CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



751 



containing glass beads. Shake to 

 defibrinate the blood. 



(2) Mix an equal volume of (1) and N/1 

 KOH solution. 



(3) Prepare nutrient gelatin. It may be 

 necessary to increase the gelatin 

 content of the medium to 20 or 30.0% 

 in order to obtain a solid medium 

 after the addition of the alkaline 

 blood. 



(4) Neutralize (3) to litmus. 



(5) To 700.0 cc. of the melted neutral 

 nutrient gelatin add 30.0 cc. of sterile 

 (2) and mix. 



(6) Distribute into sterile tubes or plates. 

 Sterilization: Sterilize (2) by heating in 



the Koch steamer for 30 minutes. Steri- 

 lization of gelatin not specified. 



Use: Enrichment and detection of cholera 

 vibrio. The author reported that the 

 cholera vibrio liquefied the gelatin. 



Reference: Pergola (1910 p. 493). 



2320. Miiller's Blood Gelatin 



Constituents : 



1. Nutrient gelatin 1000.0 cc. 



2. Blood, goat 100.0 cc. 



Preparation : 



(1) Distribute nutrient gelatin in 240.0 

 cc. lots in sterile 400.0 cc. Erlenmeyer 



(2) Allow about 25.0 cc. of blood to pass 

 directly from the juglar vein of a 

 goat into melted gelatin, cooled to 

 about 45°C. 



(3) Mix well and pour into sterile Petri 

 dishes. 



Sterilization: Method not specified. 

 Use: Cultivation of fowl diphtheria bacilli. 



Ito and Matsuzaki cultivated Spiro- 



chaeta icterohaemorrhagiae on a similar 



medium. 

 Variants: Ito and Matsuzaki prepared 



their medium as follows: 



(1) Prepare nutrient gelatin. 



(2) Melt (1) and cool to 25-30°C. 



(3) Add from 2 to 4 parts of guinea pig 

 blood or human blood to one part of 

 (2). The erythrocytes settle to the 

 bottom of the gelatin before solidifi- 

 cation giving it an opaque and deeper 

 color. 



(4) Inoculate with a drop of infected 

 blood and mix well by stirring. 



(5) May be covered with a layer of 



paraffin, but not necessary in order to 



obtain growth. Incubate at 15 to 



37°C. 



References: MuUer (1906 p. 519), Ito and 



Matsuzaki (1916 p. 558). 



2321. Miiller's Serum Gelatin 



Constituents : 



1. Nutrient gelatin 1000.0 cc. 



2. Serum, goat 1000.0 cc. 



Preparation : 



(1) Prepare nutrient gelatin. 



(2) Melt (1) and cool to about 60°C. 



(3) Mix equal parts of (2) and goat serum 

 that has been obtained under aseptic 

 conditions and heated to 60°C. 



(4) Pour into sterile plates or sterile 

 tubes. 



Sterilization: Sterilization of gelatin not 



specified. See step (3) for sterilization 



of serum. 

 Use : Cultivation of fowl diphtheria bacilli. 



Miiller reported that the gelatin was 



liquified. 

 Reference: Muller (1906 p. 520). 



2322. Beijerinck's Trypsinized Gelatin 



Constituents : 



1. Water 900.0 cc. 



2. Gelatin (liquefied) 10.0 cc. 



3. (NH4)N03 5.0 g. 



4. Potassium phosphate 5.0 g. 



5. Gelatin 80.0 g. 



Preparation : 



(1) Liquefy gelatin with pancreas extract 

 (method not given). 



(2) Dissolve 10.0 cc. (1), 3, 4 and 5 in 

 900.0 cc. of water. 



(3) Neutralize, or adjust to a slight 

 acidity. 



Sterilization: Not specified. 



Use: To study liquefaction of gelatin and 



pigment production by Chlorococcum. 



Beijerinck reported that chlorococcum 



did not liquify the gelatin. 

 Reference: Beijerinck (1890 p. 461). 



2323. Jensen's Pepsinized Milk Gelatin 



Same as medium 1112 but solidified by the 

 addition of gelatin. 



