The foregoing experiments conclusively show that a broth medium is quite unsuitable as a test culture medium to determine the vitality of anthrax spores in disinfection experiments, whereas agar is a suitable and delicate medium for the purpose, even when considerable traces of the disinfectant are carried over with the inoculation.
The reason for this inefficiency of broth is not obvious. We thought that it might be due to the absence of bacillar forms in the sporing material, but the emulsion of spores heated to 80°C. for 15 minutes and then inoculated directly into broth gave good growths.
Absence of oxygen might be another factor, but the results were the same when splinters of sterilised wood infected with anthrax spores were treated. The wood floated on the surface of the broth and so was subjected to a free supply of oxygen, yet no growths were obtained in broth when the splinters were soaked in the disinfectants, while good growths were obtained on agar. The control splinters gave good growths in broth. It may be that the anthrax spores are partially de-vitalised by the action of the disinfectant and that in this condition broth is a comparatively unsuitable culture medium for them. Prolonging the time of incubation of the broth cultures up to 10 or 14 days makes no difference. If a culture in broth shows no growth in 48 hours, a growth hardly ever appears with more prolonged incubation. Nor is this superiority of agar over broth as a culture medium confined to the emulsified disinfectants employed in these experiments, for similar results have been obtained with phenol, and with formaldehyde, the latter both in the fluid (formalin), and in the gaseous, conditions.