LT3985B - Vaccine against animals trichophytosis - Google Patents

Abstract

This invention applies to the veterinary field, namely for the prevention and treatment ringworm in animals. The proposed vaccine, against the ringworm in animals, differs from known vaccines, because it additionally has a quantity of stem VGNKI No. 27 antigen of fungus Trychophyton mentagrophytes, which is enough to initiate the immune reaction of the animal.

Description

Wed. verrucosum, whereas according to Georg (1957) (see ibid., p. 34) the main name of the pathogen of the animal trichophytic is Tr. verrucosum.

The object of the invention is the development of a highly immunogenic vaccine against animal trichophytes called Tr. verrucosum, Tr. verrucosum var autotrophicum, Tr. mentagrophytes, Tr. eąuimen, Tr. sarcovic.

This object is achieved by providing the vaccine with an additional antigen from the strain Tr. mentagrophytes VGNKI No.27 at the following component ratios (% w / v):

Antigen from strain Tr. vemcosum 130L. 300-600 million microconidia enF

Antigen from strain Tr. Mentagrophytes VGNKI No.27 300-600 million. microconidia / cnF Sucrose. 10.0 - 20.0

Gelatin In, 5 - 4.0

Water remaining quantity

Uses strain Tr. mentagrophytes VGNKI No.27.

Trunk Tr. mentagrophytešVGSYA No.27 is derived from an epizootic isolated from a white mouse using a directed graded screening method for the fastest growing colonies with active microconidia aggregation.

Trunk Tr. rnentagrophytes No.27 is deposited with the Russian State Institute of Veterinary Medicinal Products Control, Standardization and Specification collection of microorganisms and its registration number is 27.

Mushroom Tr. The strain mentagrophytes VGNKI No.27 is characterized by the following characteristics and properties:

Morphological features. The micelle is monochrome, smooth, branched, septic, can break into fragments, the terminal gifs do not always form spirals (usually 3-5 curves), and have a micelle diameter of 2.0-3.0 μιη. Many microconidia, round, pear-shaped, elongated, close to rod-shaped, with micelles on the surface of the bunches, more often individually, size 1.5 - 5.2 µm. Microconidia unitary. Number of partitions 1 - 5, more often 1-2, size 13-30 x 3-8 μπι.

- In 30-day-old cultures, a round-shaped, two-contoured chlamydosporus, always discrete, with a diameter of 3.0 to 5.0 pm can be observed. Artospores appear on individual portions of the micelle, its thin-walled shape resembling oval or rod-shaped microcondions of 3.0 to 9.0 pm.

Properties of cultures. The must agar grows as a flat, white, velvety colony with a slight prominence in the middle, the growing edge is smooth, the bottom half reddish-brown, the colony diameter is 50 - 90 mm (15 - 20 days culture).

Sabur agar grows as a flat, white, velvety, cream-colored colony with a slight protrusion in the center, the growing side is smooth, the underside pale yellow, sometimes darker in the center, colony size 50 - 90 mm (15 - 20 days culture).

Henneberg's medium grows as a white, velvety, radial-pleated colony, with a growing spider-like edge, a red-purple underside, a colony diameter of 50-80 mm (15-20 days culture).

Spore formation. Maximum spore formation on must agar 198,4 - 259,2 million / em ³ .

Virulent properties. After intramuscular injection of Tr. mentagrophytes No.27, guinea pigs, rabbits, sheep, goats, calves develop a specific localized superficial 2.0 - 2.5 cm focal site at the injection site within 2 to 3 weeks, which self-heals within 15 to 20 days. Mycologically, the agent cannot be isolated. Applying the strain culture to non-immunized animals on scarified skin (at a dose of 0.25 million / cm ³ microconidia) develops a typical grichophytic foci at the application site within 10 to 20 days. The characteristics of the strain isolated from the pathological material correspond to those of the parent strain.

Antigenic properties. When strain cultures are injected, specific agglutinins with titers of 1:40 to 1: 1280 are formed in calves, sheep and rabbits.

Immunogenic properties. Twice the intramuscular injection in calves and rabbits with a dose of 10 to 20 million doses of this strain. live} microconidia, the animals develop immunity to trichophytes for at least 5 years (observation time).

Trunk Tr. rnentagrophytes VGNKI No.27 is different from epizootic with weak virulent necrosis, non-infectious contact between animals, high activity of microconidia aggregation and immunogenicity when administered intramuscularly in animals.

Examples of preparation of a vaccine according to the invention

The vaccine received strains of 15 to 20 days Tr. vemicosnm 130L and Tr. Cultures of rnentagrophytes VGNKI No.27 are grown individually in matrix flasks for 12 to 20 days on must agar (pH

6.2 - 6.8) at 26 - 28 ° C.

The cultured mycelium is aseptically removed from the culture medium using special scrapers and placed in sterile jars. The mass of mycelium from each strain is homogenized in a mixer or colloidal mill and then using sterile water from Tr. verrucosurn resuspends the 130 L homogenate to a concentration of 300 million microns / cm ³ and the homogenate Tr. rnentagrophytes VGNKI No.27 - up to 600 million / cm ^J. Resuspend in the presence of penicillin or streptomycin at 100 units. Of 1 cm ^{J of} homogenate. Mix equal volumes of mushroom Tr. vemicosnm 130L Strain and Fungus Tr. rnentagrophytes VGNKI No.27 strain homogenates. To the resulting fungal mass suspension is added an equal amount of a stabilizer (pH 6.2-6.8) containing 10% sucrose and 1.5% gelatin in water. It is thoroughly mixed, blown into sterile vials and lyophilized.

example

The vaccine is obtained in the same manner as described in Example 1. For its preparation is used mushroom Tr. vemicosnm 130L and Tr. rnentagrophytes VGNKI No.27 strains homogenates containing inikrokonidijų mln.'cm strength of 450 ³ and a stabilizer using an aqueous solution containing 15% sucrose and 2.5% gelatin.

example

The vaccine against trichophytic livestock is prepared as described in Example 1. It is used to obtain it by the fungus Tr. verrocosutn 130L strain homogeneity with a microconidia concentration of 600 million / cm ³ , ii 'strain Tr. rnentagrophytes No.27 homogenate having a concentration of 300 microns / cm. The stabilizer used is an aqueous solution containing 20% sucrose and 4% gelatin.

The reconstituted vaccine is a homogeneous honeycomb dry mass, which is a gray tablet. The vaccine retains its biological properties when stored indoors in the dark at 2 - 10 ° C. Shelf life -12 months. Vials containing sterile ¹ diluent are supplied with the vaccine.

example

A series of vaccines against animal trichophytes prepared according to Examples 1, 2, 3, before o

use dissolves in a diluent at a rate of 1 dose per cm. It produces a series of vaccines containing these antigen concentrations at a dose of million / cm (Table 1).

table

The name of the antigen Vaccine according to Example 1 Vaccine according to Example 2 Vaccine according to Example 3 Wed. vemicosurn 130L S 7.5 10th Wed. rnentagrophytes VGNKI No.27 10th 7.5 5

Examples 1, 2, 3 use the vaccine to immunize calves by administering a dose of 1 cnT intramuscularly twice at intervals of 10 to 14 days.

Analogously produced strain Tr. vemicosurn 130L and Tr. rnentagrophytes VGNKI No.27. containing antigen microconidia at a vaccination dose of 7.5 million / cm ³ . 35 days after the second immunization, calves are infected with Tr. vemicosurn No.153 (TB-153), Tr. rnentagrophytes No.251 (TC-251), Tr. sarcovov 724 (TC-724), Tr. vemicosurn var.

antothrvpiern 118 (TB-118). At the same time, 6 control (unimmunized) calves are inoculated with the same cultures.

The results of experimental calf challenge are shown in Table 2.

table

Groups No. Livestock number in the group Biopreparation, Immuni- Number of diseased animals infected experimentally used in ver- sion pigs for immunization of calves TB-153 TM-251 TC-724 TB-118 TB-20 1 9th Vaccine according to Example 1 9th 0 0 0 0 0 T 10th Vaccine according to Example 2 10th 0 0 0 0 0 3 7th Vaccine according to Example 3 7th 0 0 0 0 0 4 10th Vaccine from strain Tr.verrucosum 130L 10th 0 4 2 0 1 5 9th Strain Vaccine Tr. mentagmphytes VGNKI No.27 9th 5 0 1 2 2 6th 6th Control (not immunized) 2 O O 2 O is

The table shows that 1, 2 ii '3 series vaccines develop a high degree (100%) of immunity against immunized calves in the case of experimental challenge. Immunization of calves with monovaccines does not develop sufficient immunity if subsequently heterologous to trichophytic pathogen cultures. Previously unimmunized and untreated trichophytes calves are prone to experimental infection with virulent trichophytic pathogen cultures.

example

Similarly, for example 4, 14 goats are immunized in three test vaccine series and then challenged with Tr. vemtcosurn No.153 and Tr. mentagruphytes No.251 in virulent cultures. Control animals (2 animals) should not be unified.

All vaccinated animals did not develop trichophytes, and control animals showed all signs of disease, and strains identified by virulent cultures were isolated from lesion foci.

Example 2 The vaccine used in Example 2 is used to immunize chinchillas rabbits weighing 2.5 to 3.0 kg. The vaccine is diluted in a diluent with 1 dose per cm ^J before use. Gets the following antigen concentration per vaccination dose: 7.5 million micronidia Tr.

vemicosum 130L and 7.5 million. Microconidiae Tr. mentagrophytes VGNKI No.27.

The vaccine is given twice by intramuscular injection to rabbits at a single dose of 0.5 cm ³ . For the control of rabbits, the vaccine is derived from the strain Tr.mentagrophytes No.135 containing 20 million doses per vaccine dose. microconidia 1 cnl, this vaccine is injected into the muscle of the buttocks. After 30 days, rabbits are infected with Tr. mentagrophytes No.251 and Tr. mentagrophytes No.18 in virulent cultures. The results of the infection are recorded within 25 days. The use of the vaccine has been shown to facilitate the development of immunity in all immunized rabbits, while the use of the known vaccine Mentavak does not produce a reliable effect - 2 out of 10 rabbits develop trichophytes and virulent strains are isolated from lesions.

Example 1 immunizes calves with the vaccine variants of Examples 1 and 3, wherein the antigen concentration at the vaccination dose is as in Example 4. In each calf group (10 animals), the vaccine is administered twice by intramuscular injection of 1 cm ^{3 at} intervals of 10 to 14 days. At 4 and 9 months after the second immunization, both groups of calves are infected with Tr. vemicosum No.153 and Tr. mentagrophytes No.251 in virulent cultures. No calf became ill. Thus, the vaccine of the invention retains persistent immunity 4 and 9 months after challenge.

The vaccine is highly effective against trichophytic livestock. For prophylactic use, the vaccine is administered twice with an interval of 10 to 14 days at the following doses:

calves from 14 days to 4 months - 1 cm ³ , over 5 months - 2 cm ', and small cattle (sheep, goats) from 3 to 6 months - 0.5 cynic, over 1 month - 1 cm. In the case of rabbits weighing between 2.5 and 3.0 kg, 0.3 - 0.5 cm is used for prophylactic purposes. For treatment purposes, the vaccine is given twice at a dose of 10 to 14 days with a double dose.

The use of a vaccine according to the invention in veterinary practice will increase the efficacy of the means for combating animal trichophytes.

Claims (1)

DEFINITION OF INVENTION Vaccine against trichophytic livestock. containing antigen from strain Ttychop f lytoti vermeosum 130L, sucrose, gelatin and water, characterized in that it additionally contains antigen from strain Trychophvtoti nictitagtvphytes VGNKI No.27, wherein the component ratios (w / v) are as follows: antigen from the strain Ttychophyton vermeosum 130L 300-600 million. microconidia / cm ³ antigen from the strain