CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



727 



6. Gelatin 100.0 to 150.0 g. 



7. Blood 



8. KOH (normal) 

 Preparation : 



(1) Dissolve 2, 3, 4, 5 and 6 in 1. 



(2) Filter. 



(3) Mix blood with equal amounts of 

 N/1 KOH. 



(4) Steam (3) for 30 minutes to sterilize. 



(5) Mix 30.0 parts sterile (4) with 70.0 

 parts sterile (2). 



(6) Pour into sterile Petri dishes. 

 Sterilization : Method of sterilization of (2) 



not given. See step (4) for sterilization 



of alkaline blood mixture. 

 Use: Enrichment medium for cholera 



vibrio. 

 Reference: Pergola (1911 p. 85). 



2273. Matzuschita's Peptone Milk 

 Gelatin 



Constituents : 



1. Milk 1000.0 cc. 



2. Gelatin 100.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



Preparation: (1) Dissolve 2, 3 and 4 in 1. 

 Sterilization: Method not specified. 



Use: Cultivation of mammalian and 

 chicken tubercle bacilli. The author 

 reported that chicken type grew slightly 

 more luxuriantly than did the mam- 

 malian types. Utz cultivated organisms 

 found in milk on a similar medium. 



Variants: Utz prepared a similar medium 

 as follows: 



(1) Dissolve 100.0 g. gelatin and 10.0 g. 

 peptone in 1000.0 cc. milk. 



(2) Neutralize. 



(3) Mix with egg white. 



(4) Boil one hour. 



(5) Filter. 



(6) Sterilization not specified. 

 References: Matzuschita (1899 p. 128), 



Utz (1903-04 p. 610). 



2274. Raskin's Whey Peptone Gelatin 

 Constituents : 



1. Whey 1000.0 cc. 



2. Gelatin (6.0 to 



10.0%) 60.0 to 100.0 g. 



3. Peptone (1.0%) 10.0 g. 



Preparation : 



(1) Heat 1000.0 cc. of milk in a porce- 

 lain dish at 60 to 70 °C. 



(2) Add 60.0 to 100.0 g. (6.0 to 10.0% 

 gelatin and heat until the gelatin 

 has become melted. 



(3) Boil until the casein is completely 

 coagulated. 



(4) Pass thru four folds of linen cloth 

 separating the casein and liquid. 



(5) The reaction of the fluid is slightly 

 acid. 



(6) Pour into a tall glass cylinder and 

 place in a warm thermostat. 



(7) After about 20 or 30 minutes two 

 layers are formed, the lower layer 

 being transparent and fat free, the 

 upper layer yellowish white, con- 

 taining the fat. 



(8) Allow to cool and remove the fat 

 by means of a spoon. 



(9) Heat the fat free portion to boiling 

 and add 1.0% peptone. 



(10) Add soda to neutralize the reaction. 



(11) Filter until clear thru a paper in a 

 hot water funnel. 



Sterilization: Not specified. 



Use: CultivMion of pathogenic organisms. 



Variants : 



(a) Raskin added 0.5% NaCl. 



(b) Klimmer prepared the medium as 

 follows: 



(1) Coagulate the casein from milk 

 by heating at 40 °C. in the presence 

 of rennet. 



(2) Separate the whey from the casein. 



(3) Dissolve 1.0% peptone, 0.5% NaCl 

 and gelatin to solidify in the whey. 



(4) Sterilization not specified. 



(c) Cunningham gave the following 

 method of preparation: 



(1) Warm 2 liters of clean fresh skim 

 milk in a large pot at 37°C. 



(2) Add sufficient rennet to curdle. 



(3) Allow to settle for 10 minutes. 



(4) Bteak the curd into large pieces 

 by means of a stirring rod. 



(5) Heat to 80°C. to contract the clot 

 and to express the whey. 



(6) Strain thru a cheese cloth. 



(7) Add 1.0% peptone and 0.5% NaCl. 



(8) Steam for 30 minutes. 



(9) Neutralize to turmeric paper. 



(10) Steam for one hour. 



(11) Dissolve 10.0% gelatin in (10). 



(12) Filter thru paper until clear. 



(13) Add 1.0% Andrades indicator. 



(14) Sterilize intermittently in steam. 



