Effect of a sputum digestant on the viabiltiy of Mycobacterium fortuitum

Abstract

A microcolony technique has been demonstrated as being useful for the rapid determination of the viabilities of single cells of Myocbacterium fortuitum. Cultures of M. fortuitum grown to early logarithmic phase in broth were treated with the sputum digestant N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH) for periods of 10 to 40 s. After growth for three generations (7.5 h) on agar films, viabilities were determined by counting under a phase contrast microscope. The viable mycobacteria grew into microcolonies that exhibited extensive branching, whereas the nonviable mycobacteria remained as single cells. Sputum was mixed with some broth cultures before treatment to stimulate the digestion process in a clinical laboratory. When broth cultures were treated with sputum digestant for 40 s, only 2.8% of the cells remained viable. When the broth cultures were mixed in a ratio of 1:4 with sputum and then treated for 40 s, 16.4% of the cells remained viable. The results also indicate M. fortuitum is very sensitive to the digestant. The results also indicate that a microcolony technique could be used for the assessment of the viability of mycobacteria.