FACTORS INFLUENCING THE SURVIVAL OF SPIROCHETES IN THE FROZEN STATE

TITRATION EXPERIMENTS MADE WITH RELAPSING FEVER SPIROCHETES BEFORE AND AFTER FREEZING SHOWED THE FOLLOWING: 1. With each freezing and thawing there is a slight but regular decrease in virulence, which decrease bears no relation to the duration of storage at - 78 degrees C. Ordinarily infectivity is destroyed by more than 4 refreezings. 2. There was not always close correlation between motility and infectivity. 3. Cooling spirochetes from 0 degrees C. to -78 degrees C. over a 2 to 6 hour period damages them only slightly more than does rapid cooling, but warming from - 78 degrees C. to 0 degrees C. over a 2 to 6 hour period kills most of the organisms. Rapid thawing, as in a water bath, damages the spirochetes less than thawing more slowly, as at room temperature. 4. At storage temperatures of -12 degrees C. and -20 degrees C. there is a gradual decrease in virulence over a period of days or weeks, and by the 6th week the infectivity of the material is markedly reduced.