RU-2861563-C1 - RABIES VACCINE FOR ORAL IMMUNISATION OF WILD CARNIVORES AND METHOD FOR ITS PRODUCTION

RU2861563C1RU 2861563 C1RU2861563 C1RU 2861563C1RU-2861563-C1

Abstract

FIELD: veterinary virology. SUBSTANCE: rabies vaccine includes the vaccine strain of rabies virus "TriGAS" 1251/2, grown in a continuous cell culture BHK-21/13-13, and food and shaping components. The vaccine contains, as a vaccine strain, a culture liquid containing an avirulent strain of rabies virus "TriGAS" 1251/2 with an infectivity titre of 6.0-8.5 lg FFU50/cm 3 . As food and shaping components, the vaccine contains fish meal, beef fat, paraffin, additionally contains a tetracycline marker, with a certain ratio of components. The method for producing the vaccine consists in that the vaccine strain of rabies virus is grown in continuous monolayer-suspension sublines of newborn Syrian hamster kidney cells BHK-21/13-13, wherein the culture liquid additionally contains L-serine and iron nitrate nonahydrate at the rate of 0.035-0.045 g/l and 0.00005-0.0001 g/l, respectively. EFFECT: creation of an effective vaccine for oral immunisation of wild carnivores. 2 cl, 1 tbl, 4 ex

Inventors

Litenkova Irina YUrevna
Babina Ekaterina Aleksandrovna
Elnikov Vasilij Viktorovich
Surnev Dmitrij Sergeevich
Kruglov Aleksandr Aleksandrovich
Kolodina Kseniya Sergeevna
Stepanyuk Vladimir Leontevich
Stoyanov Ilya Valerevich
Khaustova Natalya Vladimirovna
MELNIK NIKOLAJ VASILEVICH

Dates

Publication Date: 20260506
Application Date: 20250221

Claims (3)

1. A rabies vaccine for oral immunization of wild carnivores, comprising a rabies virus vaccine strain grown in a continuous cell culture and food and form-forming components, a tetracycline marker, characterized in that it contains, as a vaccine strain, a culture fluid containing the rabies virus strain "TriGAS" 1251/2 with an infectivity titer of 6.0-8.5 lg FFU 50 /cm 3 , and contains fish meal, beef fat, paraffin or a food polymer as food and form-forming components, in the following ratio of components, wt. %:
Culture fluid containing the avirulent rabies virus strain "TriGAS" 1251/2 with an infectivity titer of 6.0-8.5 lg FFU 50 /cm 3 1.0-4.0 Tetracycline 0.3-1.2 Paraffin 5.0-10.0 Fish meal 30.0-40.0 Beef fat the rest
2. A method for producing a rabies vaccine for oral immunization of wild carnivores, comprising cultivating a rabies virus strain in a continuous cell culture in a medium containing sodium chloride, potassium chloride, magnesium sulfate, dibasic sodium phosphate, monobasic potassium phosphate, calcium chloride, sodium bicarbonate, L-arginine, L-glutamine, L-tyrosine, L-tryptophan, calcium pantothenate, pyridoxal HCl, thiamine, meso-inositol, nicotinamide, riboflavin, folic acid, glucose, enzymatic hydrolysate of muscle proteins, enzymatic hydrolysate of lactalbumin, benzylpenicillin sodium salt with an activity of 0.09×10 5 -1.1×10 5 , kanamycin sulfate with an activity of 0.09×10 5 -1.1×10 5 , nystatin with an activity of 6×10 4 to 6.5×10 4 U/l, choline chloride, succinic acid and distilled water in a given ratio of components, a culture fluid containing an avirulent strain of the rabies virus are placed in containers and frozen at minus 20-40 ° C, then tetracycline, paraffin, fish meal, beef fat are mixed for 25-30 minutes at a temperature of 55-60 ° C, then a mixture of tetracycline, paraffin, fish meal and beef fat is poured into plastic molds at a temperature of 40-45 ° C, a culture fluid containing an avirulent strain of the rabies virus is placed in each cell of the bait mold, and frozen at a temperature of minus 20-40°C, characterized in that the rabies virus vaccine strain "TriGAS" 1251/2 is grown until an infectivity titer of 6.0-8.5 lg FFU 50 /cm 3 is achieved in transplantable monolayer-suspension sublines of newborn Syrian hamster kidney cells BHK-21/13-13, while the culture fluid additionally contains L-serine and iron nitrate nonahydrate at a rate of 0.035-0.045 g/l and 0.00005-0.0001 g/l, respectively.

Description

The invention relates to veterinary microbiology and biotechnology and can be used in the development of specific prophylactic agents and, in particular, for producing a vaccine against rabies in animals. One of the most important factors in the fight against rabies is oral vaccination of wild carnivores, which are the natural reservoir of the disease. A method for producing an anti-rabies vaccine for animals is known, which includes preparing seed material from a rabies virus strain, infecting a culture of transplantable cells with the seed material, cultivating the rabies virus, collecting the virus-containing fluid, and then preparing the target product in liquid form (RU Patent No. 2287343, “Method for Producing an Anti-rabies Vaccine,” Bulletin No. 26, May 31, 2005, IPC A61K 39/205). Also known is a rabies vaccine for oral immunization of wild carnivores, which includes a vaccine strain of the rabies virus grown in a continuous cell culture and food and formative components (RU Patent 2311924, IPC A61K 39/205, Bulletin 16, 2007.12.10), in particular, it contains a lyophilized vaccine strain TS-80 of the rabies virus with protective components, skim milk powder, lactose, egg powder, finely dispersed chalk T-60, finely dispersed chalk T-90 and calcium stearate in a certain ratio of components. The disadvantage of the analogue is that the resulting vaccine has low target product quality, as determined by oral administration to animals, and has low immunogenicity, resulting in limited protection in wild animals. It is not homogeneous, the briquettes are not standardized, are not moisture-resistant, and may crumble during storage. The closest technical solution is a rabies vaccine for oral immunization of wild carnivores and a method for producing it, in particular, the vaccine contains, as a vaccine strain, a culture fluid containing an avirulent strain of the rabies virus RV-97 or ERA G333 with an infectivity titer of 6.0-8.5 lg FFU 50 /cm 3 , and contains fish meal, beef fat, paraffin or food polymer and unrefined cereal grain as food and form-forming components, and additionally contains a tetracycline marker, at a certain ratio of components (RU Patent No. 2440139, “Rabies Vaccine for Oral Immunization of Wild Carnivores and a Method for Preparing It,” Bulletin No. 2, 20.01.2012, MKI A61K 39/205, A61P 31/12). The disadvantages of this technical solution are that the resulting vaccine has a low quality of the target product, which is revealed when administered orally to animals, and has low immunogenicity, which results in low protection of the body of wild animals. The technical result of using the invention consists in increasing the effectiveness due to an increase in the antigenic and immunogenic activity and harmlessness of the vaccine. The stated objective is achieved in that the rabies vaccine for oral immunization of wild carnivores, including a vaccine strain of the rabies virus grown in a continuous cell culture, and food and form-forming components, the tetracycline marker contains, as a vaccine strain, a culture fluid containing the rabies virus strain "TriGAS" 1251/2 with an infectivity titer of 6.0-8.5 lg FFU 50 /cm 3 , and contains fish meal, beef fat, paraffin as food and form-forming components, in the following ratio of components, wt. %: Culture fluid containing the avirulent rabies virus strain "TriGAS" 1251/2 with an infectivity titer of 6.0-8.5 lg FFU 50 /cm 31.0-4.0Tetracycline0.3-1.2Paraffin5.0-10.0Fish meal30.0-40.0Beef fatthe rest The stated objective is also achieved in that the method for producing a rabies vaccine for oral immunization of wild carnivores comprises cultivating a rabies virus strain in a continuous cell culture in a medium containing sodium chloride, potassium chloride, magnesium sulfate, sodium phosphate dibasic, potassium phosphate monobasic, calcium chloride, sodium bicarbonate, L-arginine, L-glutamine, L-tyrosine, L-tryptophan, calcium pantothenate, pyridoxal HCl, thiamine, meso-inositol, nicotinamide, riboflavin, folic acid, glucose, enzymatic hydrolysate of muscle proteins, enzymatic hydrolysate of lactalbumin, benzylpenicillin sodium salt with an activity of 0.09×10 5 -1.1×10 5 , kanamycin sulfate with an activity of 0.09×10 5 -1.1×10 5 , nystatin with an activity of 6×10 4 to 6.5×10 4 U/l, choline chloride, succinic acid and distilled water in a given ratio of components, the culture fluid containing the avirulent strain of the rabies virus is placed in containers and frozen at minus 20-40 ° C, then tetracycline, paraffin, fish meal, beef fat are mixed for 25-30 minutes at a temperature of 55-60 ° C, then the mixture of tetracycline, paraffin, fish meal and beef fat is poured into plastic molds at a temperature of 40-45 ° C, in each cell of the bait form the culture fluid containing the avirulent strain of the virus is placed rabies, and frozen at a temperature of minus 20-40°C, wherein the rabies virus vaccine strain "TriGAS" 1251/2 is grown until an