of American grains, media were prepared from the following: wheat, cracked
wheat, yellow corn and white corn. These media were tested for their ability.
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KLUYVER, A. J., AND HOPPENBROUWERS, W. J. 1931 Ein merkwiurdiges Garungsbakterium, Lindners Termobacterium mobile. Arch. Mikrobiol., 2, 245-260. KLUYVER, A. J., AND VAN NIEL, C. B. 1936 Prospects for a natural system of classification of bacteria. Zentr. Bakt. Parasitenk., II, 94, 369-403. KftHR, C. A. H. VON WOLZOGEN. 1932 Ueber eine GArungsmikrobe in Fakalien von Mickenlarven. Zentr. Bakt. Parasitenk., II, 85, 223-250. KULP, W. L., AND BORDEN, D. G. 1942 Further studies on Proteus hydrophilus, the etiological agent in "red leg" disease of frogs. J. Bact., 44, 673-685. ZImmERMAN, 0. E. R. 1890 Die Bakterien unserer Trinkund NutzwAsser. Chemnitz.
L. S. McCLUNG Bacteriological Laboratories, Indiana Univers8ty, Bloomington, Indiana
Received for publication June 24, 1943
Rubbo et al. (1941) suggested a wheat-mash medium for the selective cultivation of Clostridium acetobutylicum. Most other anaerobes and the sporing aerobes failed to grow on the medium when anaerobic conditions were provided. The constant need for new strains of this organism for the biological production of acetone and butyl alcohol on industrial scale suggested immediately a possible use of the medium as a selective enrichment medium. Following exactly the technique of Rubbo et al., with the exception of the use of American grains, media were prepared from the following: wheat, cracked wheat, yellow corn and white corn. These media were tested for their ability to support the growth of authentic strains of Clo8tridium acetobutylicum, C. roseum, C. fetsneum, and three butyric-acid-producing clostridia. C. roseum and C. fetineum were included because of their close affinity to C. acetobutylicum in physiological reactions. The results were not encouraging. All types of media either failed to support growth or did so only when the inoculum was relatively large-0.2 to 0.5 ml. of an active culture. Usually, if the medium supported growth of C. acetobutylicum, other cultures, including butyrics, also developed. In view of the negative results several different batches of the media were tested and these were prepared by different individuals to eliminate, if possible, personal factors in the technique. These all failed to be selective for C. acetobutylicum. The media were tested by seeding plates and/or deep columns in tubes, or both. Yeast-water glucose agar, included as a control, gave consistently positive cultures with smaller inocula than those which were positive with the hydrolyzed mash media. The pH deterninations were made with a Beckman electrode. All plates were incubated in an oat jar (McClung, ' This problem has been supported, in part, by a grant from the Research Fund of the Graduate School of Indiana University.
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ON THE USE OF HYDROLYZED WHEAT MASH FOR THE ENRICHMENT OF CLOSTRIDIUM ACETOBUTYLICUMI
DRIED TISSUE IN BEEF HEART MEDIUM
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McCoy and Fred, 1935) and negative results were discarded unless yeast-water glucose agar plates within each jar were positive. On the basis of the above results it is concluded that the hydrolyzed wheat medium, prepared from American grains, may not prove to be a successful enrichment medium for C. acetobutylicum.
USE OF DRIED TISSUE IN BEEF HEART MEDIUM FOR ANAEROBIC BACTERIA' L. S. McCLUNG Bacteriological Laboratories, Indiana University, Bloomington, Indiana
Received for publication June 24, 1943
Although the thioglycollate medium of Brewer (1940) has been shown to be satisfactory for the enrichment of Clostridium welchii and other spore-forming anaerobes (McClung, 1940, 1943; Marshall, Gunnison and Luxen, 1940 and others), it may not replace entirely the time-honored beef-heart infusion medium, due to unavailability of the dehydrated medium in certain regions or personal preferences of some investigators. The common practice of preparing each batch of beef-heart medium from fresh tissue has many obvious disadvantages. Following an observation by W. G. Roessler, in these laboratories, that the tissue could be dried 'and used in a medium destined for routine transfer of certain proteolytic anaerobes, a short study has been made of the possibilities of this technique. The results show that it is possible to dry the tissue after extraction and use it with the broth (sterilized and stored in screw-capped bottles) at a later time, as needed, to prepare a medium comparable to that made from the tissue before drying. No unusual equipment is needed and the convenience of the technique is suggested to those using beef-heart medium in civilian or military clinical laboratories. The medium we have tested was prepared according to the method previously published (McClung, 1940). In these experiments the tissue was squeezed dry by hand pressure, spread in a thin layer on two thicknesses of cheese cloth, and 1 This problem has been supported, in part, by a grant from the Research Fund of the Graduate School of Indiana University.
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REFERENCES MCCLUNG, L. S., McCoy, E., AND FRED, E. B. 1935 Studies on anaerobic bacteria. II. Further extensive uses of the vegetable tissue anaerobic system. Zentr. Bakt. Parasitenk., II, 91, 225-227. RUBBO, S. D., MAXWELL, M., FAIRBRIDGE, R. A., AND GILLESPIE, J. M. 1941 The bacteriology, growth factor requirements and fermentation reactions of Clostridium acetobutylicum (Weizmann). Australian J. Exptl. Biol. Med. Sci., 19, 185-198.
