178 DIFGO MANUAL 



means in schema for identification of organisms. Hajna^ has used "H" and "O" 

 agglutination together with other characteristics and serological reactions for 

 the identification of Salmonella. In this method, typical colonies are picked from 

 the primary plating medium, S S Agar, or Bismuth Sulfite Agar plates, and 

 inoculated into tubes of semisolid agar, Triple Sugar Iron Agar and finally into 

 H Broth, without securing further inoculum. The inoculated tubes are then 

 incubated at 30 °C. overnight. Cultures showing reactions on the semi-solid agar 

 and Triple Sugar Iron Agar suggestive of Salmonella typhosa or other "Sal- 

 monella like organisms" are examined for "H" antigen. Place 0.02 ml. of a 1-5 

 dilution of Edwards mixed polyvalent Salmonella "H" Serum in the bottom of 

 an agglutination tube. Then remove 1 ml. of the H Broth culture and add 1 ml. 

 of 0.6 per cent formalinized saline, mix thoroughly and place in a water bath at 

 50° C. for 1 hour, but not to exceed two hours. A flocculant agglutination in the 

 tube is an indication of the presence of Salmonella "H" antigen. The individual 

 "H" antigens are detected by specific H antisera, using the tube agglutination 

 technique. The presence of "O" antigens is determined by an initial slide 

 agglutination test using the H Broth and polyvalent "O" antiserum followed by 

 specific "O" antiserum. The balance of the H Broth is used for the indole test 

 by adding 0.5 ml. of Kovacs' reagent to the tube. A dark red color in the surface 

 layer indicates a positive test; a yellow color indicates a negative test. 



To rehydrate the medium, dissolve 21.5 grams Bacto-H Broth in 1000 ml 

 cold distilled water. Distribute in 4 ml. amounts in 13 x 100 mm. tubes. Sterilize 

 in the autoclave for 15 minutes at 10 pounds pressure (117°C.). The final re- 

 action of the medium will be pH 7.2. 



One pound of Bacto-H Broth will make 21.1 liters of medium. 



iPub. Health Rep., 65:116:1950. 

 2 Pub. Health Lab., 9:23:1951. 

 8 Personal Communication, 1951. 



FRIEWER SHAUGHNESSY MEDIUM 



BACTO 



FRIEWER SHAUGHNESSY BASE NO. 1 (B310) 



DEHYDRATED 



Bacto-Beef Extract 3 g. 



Bacto-Peptone 30 g. 



Bacto-Agar 3 g. 



Bacto-Brom Thymol Blue 0.01 g. 



BACTO 



FRIEWER SHAUGHNESSY BASE NO. 2 (B311) 



DEHYDRATED 



Bacto-Lactose 10 g. 



Lead Acetate 0.05 g. 



Bacto-Friewer Shaughnessy Medium prepared according to the formula of 

 Friewer and Shaughnessy^ is recommended for use as a primary screening 

 medium in the laboratory diagnosis of enteric infections. The medium is prepared 

 in two parts, Bacto-Friewer Shaughnessy Base No. 1 and Bacto-Friewer 

 Shaughnessy Base No. 2. A solution of each base is prepared separately and mixed 

 before sterilization to secure the complete medium. 



