Monitoring and characterization of Mycobacterium avium subsp. рaratuberculosis cultures isolated on the territory of Western Siberia

   The results of microbiological studies of a standardized strain and isolates related to Mycobacterium avium subsp. paratuberculosis (MAP) isolated in Western Siberia are presented. Experimental paratuberculosis infection was reproduced on nonlinear white mice using intraperitoneal and intravenous methods of infection with experimental doses of 0.1;0.25; 0.5 and 1.0 ml of suspensions of standardized strains and isolates. Microbiological studies of strains and isolates included microscopic, cultural and biochemical methods: Ziehl-Neelsen staining of smears; inoculation of mycobacteria suspension on thick egg culture medium Lowenstein-Jensen with mycobactin growth factor; pre-sowing treatment of biomaterial obtained from laboratory animals with subsequent inoculation of the resulting sediment on egg culture medium with mycobactin. Biochemical tests: pathogen growth at 30, 37 and 42 ºC, determination of amidase activity; presence of colony growth or its absence on medium with sodium salicylate, determination of catalase and arylsulfatase activity, nitrate reduction, hydrolysis of tween-80, resistance to 5 % sodium chloride. According to the results of the studies, the cultures studied were classified as mycobacteria group 3 according to Runyon's classification. Mice infected with 0.1; 0.25; 0.5 and 1.0 ml of standardized strain suspension and isolates intraperitoneally showed enlargement of lungs, spleen and liver, single purulent foci on liver, spleen and mesentery. The isolation rate index was (++) 2 to (+++)3 to (++++) 4. Mice infected with 0.1; 0.25; 0.5 and 1.0 ml of standardized strain suspension and intravenous isolates showed enlargement of lungs, spleen and liver, marbled liver, suppurative foci in liver. The isolation rate index was (++)2 to (+++)3 to (++++) 4. Scientific data on the cultural, biochemical and biological properties of isolates of paratuberculous mycobacteria coinciding with identical parameters of the standardized strain Mycobacterium avium subsp. paratuberculosis (MAP) were obtained, which allows to refer them to mycobacteria of paratuberculosis. Biological method of research on nonlinear white mice using intraperitoneal and intravenous methods of infection with experimental doses of suspensions containing a standardized strain and isolates of
the paratuberculosis pathogen, allowed to establish the effectiveness of these methods and doses for reproduction of experimental paratuberculosis infection on nonlinear white mice.

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