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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



1970. Esch's Hydrolyzed Meat Agar 

 (Kohlisch and Otto) 



Constituents : 



1. Nutrient agar 



2. Beef 500.0 g. 



3. NaOH (normal) 250.0 cc. 



Preparation : 



(1) Dissolve 500.0 g. of beef in 250.0 cc. 

 of N/1 NaOH solution in an aluminum 

 kettle by heating. Stir constantly. 

 Solution is complete after from about 

 10 to 15 minutes. 



(2) When cool filter thru a towel. 



(3) Mix 3 parts sterile (2) with 7 parts 

 neutral nutrient agar. 



(4) Pour into plates. 



(5) Dry at 60° for 30 minutes. 



(6) Allow the plates to stand open at 

 room temperature for 24 hours. 



Sterilization: Sterilize in streaming steam 

 for one hour. Method of sterilization of 

 agar not given. 



Use: Elective medium for cholera vibrio. 



Variants : 



(a) The author used pike instead of beef. 



(b) Stitt prepared the medium as follows: 



(1) Treat 500.0 g. chopped beef with 

 250.0 cc. of normal NaOH until dis- 

 integrated. 



(2) Filter thru cloth. 



(3) Sterilize (method not given). 



(4) Add about one part of (3) to about 

 2.5 or 2 parts of nutrient agar. 



(5) Dry the plates. 



References: Kohlisch and Otto (1915 p. 

 435), Stitt (1923 p. 50). 



1971. Wellman's Placenta Infusion Agar 

 Add Wellman's Placenta Infusion (see 

 medium 1377) to 2.0% nutrient agar. 



1972. Orcutt and Howe's Fat Blood Agar 

 Same as medium 960 but solidified by the 

 addition of agar. 



1973. Dieudonne's Alkaline Blood Agar 



Constituents : 



1. Nutrient agar 700.0 cc. 



2. Blood, defibrinated beef 150.0 cc. 



3. KOH N/1 150.0 cc. 



Preparation : 



(1) Prepare nutrient agar and neutralize 

 to litmus. 



(2) To 150.0 cc. of defibrinated beef blood 



add an equal part of N/1 KOH. 



(3) To 7 parts melted (1) add 3 parts 

 (2), mix well and pour into plates. 



(4) Dry the plates for several days at 

 37°C. or for 4 minutes at 60°C. 



Sterilization: Sterilization of agar not 

 specified. The laked blood alkali mixture 

 maj^ be sterilized in the autoclave. 



Use: Enrichment medium for cholera 

 vibrio. Author reported that cholera 

 vibrio grew very luxuriantly on this 

 medium. B. coli was inhibited. 



Variants : 



(a) Sineff and Drosdowitsch gave the 

 following method of preparation: 



(1) Collect beef blood in a sterile 

 enamel container and defibrinate it 

 with a sterile egg beater. 



(2) Strain thru a fine tin sieve. 



(3) Mix equal parts of the filtrate from 

 (2) and a normal solution (5.61%) 

 of KOH. 



(4) Sterilize for one-half hour. 



(5) Mix 7 parts sterile agar and 3 

 parts (4) (sterile alkaline defibri- 

 nated blood). 



(6) Pour into sterile Petri dishes. 



(7) Dry at 37°C. for 24 hours. 



(b) Hofer and Hovorka prepared a sim- 

 ilar medium as follows: 



(1) To 3.0% melted nutrient neutral 

 agar add 4.0 cc. of defibrinated 

 beef blood, and 16.0 cc. of N/1 

 KOH. 



(2) Boil. 



(3) Distribute into 10.0 cc. lots. 



(4) Prepare a 0.1% solution of crystal 

 violet in distilled water. 



(5) To each 10.0 cc. of (3) add 0.5 cc. 

 of (4). 



(6) Pour into Petri dishes. 



(7) Place the plates in the incubator 

 for 24 hours with covers partly re- 

 moved and then at room tempera- 

 tures for 12 hours with covers on 

 before use. 



(c) Lentz prepared the medium as 

 follows: 



(1) Mix fresh defibrinated beef blood 

 with equal amount of N/1 alkali. 



(2) Heat 30 minutes. 



(3) Dry in Faust-Heim drying appa- 

 ratus. 



