CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



375 



finished medium in the steamer, using the 

 fractional method. 



Use: To study decomposition of organic N 

 materials by soil forms. Author reported 

 little change in total nitrogen content 

 after 12 days when inoculated with soil. 



Reference: Remy and Rosing (1911 p. 39). 



1282. Klimmer's Hemoglobin Solution 



Constituents: 



1. Blood 1000.0 cc. 



2. Physiological salt solution . . 1000.0 cc. 

 Preparation: 



(1) Collect pigeon blood under aseptic 

 conditions and add to an equal 

 volume of physiological salt (0.85%) 

 solution. 



(2) Mix well and centrifuge, or allow to 

 stand in the cold for 24 hours. 



(3) Thoroly wash the red residue twice 

 more with sterile NaCl solution. 



(4) Dissolve the hemoglobin with a trace 

 of ether. 



(5) Evaporate the ether at 30°C. 



(6) Separate the hemoglobin from the 

 stromata by passing thru a bacterial 

 filter. 



Sterilization: Method of sterilization of 

 salt solution not specified. 



Use: This hemoglobin solution is added to 

 other media which is used for the cultiva- 

 tion of pneumococci, vibrio, etc. One 

 cubic centimeter of this solution is added 

 to 8.0 cc. of media, or this solution is 

 streaked on solid media. 



Variants: The author also used commercial 

 hemoglobin. This solution was prepared 

 as follows: 



(1) Mix 10.0 g. of commercial hemoglobin 

 with 90.0 cc. of distilled water. 



(2) Add about 10.0 cc. of 10.0% KOH. 



(3) Sterilize in the steamer. 

 Reference: Klimmer (1923 p. 196). 



1283. Meyerstein's Bile Medium 



Constituents: 



1. Bile, beef. 

 Preparation: 



(1) Evaporate beef bile with animal char- 

 coal to dryness on a water bath. 



(2) Take up the residue in alcohol. 



(3) Filter. 



(4) Add quite a lot of ether to the some- 

 what concentrated filtrate. This 



causes the bile acid to precipitate out 

 in silky crystals — so-called crystalline 

 bile. 



(5) Dry and pulverize the crystals. 



(6) Add one or two knife points of (5) 

 to sterile test tubes. 



Sterilization: Medium sterilized during the 

 preparation. 



Use: Enrichment of typhoid bacilli from 

 patients blood. Diagnosis of typhoid 

 fever. The author added 2.0 to 4.0 cc. of 

 the patients blood to each tube of (6). 



Variants: The following authors have pre- 

 pared media in the manner indicated. 



(a) Kayser 



(1) Secure normal beef bile immedi- 

 ately after the death of an animal in 

 large sterile flasks. 



(2) Distribute in amounts of at least 

 5.0 cc. in test tubes. 



(3) Sterilize at 110°. 



(4) Put 2.5 cc. of blood of patient into 

 this at bedside. 



(b) Bezangon. 



(1) Collect bile at a slaughter house. 



(2) Sterilize at 100°C. 



(3) Allow to settle. 



(4) Decant the clear liquid. 



(5) Tube in 3.0 cc. quantities in sterile 

 tubes. 



(6) Sterilize at 105°C. for 20 minutes. 



(c) Klimmer. 



(1) Puncture the gall bladder of a beef 

 with a knife, and collect the bile. 



(2) Boil (1) in a steamer for 15 minutes. 



(3) Distribute in 5.0 cc. quantities in 

 test tubes. 



(4) Steam on each of 3 successive days 

 for 20 to 30 minutes. 



References: Meyerstein (1906 p. 1864), 

 Kayser (1906 pp. 823-826), Harvey 

 (1921-22 p. 89), Bezangon (1920 p. 122), 

 Klimmer (1923 p. 202), Stitt (1923 p. 47). 



1284. Ottolenghi's Nitrate Bile Solution 



Constituents: 



1. Bile 1000.0 cc. 



2. NasCOa (10.0% soln.) 



(3.0%) 30.0 cc. 



3. KNOj (1.0%) 1.0 g. 



Preparation: 



(1) Filter fresh ox bile thru a paper. 



(2) Add 3.0% of a 10.0% watery solution 



