Abstract
A technique modeled on the Lee tube was developed for enumeration of anaerobes. It consists of two tubes. The larger outer tube contains molten culture medium tempered at 45–50°C. After the organisms are introduced and distributed in the medium, a sterile smaller inner tube is inserted centrally into the outer one; the outer tube is then plugged with cotton. This technique was compared to the petri plate and Lee tube methods for the enumeration of various clostridia. There was no significant difference among the three methods. A range of 20 to 300 colony forming units per modified Lee tube could be counted with accuracy. Heat-stressed spores of Clostridium botulinum 62A were enumerated by the modified Lee tube and plate methods. The D and z values obtained with either technique were not significantly different and they compared favorably to data reported in the literature.
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