RU2506093C1 - Method for specific prevention of herd equine rhinopneumonia, salmonella abortion and strangles with using associated vaccine - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to veterinary science. A method involves immunising with using an associated inactivated vaccine prepared of equal portion of a salmonella abortion vaccine prepared of the bacterial strain Sal. abortus equi BN-12, an inactivated equine rhinopneumonia vaccine prepared of the rhinopneumonia viral strain CB/69, an inactivated strangles vaccine prepared of the bacterial strain Six. equi H-34.

EFFECT: method enables more effective vaccination ensured by creating the high-stress immunity and reducing the labour intensity.

3 cl

Description

The invention relates to the field of veterinary medicine, in particular to methods for the prevention of infectious diseases of horses.

Analogs of the invention. Known methods for the prevention of salmonella abortion in horses, which include immunization of horses with a vaccine from a bacterial strain Salmonella abortus equi BN-12 with the addition of polyribonate or a culture of bacterial strains of Bacillus subtilis TNP-3-DEPT (RF patents for inventions No. 2178713, publ. 01.27.2002, , bull. No. 3 and No. 22198925, publ. 12/20/2003, bull. No. 35). However, these methods are not used to prevent rhinopneumonia and wash. Vaccines are administered separately from other preventative drugs (for example, from the rhinopneumonia vaccine and wash).

Known live vaccine against rhinopneumonia of horses, dry culture of strains CB / 69 (instructions for use approved by the Rosselkhoznadzor 12/20/2006). However, the instructions for use of the indicated virus vaccine provides for double administration, which causes certain difficulties in its use in herd horse breeding. Also, as preliminary studies have shown, it is undesirable to use live vaccines in extreme conditions of herd content due to their increased reactogenicity.

A known method of specific prophylaxis of rhinopneumonia and salmonella abortion in horses, comprising the simultaneous introduction of inactivated vaccines against salmonella abortion from different bacteria strain Sal. abortus equi BN-12 and a live virus vaccine against rhinopneumonia of horses from a strain of the CB / 69 virus (Petrova S.G. Immunoprophylaxis of horse abortion of salmonella and rhinopneumonia etiology in Yakutia. Abstract of thesis for the degree of Candidate of Veterinary Science . Sciences, Yakutsk, 2006).

However, this method involves the introduction of two different vaccines in different places. For the prevention of rhinopneumonia, a live rather than an inactivated vaccine is used. Therefore, the method is ineffective in conditions of herd keeping horses.

The above vaccines are administered separately in different places, which greatly complicates the immunization process and increases its complexity.

Known methods for the prevention of horse washing, which include immunization of horses with a vaccine from bacterial strains of Streptococcus equi H-34 with the addition of polyribonate or a culture of the bacterial strain Bacillus subtilis TNP-3-DEP (RF patents for inventions No. 2122428, published on November 27, 1998, bull. No. 33 and No. 2143279, publ. 12/27/1999, bull. No. 36). However, these methods are not used to prevent equine salmonellosis and rhinopneumonia.

There is a method of specific prophylaxis of rhinopneumonia and salmonella abortion in horses with an associated vaccine in the conditions of herd keeping (RF patents for inventions No. 2424823, publ. July 27, 2011, bull. No. 21). However, the vaccine is not used to prevent horse washing.

The basis of the invention is the task of developing the prevention of salmonella abortion, rhinopneumonia and washing of horses with an associated inactivated vaccine.

The technical result of the invention is a method for specific prophylaxis of salmonella abortion, rhinopneumonia and wash associated inactivated vaccine. Equal parts of the inactivated vaccine from the Sal strain are used to make the associated vaccine. abortus equi BN-12, an inactivated vaccine from strain CB / 69 and an inactivated vaccine from strain of bacteria Streptococcus equi H-34. The vaccine is added the culture fluid of the bacterial strain Bacillus subtilis TNP-3-DEPT, which increases the immunogenicity of the vaccine and the immunobiological reactivity of the body. The introduction of an associated vaccine with the culture fluid of the bacterial strain Bacillus subtilis TNP-3-DEPT causes high tension immunity in at least 88.8% of vaccinated laboratory animals and reduces the incidence of washes (prevents 95.1% of young animals from the disease).

The Sal strain is used as a component of the associated vaccine. abortus equi BN-12 and a strain of bacteria Str. equi H-34, deposited in the collection of microorganisms of the All-Russian State Research Institute of Control, Standardization and Certification of Veterinary Medicines (VGNKI) (certificates of deposit of the strain of 01/20/1997, No. 63/27 and of 12/17/1993, No. 988/27). Strains can be obtained in the laboratory for the development of microbial preparations of the Yakutsk Research Institute of Agriculture of the Russian Agricultural Academy and VGNKI.

As another component of the associated vaccine, a CB / 69 rhinopneumonia virus strain is used.

To increase the efficiency of the associated vaccine and increase the immunobiological reactivity of the organism, the culture liquid of the broth culture of the bacterial strain Bacillus subtilis TNP-3-DEPT is added to the vaccine as an immunomodulator. The bacterial strain Bacillus subtilis TNP-3-DEPT was isolated from permafrost-transitional loamy soil of Yakutia. Deposited in the collection of microorganisms of the All-Russian State Research Institute for Control of Standardization and Certification of Veterinary Medicines (VGNKI). The certificates of deposit were issued on October 17, 1994 for No. 002/9 and on February 6, 2001 for No. 181/15. Obtained RF patent No. 2105810 “Bacillus subtilis bacterial strain used for disinfection of bird droppings and manure from pathogenic microorganisms” dated February 27, 1998. Laboratory experiments confirmed the ability of the Bacillus subtilis TNP-3-DEPT bacterial strain to stimulate the body's immunological reactivity.

Bacterial Strain Sal. abortus BN-12 is grown on meat and peptone broth until 8 billion microbial cells are accumulated in 1 ml. The bacterial mass is inactivated by formalin for 72 hours, then an aluminum hydroxide gel is added to a concentration of 6%.

Bacterial strain Str. equi H-34 is grown in meat and peptone broth containing 10% horse serum and 1% glucose until 3 billion microbial cells are accumulated in 1 ml. The bacterial mass is inactivated with formalin for 72 hours, then an aluminum hydroxide gel is added to a concentration of 6%.

Strain CB / 69 was cultured in primary cultures (or subculture) of kidney cells of a piglet, calf, rabbit or transplantable, virus-sensitive cultures (PT, SPEV, etc.) for 48-72 hours. The titer of the culture fluid virus before it is inactivated with formaldehyde should be at least 7.0-7.51 g ₅₀ / Mn.

The bacterial strain Bacillus subtilis TNP-3-DEPT is grown in meat and peptone agar pH 7.0-7.2 for 5-7 days at 37 ° C, then suspended in a physiological solution of sodium chloride at a concentration of 1 billion CFU in 1 ml. Filter through millipore filters.

Then, to 3 ml of the associated vaccine consisting of 1 ml of inactivated salmonella abortion vaccine, 1 ml of inactivated rhinopneumonia vaccine, 1 ml of inactivated wash vaccine and 1 ml of culture fluid of the Bacillus subtilis TNP-3-DEPT bacterial strain.

To determine the immunity, 40 heads of white BALB / c mice aged 10-40 days weighing 5-7 g were selected. The first group (9 goals) was immunized with a single vaccine associated with the culture fluid of the bacterial strain Bacillus subtilis TNP-3-DEPT, the second group ( 8 goals) are vaccinated twice with an interval of 14 days, and the third group (22 heads) are not vaccinated and are subsequently used as negative and positive controls. The vaccine is administered subcutaneously in the back at a dose of 0.3 ml.

14 days after a single and 12 days after re-immunization, the experimental and control white mice are infected with an intracerebrally adapted neurotropic strain PP1 / 1 of the horse rhinopneumonia virus at a dose of 0.02 · 6.0 lg TSD ₅₀ / ml. The effectiveness of immunization is determined by the number of mice resistant to morbidity and mortality. Within 10 days of observing animals, the protective effect of experimental infection with rhinopneumonia virus with a single vaccine is 44.4%, and with a double - 88.8%.

80 white mice weighing 18-20 grams were selected. The first group (40 animals) of mice is immunized with an associated vaccine at a dose of 0.2 ml, and the second group (40 animals) is not immunized. Drugs are administered subcutaneously in the back.

14 days after vaccination, 20 heads of mice of the first group and 20 goals of the control group are infected with a daily culture of a pathogenic strain of washed streptococcus at a dose of 5 LD ₅₀ . 20 goals of the first group and 20 goals of the control group are infected with a daily culture of a pathogenic Salmonella strain at a dose of 5 LD ₅₀ . Within 12 days of observation of animals in the first group (in the subgroups infected with streptococci and Salmonella), the disease and mortality were not noted. In the control group, all infected mice got sick. Mortality after infection with streptococci - 6 goals, and after the introduction of Salmonella - 8 goals.

265 heads of young horses of 6-7 months of age are selected. The first group (81 heads) is immunized with an associated vaccine with culture fluid from a bacterial strain of Bacillus subtilis TNP-3-DEPT in a dose of 4 ml. The second group (111 animals) is immunized with an inactivated horse wash vaccine. The third group (73 heads) is not vaccinated and is used as a control. Vaccines are administered intramuscularly in the upper third of the neck.

During 6 months of monitoring the young horses from the first group, 4 heads fell ill with washing, and the second group - 8 goals. The effectiveness of immunization in the first group vaccinated with the associated vaccine is 95.1%, and in the group vaccinated with the vaccine against the wash - 92.7%. In the group of non-immunized young animals, the incidence is 84.9%.

Vaccination of young animals with a trivalent vaccine causes the production of hemagglutinizing antibodies to the rhinopneumonia virus (1: 800-1: 6400), agglutinins to the causative agent of salmonellosis (1: 200-1: 1600) and precipitating antibodies to streptococci (1: 100) of fairly high titers.

Thus, immunization with an inactivated associated vaccine from Sal. abortus equi BN-12, Str. equi H-34 and strain CB / 69 with the culture fluid of the bacterial strain Bacillus subtilis TNP-3-DEPT causes high immunity and can be used as a method for the specific prevention of rhinopneumonia, salmonellosis and horse washing.

Claims (3)

1. A method for the specific prophylaxis of rhinopneumonia, salmonella abortion and washing of horses by immunization with an inactivated vaccine, characterized in that the associated inactivated vaccine from equal parts of the salmonella abortion vaccine from the bacteria strain Sal. abortus equi BN-12, an inactivated vaccine against rhinopneumonia of horses from a strain of rhinopneumonia virus CB / 69 and an inactivated vaccine against wash from a strain of bacteria Str. equi H-34. 2. The method according to claim 1, characterized in that, in addition, as the immunomodulator that enhances the immune response, the culture fluid of the broth culture of the bacterial strain Bacillus subtilis TNP-3-DEPT is introduced into the vaccine. 3. The method according to claim 1, characterized in that the intense immunity against rhinopneumonia, salmonella abortion of mares and wash is achieved as a result of a single vaccination.