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CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



blood and mixed with nutrient agar 

 cooled to 45 to 60°C. The medium 

 was flaky and of a brownish color and 

 used for the cultivation of influenza 

 bacilli. 



(d) Miiller cultivated fowl diphtheria 

 bacilli on a medium prepared as 

 follows: 



(1) Prepare nutrient agar. 



(2) Liquify (1) and place into sterile 

 400.0 cc. Erlenmeyer flasks in 

 250.0 cc. lots. 



(3) Allow about 25.0 cc. of blood to 

 pass directly from the jugular vein 

 of a goat into the melted agar or 

 gelatin which has been cooled to 

 about 45°C. (Dog and chicken 

 blood gave good results also.) 



(4) Pour the blood medium into sterile 

 Petri dishes. 



(e) Savini and Savini-Castano cultivated 

 influenza bacilli on a hematin agar 

 prepared as follows: 



(1) Allow about 100.0 cc. of blood to 

 coagulate. 



(2) Pour off the serum. 



(3) Cut the blood clot into fine pieces 

 and place into a flask containing 

 100 to 150.0 cc. of N/1 NaOH or 

 N/10 soda solution. 



(4) Boil for a long time over a free flame 

 until all is well dissolved. 



(5) Filter and sterilize (method not 

 given). 



(6) Prepare nutrient agar. 



(7) Melt sterile (6) and cool to about 

 50°C. 



(8) To each 10.0 cc. of (7) add 1.0 cc. 

 of (5) and mix well without shaking 

 violently. 



(9) Allow the agar to solidify over night 

 in a cool place. 



Authors reported that influenza 

 bacilli developed upon this medium 

 when another organism grew side by 

 side with it. Staphylococcus aureus 

 gave best results for this purpose, 

 (f) Ito and Matsuzaki cultivated Spiro- 

 chaeta icterohaemorrhagiae on the 

 following medium : 



(1) Prepare nutrient agar. 



(2) Mix one part normal guinea pig 

 blood or human blood with one or 

 two parts melted agar at 50°C. 



(3) Add a drop of infected blood to the 

 tubes before solidification and dis- 

 tribute by stirring the tubes. 



(4) Incubate at 15° to 37°C. Paraffin 

 may be poured over the medium but 

 growth may be obtained without it. 

 Authors reported that the erythro- 

 cytes settled to the bottom of the 

 agar before solidification, giving it an 

 opaque and deeper color. 



(g) WoUstein cultivated Pfeiffer's bacil- 

 lus on the following media: 



(1) Prepare nutrient agar. 



(2) Adjust the agar to pH = 7.5. 



(3) Boil fresh rabbit blood for two 

 minutes in a water bath. 



(4) Centrifuge. 



(5) Add 0.5 cc. of the resulting clear 

 pale pink or yellow fluid to 20.0 cc. 

 of the broth or two or three drops 

 to 5.0 cc. of the melted agar. 



(h) Levinthal (Klimmer) cultivated in- 

 fluenza bacilli on a medium prepared 

 as follows. Klimmer reported that 

 best results were obtained using a 

 medium 24 hours old. 



(1) Melt 1000.0 cc. of neutral 2.0 or 

 3.0% agar and cool to 70°C. 



(2) Mix 50.0 cc. of fresh human or dog 

 blood with (1). The blood may be 

 defibrinated and stored on ice, if 

 desired. 



(3) Heat (2) to boiling over a wire gauze 

 until the agar begins to raise in the 

 neck of the flask. 



(4) Repeat the boiling twice more. 



(5) Separate the agar from the coagu- 

 lum by filtering thru sterile cotton 

 or gauze filter. Do not heat again 

 in steam. 



(6) Agar is alkaline to litmus. 



(7) Tube or pour in plates. 

 References: Bezangon, Griffon and Le- 



Sourd (1901 p. 3), Thoinot and Masselin 

 (1902 p. 36), Davis (1903 p. 405), Cza- 

 plewski (1902 p. 668), Heinemann (1905 

 p. 128), MuUer (1906 p. 519), Savini and 

 Savini-Castano (1911 p. 493), Ito and 

 Matsuzaki (1916 p. 558), Roddy (1917 

 p. 45), Wollstein (1919 p. 556), Ball (1919 

 p. 82), Bezangon (1920 p. 120), Kristensen 

 (1922 p. 223), Klimmer (1923 pp. 225, 

 227), Stitt (1923 p. 44), Park, Williams 

 and Krumwiede (1924 p. 125). 



