INOCULATION EXPERIMENTS. 149 



suspended in water caused no symptoms, but a culture from it, 

 six days old, contained the bacilli, and when injected subcutaneously 

 produced the disease. From the absence of the bacilli in the 

 internal organs, he concluded that this microbe does not permeate 

 the whole body, and that the disease owes its origin to absorption 

 of toxic agents from the wound. 



Ohlmuller and Goldschmidt ( u Ueber einen Bakterienbefund 

 bei Menschlichem Tetanus," Centrcdblatt f. die ges. Medicin, 1887, 

 No. 31) made a thorough bacteriological examination of a case of 

 tetanus following complicated fracture of the right thumb. The 

 disease appeared the day following the injury and resulted in death 

 after not more than seventeen hours. Soon after death inoculation 

 experiments were made with blood taken from the heart and spleen 

 and pus from the seat of fracture, according to directions given by 

 Bumm. The cultivations were made in solid blood-serum and 

 kept at a temperature of 38 C. (100.7 F.). The tubes con- 

 taining blood from the heart and spleen remained sterile, but the 

 nutrient media infected with pus showed signs of growth by lique- 

 faction of the solid serum. The bacilli which were detected 

 resembled those of mouse septicaemia, only somewhat larger in size. 

 In addition to these microbes streptococci and a thicker bacillus 

 were found. Twenty-four hours later liquefaction had increased, 

 but the streptococci had diminished in number. The characteristic 

 bacilli were pin-rods with globular ends, and club-shaped rods with 

 colorless terminal spores. On the third day the serum had under- 

 gone more advanced liquefaction and at the same time a fetid odor 

 was noticed. A slide compared with one prepared by Nicolaier 

 showed the identity of the two microbes. In order to prove still 

 further their identity two mice were inoculated with the mixed cul- 

 tivation. Twelve hours after infection tetanus had made its 

 appearance, followed by death in seventeen hours. It should be 

 remarked that the spasms commenced in the tail, extended to the 

 posterior extremities, and then gradually forward. From these 

 animals blood-serum was taken with which other mice were infected. 

 Again tetanus was produced and successful cultivations were 

 made. Successive cultivations appeared to diminish the intensity 

 of the virus. Of two mice of equal size and age, one, which 

 received one portion, died of tetanus on the ninth day, while the 

 other, which received a dose three times as large, died on the third 

 day. Cultivations on agar-agar always remained sterile. Cultiva- 

 tions in sterilized coagulated albumen from chicken's and goose- 

 eggs showed that the bacilli retained their properties for about a 

 week, but later they were displaced by other organisms. An 

 attempt was made to destroy the other microorganisms by heating 

 the mixed cultivation to 100 C. (212 F.) for five minutes. The 



