648 Typhoid Fever 



Adami and Chapin* have suggested a method for the isolation of 

 typhoid bacilli from water, in which use is made of the agglutination 

 of the bacilli by immune serum. 



Two quart bottles (Winchester quarts) are carefully sterilized and filled with 

 the suspected water with an addition of 25 cc. of nutrient broth and incubated 

 for eighteen to twenty-four hours at 37°C. By this time the t3rphoid bacillus 

 grows abundantly in spite of the small amount of nourishment the water contains. 

 At the end of the incubation, 10 cc. of the fluid is filled into each of a number of 

 long narrow (7 mm.) test-tubes made by sealing a glass tube one-half meter long 

 at one end. About i inch from the bottom the tube is filed completely round so 

 as to break easily at that point. The different tubes next receive additions of 

 typhoid immune serum sufficient to make the dilutions i : 60, i : 100, i :i5o, and 

 I : 200. If typhoid bacilli are present, within a quarter of an hour beginning 

 agglutination can be seen, and by the end of two to five hours flocculent masses 

 collect at the bottom of the tube, forming a flocculent precipitate. The next 

 procedure should be with the tube showing agglutination with the greatest dilu- 

 tion, as the more concentrated preparations carry down not only the typhoid 

 bacilli, but also closely related organisms. After the sedimentation of the agglu- 

 tinated bacilli is complete, the tube is broken at the file mark, and the sediment 

 contained in the short tube washed with two or three changes of distilled water, 

 being allowed to settle each time. This removes many of the organisms not 

 agglutinated. A loopful of the washed sediment is transferred to a tube of 

 nutrient broth, and finally from this tube plate cultures are made upon Eisner's or 

 Hiss' media. 



A culture-medium for isolating the typhoid bacUlus from feces is 

 recommended by Drigalski-Conradif and by Petkowitsch. J It is 

 made as follows: 



Horse-meat infusion (3 pounds of horse meat to 2 liters 



of water) 2 liters 



Witte's peptone 20 grams 



Nutrose 20 grams 



Sodium chlorid 10 grams 



Agar-agar 60 grams 



Litmus solution (Kubel and Tiemann) 260 cc. 



Lactose ; 30 grams 



Crjrstal-violet solution (o.oi per cent.) 20 cc. 



Before adding the crystal-violet solution render feebly alkaline to litmus 



(about 0.04 per cent, of pure soda). 

 Colon colonies upon this medium appear in fourteen to sixteen hours to be red 

 and opaque. Typhoid colonies blue or violet, transparent and drop-like. 



Beckman§ modifies the preparation, making it as follows: 



(a) Add I liter of water to 680 grams of finely chopped lean beef and place in 

 the cold for twenty-four hours. Express the juice and make up to i liter. Coagu- 

 late the albumin, either by boiling for ten minutes or by heating to i2o°C. in the 

 autoclave. Filter. Add 10 grams of Witte's peptone, 10 grams of nutrose, and 5 

 grams of sodium chlorid. Heat in the autoclave at a temperature of i2o°C. for 

 thirty minutes, or boil vigorously for fifteen minutes. Render slightly alkaline 

 to litmus paper. Filter. Add 30 grams of agar. Heat in the autoclave 

 at a temperature of i2o°C. for one-half hour, or heat over the gas-flame until the 

 agar is dissolved. Render slightly alkaline to litmus paper whfle hot, if necessary. 

 Filter through glass wool into a sterile vessel. 



(6) To 130 cc. of litmus solution (Kubel and Tiemann's) add 15 grams of chem- 

 ically pure lactose. Boil for ten minutes. 



* "Berliner klin. Wochenschrift," Feb. 13, 1899. 

 t "Zeitschrift f. Hygiene," Bd. xxrx. 



t " Centralbl. f . Bakt.," etc., May 28, 1904, Bd. xxxvi. No. 2, p. 304. 

 § See F. F. Wesbrook, "Jour. Infectious Diseases," May, 1905, Supplement, 

 No. I, p. 319. 



