Isolation and Pure Cultivation of the Smaller Mouth Spirochetes by an Improved Method

Abstract

and probably other factors of the vitamin B complex for norma: growth under the conditions of these studies. Possible factors altering the requirement of choline for dogs on synthetic rations are discussed.The characteristics and properties of the so-called commensal spirochetes of mucous menibraiies, and their individual or contrihuting rbles in fuso-spirochetal infections, remain inadequately understood largely because of a lack of effective nieans for their isolation and maintenance in pure culture. Treponema vimenti, the large delicate loosely wound spirochete of the mouth, seems never to have been obtained in pure culture; 7'. buccale, the large? thick, loosely wound form, has been reported in culture rarely,'. 2* apparently under conditions which could not be duplicated. T. ,microdentiurn, the small mouth spirochete which resembles T . pallidurtz in morphology and is cultivable with the least difficulty,*-* seems for that reason to be the only member of the group which can be accepted with little doubt as a distinct species. Studies of pure cultures of T. microdentkn, moreover, have yielded interesting information on the pathogenesis of fuso-spirochetal infection^.^? 6p ' 9On the other hand, none of the other varieties which have been described and named, such as Noguchi's T . ynncrodentiim~~ or the several species named by Seguin and Viiizent," is so clearly defined.

Cultivation and Isolation of Mouth Spirochetes

“…Robinson (20) has reported isolation of mouth spirochetes by growing them through filter paper, and Kolmer and Kast (19), using Robinson's method for purification of their strains have isolated mouth spirochetes in liquid media by using high dilutions and selective media. Recently Rosebury and Foley (14) have obtained spirochete growth in small Petri plates, containing a very thick layer of a solid medium.…”

“…All authors agree that growth of the mouth spirochetes can be obtained only under anaerobic conditions in media containing some enriching substance such as ascitic fluid, serum or blood, whereas the addition of fresh tissue does not seem to be an absolute requirement for all strains. The isolation of the spirochetes has been attempted by many different methods such as centrifugation (21), prolonged incubation (10), selective media (10,18), and most frequently by growth in solid media (10,11,12,13,14,15,16,17). Miihlens (11,12) and Shmamine (15) preferred to cultivate the spirochetes in deep agar shake tubes.…”

Genetic characterization of an oral treponeme isolated from human subgingival plaque

“…A host-associated treponeme, strain G7201, was isolated from the subgingival plaque of a 52-year-old man with adult periodontitis by a well-plate method (16,24,25). It has been shown that the organism was moiphologically and serologically different from any of the previously established oral Treponema species by the studies using scanning electron microscopy (24,25).…”

The American perspective.

“…Much work has also been done on oral and other non-pathogenic treponemes, e.g. by Rosebury and Foley (1941).…”