Clostridium acetobutylicum McCoy et al. emend. Keis et al.
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2. Under anaerobic conditions, withdraw 0.5 ml of #2107 broth from a single test tube (5 to 6 ml), with a Pasteur or 1.0 ml pipette. Rehydrate the entire pellet.
3. Aseptically transfer this aliquot back into the broth tube. Mix well.
4. Use several drops of the suspension to inoculate a pre-reduced #260 blood plate and a #260 slant. An aerobic blood plate may also be streaked to check for purity.
5. Incubate the tubes and plate under anaerobic conditions at 37°C. Incubate the purity plate aerobically at 37°C. Turbid growth is evident in the broth at 24 to 48 hours.
Anaerobic conditions for transfer may be obtained by either of the following:
· Use of an anaerobic gas chamber, or
· Placement of test tubes under a gassing cannula system hooked to anaerobic gas.
Anaerobic conditions for incubation may be obtained by any of the following:
· Loose screw caps on test tubes in anaerobic chamber,
· Loose screw caps on test tubes in an activated anaerobic gas pack jar, or
· Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.
This strain is cited to produce acetone and 1-butanol (2).
Always use freshly prepared pre-reduced media or pre-reduced media that has been previously prepared but stored under anaerobic conditions. Resazurin in the media is a color indicator for anaerobic conditions. Observance of pink color in medium before use or during incubation shows anaerobic conditions have not been met and oxidation has occurred. Medium should be discarded.
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British Patent 4845
Weizmann C. Production of acetone and alcohol by bacteriological processes. US Patent 1,315,585 dated Sep 9 1919
J. Infect. Dis. 56: 333-346, 1935.