ISOLATION OF THE BACILLUS 



419 



habitats outside the body and in the pus of wounds, other spore- 

 forming obligatory and 



facultative anaerobes 



occur, which grow faster 



than the tetanus bacillus, 



and thus overgrow it. 



(2) If in any discharge / 



the spore-bearing tetanus , ' 



bacilli be seen on micro- * 



scopic examination, then 



a method of isolation 



based on the same prin- 

 ciple as the last may 



be adopted. Inocula- ?"*"* * 



tions with the suspected 



material are made in 



half a dozen deep tubes 



of glucose bouillon, pre- FIG. 122. Spiral composed of numerous 



viously raised to a tem- twisted tiagella of the tetanus bacillus. 



perature of 100 C. After Staiue(l b * R(L Muir ' s raethod ' x 100 - 



inoculation they are 



again placed in boiling water and kept for varying times, say 



for half a minute, for 



one, three, four, five, and 



six minutes respectively. 



^ They are then plunged 



-^ ' in cold water till cool, 



and thereafter placed in 



j.1, ^ ' .-1* j_^__ 



\ 





Fi<;. l'J:J. Trtiinus liacilli ; some of which 

 possess sjxnvs. From a culture in glucose 

 agar, incubated for three days at 37 C. 

 See also Plate IV., Fig. 20. 



Stained with carbol-fuchsin. x 1000. 



Bulloch, may be employed. The 



the incubator at 37 C., 

 in the hope that in one 

 or other of the tubes all 

 the organisms present 

 will have been killed, 

 except the tenanus spores 

 which can develop in 

 pure culture. A series 

 of deep glucose agar 

 tubes may also be in- 

 oculated from the series 

 of bouillon tubes. 



(3) Some method of 

 anaerobically making 

 plates, such as that of 

 isolation of the tetanus 



