CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



749 



methyl green, 4.0 cc. of chrysoidine 

 and 3.0 cc. of fuchsin. 



(3) Add 200.0 cc. of distilled water to 

 (2) and allow to stand several 

 hours. The color should be green- 

 ish blue. If not obtain the origi- 

 nal color by adding either blue, 

 green or red dye. 



(4) Sterilize at 100°C. 



(5) Add 7 to 10 drops of (4) to a tube 

 of sterile melted nutrient gelatin. 



References: Heller (1905 p. 118), Calandra 

 (1910 p. 570), Signorelli (1912 p. 472), 

 Besson (1920 p. 60). 



2310. Matzuschita's Glucose Gelatin 



Constituents : 



1. Bouillon 1000.0 cc. 



2. Gelatin 100.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



5. Glucose 20.0 g. 



Preparation: (1) Dissolve 2, 3, 4 and 5 in 



1000.0 cc. bouillon. 



Sterilization: Not specified. 



Use: Cultivation of mammalian and 

 chicken tubercle bacilli. The author re- 

 ported that the Mammalian and chicken 

 types gave same kind of growth. A 

 white precipitate was formed in the 

 liquid gelatin (at 30°C.). Other in- 

 vestigators used similar media for pur- 

 poses as indicated below. 



Variants : 



(a) Rivas solidified medium 923 by the 

 addition of gelatin. This medium 

 was used to cultivate anaerobes. 



(b) Frost added 1.0% glucose to nutrient 

 gelatin. 



(c) Worth cultivated members of the 

 colon-typhoid group on nutrient 

 gelatin of a reaction 0.4% or 1.0% 

 acid to which was added 2.0% glu- 

 cose. He also prepared a medium 

 adding calcium carbonate. 



References: Matzuschita (1899 p. 128), 

 Rivas (1902 p. 836), Frost (1903 p. 64), 

 Worth (1919 p. 604). 



2311. Ramond's Rubine Acid Lactose 



Gelatin 



Same as medium 1796 but solidified by the 

 addition, of gelatin instead of agar. 



2312. Wurtz's Litmus Lactose Gelatin 



Constituents : 



1. Nutrient gelatin 1000.0 cc. 



2. Lactose (2.0%) 20.0 g. 



3. Litmus 

 Preparation : 



(1) Add 20.0 g. lactose to sterile neutral 

 nutrient gelatin. 



(2) Tube. 



(3) Add sufficient tincture of litmus to 

 each tube to give a violet color. 



(4) Pour sterile (3) into plates. 

 Sterilization: Sterilize (3) at 100°C. 



Use: Differentiation of Bad. colt. The 

 author reported that Bad. coli fermented 

 lactose with the production of acid. 

 Typhoid bacilli colonies were colorless, 

 colon colonies red. 



Variants : 



(a) Heinemann gave the following 

 method of preparation: 



(1) Dissolve 1.0% lactose in ordinary 

 10 or 12.0% gelatin. 



(2) Distribute in 8.0 cc. quantities in 

 tubes. 



(3) Add 1.0% sterile litmus solution to 

 each tube before using (amount of 

 solution not specified). 



(b) Abbott prepared the medium as 

 follows: 



(1) Prepare nutrient gelatin so the 

 alkalinity is such that it requires 

 0.1 cc. of a 1:20 normal H2SO4 

 solution to neutralize 1.0 cc. of 

 medium. Indicator not specified. 



(2) Add 2.0 to 3.0% lactose. 



(3) Decant into test tubes. 



(4) Sterilize in the usual way (method 

 not given). 



(5) Add sufficient sterile litmus tincture 

 to each tube to give a decided but 

 not intense blue color. Add the 

 litmus under aseptic conditions. 



References: Wurtz (1897 p. 43), Heinemann 

 (1905 p. 127), Tanner (1919 p. 55), Abbott 

 (1921 p. 142). 



2313. Wurtz's Glycerol Gelatin 



Constituents : 



1. Nutrient gelatin 1000.0 cc. 



2. Glycerol (6.0%) 60.0 g. 



Preparation: (1) Add 6.0% sterile glycerol 



to sterile nutrient gelatin under aseptic 

 conditions. 



