CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



545 



(i) Krumwiede, Kohn, Kuttner and 

 Schumm. 



(1) Dissolve 25.0 g. agar, 10.0 g. pep- 

 tone and 5.0 g. meat extract in 

 1000.0 cc. water by heating over a 

 gas stove. 



(2) Adjust reaction neutral to litmus 

 using 10.0% Na2C03 solution. 



(3) Add 10.0 cc. of 10.0% NajCOa. 



(4) Cool to 45°C. and add one egg. 



(5) Autoclave for 30 minutes at 15 

 pounds pressure. 



(6) Filter and add 10.0 g. lactose 

 (boil to dissolve). 



(7) Add 10.0 cc. of 10.0% Na2C03 

 solution, giving a reaction of 

 -0.3 to phenolphthalein (hot 

 titration). 



(8) Distribute in bottles in 100.0 cc. 

 lots. 



(9) Autoclave for 10 minutes at 10 

 pounds pressure. 



(10) To melted (9) just before use add 

 0.5 cc. of a saturated alcoholic 

 solution of fuchsin and 1.0 cc. of a 

 10.0% solution of amorphous so- 

 dium bisulphite (per 100.0 cc.?). 

 (j) Tanner (Hygienic Laboratory). 



(1) Add 3.7 cc. of a 10.0% anhydrous 

 Na2C03 solution to 1000.0 cc. of a 

 3.0% extract agar with a reaction 

 of +0.5 to phenolphthalein. 



(2) Flask in 200.0 cc. quantities, 



(3) Dissolve 2.0 g. of chemically pure 

 lactose in 25.0 to 30.0 cc. of dis- 

 tilled water with the aid of gentle 

 heat. 



(4) Dissolve 0.5 g. anhydrous sodium 

 sulphite in 10 to 15.0 cc. distilled 

 water. 



(5) Add 1.0 cc. of a saturated solution 

 of basic fuchsin in 95.0% alcohol 

 to (4). 



(6) Add (5) to (3). 



(7) Add (6) to 200.0 cc. of melted agar. 



(8) Pour into plates. 



(9) When hard, dry for 15 minutes in 

 the incubator. 



(k) Kendall (Tanner). 



(1) Prepare plain sugar free extract 

 agar using 15.0 g. agar per liter. 



(2) Adjust to slightly alkaline to 

 litmus. 



(3) Distribute in 100.0 cc. quantities 

 in flasks. 



(4) Prepare a 10.0% basic fuchsin 

 solution in 96% alcohol. 



(5) Prepare a 10.0% solution of chem- 

 ically pure anhydrous sodium 

 sulphite (1.0 g. in 10.0 cc. water). 



(6) Add 1.0 cc. of (4) to 10.0 cc. of (5), 

 freshly prepared. 



(7) Heat in the Arnold sterilizer for 20 

 minutes. 



(8) Add 1.0 g. lactose to 100.0 cc. of 

 (3) and place in the autoclave until 

 melted and the lactose is dissolved. 



(9) Add about 1.0 cc. of (7) to each 

 100.0 cc. flask. 



(10) This should give a pink color. 



(11) Pour into sterile Petri dishes. 

 (1) Bezangon. 



(1) Add 120.0 cc. of a 10.0% soda 

 solution to a liter of neutral 3.0% 

 extract agar. 



(2) Add 120.0 cc. of a 10.0% lactose 

 solution to (1). 



(3) Sterilize at 100° for one hour. 



(4) Add 10.0 g. of crystalline fuchsin 

 to 100.0 cc. of 96% alcohol and 

 allow to stand for 2 hours. 



(5) Decant (4) and add 2.5 cc. of the 

 supernatant fluid to (3). 



(6) Sterilize a 10.0% solution of 

 Na2SO3at80°for one hour. 



(7) Add25.0cc.of (6)to(5). 



(8) Pour in plates, 

 (m) Giltner. 



(1) Prepare an ordinary agar from 

 5.0 g. Liebig's meat extract, 5.0 g. 

 NaCl, 10.0 g. peptone, 30.0 g. agar 

 and 1000.0 cc. water. 



(2) Adjust the reaction to or +0.2%. 



(3) Store in 100.0 cc. quantities in 

 Erlenmeyer flasks. 



(4) Melt a flask of (3), and add 6 drops 

 of a saturated alcoholic solution 

 of basic fuchsin (4.0 g. fuchsin to 

 100.0 cc. of 95% alcohol). 



(5) Mix well. 



(6) Add about 18 to 20 drops of a 

 freshly prepared 10.0% solution of 

 NazSOa. 



(7) Add 5.0 cc. of a freshly hot 20.0% 

 aqueous solution of lactose to the 

 hot agar. 



(8) Pour into plates. 



