CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



645 



Use : Cultivation of diphtheria bacilli. 

 Reference: Czaplewski (1920 p. 828). 



2013. Klein's Alkaline Serum Agar 



Constituents : 



1. Nutrient agar 400.0 cc. 



2. Serum 90.0 cc. 



3. NaOH (15.0%) 10.0 cc. 



Preparation: 



(1) Mix 90 parts serum with 10 parts 

 15.0% NaOH. 



(2) Incubate 2 days at 37'=C. 



(3) Neutralize by the addition of HCl, 



(4) Mix one part (3) with four parts 

 nutrient agar. 



Sterilization: Heat (4) for 30 minutes 



at 105°C. 

 Use: Cultivation of diphtheria bacillus. 

 Reference: Klein (1920 p. 297). 



2014. Frost, Charlton and Little's Milk 

 Serum Agar 



Constituents: 



1. Distilled water 100.0 cc. 



2. Nutrient agar 200.0 cc. 



3. Milk 200.0 cc. 



4. Serum 100.0 cc. 



Preparation: 



(1) Prepare ordinary nutrient agar (1.0 to 

 1.5%) with a reaction of +1.0. 



(2) Tube in 1.0 to 3.0 cc. lots. 



(3) Tube well separated milk in 1.0 to 

 3.0 cc. quantities. 



(4) Mix one part serum (sheep, beef, 

 horse or rabbit) with three parts dis- 

 tilled water. 



(5) Tube (4) in 1.0 to 3.0 cc. quantities. 



(6) When ready for use melt the sterile 

 agar tubes and cool to 45° to 50°C. in 

 a water bath. 



(7) Heat sterile tubes of (3) and (4) in 

 the water bath and add one tube of 

 (3) and one tube of (4) to a tube of 

 agar (equal parts agar, milk and 

 serum). 



(8) Mix thoroughly. 



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



the autoclave. 

 Use: Isolation of B. diphtheriae. The 



authors used this medium in preparing 



"little plate" cultures on slides. 

 Reference: Frost, Charlton and Little 



(1921 p. 30). 



2015. Salomon's Basal Ascitic Fluid Agar 



Constituents : 



1. Nutrient agar (3.0%) 100.0 cc. 



2. Ascitic fluid 50.0 cc. 



3. Litmus tincture 

 Preparation : 



(1) Prepare 3.0%, nutrient agar. 



(2) Distribute (1) into 10.0 cc. lots. 



(3) Prepare a 10.0% solution of one of 

 the added nutrients in litmus tinc- 

 ture. 



(4) Add 1.5 cc. of one of (3) to 10.0 cc. of 

 melted (2) cooled to 58°C. 



(5) Add 5.0 cc. of ascitic fluid, heated to 

 58°C. to each tube of (4). 



Sterilization: Not specified. 



Use: To study fermentation of strepto- 

 cocci. Author gave the following re- 

 actions: 



A. Strept. pyogenes group. 



I. Strept. pyogenes: Acid formation 

 from soluble amylum. Mannitol 

 and raffinose unchanged. 

 II. Strains from blood. Acid forma- 

 tion from glycerol and mannitol. 



B. Strept. mucosus. 



I. Acid from glycerol, arabinose and 

 mannitol. Raffinose and soluble 

 amylum unchanged. 

 II. Do not attack any of the sugars in 

 24 hours and rarely after 48 hours. 



C. Pneumococci formed no acid on carbo- 

 hydrate litmus ascitic agar. 



Similar media were used by other in- 

 vestigators to study fermentation of other 

 cocci. 

 Added nutrients: The author used one of 

 the following materials: 



glycerol levulose 



erythritol dulcitol 



arabinose lactose 



adonitol raffinose 



isodulcitol inulin 



mannose maltose 



glucose sucrose 



mannitol dextrin 



galactose amylum (soluble) 



Variants : 



(a) Symmers and Wilson cultivated 

 meningococci on a similar medium 

 prepared as follows: 

 (1) Prepare nutrient agar containing 

 3.0% Chapoteaut's peptone. 



