RU2261109C1 - Strain of bacterium streptococcus agalactiae 71 for preparing vaccine against streptococcosis in fur animal - Google Patents

Abstract

FIELD: biotechnology, veterinary science.

SUBSTANCE: invention relates to the strain that is deposited in the Collection of strain-pathogens of diseases in fur animals in the Museum of bacterial and viral structures of the Science-Research institute of fur farming and rabbit farming (NIIPZK) named for V. A. Afanasiev and has the registration number 71. The strains relates to streptococci of the group B. The strain is cultures in Todd-Hewite beef-extract broth, Pike medium at 37°C for 18-20 h. After control of the grown culture for virulence the separated biomass is inactivated by addition of 0.5% formaldehyde followed by addition of aluminum hydroxide as an adjuvant and packaging. Invention provides preparing the effective vaccine against streptococcosis in fur animals based on this strain.

EFFECT: valuable properties of strain.

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Description

The invention relates to the field of biotechnology and veterinary medicine and is intended for the production of vaccines against streptococcal infections of fur animals, mainly arctic foxes and foxes.

Streptococcal infection is widespread, causing significant economic damage to animal husbandry and poses a risk to human health. In fur farms, the disease of foxes and foxes, rabbits, nutria, trochees is annually recorded.

The pathology of streptococcal infection in both humans and animals is very diverse - damage to the reproductive system, septic processes in the body, damage to the upper respiratory tract and lungs, and damage to the mammary glands. In young foxes and arctic foxes, it causes septicemia and early withdrawal in the early days of life.

The causative agent of Streptococcus agalactiae group B has an antigenic structure - the C-polysaccharide, which is associated with the virulence and immunogenicity of the microbial cell. Therefore, the effectiveness of specific prevention is determined by the presence of the C-factor in the antigenic composition of the components included in the vaccine.

Known vaccine strain "K-DEPT" of serogroup C streptococcus used for the manufacture of inactivated GOA - formovirus against streptococcus nutria. Patent No. 2097422 of 11/27/1997, the strain is isolated from nutria, adapted to this type of animals and is used for the manufacture of prophylactic drugs against streptococcosis of nutria. However, vacuum strains of streptococcus isolated from arctic foxes and foxes and used to produce a vaccine against streptococcosis of arctic foxes and foxes have not been registered.

The technical result of the invention is to obtain a strain of Streptococcus agalactiae No. 71, positive for polysaccharide-C and allowing to produce a vaccine against Streptococcus agalactiae type B based on the C-pathogenicity factor.

The inventive bacterial strain Streptococcus agalactiae No. 71 isolated from a puppy of a fox and deposited in a collection of pathogen strains of diseases of fur-bearing animals of the Museum of Bacterial and Viral Cultures of the Scientific Research Institute of Fur Breeding and Rabbits (NIIPZK) them. V.A. Afanasyev and has a registration number 71.

C-positive strain Streptococcus agalactiae No. 71 is intended as a producer of streptococcal polysaccharide C antigen in the manufacture of a specific immune preparation for streptococcal infection.

The bacterial strain Streptococcus agalactiae NIIPZK No. 71 is characterized by the following features and properties:

Genetic markers

- obtaining group-specific antibodies to polysaccharide-C is achieved by immunization with a whole microbial cell.

Morphological properties

- hemolytic streptococcus type B,

- industrial culture is a motionless spherical cocci with a diameter of 0.8-1.0 μ, forming short chains (2-4 cells),

- positively stained by Gram.

Cultural properties

- anaerobic, optimal growth temperature of 37-38 ° C, the upper and lower temperature border of 20 and 42 ° C,

- on blood agar forms round slightly bluish colonies surrounded by a hemolysis zone,

- on the environment Endo forms white-grove colonies with a hemolysis zone of type B,

- on Levin, Ploskereva’s media it forms cloudy grayish colonies with a hemolysis zone,

- on the determination medium of streptococci (Medium nutritium pro isolatio streptococcus siccum) gives a bottom sediment with diffuse turbidity of the medium.

- Conventional media for growing MPB, CT, MPA with 5% defibrated ram blood, with 1% glucose, pH 7.2-7.6.

Example No. 1

- It is well preserved under the condition of freeze drying with a sucrose-gelatinous stabilizer, as well as with rapid freezing and storage at 40 °.

Enzymatic properties

Bacterial strain Streptococcus agalactiae

- does not thin the gelatin,

- coagulates milk

- coagulates and provides litmus milk,

- ferments glucose, maltose, sucrose, lactose, mannitol,

- forms turbidity on sorbitol, dulcite.

Virulent properties of strain No. 71

- with intraperitoneal administration to white mice, LD / 50 is 10 ⁶ × 0.2 ml.

Antigenic properties

- the strain contains a C-polysaccharide, has antigenic properties and stimulates the production of tino-specific antibodies in the blood serum that have a protective effect against streptococcus.

Toxic properties

- In a liquid and solid nutrient medium it produces erythrogenic exotoxin, hemolysin, leukocidin, necrotic toxin.

Strain storage

Strain strain No. 71

- at a temperature of 6 ± 2 ° C in ampoules in a lyophilized state for more than 2 years,

- in ampoules of 0.5 ml of culture Str. on Todd-Hevit's medium at a temperature of 70 ° C for more than 3 years.

Strain control

- controls morphological, biochemical, cultural and virulent properties at least 1 time in 3 months.

- antigenic properties (C-factor) are checked before the production of the immune preparation.

The invention is illustrated by the following examples.

Example No. 1

A culture of a strain of Streptococcus agalactiae was cultured on Todd-Hevit broth for 24 hours at 37.5 ° C. After 12 hours, the culture was seeded on blood agar and grown 24 hours. At the end of the incubation period, the culture was checked for growth in Gram stained smears. The resulting culture was again seeded on Todd-Hevit's medium and incubated for 6 hours. The resulting culture in a logarithmic growth stage was seeded with 500 ml vials of Todd-Hevita Medium, incubated for 24 hours at 37.5 ° C. After incubation, culture vials were centrifuged at 1,500 rpm for 30 minutes. The non-residual fluid was drained. 0.5% formaldehyde was added to the precipitate, after a day it was checked for inactivation purity, the concentration of microbial cells was determined according to the turbidity standard, and it was adjusted to 2 × 10 ⁹ microbial bodies in 1 ml. As an adjuvant, a 3% solution of alumina hydrate was used in an amount of 10% of the volume of the microbial mass. The pH was adjusted to 7.0.

Claims (1)

The bacterial strain Streptococcus agalactiae NIIPZK No. 71 for the manufacture of vaccines against streptococcosis of fur animals.