With the increasing significance of group IV atypical mycobacteria as etiological agents in a variety of infections, studies were conducted to determine their growth capabilities in water and their comparative resistance to disinfectants used to decontaminate hospital equipment. Isolates of Mycobaterium chelonei (TM strains) from peritoneal fluids of patients and peritoneal dialysis machines were able to multiply in commercial distilled water, with generation times at 25°C ranging from 8 to 15 h. Levels of 105 to 106 cells per ml were attained, and these stationary-phase populations declined only slightly over a 1-year period. Results of studies to determine resistance to disinfectants showed the following. (i) TM strains of M. chelonei cultured in commercial distilled water showed survivors in 2% aqueous formaldehyde (HCHO) solutions up to 24 h; in 8% HCHO, only a 2log reduction in viable counts was observed over a 2-h sampling period. Reference ATCC strains of M. chelonei and M. fortuitum were rapidly inactivated, with no survivors after 2 h of exposure to 2% HCHO or 15 min of exposure to 8% HCHO. (ii) In 2% alkaline glutaraldehyde, TM strains survived 60 min, whereas ATCC strains showed no survivors after 2 min of contact time. (iii) All M. chelonei and M. fortuitum strains survived 60 min of exposure to concentrations of 0.3 and 0.7 ,ug of free chlorine per ml at pH 7. Until the last decade, mycobacteria other than the tubercle bacilli have only sporadically been reported as etiological agents in human infections (18, 34). However, they have been demonstrated as part of the normal flora in sputum and feces of healthy animals and humans (8, 17, 29, 34), and their widespread distribution in nature has been documented by isolations from soil sources (8, 29), wastewater effluents (12, 13), and a variety of municipal, laboratory, and other freshwater sources (2, 11, 17, 18, 32). The infective potential of the atypical mycobacteria has increasingly been recognized, and Barksdale and Kim (3) recently described the