lyier's modifications of Anthonc^'s method, but there was evidence of a 

 thin capsule in stained India ink preparations of smooth strains. The 

 organism was found to be a facultative anaerobe, groTdng best mth 

 unrestricted access to air. 



The optimum temperature for growth was 20° to 25° C. No growth 

 occurred at 35° C. and was very slight and slow at 7° C. The pH range 

 for growth was from 5.5 to 8.2 with the optimum at about 7.0. 



At 20° C, in a liquid medium, the culture remained viable for 

 less than 2 weeks. On agar slants the viability was increased to a 

 period of 5 weeks. 



Colonies on agar plates reached a size of about 1 mm. in 2 days 

 and, if well separated, attained a diameter of 2 to 3 mm. within a 

 week. Colonies were circular, smooth, entire, convex, opaque, and 

 cream colored. The colonies of "S" variants grew relatively faster 

 and were butyrous; "R" colonies were compact and tough, and could be 

 removed intact from the medium. Colonies on gelatin plates were of the 

 same appearance. There was no liquefaction. 



On agar slant, gro-'-vth was good within 2 to 3 days. The growth 

 was filiform, slightly glistening, cream colored, butyrous with "S" 

 variants, and brittle with "R" variants. There was no odor and the 

 medium remained unchanged in appearance. 



In liquid media, a scant pellicle or ring appeared within a vreek. 

 Transient clouding occurred ■■/d.thin 24 hours and eventually the grovcth 

 became granular and adhered to the walls, the medium becoming clear. 

 A fine granular sediment was produced. In "S" variants the media 

 remained uniformly clouded for several days. 



There was no growth on nutrient agar or in nutrient broth. In 

 gelatin stab cultures, growth was uniform, best near the surface, 

 filiform, and without liquefaction. 



Nitrates were not reduced and indole was not formed (Kovac's 

 method). Production of hydrogen sulfide was doubtful, mth occasional 

 slightly positive results -with lead acetate paper strips. The methyl 

 red test was oositive and the Voges-Proskauer test negative after 1 week 

 of growth in Difco V.P. and M.R. medium iTith fish extract. There was 

 no change in litmus milk and no visible growth on potato, mth or without 

 fish extract. There was a slow hydrolysis of starch as indicated with 

 Lugol'3 solution on starch agar plates. Agar plates with rabbit 

 erythrocytes showed beta-hemolysis. 



5A 



