio of 1 part of virus with 0.5 to 2 parts of the composition.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

Composition Ba is prepared from the following ingredients:

crystalline collagen 200.0

2,4,6-Tris (dimethylaminomethyl) phenol 100.0 potassium glutamate 50.0 ethylenediaminotetraacetic acid 60.0 bovine serum inactivated 400.0 distilled water ad 10 000.0

After dissolution of the components, the pH is adjusted to 7.6 to 7.9 and the solution is sterilized by filtration. Immediately prior to filling, 1 part of the composition is mixed with 1 part of rabies virus. The mixture is filled, frozen and wrapped with bait.

Example 2

Crystalline collagen solution 12 *

2,4,6-Tris (dimethylaminomethyl) phenol solution 5% Potassium glutamate solution 9 X Ethylenediaminotetraacetic acid solution Q% Horse serum inactivated

5 000 ml 2 000 ml 900 ml 2 000 ml 100 ml

Deionized sterile pyrogen-free water is used to prepare the solution. After dissolution of the individual components, these are mixed gradually, the mixture is inactivated for 2 hours at 58 ° C and the pH is adjusted to 7.6 to 7.9. The composition is cooled. The cooled composition is mixed in a ratio of 1.3 parts of virus to 1 part of the composition.

Example 3

Crystalline collagen solution 2%

2,4,6-Tris (laminomethyl) phenol solution 485 Potassium glutamate solution 6%

Ethylenediaminotetraacetic acid solution 5% Horse serum inactivated

5 000 ml 2 000 ml 900 ml 2 000 ml 1 000 ml

Deionized sterile pyrogen-free water is used to prepare the solution. After dissolution of the individual components, these are gradually mixed and the pH is adjusted to 7.6 to 7.9. The composition is cooled and mixed in a ratio of 2 parts of virus to 1 part of the composition.

Industrial usability

The beneficial effect of the composition on rabies virus titer at 5 ° C was demonstrated by a decrease in the virus titer decrease by 100-fold. The results are shown in the following table. To ensure 100% protection of the foxes, a minimum titre of 10 ⁵ → ⁹ / ml should be given in the vaccine dose (Lawson FK et al., 1989, Can. J. Vet. Res. 53, 438-444).

Effect of the composition on virus titer during storage

time Aging management consists imported nia Virus with logioPFU virus titer composition in ml Virus without composition locrioPFU virus titer in ml storage temperature storage temperature -15 ° C 0 ° C + 20 ° C 20 C 0 ° C + 20® C □ 7.3 7.3 7.3 7.3 7.3 7.3 4 days 7.4 7.1 6, 8 7.3 6.8 6.5 7 days - 7.0 6.2 - 6.0 5.5 shipping 7.0 6.5 6.0 7.0 5.0 4.0 14 days 7.3 6.0 5.5 7.3 4.0 3.0 LMES. 7.0 - - 7.3 - - 3months. 7.0 - - 6.5 - - 6mes. 7.0 - - 6.0 - -

Decrease titer during storage at -60 ° C.

Shelf life at -60 ° C in months 0 2 3 4 5 7 8 ziBtený titer 7.4 7.5 7.1 7.0 6.9 6.9 6.7

After the first vaccination for oral immunization of rabies foxes, tetracycline (marker) in bone was found in 79% of the foxes examined. In these foxes, antibodies were detected in 84% of cases. The distribution of antibody levels by titers or by the number of International Units (I.U.) in 1 ml indicates good virus survival in the vaccine, and thus a good antibody response after ingestion of the bait. The results are shown in the following table.

Detected antibody titer 20 60 180 520 Mean antibody titer I.U. in ml 1.3 2.1 2.7 3.5 X animals with given antibody titer 54 36 8 2

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Claims (1)

PATENT 7> V A rabies virus protection composition comprising 0.5 to 10 wt. 96 crystalline collagen, 0.7 to 1.5 wt. X 2.4,6, -Tris (dimethylaminomethyl) phenol, 0.1 to 0.6 wt. 0.1 to 1.8 wt.% potassium glutamate; % Of ethylenediaminotetraacetic acid, 2 to 10 wt. 96 wt. animal inactivated serum free of anti-rabies virus antibodies in distilled water.