536 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



every 100.0 cc. agar. Cool agar to 

 60°C. or precipitate will be formed. 



(6) Transfer to small test tubes, to a 

 depth of 1.5 cc. 



(7) Stock solution of 1.0% china blue in 

 distilled water is prepared. 



(8) Add 0.4 cc. N sodium hydrate to 

 10.0 cc. of (7). Heat on water bath 

 for 10 minutes at 100°C. (Color 

 changes from blue to brown.) 



(9) 1.2 cc. of (8) is added to 100.0 cc. of 

 (3) reaction -0.2 to -0.4. 



(10) Add 1.0% lactose and 0.1% glucose 

 to (9) and heat in water bath at 

 100°C. for 10 minutes. 



(11) Cool to 60°C. 



(12) Add to (6) making a second layer in 

 the tubes. 



(13) Incubate over night — make stab 

 inoculation. 



Sterilization: Not specified. 



Use: Differentiation of B. paratyphosus 



A, B. paratyphotus B, B. enteritidis and 



B. coll. 



Variants: The author prepared a similar 

 medium but used 1.0% inositol and 0.1% 

 arabinose instead of 1.0% glucose. 



Reference: Kan-Ichiro Morishima (1918 

 p. 19). 



1745. Aronson's Fuchsin Sulphite Agar 



(Harvey) 



Constituents : 



1. Infusion agar 1000.0 cc. 



2. NasCOa (10.0% anhydrous 



soln.) 60.0 cc. 



3. Sucrose (20.0% soln.) 50.0 cc. 



4. De.xtrin (20.0% soln.) 50.0 cc. 



5. Fuchsin (sat. alcoholic 



soln.) 2.5 cc. 



6. Na.SOa (10.0% soln.) 25.0 cc. 



Preparation : 



(1) Prepare a 10.0% anhydrous NaaCOs 

 solution. 



(2) Prepare a 20.0% sucrose solution. 



(3) Prepare a 20.0% dextrin solution. 



(4) Prepare a saturated alcoholic fuch- 

 sin solution. 



(5) Prepare a 10.0% NaaSOs solution 

 (must be freshly prepared). 



(6) Mix 60.0 cc. sterile (1), heated to 

 45°C. and 1000.0 cc. melted sterile 

 infusion agar (see variant (v) 

 1661 for preparation), cooled to 45 °C. 



(7) Steam 15 minutes. The mixture 

 becomes dark brown and very 

 cloudy. 



(8) Add 50.0 cc. of sterile (2), 50.0 cc. 

 of sterile (3), 2.5 cc. sterile (4) and 

 25.0 cc. of sterile (5) to hot (7). 



(9) Place the flasks in a sloping position 

 to allow the precipitate formed to 

 settle. 



(10) Pour plates carefully, avoiding as 

 far as possible the transference of 

 settled precipitate. 



(11) Place the plates to dry in the in- 

 cubator with their agar surface 

 downwards. 



(12) Preserve in the dark for three days 

 before use. 



Sterilization: Sterilize (1), (2), (3) and (5) 



at 100°C. 

 Use : Isolation of V. cholerae. 

 Reference: Harvey (1921-22 p. 93). 



1746. v. Szaboky's Glycerol Lung Agar 



Constituents : 



1. Water 2000.0 cc. 



2. Lung 1000.0 g. 



3. Agar 10.0 g. 



4. Peptone (Witte) 20.0 g. 



5. Glycerol 100.0 g. 



6. Glucose 10.0 g. 



Preparation : 



(1) Boil 1 kilogram of lung with 2 kilo- 

 grams of water. (Time not specified.) 



(2) Filter and dissolve 3, 4, 5 and 6 in the 

 filtrate. 



(3) Neutralize to litmus. 



(4) Heat again. 



(5) Distribute in 20.0 cc. lots into suitable 

 wide sterile test tubes. 



Sterilization: Sterilize in the usual manner 

 by short heating in streaming steam (time 

 or number of days not specified). 



Use: Cultivation of tubercle bacilli. The 

 author reported that colonies developed 

 in one day when reaction was 0.5% alka- 

 line to 0.5% acid. 



Variants : Tubercular lung may be used as 

 well as a normal lung. This tubercular 

 material must be sterilized on 5 days for 

 2 hours each. 



Reference: v. Szaboky (1907 p. 652), Kolle 

 and Wassermann (1912 p. 413). 



