ASCITIC FLUID AGAR 213 



6. Sterilise in the steamer at 100 C. for twenty minutes on 

 each of three consecutive days. 



Egg Albumin Broth (Lipschuetz). 



1. Weigh out 



Egg albumin (extra fine powder, Merck) . . 4 grammes 



and place in a 2 -litre flask with a number of sterile glass beads. 



2. Measure out distilled water 200 c.c. into a half-litre flask 

 and warm to 37 C. in the incubator. 



3. Add the water to the flask containing the albumin and beads 

 and dissolve by shaking. 



4. Add -^ NaOH, 40 c.c. Allow the mixture to stand for 

 thirty minutes with frequent shaking. 



5. Filter through Swedish filter paper. 



6. Sterilise by boiling two or three times at intervals of two 

 hours. 



7. Add ordinary nutrient bouillon 600 c.c. 



8. Fill into small Erlenmeyer flasks in quantities of 50 c.c. 



9. Incubate for forty-eight hours at 37 C. discard any con- 

 taminated flasks and store the remainder for future use. 



Egg Albumin Agar. 



1. Prepare egg albumin solution as above 1-6. 



2. Liquefy and measure out ordinary nutrient agar 600 c.c. and 

 add to the egg albumin solution (in place of the nutrient broth). 



3. Complete as above 8-9. 



Diplococcus Meningitidis Intracellularis. 



Ascitic Fluid Agar (Wassermann) Synonym N=as=gar (Mervyn 

 Gordon) . 



1. Liquefy and measure out into a sterile flask: 



Nutrient agar 600 c.c. 



2. Measure out into a half litre flask 



Distilled water 210 c.c. 



and add to it 



Ascitic fluid 90 c.c. 



Nutrose 6 grammes 



3. Heat over a bunsen flame, shaking constantly until the fluid 

 boils, and the nutrose is dissolved, 



4. Add the nutrose ascitic solution to the fluid agar. 



5. Heat in the steamer for thirty minutes, then filter. 



6. Tube and sterilise as for nutrient agar. 



NOTE. The finished medium in this case measures 900 c.c. only 

 since inconvenient fractions would be introduced in making up to 

 one litre exactly. 



