RU2166329C2 - Mixed vaccine against plague and salmonellosis in fox and polar fox - Google Patents

Abstract

FIELD: veterinary science, microbiology. SUBSTANCE: mixed vaccine has viral and bacterial components balanced with antigenic and immunogenic relationship. Among plague viruses of carnivores vaccine has cultural viral suspension of the strain Virus distemper canine ЭПМ with titer of infectious activity 10³-10⁴ TCD/0.5 ml. Among salmonella antigens vaccine has cell suspensions of strains Salmonella typhimurium N 3, Salmonella dublin N 6, Salmonella cholerasuis N 9. Concentration of each strain is 10 billion cells/1 ml of physiological solution. Vaccine has stabilizing agent also. Mixed vaccine shows enhanced antigenic and immunogenic activity. Mixed vaccine can be used for specific prophylaxis of carnivorous plague and salmonellosis in foxes and polar foxes. EFFECT: enhanced effectiveness of vaccine. 2 cl, 5 tbl, 4 ex

Description

 The invention relates to the field of biotechnology, in particular to means for the specific prophylaxis of plague of carnivores and salmonellosis of fur-bearing animals.

 For the prevention of plague and salmonellosis in this type of fur-bearing animals, single vaccines are used.

 A known vaccine for the prevention of carnivore plague containing a culture suspension of an attenuated strain "EPM" (RF Patent N 2067002, A 61 K 39/175, 1996).

 Known polyvalent formolthiomersal vaccine against salmonellosis and Escherichiosis of young farm animals and fur animals, containing a suspension of Escherichia and Salmonella S. typhimurium, S.dublin, S. choleraesuis, inactivated formalin and thiomersal, and adjuvant alum potassium chloride. Edited by D.F. Ozidze, Moscow: Kolos, 1981, p. 223-224).

 The disadvantage of monovaccines is the need for preventive treatments against each infection separately, in addition, the well-known vaccine against salmonellosis is administered to fur animals three times. All this affects labor costs, stretches the formation of immunity in animals, contributes to the occurrence of stressful phenomena and complications.

 The vaccine associated with plague and salmonellosis of foxes and arctic foxes is not known.

 The objective of the invention is to develop an associated vaccine against plague and salmonellosis of fur animals, characterized by high antigenic and immunogenic activity.

 The technical result of the invention is to obtain an associated vaccine against plague and salmonellosis of fur animals, which has a wide spectrum of protective action, capable of providing long-term and reliable immunity against these infections.

The essence of the invention is as follows: the associated vaccine against plague and salmonellosis of foxes and arctic foxes contains from the antigens of the plague of carnivorous plague virus (VChP) a virus culture suspension of the Virus distemper canine strain EPM with an infectious activity titer of 10 ³ -10 ⁴ TCD 50 / 0.5 ml, from Salmonella antigens - cell suspensions of strains of Salmonella typhimurium N 3, Salmonella dublin N 6, Salmonella cholerasuis N 9 with a concentration of each strain of 10 billion cells in 1 ml of physiological saline, and any known stabilizer, for example, based on sorbitol and gelatin.

 The vaccine strains included in the vaccine are Salmonella typhimurium N 3, Salmonella dublin N 6, Salmonella cholerasuis N 9 are known production strains, but were not previously used for the manufacture of vaccines for fur animals.

 Young foxes and arctic foxes are vaccinated from 45-50 days of age twice with an interval of 10-14 days. Adult animals are vaccinated once until rutting in December-January. In farms that are permanently unsuccessful for salmonellosis, vaccination of females is recommended in the second half of pregnancy, but no later than 14 days before whelping. The vaccine is administered intramuscularly in a volume of 1 ml.

 The invention is illustrated by the following examples.

 Example 1. Obtaining an associated vaccine against plague and salmonellosis of foxes and arctic foxes is carried out by separately preparing a culture virus suspension of Virus distemper canine strain EPM and cell suspensions of strains of Salmonella typhimurium N 3, Salmonella dublin N 6, Salmonella cholerasuis N 9.

To obtain virus-containing material, the carnivore plague virus is grown in a primary trypsinized cell culture of Japanese quail embryos in a growth medium containing 0.5% lactalbumin hydrolyzate (GLA) in Hanks solution, 25-50% medium 199 and 8-10% cattle serum with the addition of antibiotics. The virus is introduced into the cell suspension and grown by roller method at 37 ^o C. After 4-6 hours, the growth medium is drained. Cultivation is continued in a support medium containing 60-70% of 199 medium and 30-40% GLA in Earle's solution with the addition of antibiotics. After the full development of the cytopathic effect (CPP), the virus-containing culture fluid is drained, frozen at a temperature of minus 10-20 ^o C, thawed at room temperature and combined with the bacterial component.

To obtain the bacterial component, strains of Salmonella typhimurium N 3, Salmonella dublin N 6, Salmonella cholerasuis N 9 are grown separately in the reactors on Hottinger broth for 10-12 hours at a temperature of 37 ^o C with constant stirring and continuous aeration. Cultivation is stopped at the beginning of the stationary growth phase. The grown cultures are cooled to a temperature of 6-8 ^o C and subjected to verification. Then they are concentrated in a supercentrifuge. The bacterial masses of each strain separately diluted with sterile saline to a concentration of 10 billion microbial cells in 1 ml according to the optical standard GISK them. Tarasevich, combine in equal volumes and mix thoroughly.

 To the combined bacterial mass is added an equal volume of the cultural viral suspension containing at least 400-500 TCD 50 / 0.5 ml. After thorough mixing, an equal volume of stabilizing drying medium is added. For lyophilization of the vaccine, a stabilizing drying medium is used, which is usually used in the production of a vaccine against carnivore plague from an EPM strain, for example, based on sorbitol and gelatin. The resulting mass is packaged with continuous stirring in sterile ampoules or vials and lyophilized.

 Before use, the lyophilized associated vaccine is diluted in a special solvent, which is used to dissolve vaccines against carnivorous plague.

 Get three series of vaccines. The vaccine associated against plague and salmonellosis of foxes and arctic foxes has the following component composition, in vol.% (Table. 1a).

 Example 2. The associated vaccine obtained in Example 1 was tested for harmlessness on white mice weighing 14-16 g, guinea pigs weighing 300-350 g and thorzofrettes 4-5 months of age with the introduction of various doses. Generalized safety results are presented in table. 1.

 The test vaccine is harmless to animals that remained clinically healthy for 21 days of observation after vaccination.

 Example 3

 Assessment of the immunogenic activity of the associated vaccine against plague and salmonellosis of foxes and arctic foxes is carried out respectively on thorzofret and guinea pigs. Animals are immunized once intramuscularly at a dose of 1.0 ml and 21 days after vaccination, they are infected with virulent strains of the carnivorous plague virus and the pathogen of salmonellosis. The results of the control are presented in table. 2 and 3.

 Example 4. The antigenic activity of the obtained associated vaccine is evaluated on arctic foxes and foxes by studying the dynamics of the accumulation of specific antibodies to the plague of carnivores in the indirect hemagglutination reaction (RNGA) and salmonella in the agglutination reaction (RA). Research data are presented in tables 4 and 5.

 From the table. 4 and 5 it follows that the vaccine associated against plague and salmonellosis in arctic foxes and foxes causes intense immunity to the plague virus of carnivores and salmonella.

 The use of the proposed vaccine in veterinary practice will improve the effectiveness of ongoing measures for the prevention of infectious diseases of foxes and foxes.

Claims (1)

1. The associated vaccine against plague and salmonellosis of foxes and arctic foxes, characterized in that it contains from the antigens of the plague of carnivorous virus culture virus suspension of the strain Virus distemper canine EPM with an infectious activity titer of 10 ³ - 10 ⁴ TCD 50 / 0.5 ml, from antigens Salmonella - cell suspension of strains of Salmonella typhimurium 3, Salmonella dublin 6, Salmonella cholerasuis 9 with a concentration of each strain of 10 billion cells in 1 ml of saline and a stabilizer in the following ratio of components, vol.%:
Culture virus suspension of the strain Virus distemper canine EPM with a titer of infectious activity 10 ³ - 10 ⁴ TCD 50 / 0.5 ml - 20 - 25
Suspension of the cells of the strain Salmonella typhimurium 3, Salmonella dublin 6, Salmonella cholerasuis 9 with a concentration of each strain of 10 billion cells in 1 ml of fiz.r-ra - 20 - 25
Stabilizer - Else
2. The vaccine according to claim 1, characterized in that it contains a stabilizer based on sorbitol and gelatose.

Images