225 



THE CITLT^VATON OF SPIROCIIAETA NOVYI* WITHOUT. THE USE 

 OF TISSUE FROM ANIMAU ORGANS. 



CiTAULES A. Rkiikf:ns. 



(riinliie University, Lafayetfe, Indiana.) 



In l!)Or) Sclioroscliewsky' liy dooply iinl)C(l(lin>: a picM-o of 1iuiii:im tissue 

 (•(.ntaiuinjj Sinrochaota Pallida in gelatinized horse serum, lirst dcuion- 

 st rated that the treponema might be cultivated m vitro. He was not able 

 to obtain a pnre culture of the organism, for bacteria grew along with the 

 pallidum, nor did he succeed in reproducing syphilitic lesions by inoculating 

 iinimals. 



In 1910. MuhkMr obtained the first generati<»n of the spirochaete by 

 utilizing the above mentioned method. By the use of horse serum agar, 

 lie furtlior succeeded in obtaining a culture devoid of bacteria. Muhlen's 

 pure cultures also were evidently non-pathogenic. 



During the same year, Bruchner and Galasesco^ succeeded in cultivating 

 "young impure cultures" by using Schereschewsk5'"s medium. But upon 

 inoculating rabbits with the material which also contained the original 

 spirochaetal ti.ssue. syphilitic lesions were produced. They were, however, 

 unable to obtain a second generation of these organisms. 



In 1911. Hoffman^ succeeded in getting pure cultures of the spirochaete 

 by the utilization of Schereschewsky's and Miihlen's methods. Although 

 his cultures were morphologically typical, he. like the above mentioned 

 experimenters, was not able to demonstrate their pathogenic properties. 



Also at this time, Sowade^ succeeded in procuring impure virulent cul- 

 tures by using the gelatinized horse serum or gelatinized ascitic fluid. This 

 inocuable material also still contained the original pallidum tissue. 



In 1911, Noguchi" was able to cultivate pure virulent cultures in vitro 

 of this organism. He accomplished this by placing a small piece of fresh 

 sterile rabbit kidney or testicle tissue into each tube containing about 

 sixteen cubic centimeters of serum water (one part of serum and three 

 parts of distilled water) which was previously fractionally sterilized. A 

 layer of sterile parattin oil was added to each sterile tube containing this 

 cultural medium and placed under strict anaerobiosis and incubated at 

 35-37° Centigrade. Under these conditions the spirochaetes which were 

 morphologically typical and virulent were obtained and cultivated for 

 many generations. 



In the following year. Noguchi" cultivated for the first time the follow- 

 ing relapsing-fever spirochaetes: Spirochaeta Duttoni, Spirochaeta Kochi, 

 Spirochaeta Obermeieri and Spirochaeta Novyi. 



In 1912. Treponema macrodentium and micr(identium\ Treponema Re- 

 fringens". Treiionema mucosum'". Spirochaeta Phagedenis^'. and Spirochaeta 

 Gallinarum'- were likewi.se successfully cultivated. 



The media u.sed in their cultivation was of a similar nature to that used 

 in the pallidum cultural work. It was not essential in some instances to 



*The author is greatly indebted to Dr. F. G. Novy, Ann Arbor. Micbigan, thru 

 whose kindness this strain of Spirochaete was obtained. 



