694 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Preparation : 



(1) Dissolve 2 and 3 in 1. 



(2) Neutralize or make slightly alkaline. 



(3) Distribute in tubes of the same 

 diameter. 



(4) Cool sterile (3) to 48 to 52°C. and add 

 one third the volume of rabbit blood 

 to each tube. Draw the blood di- 

 rectly from the heart under aseptic 

 conditions. 



(5) Slant the tubes. 



Sterilization: Sterilize (3) in the autoclave. 



Use: Cultivation of kala azar parasite. 

 Similar media were used for the cultiva- 

 tion of Prioplasma donovai, Piroplasma 

 equi, other piroplasma, trypanosomes, 

 leishmania, and protozoa. 



Variants : 



(a) Harvey cultivated trypanosomes on a 

 medium prepared as follows: 



(1) Dissolve 0.6 g. NaCI and 1.5 g. 

 agar in 100.0 cc. water. 



(2) Mix equal parts of (1) and de- 

 fibrinated rabbit blood at 45°C. 



(b) Harvey prepared a N. N. N. or whole 

 blood agar as follows for the cultiva- 

 tion of leishmania: 



(1) Dissolve 6.0 g. NaCl and 16.0 g. of 

 well washed agar in 900.0 cc. of 

 water. 



(2) Sterilize (1), method not given. 



(3) Mix one part rabbit blood with 3 

 parts (2) at 50°C. 



(4) Slope. 



(5) Test sterility. 



(6) Preserve in the dark. 



(c) Harvey cultivated the trypanosomes 



of cold blooded animals on the 

 following medium : 



(1) Dissolve 20.0 g. of agar in 1000.0 

 cc. tap water. 



(2) Tube and sterilize. 



(3) Mix equal parts (2) and sterile 

 defibrinated rabbit blood. 



(4) Solidify in the sloped position and 

 inoculate in the water of con- 

 densation. 



(d) Leiva cultivated Leishmania infan- 

 tum and Leishmania ctenocephali 

 on a medium prepared as follows: 



(1) Dissolve 20.0 g. agar and 4.0 g. 

 NaCl in 1000.0 cc. water. (May 

 use 5.0 g. agar.) 



(2) Tube. 



(3) Add 25.0% (Use only 5.0% when 

 using 0.5% agar) sterile defibri- 

 nated rabbit blood to sterile melted 

 and cooled (2). 

 (e) Stitt gave the following method of 

 preparation of N. N. N. medium for 

 the cultivation of trypanosomes, 

 leishmania and protozoa: 



(1) Dissolve 14.0 g. agar and 6.0 g. 

 NaCl in 900.0 cc. of water. 



(2) Tube and sterilize. 



(3) To one part of (2) liquified and 

 cooled to 48°C. add one third its 

 volume of defibrinated rabbit 

 blood. (Use human or rat blood for 

 the cultivation of human trypano- 

 somes.) 



(4) Mix thoroly. 



(5) Slant. 



(6) Use rubber stoppers or cover cotton 

 plugs with paraffin. 



References: Nicolle (1909 p. 397), Marzin- 

 owsky (1908-09 p. 419). Bezan^on (1920 

 p. 120), Harvey (1921-22 pp. 72, 74), 

 Leiva (1922 p. 179) taken from (1923 p. 

 342), Stitt (1923 p. 52). 



2153. Noguchi's Ringer Solution Plasma 

 Agar (Abbott) 



Constituents : 



1. Water 1000.0 cc. 



2. NaCl 10.0 g. 



3. KCl 0.2 g. 



4. CaCl2 0.2 g. 



5. NaHCOa 0.1 g. 



6. Glucose 1.0 g. 



7. Serum, rabbit 



8. Citrate plasma 



9. Agar (2.0%) 

 Preparation: 



(1) Dissolve 2, 3, 4, 5 and 6 in 1. (Ringer 

 solution.) 



(2) Add 1.5 parts serum and 1 part citrate 

 plasma to 4.5 parts (1). 



3) Stiffen the medium by the addition of 

 "sterile agar (free of peptone and 

 sugar) in the proportion of 2.0%." 



Sterilization: Not specified. 



Use: Cultivation of spirochaetes. 



Variants : 



(a) Kaneko cultivated Spirochaeta ic- 

 ier ohaemorrhagiae and Spirochaeta 

 hebdomadis on a medium prepared 

 as follows: 



