Live vaccine, "ks" against classical swine fever and method of prevention of classical swine fever

 

(57) Abstract:

The invention relates to biotechnology and the receipt of the vaccine and method of prevention of classical swine fever. The vaccine contains an attenuated strain "CS" of classical swine fever virus and milk powder as a stabilizer. The vaccine has a high activity against classical swine fever and the introduction of high doses can eliminate the disease in disadvantaged large pig farms. 2 S. and 1 C.p. f-crystals.

The invention relates to biotechnology and related to vaccines for the prevention and treatment of classical swine fever (CSF). For specific prevention CoES known vaccine "LK-Vniivvim" containing lepidosirenidae Chinese vaccine strain "C" obtained by adaptation and reproduction of it in the culture of testicular tissue of lambs (EN 2064302C, A 61 K 39/187, 27.07.96) (C. S. Romanov). A disadvantage of the known vaccine and method of its application is the inability to liquidate CoES disadvantaged farms industrial type.

The invention consists also in the fact that the vaccine is given in high doses repeatedly. Repeated introduction of the vaccine in large doses (hyperbac the acquisition is a significant increase in the activity of a live vaccine against CSF and its applicability to the elimination of disease in large pig farms.

The invention consists in the following. The vaccine contains an attenuated strain "KC" virus CoES and stabilizer (medium dry).

Vaccine "KC" is a dry porous mass of white obtained by lyophilization of the mixture vaccinated culture liquid with medium drying. Mass fraction of moisture in the dry vaccine is in the range of 1-4%. Dry vaccine in vacuum bottles at 2-6oC preserves the original activity of not less than 12 months. During preventive vaccination a single dose of the vaccine "KC" is equal to 1000 IPD50and with the elimination of CSF - 100000-1000000 IPD50.

The strain KC received from the Chinese lepidosirenidae strain "C" virus CSF through a long passage and breeding in culture cells of porcine origin. Phenotypic strain "KC" is different from the vaccine strain "C" that does not cause hyperthermic response in rabbits when administered intravenously. It belongs to the genus of pestiviruses, family flaviviruses. The virion diameter of 40-60 nm; floating density of 1.12-1.13 g/cm3; sedimentation coefficient 140S. The genome consists of 12300 nucleotides and encodes polyprotein from 4000 amino acids. The genome of strain KC" differs from the genome of other known which of the 3 virion glycoprotein with a molecular mass of 23-57 KD. Nucleocapsid contains protein with molecular weight of 25 KD. For the synthesis of neutralizing antibodies and protective immunity responsible glycoprotein with a molecular mass of 55 KD.

Safety and immunogenicity of the strain "KC" fully meets the requirements of O. I. E. (OIE) and is different from the vaccine strains ("C", "GP", "Thiverval" and others) only a higher concentration of the virus obtained by reproduction in cell culture (1 ml not less than 108doses of infecting a cell culture - 108TCID50/ml).

Strain "KC" has the following biological properties:

breeds without cytopathic effect;

detected using antibodies labeled with peroxidase (PLA) or polymerase chain reaction (PCR);

safe for pigs of any age, including pregnant sows and newborn piglets;

not reversible (6 consecutive passages of the vaccine virus on bezmoroznyj pigs);

does not cause persistence in vaccinated pigs and rapidly disappears from the blood (7-10 days);

quickly (3-4 days) creates a long stressful immunity (not less than 2 years, the period of observation);

free from contamination by viruses, bacteria, fungi and Mikayla sjonnie trials have confirmed full compliance with strain, "KC" requirements and was the basis for its Deposit in the collection of microbial cultures VGNKI. Strain "KC" assigned registration number: the strain KC-DEPT".

The invention is illustrated by the following examples.

Example 1. For the preparation of vaccines using strain "KC", which is propagated in cell culture. Culture the virus combined with medium drying in the ratio required for obtaining vaccines with a given immunogenic activity. The resulting mixture was poured into 1 ml vials with a capacity of 3 ml and subjected to lyophilization. Vials with dry vaccine under vacuum hermetically closed with rubber stoppers and running aluminum caps.

Example 2. Under optimal culture conditions (a method of growing cells and virus multiplicity of infection, duration of incubation, treatment of infected culture) accumulation of strain KC reaches a high level of 8.0 to 9.2 lg TCID50/mlthat 100-1000 times the titer of all known vaccine strains used in the manufacture of live vaccines against CSF. The high degree of correlation of biological activity of strain KC" during the titration in cell cultures PK-15 (PLA, TCID50/mland during the titration on the pigs (IPD50/ml- immunizing dose that protects 50% of the vaccinated pigs). 1 IPD50<" studying the effects of various stabilizing environments and mode lyophilization. As cryoprotectants used widely used for these purposes, ingredients: sucrose, lactose, mannitol, gelatin, peptone, hydrolyzed lactalbumin, etc. the Use of these substances in various combinations and concentrations allows to stabilize the vaccine, although its infectious (immunogenic) activity was decreased by 1.3 to 2.5 lg. The use of dry cow's milk as createsite environment was preferred. Reducing the infectious activity of the vaccine does not exceed 0,7-1,0 lg. Drying vaccine "KC" with such a protective environment showed a high reproducibility of the results. So out of 30 episodes dry vaccine 27 series had the activity of 7.2 to 7.7 lg TCID50/mlrest - 6,7-7,0 lg TCID50/ml. At 2-6oC dry vaccine almost kept the original activity within 12 months.

Example 4. Epizootic activity of the vaccine "KC" was tested in two farms of industrial type (25000-27000 pigs; reproduction-fattening) affected with CSF.

In both farms CSF flowed the same type on the background of systematic preventive vaccine immunization LK-Vniivvim according to the instruction and was mainly characterized by a high incidence (40-50%) and death (25-30%) paino "KC" in higher doses and terms provided guidance on its application. As a result of hyperventilatie for 8-10 months of the disease CoES stopped. Thus, it was proved that in large disadvantaged farms CoES can be eliminated only by hyperventilatie. This goal can be achieved if the dose exceeds the usual 100 or more times and if hyperactivate is carried out within a certain timeframe.

1. Vaccine against classical swine containing an attenuated strain of the virus swine fever and stabilizer, characterized in that the attenuated strain it contains a strain VGNKI N "COP-DEPT" swine fever virus with a titer of infectious activity of 7.0 lg TCID50/mland more.

2. The vaccine under item 1, characterized in that it contains stabilizers skimmed cow's milk.

3. Method of prevention of classical swine fever, including intramuscular vaccine, wherein the vaccine is used vaccine under item 1 and enter its pigs in dysfunctional households repeatedly in large doses 100000 - 1000000 IPD50(hyperventilate).

 

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3 ex, 3 tbl

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10 cl, 11 dwg, 1 tbl, 5 ex

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3 tbl, 3 ex

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