RU2051965C1 - Strain of bacterium corynebacterium pseudotuberculosis used as reference and for preparing diagnostic preparations at cavernous lymphadenitis in sheep - Google Patents

Abstract

FIELD: biotechnology. SUBSTANCE: preparing strain C. pseudotuberculosis ВГНКИ N 146S-ДЕП for identification acceleration of cavernous lymphadenitis pathogen in sheep at bacterilogical, allergic and serological infection diagnosis. Molecular properties of strain DNA: spectral indices: A 260/280 = 1.5, A 260/230 = 1.8, concentration is 915 mcg/ml, content of G + C = 43.0 mole%. On the basis of homology of DNA nucleotide sequence strain is concerned to the group III. For diagnosis allergen prepared of this strain is administrated to sheep by intracutaneous route into subtail wrincle. Reaction recording is carried out after 24 hr. Inflammatory reaction appeared in sick animals at the place of diagnosticum administration. No inflammation in health animals. Method ensures to detect additionally up to 80% infected animals as compared with conventional method (by palpation of surface lymphatic nodes). EFFECT: accelerated idendification of pathogen.

Description

 The invention relates to veterinary microbiology, epizotology and can be used in the practice of bacteriological laboratories.

 The widespread spread of C. pseudotuberculosis infections in sheep farms has led researchers to the need to develop intravital methods for diagnosing the disease. Currently, in our country, these studies are hampered by the absence of the causative agent of this infection in the collections of microorganisms.

 The strains of C. pseudotuberculosis available in foreign collections, based on the data of the Bergi microbe determinant, have significant variability of phenotypic properties, and information about their genotype is very limited. There is data on the content of G + C in DNA in microorganisms of this species of 67.5 mol. and all pathogens for animals and humans, microorganisms of the genus Corynebacterium in the range from 57 to 60 mol.

 The strain Corynebacterium pseudotuber-culosis N 146S was obtained on September 26, 87 from the purulent contents of an abscess in the muscles of a sheep belonging to the Victory state farm in the Novovarshavsky district of the Omsk Region.

 Morphological signs. In smears from pure cultures, microorganisms are present in the form of gram-positive small ovoids with a length of 0.908 ± 0.022 μm, a thickness of 0.826 ± 0.012 μm, the cell contour and its ends are stained more intensively than the central part, the cells are collected in large heaps.

Cultural, physiological and biochemical characteristics. NA telkuritovom blood agar after 2-3 days of culture at ³⁷ ° C colonies appear black circular spherical shape, smooth, shiny with smooth edges, pasty consistency, good suspended in saline.

 On the MPA, S-forms of gray-white colonies are formed, sometimes with a small amount of yellow pigment. They are round, spherical in shape, with smooth edges, the surface is moist, shiny, pasty consistency, when touched, smeared on the environment.

 BCH evenly grows turbid, a small amount of powdery sediment forms at the bottom, which rises with a pigtail when shaking.

 With growth on MPB, exotoxin is formed, the maximum amount of which can be fixed by lowering the pH of the medium, more often this occurs on the third day of cultivation.

 The strain reduces nitrates to nitrates, ferments glucose, lactose, sorbitol, raffinase. Contains catalase, does not contain urease and gelatinase.

 Intraperitoneal infection of guinea pigs at a dose of 10 MEM / ml causes damage to internal organs, hemorrhagic foci in the lungs, and an enlarged spleen.

Molecular characteristics of DNA strain: 1. Spectral characteristic
A 260/280 1,5
A 260/230 1.8 2. Concentration 915 μg / ml 3. The content of G + C 43.0 mol.

 In the DNA reaction, DNA hybridization with several tens of C. pseudotuberculosis N 146S strains is assigned to group III by the level of homology of the nucleotide sequences of DNA (Trofimov I.G. Borisenko I.S. Fomin B.A. and Artyushin O.K.).

 The strain is assigned the registration number C. pseudotuberculosis N 146S DEPT in the collection of microorganisms of the laboratory for the control and standardization of drugs against tuberculosis, pseudo-and paratuberculosis. VGNKI of veterinary drugs (123022 Moscow, Zvenigorodskoye Shosse, 5), where it is stored.

Claims (1)

 The bacterial stamp Corunebacterium pseudotuberculosis VGNKI N 146S-DEPT, used as a reference and for the manufacture of diagnostic drugs for caseous sheep lymphenitis.