634 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Sterilization: Method not given. 



Use: To study fermentation and hemolysis. 

 Author reported that typhoid colonies 

 did not produce hemolysis on lactose 

 plates. Colon colonies did produce 

 hemolysis on lactose plates. On raffi- 

 nose, typhoid colonies showed umbili- 

 cated colonies with lines radiating from 

 the center. Colon colonies did not show 

 these lines. On maltose typhi were 

 deeply pigmented, almost black; colon 

 colonies were white. On dextrin ty- 

 phoid colonies caused precipitation of the 

 medium and were black. Colon colonies 

 caused hemolysis and were white. 



Variants: The author added 1.0% of any 

 desired carbohydrate to nutrient agar. 



Reference: Bernstein (1909 p. 2). 



1983. Wordley's Oxalated Blood Agar 



Constituents : 



1. Nutrient agar 150.0 cc. 



2. Blood (human oxalated) 10.0 cc. 



Preparation: (1) Prepare nutrient agar. 



(2) Melt agar and pour into sterile Petri 

 dishes in 15.0 cc. lots. 



(3) Add 1.0 cc. of human oxalated blood 

 to each Petri dish. 



Sterilization: Not specified. 



Use: To study hemolysis by streptococci. 



Reference: Wordley's (1921 p. 66). 



1984. Wilson and Darllng^'s Laked Blood 

 Agar 



Constituents : 



1. Water 10.0 cc. 



2. Nutrient agar 



3. Blood, sheep 1000.0 cc. 



4. Sodium citrate 10.0 g. 



Preparation : 



(1) Dissolve 10.0 g. of sodium citrate in 

 10.0 g. of water. Place this in a 

 bottle with volume graduation marks. 



(2) Take (1) to the slaughter house and 

 .when a sheep is slaughtered, after 



the first gush of blood, collect the 

 blood in the bottle until a liter of 

 fluid has been obtained. 



(3) Shake the bottle vigorously during 

 the process. 



(4) Add 1.25 cc. of formalin to the liter 

 of citrated blood and mix thoroly. 



(5) Transfer the blood with aseptic pre- 



cautions to a ground glass stoppered 

 bottle and add 30.0 cc. of methylated 

 ether. 



(6) Stopper firmly and shake well. 



(7) Incubate over night at 37°C. 



(8) When required for use, 3.0 cc. of (7) 

 are pipetted to 100.0 cc. of melted 

 and cooled to 50°C. nutrient agar. 



(9) Keep the blood agar at 50°C. for one 

 hour before pouring into plates, if to 

 be used at once. Or pour into plates 

 and incubate at 37°C. over night. 

 This drives off the ether, and if the 

 plates are incubated over night, tests 

 sterility. 



Sterilization: Not specified. 



Use: Isolation and preservation of menin- 

 gococci. Also used for the cultivation of 

 B. influenzae and anaerobes. 



Reference: Wilson and Darling (1918 

 p. 105). 



1985. Wilson and Darling's Lactose Blood 

 Agar 



Constituents : 



1. Distilled water 10.0 cc. 



2. Nutrient agar 



3. Blood, sheep 1000.0 cc. 



4. Sodium citrate 10.0 g. 



5. Lactose 0.5 to 1.0% 



6. Crystal violet (0.1%, 

 soln.) 



7. Litmus solution (Kubel 

 and Tiemann) 



Preparation: 



(1) Dissolve 10.0 g. of sodium citrate in 

 10.0 cc. of water. Place this in a 

 bottle with volume graduation 

 marks. 



(2) Take (1) to the slaughter house and 

 when a sheep is slaughtered, after 

 the first gush of blood, collect the 

 blood in the bottle until a liter of 

 fluid has been obtained. 



(3) Shake the bottle vigorously during 

 the process. 



(4) Add 1.25 cc. of formalin to the liter 

 of citrated blood and mix thoroly. 



(5) Transfer the blood with aseptic pre- 

 cautions to a ground glass stoppered 

 bottle and add 30.0 cc. of methylated 

 ether. 



(6) Stopper firmly and shake well. 



