656 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



2038. Besredka and Jupille's Egg Stomach 



Digest Agar 

 Constituents : 



1. Distilled water 



2. Beef or veal 750.0 g. 



3. Egg 



4. Stomach (hog) 



5. Agar 2% 



Preparation : 



(1) Macerate 750.0 g. beef in 1200.0 cc. 

 water. 



(2) Heat slowly at first and then boil for 

 30 minutes. 



(3) Concentrate to one liter. 



(4) Filter and dissolve 2.0 g. of agar for 

 every 100.0 cc. of filtrate. 



(5) Make slightly alkaline and boil for 

 25 minutes. 



(6) Filter and distribute in tubes. 



(7) Add 4.0 cc. of egg stomach digest 

 solution (see medium 1007 for prepara- 

 tion) to each tube of sterile (6). 

 (Ordinary peptone agar may be used.) 



Sterilization: Sterilize (6) at 115°. 



Use: Cultivation of gonococcus, whooping 

 cough bacillus, pnemnococci, tubercle 

 bacilli and other forms difficult to cul- 

 tivate. 



Reference: Besredka and Jupille (1914 

 p. 577). 



2039. Sellards and Bigelow's Veal Digest 



Blood Agar 



Same as medium 1000 but solidified by the 

 addition of 2.0% agar. 



2040. Martin's Basal Stomach Digest Veal 



Infusion Agar (Harvey) 



Constituents : 



1. Water 1000.0 cc. 



2. Veal 500.0 g. 



3. Stomach digest solution 1000.0 cc. 



Preparation : 



(1) Mince finely fat-free veal. 



(2) Add 500.0 g. to 1000.0 cc. water. 



(3) Place 18 hours at 37°C. 



(4) Pour the mixture onto a thick, clean 

 cloth. 



(5) Collect the fluid which drains thru 

 the cloth together with that obtained 

 by squeezing the meat in the cloth. 



(6) Mix the fluid collected with an equal 

 volume of stomach digest solution 

 (see medium 998) . 



(7) Heat to 70°C. 



(8) Make the reaction neutral to litmus. 



(9) Add 7.0 cc. N/1 sodium hydroxide 

 per liter. 



(10) Filter thru well-wetted, thick filter 

 paper. 



(11) Dissolve 2.0% agar in (10). 



(12) Steam 30 minutes 



(13) Filter while hot thru well-wetted, 

 thick filter paper. 



(14) Distribute the filtrate into flasks. 



(15) Add one of the added nutrients. 

 Sterilization: Method of sterilization not 



given. 

 Use: Cultivation of meningococci. 

 Added nutrients: 



(a) Defibrinated blood. The blood was 

 prepared and added as follows: 



(1) Add three parts distilled water to 

 one part defibrinated blood. 



(2) Add 0.5 cc. of a 10.0% NaOH per 

 100.0 cc. (1). 



(3) Sterilize in the autoclave at 112°C. 

 (The alkali prevents coagulation.) 



(4) Mix one part (3) with two parts 

 basal agar at 80°C. The blood may 

 be made just slightly alkaline to 

 litmus by the addition of HCI 

 before the addition to the agar. 



(b) Serum. 



(1) Add 1.0 cc. formalin to 500.0 cc. 

 horse serum. 



(2) Add 1.0% ammonia to (1) to neu- 

 tralize to litmus. 



(3) Sterilize at 110°C. for 15 minutes. 



(4) Mix one part (3) with 3 parts 

 basal agar. 



(c) Serum (alkaline). Same as for (a) 

 above, but use serum instead of 

 blood. 



Variants: Harvey used the basal medium 



without any additions. 

 Reference: Harvey (1921-22 pp. 74, 81, 99). 



2041. Harvey's Serum Digest Agar 



Constituents: 



1. Distilled water. 



2. Veal, fat free. 



3. Minced stomach. 



4. HCI. 



5. Formalin. 



6. Ammonia. 



7. Serum. 



8. Agar. 



