ME THODS OF EX AM IN A TIOh r . 3 1 5 



tion as the symptoms of the disease (oedema, fever, etc.) 

 were established, so was the curative effect diminished, 

 even though large doses of the serum were employed. 

 These results have been in the main confirmed by other 

 observers. 



Methods of Examination. These include (a) micro- 

 scopic examination ; (/;) the making of cultures ; and (c] 

 test inoculations. 



(a) Microscopic Examination. In a case of suspected 

 malignant pustule, film preparations should be made from 

 the fluid in the vesicles or from a scraping of the incised 

 or excised pustule, and stained with a watery solution of 

 methylene-blue and also by Gram's method. By this 

 method practically conclusive evidence may be obtained ; 

 but sometimes the result is doubtful, as the bacilli may be 

 very few in number. In all cases confirmatory evidence 

 should be obtained by culture. Occasionally they are so 

 scanty that both film preparations made from different 

 parts and even cultures may give negative results, and yet 

 a few bacilli may be found when a section of the pustule 

 is examined. It should be noted that the greatest care 

 ought to be taken in handling the part, as otherwise the 

 diffusion of the bacilli into the surrounding tissues may be 

 aided and the condition greatly aggravated. The examina- 

 tion of the blood in cases of anthrax in man usually gives 

 negative results, with the exception of very severe cases, 

 when a few bacilli may be found in the blood shortly before 

 death, though even then they may be absent. 



(b) Cultivation. A small quantity of the material used 

 for microscopic examination should be taken on a platinum 

 needle, and successive strokes made on agar tubes, which 

 are then incubated at 37 C. At the end of twenty-four 

 hours anthrax colonies will appear, and can be readily 

 recognised from their wavy margins, by means of a hand 

 lens. They should also be examined microscopically by 

 means of film preparations. 



(c} Test Inoculations. A little of the suspected material 

 should be mixed with some sterile bouillon or water, and 



