RU2093178C1 - Mixed vaccine for control of trichophytosis in cattle and other artiodactyls - Google Patents

Abstract

FIELD: biotechnology, veterinary mycology. SUBSTANCE: vaccine has a mixture of antigens from culture fungus Trichophyton verrucosum VIEV N 478 and Trichophyton verrucosum VIEV N 145 taken at equal ratios at concentration 60-160 mln microconidia/1 ml 4-5% NaCl solution in the presence of aluminium hydroxide as an adjuvant. EFFECT: enhanced effectiveness of vaccine.

Description

 The invention relates to the field of veterinary mycology, to a vaccine against trichophytosis of cattle and small cattle, such as sheep, goats and reindeers.

 To date, trichophytosis of cattle, reindeer, sheep and goats has occurred in countries around the world. Monovaccines have been found for these animal species. The creation of a single associated vaccine against trichophytosis of animals is relevant.

 Known vaccine TF-130K against trichophytosis in cattle, containing an antigen from a live immunogenic culture of Trichophyton Verrucosum strain 130L, diluent saline, antibiotics penicillin and streptomycin. TF-130K additionally includes a filler: sucrose, gelatin and water.

 The drug TF-130K is administered intramuscularly at a dose of 1 8 ml, twice with an interval of 10 14 days (USSR author's certificate N 955571, class A 61 K 39/00, 1984).

 To date, vaccines against trichophytosis of reindeer and goats have not been developed. There are separate reports on the use of LTF-130 against reindeer trichophytosis, which has not been conducted sufficiently.

 The technical result of the invention is to obtain an associated vaccine that provides the creation of intense and long-lasting immunity in the body of animals against trichophytosis of cattle, sheep, goats and reindeer.

 The inventive vaccine includes two new strains of strain Trichophyton verrucosum N478, isolated from reindeer in 1993 and strain Trichophyton verrusosum N 145, isolated from goats in 1991. In addition, the vaccine contains an adjuvant gel of aluminum hydroxide.

 The strain Trichophyton verrucosum was deposited in the Museum of mushroom cultures of the laboratory of mycology and antibiotics of the All-Russian Research Institute of Experimental Veterinary Medicine named after Ya.R. Kovalenko, Moscow under N 478.

 The strain Trichophyton verrucosum VIEV N 478 has the following properties.

 Cultural and morphological properties. The colonies are grayish, leathery-velvety 1 1.5 cm in diameter (30-day-old crops), elevated, tuberous with a flat, even edge. The mycelium is even, branching.

 The microconidia are oval, elongated-oval, pear-shaped, club-shaped, rod-shaped 1 3 x 3 8 microns. Macroconidia are rare and practically absent.

 Chlamydospores are abundant in old cultures with terminal or intermediate chains and clusters. A feature of this culture is the ability to form copiously arthrospores with a diameter of 3 5 7 microns in any generation.

 Virulent properties. When a homogenized culture is applied to the scarified skin of a rabbit or guinea pig on day 8-15, hyperemia is detected, alkalizing areas with crusts, followed by self-recovery on day 20-30.

 Immunogenic properties. When a rabbit is twice intramuscularly injected with a vaccine made from the obtained strain in a dose containing 30–40 million live microconidia, the animals develop intense immunity to subsequent infection with trichophytosis lasting at least 1.5 years (observation period).

 The culture of strain N 145 of the fungus Trichophyton verrucosum was isolated from a patient with trichophytosis of dairy goats, a herd of which was brought to Russia from New Zealand in 1990 and attenuated.

 The strain was deposited in the Museum of mushroom cultures of the laboratory of mycology and antibiotics of the All-Russian Research Institute of Experimental Veterinary Medicine named after Ya.R. Kovelenko under N 145.

 The strain Trichophyton verrucosum VIEV N 145 is characterized by the following properties.

 Cultural and morphological properties. The colonies are white, dense, woolly with a smooth cobwebly edge of 1.8 2.5 cm in a monthly culture. The mycelium is a thin, sinuous thick mycelium disintegrates into arthrospores of 3.5 microns. Chlamydospores are round, regular in shape up to 30 microns. On a thin mycelium round, oval, elongated-oval and pear-shaped microconidia of 2.5–5 μm are formed.

 Immunogenic properties. Double immunization with live antigen prepared from this strain of rabbits forms an intense and long-lasting immunity in animals for a period of at least 1.5 years (observation period).

 Virulent properties. When a vaccine strain is ground in a sterile mortar culture of fungus in a concentration of 500 million / ml of fungal cells applied to the scarified rabbit skin on 3-5 days, hyperemia of the scarified area of rabbit skin is noticeable, which disappears on 7-10 days with the formation of superficial peeling and recovery on 15 25 day.

 Reactogenic properties. Intramuscular administration to rabbits of the antigen from the strain of the fungus Trichophyton verrucosum VIEV N 145 at a dose of 160,200 million microconidia does not lead to a change in the clinical condition of animals.

 The vaccine has the following composition.

 Antigen of the culture of the fungus Trichophyton verrucosum VIEV N 478 and Trichophyton verrucosum VIEV N 145 with a concentration of 60 160 million microconidia in equal proportions in 1 ml of 4 6% saline NaCl solution with 0.5 0.6 ml of normal aluminum hydroxide gel for every 100 ml mixtures of antigens.

 Example 1

 From mattresses inoculated with a culture of the strain of the fungus Trichophyton verrucosum VIEV N 478 and from the same number of mattresses seeded with the culture of the fungus Trichophyton verrucosum VIEV N 145, they remove the mycelium, which is homogenized in sterile 4 6% NaCl saline solution containing the mushroom film after homogenization from 300 to 800 million microconidia in 1 ml.

 The homogenate is diluted with the same saline to a concentration of 60,160 million microconidia in 1 ml and aluminum hydroxide gel is added at the rate of 0.5 ml gel per 100 ml of antigenic mixture. Thus obtained vaccine is poured into bottles and sealed.

 The resulting vaccine was used with a positive prophylactic effect at a dose of 1 ml for cattle, 380 animals, the efficiency was 98.1% in sheep, 2500 animals in a dose of 1 ml, the efficiency was 91.5% in goats 300 animals, the efficiency was 98% in reindeer 1500 goals, efficiency 100%. Duration of immunity 2 years (observation period).

 Example 2

 The antigen homogenate is prepared in the same manner as in example 1, and diluted to a concentration of 80 million microconidia in 1 ml of NaCl saline solution and aluminum hydroxide gel is added in the same amount as in the first example, i.e., by volume. The vaccine thus obtained was used with a positive prophylactic effect for cattle and small cattle in a dose of 0.8 ml (180 cattle, 75 goats, 200 sheep, 1,500 reindeer). Efficiency was 93 100%. Duration of immunity 2 years (observation period).

 Example 3

 The antigen homogenate is prepared as in Example 1, and adjusted to a concentration of 60 ml of microconidia in 1 ml of salt NaCl, aluminum hydroxide gel is added in the same amount as in the first example.

 The vaccine obtained in this way was used with a positive prophylactic effect for cattle and small cattle at a dose of 1.2 ml (43 cattle, 63 goats, 120 sheep, 360 reindeer). Efficiency was 95 100% duration of immunity 2 years (observation period).

 The vaccine can be used for therapeutic purposes, in which case the dose is doubled.

 Shelf life of the liquid vaccine is 14 months (observation period).

 The vaccine can be prepared in dry form. It is prepared in the usual way by mixing the antigen homogenate with the protective medium of the usual composition for trichophytosis vaccines.

 The associated vaccine provides the creation of immunity against trichophytosis of large and small cattle (sheep, goats, reindeers), is harmless and areactogenic.

 The vaccine will be widely used in state farms, private sectors and farms of Russia and the CIS.

Claims (1)

 Associated vaccine against trichophytosis of cattle and other artiodactyls, characterized in that it contains antigens from the strain of the fungus Trichophyton verrucosum VIEV N 478 and from the strain of the fungus Trichophyton verrucosum VIEV N 145, taken in equal proportions with a final concentration of 60 160 million microconidia in 1 ml 4 5% NaCl saline, and adjuvant aluminum hydroxide in an amount of 0.5 to 0.6 ml for every 100 ml of vaccine.