14 ROBERT E. COLLIER 



Table III 

 Degree of sparulation in aerated samples containing penicillin 



Percent Percent sporulation 



Hour of sampling sporulation of control in test sample 



2 



3 

 41/2 



5 20 18 

 51/2 50 57 



6 77 79 



7 96 — 



no nutrients available for further growth of the cells. If one assumes that 

 this method stops growth of the vegetative cells, then it is reasonable to 

 assimie that at the time of withdrawal of the sample the biochemical changes 

 culminating in the production of a spore have already started in every cell 

 in the sample. Such a suspension of cells can successfully complete the 

 process of sporulation in the absence of external nutrients. Presumably the 

 aeration-penicillin treatment of such samples is interferring with the suc- 

 cessful completion of the biochemical changes finally resulting in the for- 

 mation of a spore. 



If, at the end of seven hours of incubation, 96 percent sporulation has 

 occurred, samples are removed and are aerated with or without penicillin 

 at 30°C. Lysis of the sporangia then occurs within 24 hours with no ger- 

 mination of the freed spores. 



We have found that when a culture of Clostridium roseum actively grow- 

 ing in the vegetative state is placed at 4°C. for one to two w^eeks, total lysis 

 of the vegetative cell occurs. This phenomenon also has been found to oc- 

 cur when a culture with 96 percent spores in the sporangia and 4 percent 

 vegetative cells is held at 4°C. for one to two weeks. In the latter case, not 

 only do the vegetative cells lyse, but also do the sporangia. 



In conclusion, from this work we have developed a better method of 

 producing spores from Clostridium roseum. Also, by using methods given 

 here, more information can be obtained about the sporulation process of 

 this organism. 



Discussion 



William L. Brown 



My task is an easy one since no data have been published on this specific 

 subject. Mr. Collier's paper is timely as his work fits well into the general 



