CULTURE MEDIA FOR CULTIVATION OF MICRO ORGANISMS 



133 



sterilization: Not specified. 



Use: To determine production of urea and 

 study of nitrogen metabolism. Sears 

 detected urea by urease method of Van 

 Slyke. Ammonia determined by Folin's 

 aeration method, amino acid by Van 

 Slyke's method, total nitrogen by Kjel- 

 dahl-Gunning-Arnold method. 



Reference: Sears (1916 p. 132). 



473. Kappen's Cyanamide Asparagin 

 Solution 



Constituents: 



1. Water 1000.0 cc. 



2. Calcium cyanamide (crude). 15.0 g. 



3. K2HPO4 (b.5) 5.0 g. 



4. Asparagin (0.1%) 1.0 g. 



5. Glucose (0.1%) 10.0 g. 



Preparation : 



(1) Grind 15.0 g. of crude calcium cyana- 

 mide to a powder and dry sterilize for 

 3 hours at 160°C. 



(2) Dissolve (1) in sterile water. 



(3) Filter thru a sterile folded filter into 

 a sterile flask. 



(4) Mix with a sterile solution containing 

 0.5% K.HPO4, 0.1% asparagin and 

 0.1% dextrose. The final volume 

 should be 1000.0 cc. 



(5) All the flasks, filters, funnel, etc., 

 should be sterile and the entire 

 process of preparation carried out 

 under aseptic conditions. 



Sterilization: Method of sterilization of 

 solutions and apparatus not specified. 



Use: To study cyanamide decomposition 

 by bacteria. 



Variants: The author suggested the follow- 

 ing solution: 



1. Water 1000.0 cc. 



2. Asparagin (0.1%) 1.0 g. 



3. K.2HPO4 (0.5%) 5.0 g. 



4. Glucose (1.0%) 10.0 g. 



5. Cyanide (0.3%) 3.0 g. 



Reference: Kappen (1909 pp. 385, 391). 



474. Davis and Ferry's Basal Cystine 

 Tryptophane Solution 



Constituents : 



1. Water 1000.0 cc. 



2. Tryptophane 0.3 g. 



3. Cystine 0.4 g. 



Preparation : 



(1) Add 2 and 3 to 1. 



(2) Add one of the combinations given 

 under added nutrients to (1) and heat 

 in flowing steam for 15 minutes. 



(3) Add enough N/10 NaOH to give final 

 reaction pH = 8.0 to 8 2. 



(4) Steam again for 15 minutes and check 

 reaction colorimetrically (using 

 phenolsulphonphthalein and stand- 

 ardized HaBOs-KCl-NaOH solutions. 



(5) Distribute as desired. 

 Sterilization: Sterilize for 20 minutes at 



115°C. 



Use: Cultivation of Bad. diphtheriae for 

 production of to.xin. Authors report 

 better toxin production if 10.0% bouillon 

 be added. All amino acids prepared 

 specially, in pure form. Cultures must 

 become accustomed to medium by adding 

 small portions of it to bouillon. 



Added nutrients and variants: The authors 

 added the following compounds to the 

 basic solution: 



(a) tyrosine 1.0 g 



leucine 3-0 g 



glutaminic acid hydrochloride. 1.6 g 



glycocoll 0.4 g 



creatin 0.1 g 



creatinin 0.1 g 



sodium asparaginate 1.4 g 



NaCl 40g 



K2HPO4 3.0g 



(b) Used 4.0 g. tryptophane in the basal 

 solution and added the following: 

 tyrosine 2.5 



glutaminic acid hydrochloride. 1.9 g 



glycocoll 0.85 g 



creatin o.I g 



creatinin o.I g 



sodium asparaginate 1.2 g, 



histidine dichloride 0.3 g, 



NaCl 40 o- 



K2HPO4 3^0 g 



MgS04 0.5 g 



KNO3 0.2 g 



(c) Used 0.6 g. tryptophane in the basal 

 solution and added: 



tyrosine 0.8 g. 



leucine 3.0 g. 



glutaminic acid hydrochloride 2.5 g. 



creatin 0.2 g. 



glycocoll 0.8 g. 



histidine dichloride 0.5 g. 



glucosamine hydrochloride. . 1.5 g. 



