The Relation of the Composition of the Culture Medium to the Formation of Endospores by Aerobic Bacilli

Extract

1. Spore formation in eight typical members of the genus Bacillus has been studied.

2. Three of these strains, including one species of B. anthracis, have been found to be practically asporogenous under the experimental conditions. In general the following statements hold good for the sporogenous races studied.

3. Spore formation is almost, or entirely, inhibited by cultivation on media rich in amino acids, such as tryptic digests of casein or meat. Similar inhibition results following cultivation on a medium containing reasonably high concentrations of a mixture of amino acids and asparagine.

4. When such media are suitably diluted with standard inorganic salt solutions the percentage of spores formed is greatly increased, and frequently at least 99 per cent. of spores are formed if the dilution is sufficiently high.

5. When simple nitrogenous compounds such as amino acids are added to a dilute casein digest medium in which sporulation is almost complete, a definite decrease in the percentage of spores present is observed. Asparagine, which is probably readily assimilated, apparently completely hinders spore formation in most cases. Other amino acids do not exert so pronounced an effect, and ammonium phosphate does not appreciably inhibit the formation of spores.

6. The fact that the addition of glycine suppresses growth markedly when it is added to a dilute casein digest medium, but does not appreciably hinder sporulation, suggests that the formation of spores is not due to any toxic effect of added compounds, or compounds already present in the medium.

7. Sporulation is almost complete in a “synthetic” medium in which low concentrations of ammonium phosphate and sucrose represent the sources of nitrogen and carbon, respectively. However, frequent transfers in such a medium may inhibit spore formation partially or entirely in certain instances. This effect probably depends upon the enhanced ability of the culture in question to utilise sucrose as a source of carbon when cultivated constantly in its presence.

8. It is concluded, from the above data, that endospore formation in aerobic bacilli bears an inverse relationship to the amount of available nutrient material present in the culture medium.

I am indebted to Prof. Sir F. G. Hopkins and Miss M. Stephenson for their constant encouragement during the progress of this work. My thanks are due to Mr Pirie of this Department who kindly furnished me with several of the amino acids employed, and to Dr Miles of the Department of Pathology for his kindness in supplying me with certain of the cultures.