Cultivation of mycobacterium paratuberculosis

Abstract

The paper presents a new solid growth medium for the cultivation of Mycobacterium paratuberculosis consisting of organic and inorganic ingredients. The study of diagnostic informative value and effectiveness of solid growth media used for cultivation of Mycobacterium Paratuberculosis was carried out in the laboratory conditions. Extract from birch-wood ash of 3% concentration and a growth stimulant of biological origin, peat oxide, were introduced as a mineral salt bases into the developed medium. When constructing the test medium, Lоwenstein–Jensen egg growth medium with the addition of mycobactin, which is an extract from Mycobacterium. phlei and contains substances necessary for the nutrition and reproduction of Mycobacterium paratuberculosis on artificial nutrient media, was used as an analogue. A test on compatibility and solubility of the components was done in distilled water in accordance with the generally accepted guidelines. The duration of observation ranged from 60 to 90 days. A comparison was made between the time of appearance of the primary and intensive growth of mycobacteria of paratuberculosis on the experimental medium and the Lоwenstein–Jensen control medium with mycobactin. Colonies of primary and intensive growth of standardized M. paratuberculosis strain and M. paratuberculosis isolate obtained from the cattle biomaterial on experimental egg growth medium appeared 3-7 days faster than on Lоwenstein–Jensen control medium with mycobactin. When inoculating biomaterial from cattle (lymph nodes and intestine), the primary growth of M. paratuberculosis on the experimental medium was noted 7 days earlier than on the control one, and the intensive growth was 3 days earlier. The experimental growth medium is cheaper and simpler to prepare than Lоwenstein–Jensen control medium with mycobactin, whose preparation is a rather laborious technological process.