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The physiology and pathogenicity of Mycobacterium tuberculosis grown under controlled conditions in a defined medium
Author(s) -
James B.W.,
Williams A.,
Marsh P.D.
Publication year - 2000
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2000.01020.x
Subject(s) - chemostat , microbiology and biotechnology , biology , mycobacterium tuberculosis , chemically defined medium , growth medium , mycobacterium , bacteria , glycine , biochemistry , amino acid , tuberculosis , in vitro , medicine , genetics , pathology
A chemically‐defined culture medium was developed which supported batch growth of Mycobacterium tuberculosis , strain H37Rv, at a minimum doubling time of 14·7 h. This medium also facilitated chemostat culture of M. tuberculosis at a constant doubling time of 24 h. Chemostat growth was optimized at a dissolved oxygen tension of 20% (v/v) and 0·2% (v/v) Tween‐80. Chemostat cultures were dispersed suspensions of single bacilli (1·5–3 µm long), or small aggregates, at a mean density of log 10 8·3 cfu ml −1 . A limited number of amino acids was utilized (alanine, asparagine, aspartate and serine were depleted by >50%; glycine, arginine, isoleucine, leucine and phenylalanine, by approximately 40%). Chemostat‐grown cells were pathogenic in aerosol‐infected guinea pigs, producing disseminated infection similar to that caused by plate‐grown cells. Cells from chemostat culture were significantly more invasive for J774A.1 mouse macrophages than agar‐ or batch‐grown cells. This study demonstrates the suitability of chemostat culture for the growth of pathogenic mycobacteria in a defined physiological state with potential applications for the controlled production of mycobacterial components for therapeutic and vaccine applications.