THE CULTURE OF ANAEROBIC BACTERIA. 



315 



be necessary to pour on top, after inoculation, the contents of another 

 agar tube, taking- care to sterilize the mouths of both tubes. The drop 

 or two of liquid which sometimes accumulates on the surface of the 

 agar, and invariably on the bottom of the tubes can 

 be stained for ordinary flagella and for giant-whips. 



Likewise inoculate litmus glucose gelatin tubes 

 and place at 37. 



The four organisms are also planted in glucose 

 bouillon and these tubes are then placed in bottles in 

 hydrogen at 37. In about 3 days the tubes can be 

 taken out and examined for spores. It is best to re- 

 move the bacterial deposit from each tube by means 

 of a sterile drawn-out pipette (Fig. 61). The growth 

 can then be placed in a small Esmarch dish, or salt 

 cellar, and after dilution with water it can be used for 

 making cover-glass preparations. When preparations 

 are made direct from the bouillon they are likely the 

 give a bad back-ground owing to the organic matter 

 present. The spores can be readily double-stained. 



Inclined glucose agar should also be inoculated 

 with the four organisms, and the culture should be de- 

 veloped in hydrogen at 37 for 24 hours. The growths 

 should then be examined carefully in hanging-drops 

 for motion, spores, giant-whips, etc. The latter will 

 be found especially abundant in the liquid of conden- 

 sation on the bottom of the tube. With this material 

 cover-glass preparations will be made, as presently 

 described, and employed for demonstrating the pres- 

 ence of flagella or whips. 



FIG. $4. Tube 

 cultivation, on po- 

 tato, of the tuber- 

 cle bacillus; with 



Glucose agar Petri dishes should also be prepared, open capillary. 

 Some of these should be placed in the plating appara- 

 tus (Fig. 52) and cultivated in an atmosphere of hydrogen. The 

 pyrogallate method (p. 313) should be used to develop another set of 

 the plates. A third set of plates should be grown in a vacuum (Fig. 

 53) which is brought about by means of a Chapman aspirator. 



Make the following cultures at the same time as the preceding: 



1. Streak cultures of the tubercle bacillus on inclined glycerin 



agar and on glycerin potato (Roux tube, Fig. 54). A pure culture, or 



