Vital Resistance Isolation 367 



Concentrated sulphuric acid 2 drops 



Five per cent, oxalic acid solution 1 drop 



Distilled water 10 c.c. 



for five seconds, then to absolute alcohol, xylol, etc. The 

 bacilli appear dark blue upon a paler ground. This method 

 gives very good results, but has been largely superseded by 

 the use of Ktihne's carbol-methylene-blue : 



Methylene-blue 1.5 



Alcohol 10.0 



Five per cent, aqueous phenol solution 100.0 



Ktihne stains the section for about half an hour, washes it 

 in water, decolorizes it carefully in hydrochloric acid (10 

 drops to 500 c.c. of water), immerses it at once in a solution 

 of lithium carbonate (8 drops of a saturated solution of 

 lithium carbonate in 10 c.c. of water), places it in a bath of 

 distilled water for a few minutes, dips it into absolute 

 alcohol colored with a little methylene-blue, dehydrates it 

 in anilin oil containing a little methylene-blue in solution, 

 washes it in pure anilin oil, not colored, then in a light 

 ethereal oil, clears it in xylol, and finally mounts it in balsam. 



Vital Resistance. The organism grows only between 

 25 and 42 C. It is killed by exposure to 55 C. for five 

 minutes. 



Isolation. Attempts at the isolation of the glanders 

 bacillus from infectious discharges, by the usual plate method, 

 are apt to fail, on account of the presence of other more 

 rapidly growing organisms. 



The best method of isolation seems to be by infecting an 

 animal and recovering the bacillus from its tissues. 



The guinea-pig, being a highly susceptible as well as a 

 readily procurable animal, is appropriate for the detection 

 and isolation of the bacillus. When a subcutaneous inocu- 

 lation of some of the infectious pus is made, a tumefaction 

 can be observed in guinea-pigs in from four to five days. 

 Somewhat later this tumefaction changes to a caseous 

 nodule, which ruptures and leaves a chronic superficial ulcer 

 with irregular margins. The lymph-glands speedily become 

 invaded, and in four or five weeks signs of general infec- 

 tion appear. The lymph-glands, especially of the inguinal 

 region, suppurate, and the testicles frequently undergo the 

 same process. Later the joints are affected with a suppura- 

 tive arthritis the pus from which contains the bacilli. The 



