Biochemical properties and fatty acid composition of Mycobacterium haemophilum: study of 16 isolates from Australian patients.

The biochemical properties and fatty acid compositions of 16 strains of Mycobacterium haemophilum from Australian patients were studied. The strains proved to be indistinguishable from each other but could readily be differentiated from other slowly growing mycobacteria with similar cultural features. Mycolic acid analyses revealed the presence of alpha-, methoxy-, and ketomycolates. The fatty acid composition supports the validity of the fact that M. haemophilum is a distinct species. The fatty acid composition was consistent among the 16 strains, but it was unusual in that there was some resemblance to the fatty acid composition of M. leprae. The wide range of pHs (5.4 to 7.4) that supported growth of M. haemophilum on artificial medium is in keeping with suggestions that M. haemophilum exists in an environmental habitat.

Two-dimensional gas chromatography with electron capture detection for the sensitive determination of specific mycobacterial lipid constituents.

A method was developed for determining two characteristic mycobacterial lipid constituents, tuberculostearic acid (as its pentafluorobenzyl ester) and 2-eicosanol (as its pentafluorobenzoyl ester), by using gas chromatography with electron capture detection. A microprocessor-controlled column-switching system (two-dimensional gas chromatography) facilitated sample preparation and increased specificity. The usefulness of the technique was illustrated by its ability to reveal picogram amounts of tuberculostearate in a suspension of Mycobacterium leprae isolated from a naturally infected armadillo. Two-dimensional gas chromatography with electron capture detection may in some instances provide a convenient alternative to gas chromatography-mass spectrometry for use in demonstrating the presence of mycobacteria in a complex environment.

Biochemical characteristics and fatty acid compositions of some armadillo-derived mycobacteria and their relation to Mycobacterium gordonae.

The long-chain components of 75 strains of mycobacteria, cultivated from Mycobacterium leprae-infected or non-infected armadillos, and of eight clinical and 15 environmental isolates of M. gordonae, were compared. Four major groups could be distinguished based on the presence of 10-methyloctadecanoic (tuberculostearic) and 2-methyl 3-hydroxyeicosanoic acids and secondary alcohols (2-octadecanol and 2-eicosanol). Some heterogeneity was found in strains assigned to M. gordonae: the characteristic absence of tuberculostearic acid and secondary alcohols and the presence of the branched C14 and the hydroxylated C20 acids were seen in only 34 of the 49 strains studied. Three strains were identified as M. malmoense, one as M. kansasii, ten as belonging to the M. avium-M. intracellulare-M. scrofulaceum complex and eight as belonging to new groups of armadillo-derived mycobacteria (ADM 1, ADM 2 and ADM 3) by conventional bacteriological tests and fatty acid compositions, though M. malmoense was heterogeneous in its fatty acids composition. Four strains, identified as M. avium by conventional tests, differed from this species by their fatty acid compositions. Thirteen strains showed some similarity to M. simiae and ten strains differed from all other known mycobacteria.

Use of gas chromatography to differentiate Mycobacterium leprae from cultivable armadillo-derived mycobacteria, M. avium/intracellulare, and M. lepraemurium by analysis of secondary alcohols.

Two long-chain secondary alcohols, 2-octadecanol and 2-eicosanol, were demonstrated by gas chromatography in hydrolysates of Mycobacterium avium/intracellulare, in cultivable, armadillo-derived mycobacteria, and in M. lepraemurium grown in vivo, but they were not found in purified suspensions of M. leprae isolated from experimentally infected armadillos. Gas chromatographic analysis of these alcohols constitutes a method for rapid detection and quantification of contaminating mycobacteria in preparations of M. leprae intended, for example, for vaccine use. The technique may also be of value for critical evaluation of cultures of "in vitro-grown" M. leprae.