156 PATHOGENIC BACTERIA, 
glands, spreading a large quantity of the material 
on the surface of the tubes. 
(d@) Seal air-tight, place in the incubator, and 
examine the growth that occurs. 
4. Sections. 
(a) Prepare sections of tubercular lesions from 
the inoculated animals or from the human lung in 
acute phthisis. 
(6) Stain by the Ziehl-Neelson, Koch-Ehrlich, or 
Rosenberger methods. 
5. Detection of Tubercle Bacteria in Urine. 
(a) Make smears from the deepest layer of sedi- 
ment thrown down by the centrifuge, or in the 
sedimenting glass. 
(4) Stain in hot (steaming but not boiling) 
Ziehl’s carbol-fuchsin for one minute. 
(¢) Wash in water. 
(2) Decolorize in 20 per cent. sulphuric acid until 
pink. 
(e) Wash in water, 95 per cent. alcohol for thirty 
seconds, and again in water. 
(f) Stain in Loffler’s methylene-blue for twenty 
seconds. 
(g) Wash in water, dry, and mount. 
The smegma bacillus, which frequently occurs in 
urine, does not stain by this method, as it is 
decolorized by alcohol. Rosenberger’s method also 
gives excellent results with urine sediment, 
