534 THE ANTHRAX BACILLUS 



2. In the blood and urine, and in smears and sections of organs, in both 

 man and the lower animals. 



The clinical diagnosis of malignant pustule in man should always be con- 

 firmed by bacteriological investigation ; and in any suspected or ascertained 

 case of the disease the blood and urine should be immediately examined 

 microscopically since in both man and the lower animals the entrance of the 

 bacillus into the blood stream denotes generalization of the infection and a 

 fatal termination, death occurring soon after its appearance. 



The blood in the Jiving human subject should be obtained by pricking the finger 

 or the lobe of the ear. In the case of animals the blood is best collected from 

 the ear. 



Serum from the malignant pustule may be obtained by scratching the surface 

 of the pustule with a lancet after sterilizing the skin with antiseptics. 



Other material can be collected in the ordinary way. 



To make a complete bacteriological examination it is necessary to confirm 

 the microscopical examination by sowing cultures and inoculating animals. 



(a) Cultures. Sow the material (blood, scraping from organs, exudates 

 etc.) on ordinary media and incubate aerobically. 



(6) Inoculations. Inoculate a guinea-pig (for preference) or a mouse sub- 

 cutaneously with a few drops of blood, or with a scraping from one of the 

 internal organs rubbed up in sterile water, or preferably with a 24-hour 

 growth obtained by sowing the medium with blood or other material. 



(c) Microscopical examination. For microscopical examination prepare : 



1. Blood films. 



2. Smears of the internal organs and especially of the spleen. 



3. Smears from the gelatinous cedema or exudate from the malignant pustule. 



4. Mesentery. This is best examined in the mouse. Remove a small 

 piece of the mesentery, spread it out on a slide with needles leaving the 

 edges to dry a little so as to assure its adhering to the glass, then pull on it 

 in such a manner as to stretch the membrane, treat with alcohol-ether then 

 stain. 



5. Sections of tissues. Cut off small pieces of the liver, spleen, lungs, and 

 kidneys, fix in absolute alcohol and embed in paraffin. Stain as indicated 

 above (p. 522). 



The examination of carcases dead of the spontaneous disease. 



When examining material from a dead body it should be remembered that 

 anthrax carcases very soon after death become invaded by the bacillus of 

 malignant oedema (Chap. XXXVIII.), a micro-organism which on superficial 

 examination is liable to be confounded with the anthrax bacillus ( Jaillard and 

 Leplat). 



[In England the carcase of an animal suspected to have died from anthrax 

 may not be opened. For bacteriological examination the Board of Agriculture 

 require that an ear shall be cut off. 



[The ear should be pinned out on a board and the outer surface washed with 

 2 per cent, lysol. An incision should then be made with a sterile scalpel from the 

 base to the tip and the skin reflected from the subjacent cartilage. With another 

 sterile knife one of the small veins thus exposed should be cut across an inch or 

 so from the base to avoid organisms which may have contaminated the cut sur- 

 face and the blood squeezed out. Films are then prepared and stained (p. 522) 

 and cultures sown. 



[It is of importance to recognize that the mere microscopical examination 

 of blood films cannot be relied upon for the recognition of anthrax in animals 

 suspected to have died of the disease. It is true that in the great majority 

 of cattle which have died of anthrax, bacilli can be seen in large numbers in 



