THE PREPARATION OF CULTURE MEDIA 167 



Teayue and Travis Medium for the Cholera Spirillum.** They prepare 

 their medium as follows: 



Two pounds of chopped beef are soaked in 2 liters of distilled water in 

 the ice box over night. The fluid is squeezed out, heated in the Arnold, 

 filtered through filter paper and made neutral to litmus with sodium hydrate, 

 and is heated again. After being allowed to cool, it is inoculated with colon 

 bacillus, and incubated for 2 or 3 days to make it sugar free. Agar is then 

 prepared from it by adding 1 per cent pepton, 0.5 per cent sodium chlorid 

 and clearing with egg. The reaction is adjusted to 0.5 phenolphthalein, 

 and after the agar has been cleared and filtered, 0.25 per cent nutrose is 

 added. A stock aqueous solution of 3 per cent bluish eosin is kept on hand 

 in the dark, also a 1 per cent stock solution of Bismarck-brown. The Bis- 

 marck-brown solution must be made up in water containing 10 per cent of 

 lactose, because it is entirely soluble to 1 per cent in distilled water alone. 



To 50 c.c. of this nutrose agar add 1 per cent saccharose, 1 c.c. of the 

 3 per cent eosin solution, and 2 c.c. of the 1 per cent Bismarck-brown solution. 

 After this mixture has been shaken until the stains are uniformly distributed, 

 pour plates. These plates are uncovered and placed on a shelf, face down, 

 in the incubator for 20 minutes to remove excess of moisture before smearing. 

 On this medium the differentiation of the cholera colonies is striking, with 

 large and with red centers, while the colon colonies are uniformly pink. 



Rabbit's Blood for Ducrey Bacillus Cultivation. Rabbits are bled from 

 the heart with a sterile syringe and about IVk to 2 c.c. placed into small 

 test tubes. The blood is allowed to clot and inactivated at 56 for y% hour. 

 This makes an excellent medium for the cultivation of the Ducrey bacillus, 

 for streptococci and some other organisms. It is also excellent for the 

 preservation of streptococci and pneumococci in a virulent condition. 



The preparation of Anaerobic Tissue Tubes for the Cultivation of Spiro- 

 chaetes and other Anaerobes. This is the Tarrozi and Smith method of 

 using tissue for anaerobic purpose, adapted by Noguchi for the cultivation 

 of various spirochaetes. The proportions of broth, serum and agar are 

 adapted to the particular purpose for which it is to be used. It is necessary, 

 therefore, in this place only to describe the best method of putting up the 

 tissue tubes. High, narrow test tubes are used, about 7 to 8 inches in length, 

 with a diameter not larger than that of a Wassermann tube. One-half inch 

 tubes are convenient. A rabbit is rapidly killed by ether anesthesia. It is 

 best to bleed him from the carotid at the same time, in order not to waste 

 the blood. The animal is immediately opened with sterile precautions. It 

 is best to dissect off the fur and wash the abdominal wall with alcohol before 

 opening the abdomen. The abdomen is then opened carefully and widely, 

 so that tlio organs can T>c easily reached without unnecessary poking about. 



44 Teague ami Travis, Jour, of Infec. Dis., 18, 1916, 601. 



