TUBERCLE BACILLUS IN URINE 329 



in warm carbol-fuchsin for two to five minutes, rinsed in the 

 sulphuric or nitric acid solution, washed, dried, and mounted. They 

 can also be stained by Gram's method, which usually brings out 

 the beaded appearance very markedly, or by any of the other 

 methods mentioned under Sputum. Differentiation from the 

 leprosy bacillus will be found at p. 337, and from the smegma 

 bacillus and other acid-fast organisms at p. 339. 



3. Urine. The tubercle bacillus is often very difficult to 

 demonstrate in urine. The urine must be allowed to stand in a 

 conical glass for twenty-four hours or centrifuged, and film 

 specimens are prepared with the sediment and treated by one of 

 the methods for sputum given above. Several specimens should be 

 made and must be very carefully examined. The sediment may 

 also be treated by the antiformin method. It is important to 

 exclude the smegma bacillus, and the urine is preferably drawn off 

 by a catheter. Staining may be carried out by Housell's method, 

 by which the smegma bacillus is decolorised, viz. after staining in 

 warm carbol-fuchsin the specimen is washed and dried. It is then 

 immersed in acid alcohol (alcohol + 3 per cent, hydrochloric) for 

 ten minutes, washed in water, counter-stained for a few seconds in 

 a saturated alcoholic solution of methylene blue, washed, dried, 

 and mounted (see also p. 339). An electrolytic method for the 

 concentration of the tubercle bacilli has been devised by Russ. 1 



If a diagnosis is of importance inoculation should be resorted to. 

 Two guinea-pigs are inoculated subcutaneously in the thigh or 

 abdomen with 0-5 to 1 c.c. of the deposit from the sedimented or 

 centrifuged urine, or one may be inoculated subcutaneously, the 

 other intra-peritoneally. If tubercle bacilli are present the animals 

 may show signs of tuberculosis as early as two to three weeks after 

 inoculation. Sometimes, of course, the animals may die from some 

 intercurrent infection before the tuberculous infection has had time 

 to develop Delepine 2 recommends the inoculations to be made 

 on the inner aspect of the leg about the level of the knee. The 

 order of infection after inoculation is as follows : the popliteal, 

 superficial and deep inguinal, and sub-lumbar glands, the retro- 

 hepatic, mediastinal and bronchial, deep servical, and subscapular 

 glands, the spleen, liver, and lungs. The inoculated animals are 

 killed in two to three weeks, dissected, and the lesions examined 

 microscopically. Others inoculate two guinea-pigs, one sub- 



1 Proc. Roy. Soc. Lond., B. 1909. 



2 Brit. Med. Journ., 1893, vol. ii, p. 664. The results only apply to 

 ordinary forms of tuberculosis, and not to certain modified forms such 

 as lupus and the avian variety. 



