THE NITRIFYING BACTERIA. 513 



BACILLUS TETANI, NICOLAIER, 1884. 



In 1884 Nicolaier produced tetanus in mice and rab- 

 bits by the subcutaneous inoculation of particles of 

 garden-earth, and demonstrated that the pus produced 

 at the point of inoculation was capable of reproducing 

 the disease in other mice and rabbits. He did not suc- 

 ceed in isolating the organism in pure culture. In 1884 

 Carle and Rattone, and in 1886 Rosenbach, demon- 

 strated the infectious nature of tetanus as it occurs in 

 man by producing the disease in animals by inoculating 

 them with secretions from the wounds of individuals 

 affected with the disease. In 1889 Kitasato obtained 

 the bacillus of tetanus in pure culture, and described 

 his method of obtaining it and its biological peculiarities 

 as follows : 



METHOD OF OBTAINING IT. Inoculate several mice 

 subcutaneously with secretions from the wound of a 

 case of typical tetanus. This material usually contains 

 not only tetanus bacilli, but other organisms as well, so 

 that at autopsy, if tetanus results, there may be more or 

 less suppuration at the seat of inoculation in the mice. 

 In order to separate the tetanus bacillus from the others 

 that are present the pus is smeared upon the surface 

 of several slanted blood-serum or agar-agar tubes and 

 placed at 37 to 38 C. After twenty-four hours all 

 the organisms will have developed, and microscopic 

 examination will usually reveal the presence of a few 

 tetanus bacilli, recognizable by their shape, viz., that of 

 a small pin, with a spore representing the head. After 

 forty-eight hours at 38 C. the culture is subjected to a 

 temperature of 80 C. in a water-bath for from three- 

 quarters to one hour. At the end of this time series of 



