Right-angle branched filaments and rods micromanipulated from activated sludge foam and mixed liquor were identified as putatively novel members of the genera Gordonia, Mycobacterium and Rhodococcus using a combination of chemical, molecular and morphological data. Pyrolysis mass spectrometric analyses of gordoniae isolated in both the present and a previous study revealed pyro-groups, distinct from validly described Gordonia species, which could be equated with those based on morphological properties and 16S rDNA data. Putative gordoniae assigned to one of these groups were found to be closely related to strains currently identified as "Rhodococcus australis". These strains were also found to have properties consistent with their classification in the genus Gordonia. The results of this study highlight the limitations of the microscopic approach to filament identification and cast further doubt on the view that foaming can be attributed to members of one or a few Nocardia species.