An anaerobic film-pouch method has been developed for the isolation and enumeration of clostridia. Fabrication of the pouch is described. Counts of spore suspensions of Putrefactive Anaerobe 3679 and of Clostridium botulinum strains 41-B and 33-A in pouches were compared with those obtained by anaerobic-jar and agar-deep techniques. Statistical analysis revealed a significant difference in favor of the pouch over the tube and anaerobic-jar methods. Tests performed with C. welchii, both in spore suspension and added to chicken pot pie in culture form, also demonstrated the pouch to be at least as proficient as the other, more cumbersome, techniques.
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