EA029938B1 - Vaccine against strangles in horses - Google Patents

Abstract

The invention relates to the field of veterinary, in particular to prophylaxis and treatment of strangles in horses. Provided is a stable, harmless and highly effective vaccine against stangles comprising inactivated strain Streptococcus equi BNK; antibiotics with a wide spectrum bicillin-5, sulfadimezin and trimethoprim; polyoxidonium as an adjuvant and immunomodulatory agent.

Description

The invention relates to the field of veterinary medicine, in particular to the prevention and treatment of myta horses. A stable, harmless, and highly effective vaccine against myta horses, including the inactivated strain §1ger1osossi8 ΒΝΚ; broad-spectrum antibiotics bicillin-5, sulfadimezin and trimethoprim; polyoxidonium as an adjuvant and immunomodulatory agent.

029938 Β1

029938

The invention relates to the field of veterinary medicine, in particular to the prevention and treatment of myta horses.

Known vaccine against myta horses, including mytny antigen, T-activin and bicillin [patent of the Republic of Kazakhstan No. 3681, cl. A61K 39/00, 1996].

A disadvantage of the known vaccine is that, in connection with the withdrawal from production of individual components of the drug, its further production is not possible.

The objective of the invention is to obtain a vaccine against myta horses with the use of new modern components, allowing to increase immunogenicity, accelerate recovery time and eliminate post-vaccination complications.

The technical result provided by the invention, is expressed in obtaining a stable, harmless and highly effective vaccine against the specified infection.

A vaccine against horse myta has been proposed, which includes an inactivated strain 31ger1osossi8 ECPR, containing broad-spectrum antibiotics bicillin-5, sulfadimesin and trimethoprim, and polyoxidonium as an adjuvant and immunomodulating agent, with the following ratio of components, in 1 l:

strain 81ger1osossi§ βςιιΐ ΒΝΚ 1.8-2.0 χΐ 0 ¹⁰ CFU

bicillin-5 30-40 million IU

sulfadimezin 1800-2000 mg

Trimethoprim 300-400 mg

polyoxidonium 80-120 mg

distilled water up to 1 liter.

The strain of bacteria used for the manufacture of inactivated vaccine against myta horses, deposited in the laboratory LLP "Akintay" and has the following properties.

The strain of bacteria 81ger1osossi§ etsp.

Cultural properties.

On MPA with the addition of 10% serum and 1% glucose, fine rounded, transparent, smooth, viscous consistencies form confluent colonies that are difficult to remove by a loop. When viewed into the light, transparent with a bluish tint when stored for 2-4 days, the colonies become grayish-white, transparent, but retain smooth outlines.

On a BCH with the addition of 10% serum and 1% glucose at a pH of 7.4, it grows in the form of cloudiness of the medium with the formation of a small precipitate at the bottom of the tube, during storage for 3-5 days the broth is clarified with the formation of abundant grayish draft.

Biochemical properties.

It has poorly expressed biochemical properties, does not grow in BCH with 40% of bile, does not change milk with methylene blue, does not coagulate milk, does not reduce litmus, does not ferment sorbitol, mannitol, glycerin, arabinose, raffinose, lactose, does not thin the gelatin, but decomposes glucose , sucrose and maltose to form acid without gas.

Marker signs. Liziruet erythrocytes of horses, white mice, weakly lyses rabbit erythrocytes, does not lyse erythrocytes of golden hamsters, does not ferment lactose and attracts, does not produce streptolysin O, produces hemolysin, leukocidin, toxin of general action and agarin.

Antigenic properties.

The strain precipitates specific streptococcal serum of serum group C.

Antigenic activity.

When administered parenterally into the horse's body, it causes the production of antibodies detected in the phagal genotype, titer 1: 2800.

Belonging to the trophic group. Prototroph

Genetic characteristics of the strain.

Lyse erythrocytes of horses, white mice, weakly lyses erythrocytes of rabbits, does not lyse erythrocytes of golden hamsters, does not ferment lactose and attracts, does not produce streptolysin-O, produces hemolysin, leukocidin, general action toxin and agarin. The product synthesized by the strain is hemolytic toxin.

Activity (productivity).

Lysis of the red blood cells of a horse and a white mouse.

Reactogenicity.

It has moderate to moderate reactogenicity for horses.

Immunogenicity

When administered intramuscularly at a dose of 2x10 ⁶ cfu, the duration of the strain of immunity in white mice is 12 months.

Stability when passaged.

The main biological properties of the strain are stably maintained for 2 years (observation period).

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The strain is stored in a dark place, in a lyophilized state and in glass ampoules under vacuum at a temperature of 4 to 10 ° C for 12 months, or in a cryogenic state in a sterile glycerol solution at a temperature of -75-80 ° C for 7 years.

Example 1

An inactivated vaccine against horse myta is prepared on the basis of the strain 31ger1osossig evtsh

ΒΝΚ.

For the preparation of a vaccine against horses, the culture of mytny streptococcus obtained by passages in BCH, MPA, agar and Hottinger broth with respect to the conditions of sterility is washed into a collection with a capacity of 10,000 cm ³ (10 l), then a microbial concentration of 0.85% sodium chloride solution cells are brought to a content of 1 billion CFU for mytnyh streptococci in 1 cm ³ and poured under sterile conditions into 5-10-liter bottles.

Inactivation of bacterial suspensions of streptococcus mytnogo in bottles with a concentration of 1 billion CFU in 1 cm ^{3 is} carried out by autoclaving at 120 ° C for 30 min at 1.2 atm. The resulting inactivated bacterial mass, recognized as sterile and harmless, is sterilely poured into 10 cm ³ penicillin vials in a volume of 1 cm ³ (for 10 doses of vaccine) and 20 cm ³ penicillin vials in a volume of 2 cm ³ (for 20 doses of vaccine).

To 10-20 cm ³ penicillin vials with inactivated bacterial mass add 1-2 cm ³ respectively ^{of the} sucrose-gelatin medium (10% sucrose and 0.5% gelatin), and freeze-drying is carried out. Date of completion of lyophilization is considered the date of manufacture of the vaccine.

For the preparation of 10 doses of vaccine to the antigen (1 billion CFU in a vial), 1.5 million U of bicillin-5, 100 mg of sulfadimezin, 20 mg of trimethoprim, 6 mg of polyoxidonium are added under sterile conditions, tightly closed with a rubber stopper and rolled around with an aluminum cap .

Example 2

Inactivated vaccine against horse myta is used intramuscularly once with the following ratio of components:

strain ZerUssossiz etsh ΒΝΚ bitsillin-5 sulfadimezin trimethoprim

polyoxidonium

1,8χ10 ^1θ ΚΟΕ 20-25 million ED 1800 mg 300 mg

80 mg

up to 1 liter.

distilled water

Example 3

Inactivated vaccine against horse myta is used intramuscularly once with the following ratio of components:

strain 31ger1osossi8 βςιιϊ ΒΝΚ 1.9 x 10 ¹⁰ CFU

bicillin-5 25-30 million IU

sulfadimezin 1900 mg

Trimethoprim 350 mg

polyoxidonium 100 mg

distilled water up to 1 liter.

Example 4

Inactivated vaccine against horse myta is used intramuscularly once with the following ratio of components:

strain 31ger1osossi5 test ΒΝΚ 2x10 ¹⁰ cfu

bicillin-5 30-40 million IU

sulfadimezin 2000 mg

Trimethoprim 400 mg

polyoxidonium 120 mg

distilled water up to 1 liter.

Example 5

Immunogenic activity of the vaccine is determined on 1000 white mice, selected according to the principle of analogues, with an average weight of 18-20 g, divided into 10 groups of 100 animals each: 9 experienced, and the 10th control.

The drugs are administered intramuscularly once at a dose of 0.1 cm ³ per head. At 21, 30 and 60 hours immunized and control mice were not immunized white administered virulent culture 31ger1osossi§ edsh at 3 ΕΌ _50. The test results are presented in Table. one.

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Table 1

The results of the determination of the immunogenic activity of a vaccine against horse moth on white mice

I ’ruppa stomach of Preparations: ratio components in 1 liter of vaccine, frequency rate and dose per 1 head Colic your whites mice Goal. Term checks immunize that days Result contamination Percent immune of animals Survived Goal. Palo Goal one strain 81ger1 osssh ш 1.8x10 ¹⁰ CFU, bicillin-5 - 20-25 100 21 98 2 98 2 million units, sulfadimezin - 1800 mg, trimethoprim - 300 mg, 100 thirty 98 2 98 3 polyoxidonium - 80 mg, distilled water - up to 1 liter, once, 0.1 cm ³ 100 60 97 3 97 four strain ZierUssossiz etsh ΒΝΚ 1,9x10 ¹⁰ CFU, bitsillin-5 - 25-30 100 21 98 2 98 five million units, sulfadimezin - 1900 mg, trimethoprim - 350 mg, 100 thirty 98 2 98 6 polyoxidonium - 100 mg, distilled water - up to 1 liter, once, 0.1 cm ³ 100 60 97 3 97 7 strain 81ger1ossossiz εςιιϊ ΒΝΚ 2x10 * ° CFU, bitsillin-5 - 30-40 100 21 98 2 98 eight million units, sulfadimezin - 2000 mg, trimethoprim - 400 mg, 100 thirty 98 2 98 9 polyoxidonium - 120 mg, distilled water - up to 1 liter, once, 0.1 cm ³ 100 60 97 3 97 ten Control group, vaccine not was introduced 100 21 0 100 0

From tab. 1 shows that on the 60th day after vaccination in the 3rd group, 97 white mice survived — 97%; in the 6th and 9th groups, similar results were obtained with 100% death of the mice in the control group.

Example 6

To determine the therapeutic efficacy of the vaccine, experiments are performed on 4 groups of white mice previously infected with the virulent culture of mytnogo streptococcus. Animals are divided into four groups: three experimental and fourth control 100 animals each.

On the 2nd day after infection, the animals of group 1 are administered the drug according to the recipe described in example 2, the animals of the 2nd group are administered the drug according to the recipe described in example 3, and the animals of the 3rd group are administered the drug as described in example 4.

Mice drug is administered intramuscularly once at a dose of 0.1 cm ³ . On the 2nd and 3rd day after the injection, the appetite is restored in the mice, the animals recover from the death of all animals in the control group.

Example 7

The therapeutic efficacy of the drug is checked in a horse housekeeping dysfunctional for horses. For this purpose, 40 heads of foals are selected, patients with mytoma, 20 animals are administered to the first group of animals - the drug is administered at a therapeutic dose of 10 cm ³ , and 20 animals are administered to the second control group - the drug is not administered. On the 4-5th day after the administration of the drug, the foals decrease body temperature to normal, the appetite is restored, the swelling of the submandibular lymph nodes is completely absorbed, and the animals recover, while the foals of the second control group continue to suffer from the clinical form of myta.

Example 8

The intensity of immunity after the introduction of the vaccine is determined on a deliberately healthy 90 foals. In the experiment, 6 groups of horses with 15 heads each, three experimental and three control ones were used. The vaccine is administered intramuscularly in the area of the croup once at a dose of 5.0 cm ³ . After 2, 6 and 12 months, the immunized and control foals are infected with a virulent culture of mytnogo streptococcus at a dose of 31 T) ₅₀ and take into account the results. The test results are presented in Table. 2

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table 2

The results of the test of immunity in foals immunized with a vaccine against myta horses

Group animals, payment order Myta Vaccine horses Term checks immunity, month results contamination Percent immune foals got sick Goal not got sick Goal one Vaccinated experienced group 2 0 15 100 2 Not vaccinated control group 2 15 0 0 3 Vaccinated experienced group 6 0 15 100 four Not vaccinated control group 6 15 0 0 five Vaccinated experienced group 12 one 14 93.33 6 ... Not vaccinated control group 12 15 0 0

In the case of a control infection, in the 1st and 3rd groups of diseased animals, 100% of immune animals are not registered, and in the 5th experimental group 1 animal gets sick with mild form (93.33% of immune animals). Whereas all animals in the control groups develop clinical form of myta. All diseased animals are treated with the same vaccine in a therapeutic single dose of 10 cm ³ .

The use of a vaccine against horse myta intramuscularly once at a dose of 10 cm ³ leads to a complete clinical recovery of foals from myta within 4-5 days, and at a dose of 5 cm ^{3 it} protects 93.3% of vaccinated animals from infection for 12 months, and sick animals recover mild in mild form.

The proposed vaccine is harmless, does not cause complications in vaccinated animals, has a high activity.

Claims (1)

CLAIM A vaccine against myta of horses, including the inactivated strain of mytnogo streptococcus 81ger1osossiz etschΒΝΚ, broad-spectrum antibiotics bicillin-5, sulfadimezin and trimethoprim; polyoxidonium as an adjuvant and immunomodulating agent and distilled water in the following ratio of components (1 liter): strain 31ger1osossi5 ötsch ΒΝΚ - 1.8-2.0x10 ¹⁰ CFU, bitsillin-5 - 30-40 million IU, sulfadimezin - 1800-2000 mg, Trimethoprim - 300-400 mg, polyoxidonium - 80-120 mg, distilled water - up to 1 l.