628 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



4. Agar 5.0 g. 



5. Kidney (dog) 



6. NaoCOsdO^o) 5.0 cc. 



Preparation : 



(1) Prepare bouillon. 



(2) Mix equal parts bouillon and water. 



(3) Dissolve 2.0% peptone and 0.5% 

 agar in (2). 



(4) Add 5.0 cc. of a 10.0%, NaoCOs solu- 

 tion to give a slightly alkaline 

 reaction. 



(5) Distribute in tubes. 



(6) Cool sterile (5) to 45°C. and to each 

 tube add a small piece of sterile dog 

 kidney under aseptic conditions. 



Sterilization: Sterilize (5) in the autoclave. 



Use: Cultivation of flagellates. Author re- 

 ported that the flagellates grown in this 

 medium were larger than those grown in 

 blood culture. 



Reference: G6zony (1920 p. 566). 



1965. Duval's Trypsinized Tissue Agar 

 Constituents : 



1. Nutrient agar. 



2. Tissue. 



3. Trypsin (1.0% solution). 

 Preparation : 



(1) Pour nutrient agar in sterile Petri 

 dishes and allow to solidify. 



(2) Cut an excised leprous nodule into 

 thin slices, two to four millimeters in 

 breadth and 0.5 to 1.0 mm. in thick- 

 ness and distribute over the surface 

 of the plate. 



(3) Bathe the medium with a 1.0% tryp- 

 sin solution, taking care not to sub- 

 merge the pieces of leprous tissue. 

 Add sufficient fluid to moisten thoroly 

 the surface of the medium. 



(4) Incubate in a moist chamber at 37°C. 

 for a week to 10 days. Remove the 

 plates from time to time and add more 

 trypsin as evaporation necessitates. 



Sterilization: Not specified. 



Use: Cultivation of B. leprae. The author 

 reported that the colonies were grayish 

 white, but after several days they as- 

 sumed a distinct orange yellow tint. 



Reference: Duval (1911 p. 369). 



1966. Thoinot and Masselin's Gelatin Agar 

 Constituents : 



1. Bouillon 1000.0 cc. 



2. Gelatin 100.0 g. 



3. Agar (0.5%) 5.0 g. 



Preparation : 



(1) Dissolve 100.0 g. of gelatin in a liter 

 of ordinary bouillon. 



(2) Make alkaline. 



(3) Add 0.5% agar and dissolve by 

 boiling. 



(4) Cool to 55°C. and add the white of 

 an egg. Mix well. 



(5) Boil 20 minutes at 115°C. 



(6) Distribute as desired. 

 Sterilization: Sterilize for 20 minutes at 



113 to 114°C. 

 Use: General culture medium. 

 Variants: Besson (Tanner) prepared the 



medium as indicated: 



(1) Dissolve 80.0 g. of gelatin in bouillon. 



(2) Neutralize. 



(3) Dissolve 5.0 g. of agar in (2). 50.0 g. 

 of gelatin and 8.0 g. agar may be used 

 instead of 80.0 g. of gelatin and 5.0 g. 

 of agar. 



(4) Sterilize. Do not heat over 115°C. 

 References: Thoinot and Masselin (1902 



p. 36), Tanner (1919 p. 56). 



1967. Fremlin's Phosphate Gelatin Agar 



Constituents: 



1. Water 1000.0 cc. 



2. Agar 15.0 g. 



3. Potassium phosphate 10.0 g. 



4. Bouillon gelatin 50.0 g. 



Preparation : 



(1) Exact composition or method of 

 preparation of bouillon gelatin not 

 given. 



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

 Sterilization: Not specified. 



Use: To study nitrification by nitroso bac- 

 teria. Author reported that sub-cultures 

 from this medium to other plates did not 

 show nitrification, proving that this is not 

 a suitable medium for continued activity. 



Reference: Fremlin (1914 p. 154). 



1968. Vierling's Fat Agar 



Constituents : 



1. Nutrient agar 100.0 cc. 



2. Lanolin 10.0 g. 



Preparation : 



(1) Prepare nutrient agar. 



(2) Prepare an emulsion of 1.0 g. lanolin 

 in 10.0 cc. nutrient agar. 



Sterilization: Not specified. 

 Use: To study lipase production by Myco- 

 bacteria. Author reported that good 



