The pipette method has proved a feasible method of obtaining one-cell pure cultures of anaerobes. Both bacilli and spores may be used as seeding material, but spores give a much higher percentage of positives. Boiling alone affords a sufficient degree of anaerobiosis to the medium for initiating one-cell growths, and semisolid agar is the most convenient form of medium. Exposure to air during isolation apparently has no effect on the viability of spores of anaerobes, but young bacilli of some species suffer from a comparatively short exposure to free oxygen.
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