478 



APPENDIX 



subsequent attentive study of the biological characters of pure cultures 

 of these bacteria grown in various media. "* 



Bacillus enteritidis sporogenes of Klein. Take six tubes con- 

 taining 15 c.c. of fresh milk and sterilise them by boiling for half an 

 hour. Rapidly cool them by placing them in a beaker 

 of cold water, add to each tube 1 c.c. of a 1 in 500 

 dilution of the milk to be examined, or if it be pre- 

 ferred 0*1 c.c. of the crude milk. Heat the inocu- 

 lated tubes at 80 C. for fifteen minutes. Then remove 

 and cool, and place in Buchner tubes or cylinder con- 

 taining freshly prepared mixture of pyrogallic acid and 

 potassium hydrate solution. Seal the Buchner tubes 

 or cylinder with great care, making it absolute. Place 

 the Buchner apparatus, including the milk tubes, in the 

 incubator at 37 C. After forty-eight hours take out 

 the tubes, and examine them for the B. enteritidis 

 sporogenes. If necessary, inoculate guinea-pigs sub- 

 cutaneously with 1 c.c. of the whey, which in a few 

 hours causes swelling at the point of inoculation, and 

 extensive gangrene of the subcutaneous and muscular 

 tissues with sanguineous exudation ; the animal dies in 

 twenty-four or thirty hours. The B. butyricus of Botkin 

 may produce similar changes in milk tubes, but it has 

 no pathogenic action. Milk may be examined directly 

 by placing 20 c.c. in tubes and treating as above. For 

 the "euteritidis change" in the milk, see p. 307. 



Bacillus tuberculosis. Obtain the sediment of 

 the milk under examination and inoculate 2 c.c. of it 

 into the subcutaneous tissue of the guinea-pig. In 

 about four weeks' time, local if not general tubercu- 

 losis will have been set up. Take some of the dis- 

 charge and stain it after the Ziehl-Neelsen method. 

 The sediment of tuberculous milk may be stained 

 forthwith, without inoculation, by the same method, 

 and in some cases the tubercle bacillus may be thus 

 detected, but, generally speaking, the only sure test 

 is inoculation of animals. The pathological process is 

 slower than in pseudo-tuberculosis, and on exami- 

 nation the diseased tissues show giant cells and 

 form of Buchner Tub? numerous tubercle bacilli arranged within the giant cell 



(see Plate 23, p. 328). 



Method of Rabinowitsch for Tubercle Bacillus in Butter. The butter 

 is placed in sterile conical glass in the incubator at 37 C., where on 

 melting it will arrange itself in two layers. Three c.c. of the superna- 

 tant fatty liquid are injected into the peritoneal cavity of a guinea- 

 pig. A similar quantity of the deposit is treated in a like manner, 



Second Report of Royal Commission on Sewage Disposal, 1902, p. 411. 



