Culture and long-term storage of mycobacterial cultures
Abstract
Diagnosis of animal tuberculosis remains an urgent task of veterinary phthisiology. Despite the success achieved, it is still necessary to improve and develop modern methods of indicating and identifying mycobacteria from biological material. For this purpose, a number of nutrient media are used. The aim of the work was to comprehensively improve the system of cultivation and long-term storage of Mycobacterium avium cultures using modern lyophilization technologies and to develop a special selective nutrient medium. During experimental studies, a comparison of the growth of M. avium culture on the classical Löwenstein-Jensen egg medium and the developed “KIT” accumulation medium was carried out. It was established that the “KIT” medium allows to reduce the time of isolation of mycobacterial cultures by 5–7 days and increase the percentage of positive isolations by 27%, which significantly speeds up the process of bacteriological diagnostics. Mycobacteria were isolated from the blood of experimentally infected laboratory animals using the "KIT" nutrient medium. Thus, the highest percentage of isolation from blood was observed in guinea pigs that were inoculated with a suspension of Mycobacterium avium-intracellulare complex. It was found that the serum-glucose protective suspension medium and the drying regime effectively preserve the viability and enzymatic activity of mycobacterial strains for more than 30 years. The conducted study allowed to develop and test a comprehensive system of improved cultivation and long-term storage of mycobacteria M. avium, which includes the use of a new accumulation medium "KIT" and an optimized lyophilization regime using serum-glucose protective suspension medium. The results of the study are recommended to be implemented in the production practice of the biological industry and veterinary laboratories, which will ensure the stability and efficiency of work with Mycobacterium avium cultures.
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