144 TETANUS. 



cultivation, it was found that the last culture was still contaminated 

 by one or more additional microbes. Fliigge claimed to have 

 obtained a pure culture by heating for five minutes the mixed cul- 

 ture to 100 C. (212 F.), but after this procedure the bacillus was 

 incapable of further propagation. After many trials it was found 

 that sterilized solid blood-serum was the best soil for the propaga- 

 tion of the bacillus outside of the body. Both Nicolaier and 

 Rosenbach observed the anaerobic nature of the bacillus, as it was 

 found impossible to obtain a culture on the surface of the nutrient 

 media, or anywhere else where oxygen could not be excluded. The 

 culture appeared slowly as a delicate whitish-gray film in the track 

 of the needle puncture below the surface of the culture substance. 

 By a long series of cultivations, Rosenbach finally succeeded in 

 eliminating all other microbes with the exception of a bacillus of 

 putrefaction. The growth of the bacillus takes place most readily 

 at an equable temperature of 37 C. (98.6 F.), and becomes first 

 visible about the third day in the depth of the culture media. 



Kitasato (" Ueber den Tetanuserreger," Verh. d. Deutschen Gesell- 

 schaftf. Chirurgie, 1889) has finally succeeded in obtaining a pure 

 culture of the bacillus of tetanus from pus taken from a patient 

 suffering from tetanus. As the bacillus will only grow where 

 atmospheric air is excluded, he exposed his cultures to an atmos- 

 phere of hydrogen gas. Mixed cultures which had been kept for 

 several days in the incubator were then exposed for half an hour 

 to an hour to a temperature of 80 C. (176 F.) in a water-bath, and 

 further growth was secured upon plate cultures in closed glass vessels 

 filled with hydrogen gas. He succeeded in destroying all other anae- 

 robic bacilli found in the pus by heating the mixed culture to 80 C. 

 (176 F.), with the exception of the bacillus of tetanus, which upon 

 gelatin plates in the hydrogen atmosphere at a temperature of 18 to 

 ^0 C. (64.4 F. to 68 F.) after a week produced a visible culture. 

 Growth is more rapid at a temperature of 18 to 20 C. (64.4 F. to 

 68 F.), when a culture appears in from four to five days, and if 

 the temperature is kept at 36 to 38 C. (95.8 F. to 100.4 F.) 

 the development of spores and bacilli takes place most rapidly. 



INOCULATION EXPERIMENTS. Nicolaier (" Ueber infectiosen 

 Tetanus," Deutsche med. Wochenschrift, 1884, No. 52) produced 

 tetanus in rabbits and mice experimentally by inoculations with 

 different kinds of earth. Out of 140 experiments, in 69 a disease 

 was produced which very closely resembled tetanus in man. In 

 the pus at the point of inoculation bacilli and micrococci were con- 

 stantly found. Among the bacilli one form was invariably present ; 

 this bacillus resembled in appearance and culture the bacillus of 

 septicaemia in mice, but was more slender. This bacillus was found 

 in isolated places in the connective-tissue, but could not be found 

 in the muscles, nerves, and blood. If the earth was sterilized by 



