226 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(2) Add 3.0 to 4.5 cc. of a watery solu- 

 tion of Merck's highly sensitized 

 litmus. 



(3) Prepare a 10.0% solution of one of 

 the following in distilled water: 

 glucose, galactose, levulose, lac- 

 tose, maltose, sucrose, mannitol, 

 dulcitol, inulin and dextrin. 



(4) Sterilize (3) at 100°C. for 10 

 minutes. 



(5) Mix (2) and (4). 



(6) Tube in sterile tubes. 



(7) Incubate for 3 days to detect acci- 

 dental contamination. 



Fermentation by the meningococci 

 was studied in these media. 

 References: Buerger (1907 p. 430), Elser 

 and Huntoon (1909 p. 404). 



773. Akatsu's Basal Ascitic Fluid Bouillon 



Constituents : 



1. Bouillon 1000.0 cc. 



2. Ascitic fluid 1000.0 cc. 



3. Tissue 

 Preparation : 



(1) Composition or method of prepara- 

 tion of bouillon not given. 



(2) Mix equal parts of bouillon and 

 ascitic fluid and tube in 10.0 cc. lots. 



(3) Add to each tube of (2) a piece of 

 fresh tissue. 



(4) Add 1.0% of one of the added nu- 

 trients to each tube. 



Sterilization: Not specified. 

 Use : To study the fermentation of carbohy- 

 drates, alcohols, etc., by bacteria. 

 Akatsu used spirochetes, Treponema pal- 

 lidum, Treponema calligyrum, Trepo- 

 nema microdentium, Treponema mucosum 

 and Spirochaeta refringens. Medium was 

 covered with a layer of sterile paraffin 

 after inoculation and incubated at 36°C. 

 Added nutrients: The author added 1.0% 

 of one of the following materials: 

 amygdalin galactose 



arabinose glycogen 



beerwort glucose 



dextrin inulin 



mannitol lactose 



raffinose levulose 



sucrose maltose 



starch 

 Reference: Akatsu (1917 p. 376). 



774. Hiss' Basal Serum Bouillon 



Constituents: 



1. Bouillon (sugar free) 1000.0 cc. 



2. Serum (beef) 500.0 cc. 



Preparation : 



(1) Method of preparation of sugar free 

 broth not given. 



(2) Adjust (1) to 1.0% acid to phenol- 

 phthalein (neutral to litmus). 



(3) Mix 1 part sterile beef serum with 2 

 parts sterile (2). 



(4) Dissolve one of the added nutrients 

 in (3). 



Sterilization: Sterilize at 65-68°C. for one 

 hour on 6 consecutive days. 



Use: Differentiation of streptococci and 

 pneumococci. Pneumococci fermented 

 starch. Streptococci did not. Strepto- 

 cocci and pneumococci fermented lac- 

 tose, saccharose and dextrin, produced 

 acid and formed a yellowish white 

 coagulum. 



Added nutrients and variants: 



(a) The author added 1.0% sucrose, 1.0% 

 lactose, 1.0% glucose, 1.0% dextrin 

 or 0.66% starch. 



(b) Koch and Pokschischewsky prepared 

 similar media for the differentiation 

 of Streptococcus longus seu. erysipe- 

 latos and Streptococcus equi. They 

 reported that Streptococcus equi gave 

 a flocculent flaky sediment leaving a 

 clear bouillon. Streptococcus longus 

 seu erysipelatos gave a uniform tur- 

 bidity to the medium. (Medium 

 containing no sugars). The media 

 were prepared as follows: 



(1) Mix one part sterile horse serum 

 and two parts ordinary bouillon. 



(2) Sterilize on 3 successive days at 

 60°C. for one hour. 



(3) Prepare a 10.0% solution of one of 

 the following: 



sorbitol maltose 



dulcitol lactose 



mannitol glucose 



levulose mannose 



galactose raffinose 



sucrose 



(4) Sterilize on 3 successive days for 

 2 or 3 minutes in streaming steam. 



(5) Add 1.0 cc. of (4) to 9.0 cc. of (2), 

 under aseptic conditions. 



