BACILLUS TYPHOSUS. 43 



4. Make inoculations of Bacillus typhosus and Ba- 

 cillus coli communis on, 



(a) agar-agar tubes; 



(6) gelatine (slope) tubes; 



(c) liquefied grape sugar gelatine; 



(d) potato tubes; 



(e) peptone tubes; 

 (/) litmus milk tubes. 



Incubate all except the gelatine tubes at 37°0. Ex- 

 amine and contrast the growths from day to day. 



5. Sections of mesenteric gland, spleen and liver 

 from Typhoid fever cadaver. 



In the tissues Bacillus typhosus is found scattered 

 in clumps, so that a number of sections may have to be 

 examined before finding any bacilli. At least six sec- 

 tions of each of the tissues should be stained. 



(a) Stain in LoeflBler's methylene blue, leaving in the 

 stain from 30 minutes to 2 hours. Rinse in water 

 and then in .1% acetic acid for 2 or 3 minutes; dehy- 

 drate in ebsolute alcohol, clear in xylol and mount in 

 balsam. 



(b) Stain in Stirling's gentian violet solution. 

 Gentian violet .... 5 grammes. 

 Alcohol ..... 10 cc. 



Anilin oil 2 cc. 



Water 88 cc. 



Leave sections in this stain for 10 minutes. Rinse 

 in water and then in .1% acetic acid for 2 or 3 minutes; 

 dehydrate in alcohol, clear first with oil of cloves and 

 then wash several times with xylol before mounting in 

 balsam. 



