642 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(3) Add one-half or one-third the vol- 

 ume of sterile serum to each tube. 



(4) Mix thoroly by rolling the tube be- 

 tween the palms of the hands. 



(5) Slant or pour into a Petri dish. 



(d) Dopter and Sacquepee gave the fol- 

 lowing method of preparation: 



(1) Add 0.5 to 1.0 cc. of serum to an 

 agar slant. 



(2) Incline so that the entire surface is 

 covered with the serum. 



(3) Allow to stand for 12 to 24 hours. 



(e) Klimmer mixed one part gelatin agar 

 with one-half part serum at a tem- 

 perature of 45°C. 



References: Muller (1906 p. 520), Muhlens 

 and Hartmann (1906 p. 86), Carrel (1912 

 p. 393), Besson (1920 p. 53), Dopter and 

 Sacquepee (1921 p. 138), Klimmer (1923 

 p. 200), Kligler and Robertson (1922 

 p. 315). 



2004. Joos' Alkaline Serum Agar (Klimmer) 



Constituents : 



1. Distilled water 150.0 cc. 



2. Serum 300.0 cc. 



3. NaOH (normal) 50.0 cc. 



4. Bouillon 500.0 cc. 



5. Agar 20.0 g. 



Preparation : 



(1) Heat 1, 2 and 3 for 2 to 3 hours in a 

 water bath at 60 to 70°C. 



(2) Steam for 30 to 45 minutes. 



(3) Prepare bouillon using 2.0% peptone 

 and 1.5% NaCl. 



(4) Adjust the reaction of (3) to alkaline. 



(5) Dissolve 20.0 g. agar in (4). 



(6) Mix (5) and (2). 



(7) Filter. 



Sterilization: Method not given. 



Use : General culture medium. 



Variants: Klein (Klimmer) gave the fol- 

 lowing method of preparation for a me- 

 dium for the cultivation of diphtheria 

 bacilli. 



(1) Mix 9 parts serum with 1 part normal 

 NaOH and incubate at 37°C. for 

 2 days. 



(2) Neutralize by the addition of HCl. 



(3) Mix 1 part (2) with 4 parts nutrient 

 agar. 



(4) Sterilize at 105°C. for 1 hour. 

 Reference: Klimmer (1923 p. 222). 



2005. Muhlen's and Hoffman's Glucose 

 Serum Agar (Stitt) 

 Constituents : 



1. Nutrient agar (3.0%) 100.0 cc. 



2. Glucose (0.5%) 0.5 g. 



3. Serum (Horse) 100.0 cc. 



Preparation : 



(1) Fill sterile test tubes one-third full 

 with horse serum. 



(2) Add an equal amount of a 3.0% agar 

 containing 0.5% glucose which has 

 been melted and cooled to 50°C. to 

 sterile (1). 



(3) Keep at 55° for two hours. 

 Sterilization: Sterilize (1) on 3 successive 



days at 55°C. 



Use: Cultivation of treponemata. The 

 medium was inoculated while still liquid 

 and incubated under anaerobic condi- 

 tions. 



Reference: Stitt (1923 p. 54). 



2006. Cantanis' Glycerol Serum Agar 

 (Besson) 

 Constituents : 



1. Nutrient agar. 



2. Glycerol. 



3. Serum. 

 Preparation : 



(1) Mix equal parts of sterile serum and 

 sterile glycerol. 



(2) Allow to stand several hours. 



(3) Add 0.5 to 0.75 of (2) to tubes of 

 liquified sterile agar. 



Sterilization: Method not given. 

 Use: General culture medium. 

 Reference: Besson (1913 p. 54). 



2007. Kodama's Fuchsin Sulphite Serum 

 Agar 



Constituents : 



1. Water 40.0 cc. 



2. Nutrient agar (3.0%) 100 cc. 



3. Fuchsin (saturated alcoholic 

 solution) 0.5 cc. 



4. NasSOa (10.0% solution) .... 2.5 cc. 



5. Starch, potato 3.0 g. 



6. Serum, beef 10.0 cc. 



Preparation : 



(1) Prepare nutrient 3.0% agar, neutral 

 to litmus. 



(2) Add 1.0 cc. 10.0% soda solution, 

 0.5 cc. of a saturated alcoholic fuchsin 



