1003.] On t/ir. fr'xi'sfanw to Heed of B. anthracis. 495 



The actual procedure in all the experiments was as follows: 

 Sterilised glass tubes 3 inches long and J inch diameter were about 

 half filled with infected water, and the tubes were then sealed with 

 the blow-pipe. This was effected without warming the contents by 

 placing the tubes upright in a holder revolved by clockwork at about 

 thirty revolutions per minute ; the top of the tube was then heated by 

 an upward pointing flame until its diameter contracted to about 

 inch, and drawn to a point by means of a previously heated glass 

 tube held in the hand. 



The tubes were next placed in the holder attached to the cover of 

 the chamber, the water in the tube surrounding the thermometer was 

 brought to the same temperature as the sealed tubes and the cover 

 placed on the heating chamber. Before admitting steam, however, the 

 paper record was started and the pointer (E) brought successively to 

 90, 100, 110, 120, 130, 140, 150, on the thermometer scale, so 

 that each record might have on it a thermometer scale for reference. 

 Steam was then admitted and the requisite temperature maintained 

 for the time determined on. With a little practice in manipulating 

 the two-way cock it was found easy to keep the temperature constant 

 within a quarter of a degree Centigrade. Of course the temperature 

 which it was wished to maintain was settled beforehand, but in the 

 results the temperatures are taken from the records. 



The method of inoculating the tubes was as follows : The tubes, 

 having been plugged and sterilised by dry heat, were half filled with 

 distilled water, re-plugged, and then sterilised in the steamer for half an 

 hour on three successive occasions. On the morning of the day of experi- 

 ment the tubes were inoculated with the respective organisms, either 

 from an agar or broth culture, as stated in each instance, and again 

 plugged. Within two or three hours the inoculated tubes were 

 sealed up and submitted to the different degrees and durations of 

 heat, as detailed in the table. This having been done, the tops of the 

 tubes were filed off, and the contents sown into broth with the least 

 possible delay, generally within two hours. Microscopic examination 

 of the culture inoculated was made in each case immediately before 

 sowing into the tubes; and afterwards, as soon as growth (if any) 

 made its appearance in the inoculated broth, in order to verify the 

 purity of the cultures. . 



Care was taken in all cases to be sure that spores were present in ti 



*The oiSS growth of BaeiMus anthrads was ^*** 

 culture on agar, supplied by the kindness of Dr J. W H. Eyre, 

 Bacteriologist to Guy's Hospital; this was derived from the blood of 

 a fatal eas^ of anthrax in the wards of the Hospital m March 903 

 Sub-cultures were made from this strain, and moculated into the tubes 

 of water, as detailed in the tables. 



