CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



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(4) Grind in mortar. 



(5) Preserve in glass stoppered bottle. 



(6) For use dissolve 3.0 g. powder in 

 30.0 cc. distilled water. Add to 

 70.0 cc. neutral agar. 



(d) Roddy gave the following method of 

 preparation: 



(1) Obtain ox blood from the slaughter 

 house. 



(2) Collect the blood in small sterile 

 bottles containing glass beads. 



(3) Shake until the blood is defi- 

 brinated. 



(4) Mi.x equal volumes of (3) and 

 normal sodium hydrate solution. 



(5) Sterilize (3) in a steam sterilizer 

 for 15 minutes on each of 2 succes- 

 sive days. 



(6) Add 1 part of (5) to 7 parts sterile 

 nutrient agar. 



(7) Tube. 



(8) Slant. 



(9) Do not have the cotton plugs 

 tighter than necessary. 



(e) Stitt used normal NaOH instead 

 of KOH. 



References: Dieudonne (1909 p. 108), Sineff 

 and Drosdowitsch (1909 p. 429), Hofer 

 and Hovorka (1913 p. Ill), Lentz (1915 

 p. 425), Roddy (1917 p. 45), Ball (1919 

 p. 83), Klimmer (1923 p. 218), Stitt (1923 

 p. 50), Park, Williams and Krumwiede 

 (1924 p. 130). 



1974. Pilon's Alkaline Blood Agar 



Constituents: 



1. Nutrient agar (4.0%) 70.0 cc. 



2. Blood, defibrinated 15.0 cc. 



3. NaaCOs (crystalline 12.0% solution) 

 Preparation : 



(1) Mix defibrinated blood and a 12.0% 

 crystalline Na2C03 solution in equal 

 parts. 



(2) Prepare 4.0% nutrient agar. 



(3) Neutralize (2). 



(4) Add 3 parts non-sterilized to (1), 

 7 parts melted (3) and mix thoroly. 



(5) Pour into Petri dishes and allow the 

 plates to solidify. 



Sterilization: If the plates are to be used 

 at once sterilization is not necessary due 

 to high alkali content. If the plates are 

 to be kept several days, draw blood under 

 aseptic conditions and use sterilized agar. 



Use: Enrichment medium for cholera 



vibrio. 

 Variants : 



(a) Klimmer prepared the medium as 

 follows: 



(1) Mix 150.0 cc. of fresh defibrinated 

 beef blood, with 150.0 cc. of a 12.0% 

 soda solution. 



(2) Shake well. 



(3) Allow to stand for 1 to 6 days. 



(4) Steam for 60 to 90 minutes. 



(5) Mix 30 parts (4) with 70 parts 

 sterile neutral agar. 



(b) Park, Williams and Krumwiede gave 

 the following method of preparation : 



(1) Mix equal parts defibrinated beef 

 blood and 12.0% Crystalline soda 

 solution. 



(2) Steam in the Arnold for 30 minutes. 



(3) Prepare 3.0% agar neutral to litmus 

 (about pH 6.6). 



(4) Mix 3 parts (2) with 7 parts (3). 

 Sterilization not specified. 



(5) Pour into Petri dishes (15.0 cc. in 

 a 10 cm. dish). 



(6) Allow to harden, uncovered but 

 protected by paper. 



(7) Plates can be used after drying 

 30 minutes. 



References: Pilon (1911 p. 331), Klimmer 

 (1923 p. 218), Park, Williams and Krum- 

 wiede (1924 p. 130). 



1975. Fildes' Pepsinized Blood Agar 

 Same as 961 but using nutrient agar in- 

 stead of bouillon. 



1976. Carpano's Hemolysed Blood Agar 

 Constituents : 



1. Nutrient agar (2.5%) 1000.0 cc. 



2. Hemolysed defibrinated blood 

 Preparation : 



(1) Prepare 2.5% nutrient agar (peptone 

 broth with 2.5% agar). 



(2) Neutralize to phenolphthalein. 



(3) Adjust the reaction by the addition 

 of 4.0% normal HCl. 



(4) Add sterile, defibrinated blood natu- 

 rally hemolysed (details of prepara- 

 tion not given in the abstract) to the 

 solidified agar. 



Sterilization: Not given. 

 Use: Cultivation of gonococci. 

 Reference: Carpano (1919 p. 599) from 

 (1920 p. 176). 



