14 
which he considered the cause of the infection, but again was unable 
to obtain it in pure culture, although he carried on a mixed culture 
upon coagulated calf’s blood serum. 
Soon afterwards Rosenbach,® 1886, found a similar bacillus with a 
round terminal spore in a case complicating frost gangrene in man 
and transferred the infection to animals in the pus. 
In 1889, ^yith the aid of anaerobic technic, Kitasato^ for the first 
time grew the bacillus in pur,e culture and by successful inoculation 
experiments finally proyed that this bacillus was the real cause of 
tetanus. Kitasato succeeded in isolating the tetanus bacillus, first 
described by Xicolaier and later by Rosenbach, not alone by under- 
stanchng anaerobic methods but by taking adyantage of its resisting 
spore. Tliis bacillus appears in the pus of wounds of man and animals 
ha\dng tetanus. It often forms spores in the pus, but if the pus is 
examined early it may contain spore-free rods. He planted the 
material containing the bacillus upon agar slants, which were incu- 
bated one to two days. The mixed cultures so obtained were heated 
one hour at 80° C. and then transplanted to agar plates. Kitasato 
designed for this purpose a special shaped apparatus in wliich hydro- 
gen was used to replace the air. 
Kitasato^^ further showed that the tetanus bacillus is not found in 
the heart’s blood of mice dead of tetanus and therefore concluded 
that we are dealing with an intoxication and not ^\ith an infection. 
It was in the folloydng year, 1890, that Behring and Kitasato"^ 
published their epoch-making work upon the tetanus toxin and 
tetanus antitoxin, laying the foundation of serum therapy. On 
account of the great historical and practical importance of this docu- 
ment a transcription of its principal features is giyen: 
The immunity of rabbits and mice which have been immunized against tetanus 
depends upon the power of the cell-free serum to render the toxic substances which 
are produced by the tetanus bacillus harmless. 
Experiments show; 
1. The blood of tetanus immune rabbits possesses the property of destroHng the 
tetanus poison. 
2. This property is destroyed by the extra vascular blood and the cell-free serum 
obtained from it. 
3. This property is of such a stable nature that it is also effective in the bodies of 
other animals, so that we are in a position, by means of the transfer of blood serum, to 
accomplish noteworthy therapeutic results. 
a Rosenbach; Zur Aetiologie des \Yundstarrkrampfes beim Menschen. Arch. f. 
klin. Chir., Berk, vol. 34, p. 306-317. 
Kitasato, S.; Ueber den Tetanusbacillen. Zeit. f. Hyg., vol. 7, 1889, p. 225. 
c Kitasato, S. ; Experimentelle Untersuchungeniiber das Tetanusgift. Zeit. f. Hyg. , 
vol. 10, 1891, p. 304. 
Behring, E., and Kitasato, S.; Ueber das Zustandekommen der Diphtherie- 
Immunitat und der Tetanus- Immunitat bei Theiren. Deut. med. Woch., vol. 16, 
. 1890, p. 1113-1114. 
