RU2456021C2 - Method for preparing cattle parainfluenza-3 hyperimmune serum - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science. A method for preparing a cattle parainfluenza-3 hyperimmune serum characterised by five immunisations of 3-4-year-old donkeys with a cattle parainfluenza-3 and rhinotracheitis emulsion inactivated vaccine started with 1 ml of the vaccine and repeated every 14 days with the same vaccine however every time increasing a dose by 1 ml; aerosil or montanide is used as an adjuvant. Antibody titres in blood serum of the immunised animals as shown by indirect hemagglutination test make 1:512-1:1024 and reach 1:2048 in individual animals.

EFFECT: invention provides producing a high potent and non-reactogenic preparation for treating and preventing parainfluenza- 3 in young growths and grown cattle.

Description

The invention relates to the field of biology and agriculture, in particular the treatment and prevention of infectious diseases.

Hyperimmune serums against parainfluenza-3 virus are obtained in different types of laboratory and domestic animals.

A known method for the production and study of hyperimmune sera from laboratory animals against infectious rhinotracheitis, herpes and parainfluenza-3 viruses in cattle, where rabbits were used as test objects, and studies were carried out in reactions of neutralization, inhibition of hemadsorption and hemagglutination (Zudilina, 1970) .

The main disadvantage is the complexity of the work and the use for injection of live virus strains TK - A and TA, which under certain conditions can cause the spread of infection.

A known method for producing hyperimmune sera to infectious rhinotracheitis and parainfluenza-3 viruses in cattle (Khitrova, 2003, NPO NARVAK).

The disadvantage of this method of obtaining hyperimmune sera is associated with the immunization of expensive animals - bulls (horses), which, of course, affects the cost of sera.

The aim of the present invention is to obtain hyperimmune serums from donkeys for the treatment of calves suffering from parainfluenza-3, and the prevention of healthy young cattle. The advantages of this production method are:

- obtaining hyperimmune serums on the cheapest donors - donkeys, which reduces the cost of hyperimmune serum;

- the possibility of obtaining hyperimmune sera not on a live virus, but on an inactivated antigen in the vaccine.

For the experiment, 15 donkeys of 3-4 years weighing 150-200 kg were selected on the basis of analogues. 30 days before the start of the donkey hyperimmunization cycle, they were clinically examined: all experimental animals of average fatness, normal constitution, body temperature in the range of 37.5-38.5 ° C, pulse 30-40, breathing 8-14, wounds and other skin lesions not found.

Before the start of the experiment, blood was taken from all donkeys, serum was obtained and examined in rtga for the presence of anti-influenza antibodies. Antibodies to parainfluenza-3 virus were not found in blood serum. Donkeys were divided into 3 groups of 5 animals. All experimental animals were injected intramuscularly with 1 ml of emulsion inactivated vaccine against parainfluenza-3 and infectious rhinotracheitis of cattle, produced by the Yuryevets Biocombin in Vladimir, series No. 4, state control No. 4, manufactured on 09.05.

Every 14 days, the same vaccine was immunized with an increase in the dose of the vaccine by 1 ml. The donkey immunization schedule is presented in table 1.

The donkeys of the first group were supplemented with aerosil as equal adjuvant in the vaccine, montanide in the second group, and the third group of animals were immunized without adjuvants.

Starting with the third immunization, the total volume of vaccine and adjuvant was administered at two points.

Experiments have shown that donkeys are good test objects for obtaining hyperimmune serum against the parainfluenza-3 virus in cattle. So, of the 15 donkeys taken to work, in no case were there any complications at the site of intramuscular injection of the vaccine. In some animals, a mild depressed state was noted, an increase in body temperature by 0.5-1 ° C for 1-2 days, a short-term refusal of food.

Subsequently, blood was collected from donkeys for serum testing 14 days after the last administration of the biological product, and at the same time, the next immunization of animals was carried out according to the same scheme.

The results of the study of sera are presented in table 1.

Table 1. Donkey immunization schedule Group No. Number of living The dose of vaccine and adjuvant (in cm ³ ) Immunization 1st 2nd 3rd 4th 5th one. 5

2. 5

3. 5 one 2 3 four 5 Note: in the numerator is the dose of the vaccine, in the denominator is the dose of the adjuvant.

Table 2. Dynamics of antibody accumulation during donkey hyper immunization Group No. Antibody titers in rtga Before vacc. 14 days after immunization 1st 2nd 3rd 4th 5th one. 0 1: 8-1: 16 1: 64-1: 128 1: 128-1: 256 1: 256-1: 512 1: 512-1: 1024 2. 0 1: 8-1: 16 1: 64-1: 128 1: 128-1: 256 1: 256-1: 512 1: 512-1: 1024 3. 0 1: 4-1: 8 1: 128-1: 256 1: 32-1: 128 1: 64-1: 256 1: 256-1: 512 Note: blood sampling was performed 14 days after each immunization.

From the data in table 2 it can be seen that immunization of donkeys with an inactivated vaccine causes antibody formation in titers after 14 days that can protect animals from infection with the parainfluenza-3 virus in cattle. So, in animals to which the vaccine was administered together with Aerosil or Montanide, the antibody titers in RNGA after the second injection of the vaccine reached 1: 128. Slightly lower antibody titers in donkeys of the third group immunized without the addition of an adjuvant, although after the fourth and fifth immunizations in the blood serum of these animals, antibody titers reached up to 1: 512. In animals of the first and second groups, antibodies were within 1: 128-1: 256 14 days after the third immunization, and after the fourth and fifth immunization, the antibody level reached a dilution of 1: 256-1: 1024. These data indicate a high immunogenic activity of donkeys as producers of hyperimmune serum. The results of immunization of donkeys with an inactivated emulsion vaccine indicate the possibility of obtaining highly active hyperimmune sera using an inactivated vaccine as antigen.

Thus, our studies have shown that donkeys are good test objects for the production of hyperimmune serums against bovine parainfluenza-3.

In addition to the direct impact on the treatment and prevention of parainfluenza virus 3, donkey hyperimmune serum helps to reduce treatment time by 2-3 days and allows to reduce costs by half.

Information sources

Konopatkin A.A., Artemov B.T., Bakulov I.A. and other Epizootology and infectious diseases. Textbook for high schools. M .: Kolos, 1993, 369 p.

Zudilina Z.F. Obtaining and research of hyperimmune sera from laboratory animals against infectious rhinotracheitis, herpes and bovine parainfluenza-3 viruses in neutralization reactions, inhibition of hemadsorption and hemagglutination. Proceedings of the VIEV. Materials of the 2nd scientific conference of VIEV, 1970, 16 pp.

Khitrova A.E., Verkhovsky O.A., Nepoklonova I.V. Obtaining hyperimmune serums for infectious rhinotracheitis and parainfluenza-3 cattle viruses. (NGO NARVAK). International scientific conference dedicated. The 45th anniversary of FSI ARRIAH, “Actual problems of animal infectious pathology”, Vladimir, 2003, 222 p.

Konopatkin A.A., Bakulov I.A. and other Epizootology and infectious diseases of farm animals. Textbook for high schools. M .: Kolos, 1984, 296 p.

Claims (2)

1. The method of producing hyperimmune serum against parainfluenza virus-3 cattle, characterized in that they carry out five-fold immunization of donkeys 3-4 years of age with an inactivated emulsion vaccine against parainfluenza-3 and infectious rhinotracheitis of cattle, and begin with the introduction of 1 ml of vaccine and every 14 days immunization with the same vaccine is carried out with an increase in the dose of the vaccine at each immunization by 1 ml. 2. The method of producing hyperimmune serum against parainfluenza virus-3 cattle according to claim 1, characterized in that aerosil or montanide is used as adjuvant.