Assay of mycobactins from different mycobacterial species by its effect on growth of Mycobacterium avium subsp. paratuberculosis

2^nd International Conference on Animal & Dairy Sciences

September 15-17, 2014 Hyderabad International Convention Centre, India

Radhika S, Kollannur, J D and Das P

Accepted Abstracts: J Veterinar Sci Technolo

Abstract :

Genus Mycobacterium consists of pathogenic and saprophytic organism which meets the demand of iron by synthesizing mycobactins and exochelins. Mycobacterium avium subsp paratuberculosis (MAP) is unable to synthesize required amount of mycobactin and require exogenous mycobactin. Fast growers such as Mycobacterium fortuitum subsp fortuitum MTCC929, M. phlei MTCC 1724, M. smegmatis MTCC 940, M. vaccae and slow growers such as M. bovis AN5, M. avium D4, M. avium 1723, M. microti MTCC 1727, M. tuberculosis H37Rv and M. avium subsp. paratuberculosis ATCC 19698 obtained from Biological products division, IVRI were used for study. For studying mycobactin production Dorset and Henley?s media was prepared with iron at a concentration of 200μg/liter for mycobactin production. Fast growers were incubated for 3 weeks and slow growers were incubated for 8 weeks at 370C. After incubation mycobactin were extracted from cells by ethanol and chloroform extraction and purified by aluminium oxide chromatography. Among all the mycobacterial species studied, highest mycobactin production was noticed in M. fortuitum followed by M. tuberculosis, M. smegmatis and M. phlei. When production of mycobactin is analyzed per gram of cells, efficient mycobactin producer was M. smegmatis. MAP produced least amount of mycobactin in high iron media. MAP was unable to grow in medium and low iron medium. Assay of mycobactins was done by analyzing growth rate of MAP in iron deficient media for 2 months with extracted mycobactins. Out of all extracted mycobactins used for the growth assay, mycobactins extracted from M. tuberculosis , M. phlei and MAP had produced best growth of MAP & mycobactin extracted from M. microti was least efficienct growth promoter. Mycobactin of M. tuberculosis, M. phlei and MAP is to be used for large scale production and for primary culturing of MAP. Keywords: Mycobacterium avium subsp. paratuberculosis, Mycobactins, growth analysis.