778 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(3) Coagulate in the autoclave for 75 

 minutes at 75 to 80° or in a hot air 

 oven at the same temperature and 

 time. 

 Sterilization: Method of sterilization of 

 glucose, litmus or H2SO4 solutions not 

 given. 

 Use: Diagnosis of diphtheria. Author re- 

 ported that diphtheria colonies were red 

 Variants: Harvey prepared the litmus 

 tincture as follows: 



(1) Grind up litmus in a mortar. 



(2) Add 5 volumes 90 per cent alcohol. 



(3) Boil on a water bath. 



(4) Decant the supernatant fluid. 



(5) Add 6 parts distilled water to the 

 residue. 



(6) Boil. 



(7) Allow to cool. 



(8) Divide into two portions. 



(9) Render one portion slightly red with 

 dilute sulphuric acid. 



(10) Add to this reddened portion the 

 other untreated portion little by 

 little until the mixture becomes 

 blue again. 



(11) Filter thru paper when cool. 



(12) Distribute into test tubes. 



(13) Sterilize at 110°C. 



(14) Keep for use. 

 References: Costa, Troisier and Dauvergne 



(1917 p. 80), Harvey (1921-22 p. 81). 



2406. Rankin's Potassium-Sulphocyanide 



Glucose Serum 

 Constituents : 

 1- Water 100.0 cc. 



2. Sheep serum 300.0 cc. 



3. Glucose 40e 



4. 1.0% aqueous neutral red 

 solution 4 cc 



5. 50.0% potassium sulphocy- 



anide solution 8.0 cc. 



Preparation : 



(1) Obtain fresh serum from sheep blood. 

 Serum should contain no red blood 

 cells. 



(2) Add 100.0 cc. of water to 300.0 cc. of 

 (1). (Author used bouillon or equal 

 parts of water and bouillon and water, 

 but water gave equally good results.) 



(3) Add 1.0% glucose, 4.0 cc. of a 1.0% 

 neutral red solution and 8.0 cc. of a 

 50.0% solution of potassium-sulpho- 

 cyanide solution. 



Sterilization: Tube and sterilize m the 

 steamer for an hour on three successive 

 days at a temperature of from 180 to 

 190°F. 



Use: Show presence of B. diphtheriae in 

 throats. Author reported that B. diph- 

 theriae always gave a characteristic red 

 colony. He recommended that the me- 

 dium be used in the discharging of 

 patients. 

 Reference: Rankin (1912 p. 63). 



2407. Dubois' Glycerol Glucose Serum 

 Constituents: 



1- Serum 100.0 cc. 



2. Glucose 7.0 g 



3. Glycerol 2.O g^ 



Preparation: 



(1) Tube non-sterilized serum containing 

 7 parts glucose and 2 parts glycerol 

 per 100 in 2.0 cc. quantities. 



(2) Solidification not specified. 

 Sterilization : Not specified. 



Use : Cultivation of Koch's bacilli. 

 Reference: Dubois (de Nancy) (1896 p 

 204). 



2408. Marx's Glycerol Ragit Serum 



Constituents : 

 1- Water 1000.0 cc. 



2. "Ragit" serum 130. g. 



3. Glycerol 50.0 cc. 



Preparation : 



(1) Pour 13.3 g. "Ragit" serum in a large 

 mortar. 



(2) Add 100.0 cc. water in small lots. 



(3) When the serum has become en- 

 tangled by mixing with a pestil, add a 

 few drops of water and rub the water 

 in the powder. 



(4) As soon as the mixture has become 

 thick add more water. Always rub 

 in a circle as if mixing milk and flour 

 in cooking. Continue this until all 

 the 100.0 g. has been added. 



(5) Add 5.0 cc. of glycerol to the mixture 

 of 13.3 g. "Ragit" serum and water. 



(6) Distribute into tubes or plates. 



(7) Solidify by placing the tubes or 

 plates in flowing steam. 



Sterilization: Place the medium in the 

 autoclave for 15 minutes (pressure not 

 specified). 



Use: Cultivation of diphtheria bacilli. 

 Author reported that growth was nearly 



