272 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(5) Incubate until a faint haze is formed 

 (usually 1 to 2 hours). 



(6) Cover with a layer of sterile paraffin. 

 Sterilization: Steam (6) at 100° for 15 to 



30 minutes. 

 Use: Cultivation of Bacleriun pneumo- 



sintes. The medium was inoculated 



while the layer of paraffin was still 



liquid. 

 Reference: Olitsky and Gates (1922 p. 818), 



876. Ogata's Porphyra Infusion Broth 



Constituents : 



1. Meat infusion broth 1000.0 cc. 



2. Glucose 25.0 g. 



3. Porphyra vulgaria 50.0 g. 



Preparation : 



(1) Prepare meat infusion, using 500.0 g. 

 of meat per liter. 



(2) Dissolve 2 and 3 in (1). 



(3) Boil. 



(4) Neutralize. 



(5) Filter. 



(6) Tube. 



Sterilization: Method not given. 



Use: Isolation and cultivation of protozoa 

 (infusoria). For isolation a capillary 

 tube is filled with the medium, inoculated 

 by placing one end of the tube into the 

 culture and the tube sealed. 



Reference: Ogata (1893 p. 168). 



877. Park, Williams and Krumwiede's 

 Stomach Digest Infusion Broth 



Constituents : 



1. Water 2000.0 cc. 



2. Meat 1.0 lb. 



3. Peptone (1.0%) 10.0 g. 



4. NaCl (0.5%) 5.0 g. 



5. Stomach (hog) 200.0 g. 



Preparation : 



(1) Clean 5 pigs' stomachs, remove the 

 fat and mince finely. (A number of 

 stomachs should be used to equalize 

 the peptone content. In this way 

 an almost average composition in 

 peptone is obtained.) 



(2) Mix 200.0 g. of (1), 10.0 g. HCl, pure, 

 and 1000.0 cc. of water at 50°C. 



(3) Incubate at 50°C. for 20 to 24 hours 

 in a glass or porcelain vessel, not 

 enamel. It is most important not 

 to allow the digest to come in con- 

 tact with any metal until neutral- 

 ized. 



(4) Heat to 80°C. to stop digestion. 



(5) Pass thru a -layer of absorbent 

 cotton. 



(6) The digest may be stored without 

 sterilization. 



(7) Heat the filtrate from (5) to 70 °C. 



(8) Neutralize to litmus at 70°C. 



(9) Mi.x an equal volume of (5) and infu- 

 sion broth, prepared according to 

 Park, Williams and Krumwiede 

 (see variant (ii) medium 779). 



(10) Adjust to the desired reaction at 

 room temperature. 



(11) Autoclave for 15 minutes to clear. 



(12) Readjust the reaction as necessary. 



(13) Filter thru paper and cotton to 

 clear. 



Sterilization: Sterilize in the autoclave at 

 15 pounds pressure for 30 minutes. 



Use: General culture medium. 



Reference : Park, Williams and Krumwiede 

 (1924 p. 118). 



878. Kitchen's Yeast Autolysate Blood 

 Solution 



Constituents: 



1. Beef infusion. 



2. Blood clot 200.0 cc. 



3. Peptone (1.0%) 10.0 g. 



4. NaCl (0.5%) 5.0 g. 



5. Agar (0.07%) 0.7 g. 



6. Glucose (3.0%) 30.0 g. 



7. Maltose (3.0%) 30.0 g. 



8. Yeast. 

 Preparation : 



(1) Dissolve 200.0 cc. of strained horse 

 blood clot, 10.0 g. peptone, 5.0 g. 

 NaCl and 0.7 g. agar in sufficient 

 beef infusion to make a liter. 



(2) Adjust the reaction to pH 5.4. 



(3) Autoclave and filter while hot. 



(4) Distribute in 500.0 cc. lots in liter 

 flasks. 



(5) After sterilization, add sufficient 

 sterile solutions of glucose and mal- 

 tose to make a 3.0% solution of each 

 of the sugars. 



(6) Inoculate with yeast and keep at 

 room temperature until ready for 

 use. (One week is sufficient.) 



(7) Place the yeast cultures at 55°C. over 

 night and then bring rapidly to the 

 boiling point over a free flame. 



(8) Heat at 95°C. for 15 minutes. 



(9) The autolysate may be filtered thru 



