318 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Use: General culture medium. Substitute 

 for bouillon. Author stated that sugars 

 might be added to this medium if desired. 

 Harvey used a similar medium to demon- 

 strate the indol test. 



Variants : 



(a) The author gave a more complete 

 method of the preparation as follows: 



(1) Chop the pancreas of a hog into 

 small pieces and extract with 

 chloroform in 9 times its volume of 

 distilled water for 25 hours. 



(2) Filter on glass wool, pressing the 

 meat free from the liquid. 



(3) Add ether to preserve the extract 

 and store in a sealed flask. 



(4) Mix beef or sheep serum with an 

 equal volume of water. 



(5) Boil (4) until it assumes a milky 

 appearance. 



(6) Mix 15 volumes of (3) with 100 

 volumes of (5) and digest over night 

 at 60°C. on a salt water bath. 



(7) Filter. 



(8) Tube 



(9) Sterilize at 120° for 15 minutes. 



(b) Harvey mixed one part serum with 

 2 parts tap water and boiled until the 

 mixture was milky. Then added 

 15.0 cc. of pancreatic extract to 

 100.0 cc. of the mixture and digested 

 at 60°C. for 24 hours. Following 

 filtration thru paper and distribution, 

 the medium was sterilized by heating 

 at 120°C. for 15 minutes. This me- 

 dium was used for the detection of 

 indol production. 



(c) Harvey used equal parts serum and 

 water boiled as above and added 

 10.0% pancreatic extract. After di- 

 gestion at 60°C. over night, the me- 

 dium was filtered thru thick filter 

 paper, tubed and sterilized in the 

 autoclave. 



References: Distaso (1916 p. 600), (1917 

 p. 253), Harvey (1921-22 pp. 81, 117). 



1119. Hettinger's Trypsinized Meat 

 Solution (Klimmer) 



Constituents: 



1. Water. 



2. Meat. 



3. K2HPO4. 



4. NaCl. 



5. K3PO4. 



Preparation : 



(1) Add 1.5 liters of boiling water to 

 1000.0 g. of meat, cut in pieces the 

 thickness of a finger. 



(2) Apply heat until the mixture boils 

 again. 



(3) Remove the pieces of meat. 



(4) Pass the meat thru a grinding ma- 

 chine. 



(5) Place the liquid in a large mouthed 

 flask, add 1.5 g. dehydrated soda 

 and when the liquid is lukewarm add 

 3.0 g. pancreatin and 15-20.0 cc. 

 chloroform (and toluol). 



(6) Seal the flask and mix well. 



(7) Add the ground meat (4) and shake 

 once more. 



(8) The meat should be thoroly covered 

 with liquid. If not add water. 



(9) Incubate for 5 days at 20° or 2 days 

 at 37°. Shake daily. 



(10) Acidify slightly by the addition of 

 HCl. 



(11) Filter thru paper. 



(12) Wash the residue on the filter paper 

 until the volume of the filtrate is 

 3 liters. 



(13) Filter again after shaking thoroly. 



(14) Boil for at least 10 minutes. 



(15) Dilute to 10 to 50 liters. 



(16) Prepare a saturated solution of crude 

 salt (not purified) rich in calcium 

 and magnesium. 



(17) Dissolve 5.0 g. of K2HPO4 and small 

 knife point of K2HPO4 in a little 

 distilled water. Filter. 



(18) Add (17) to 100.0 cc. of filtered (16). 



(19) Add 20.0 cc. of (18) to each liter of 

 (15). 



(20) Filter. 



(21) Boil. 



(22) Make slightly alkaline. 

 Sterilization: Not specified. 



Use: General culture medium. Klimmer 

 reported that for bacteria producing pig- 

 ment dilute the solution to 50 liters in 

 step (15), for usual laboratory work to 

 30 liters and to 10 liters if an extremely 

 good culture medium is desired. 

 Variants : 



(a) Park, Williams and Krumwiede give 

 the following method of preparation 

 of the Hettinger medium. The phos- 

 phates are omitted. 



