The routine isolation, growth, and maintenance of the intermediate‐size anaerobic oral spirochetes from periodontal pockets

Abstract

By the combined use of membrane filters and the antibiotic rifampin, intermediate‐size anaerobic spirochetes from periodontal pockets were routinely selected and separated from other oral bacterial contaminants. Electron microscopy of six newly isolated strains showed that they belong to the genus Treponema . The organisms had generation times of about twelve hours; they produced only acetic acid when glucose was used as the carbohydrate in the medium Serum, volatile fatty acids, and thiamine pyrophosphate could be omitted individually as supplements from the growth medium, but sodium bicarbonate was absolutely essential for growth. These treponemes could be maintained by monthly transfers into fresh medium, by freezing in glycerol‐broth medium or by lyophilization.