SPORING BACILLI 311 



Pass in H or N until all the air is displaced, and then seal 

 ends of tubes in flame. 



6. Buchner's Tube. A wide tube with a constriction 

 near the closed end, so that an ordinary culture tube can 

 be inserted and is held up by the constriction. In use, 

 i grm. of solid pyrogallic acid and 20 c.c. of 10 per cent 

 KOH are put into the bulb, the previously inoculated tube 

 is inserted loosely plugged, and the Buchner tube is 

 plugged and sealed with melted paraffin or closed with 

 a rubber cap. The solution in bulb absorbs oxygen. The 

 method can be used without the special tube, with a tall 

 glass jar or a desiccator. It is better to add the alkali 

 by pipette after the inoculated tube has been inserted. 



7. Bulloch's Method. A bell jar with an inlet and an exit 

 tube, both having stopcocks. The bell jar is firmly bound 

 down to a glass plate by ung. resinae. Before fixing, 

 a glass dish is set on the plate, and 3 to 4 grm. of pyro- 

 gallic acid are heaped to one side of it. Culture plates or 

 tubes in a beaker are rested on a tripod stand placed in the 

 dish. The bell jar is now fixed on so that the inlet tube 

 will end in the dish at the side away from the pyrogallol. 

 Hydrogen is passed in, and then a solution of KOH 

 (109 grm. in 145 c.c). 



8. Vacuum Method. Desiccator with stopcock. Exhaust 

 air by burning alcohol on soaked filter-paper put into jar. 

 Stopcock is needed to release pressure when opening. 



B. Tetani. 



The cause of tetanus or lockjaw, a disease characterized 

 by the gradual onset of general stiffness and spasms of the 

 voluntary muscles, beginning in the jaw muscles and those 

 of the back of the neck. The disease is usually associated 

 with a wound received four to fourteen days previously and 

 infected with earth or dung. The majority of cases are fatal. 



Kitasato first isolated the bacillus (in 1889) and in 

 this fashion : Pus from the local suppuration of mice 

 inoculated from a human case, was smeared upon the 

 surface of agar slants. These were permitted to develop 

 at 37 C. for 24 to 48 hours. At the end of this time the 

 cultures were subjected to a temperature of 8o C. for 

 1 hour. This destroyed all non-sporulating bacteria as 



