624 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(5) Add 500.0 cc. of water to 500.0 g. of 

 fresh ox heart and immerse in a 

 56°C. water bath shaking the con- 

 tainer frequently. 



(6) When the extract has reached a 

 temperature of 39 to 42°C. place on 

 a 37°C. water bath and allow to ex- 

 tract for 2 hours. 



(7) Strain thru butter muslin. 



(8) Add (7) to (4) in five 100.0 cc. lots 

 allowing 5 minutes between each 

 addition. Keep the agar at 56°C. 



(9) Heat (8) after all the infusion has 

 been added on a water bath for 

 20 minutes. 



(10) Adjust to pH 7.6. 



(11) Cool to 56°C. and add the whites of 

 2 eggs. 



(12) Heat on a water bath for 20 minutes. 



(13) Pass thru butter muslin and filter 

 thru English Chardin paper at 55°C. 



(14) Adjust to pH 7.6. 



(15) Transfer, for storage into bottles of 

 200.0 cc. capacity, each of which con- 

 tains 10.0 to 15.0 cc. of pea extract. 



Sterilization: Sterilize for 15 minutes at 

 100 °C. It is necessary to have the 

 steamer at this temperature before plac- 

 ing the medium in the sterilizer. The 

 medium must not be over-heated, es- 

 pecially after the addition of the heart 

 extract, hence all glassware, butter mus- 

 lin, paper, etc. are to be Steamed before 

 use. 



Use: Culture medium for gonococci. 



Reference: Tulloch (1922 p. 348). 



1950. Nastukoff's Egg Yolk Nutrient Agar 

 (Recktsamer) 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Egg yolk 100.0 g. 



3. NaOH (10%) 0.5 cc. 



4. Nutrient agar 

 Preparation : 



(1) Separate the yolk of egg from the 

 white by Bunge's method (allow the 

 whites to roll from a blotting paper 

 containing the egg). 



(2) Add 0.5 cc. of a 10% NaOH solution 

 and 100.0 cc. of egg yolk to 1000.0 cc. 

 distilled water. 



(3) Prepare nutrient agar. 



(4) Mix equal parts of (2) and (3). 



(5) Evaporate to § the volume by boiling. 



(6) Allow to stand two hours in the 



steamer. 



(7) Filter the thick liquid thru a hot 

 water funnel. 



(8) Distribute into sterile test tubes. 

 Sterilization: Sterilize in the steamer. 

 Use: Cultivation of diphtheria, cholera 



influenza and other pathogenic forms. 

 Variants: Besson prepared a similar me- 

 dium as follows: 



(1) Collect egg yolk under aseptic con- 

 ditions. 



(2) Mix one volume of (1) with 3 volumes 

 of sterile water. 



(3) Melt tubes of nutrient agar. 



(4) Add 2.0 cc. of (2) to each tube of (3) 

 under aseptic conditions. 



(5) Slant. 



References: Nastiukof! (1893 ^33 and 34), 

 Besson (1920 p. 55). 



1951, Bezanfon and Griffon's Glycerol Egg 



Yolk Agar 

 Constituents : 



1. Nutrient agar 100.0 cc. 



2. Glycerol 6.0 g. 



3. Egg yolk 50.0 cc. 



Preparation : 



(1) Prepare nutrient agar. 



(2) Add 6 parts of glycerol to 100 parts (1) . 



(3) Tube. 



(4) Melt sterile (3) and cool to 50°C. 



(5) To each tube add one-half the volume 

 of sterile egg yolk under aseptic con- 

 ditions. 



(6) Mix thoroly and slant to cool. 

 Sterilization: Method of sterilization of 



(3) not given. 

 Use: Cultivation of tubercle bacilli. 

 References: Bezangon and Griffon (1903 



p. 603), Besson (1920 p. 55). 



1952. Pergola's Tellurite Egg Yolk Agar 

 Constituents : 



1. Nutrient agar (1.7%) 100.0 cc. 



2. Potassium tellurite (1.0% 



soln.) 2.0 cc. 



3. Egg yolk 1 



Preparation : Details of method of prepara- 

 tion not given in the abstract. 



Sterilization: Method not given. 

 Use: Diagnosis of diphtheria. 

 Reference: Pergola (1918 p. 101) taken 

 from (1919 p. 57). 



