474 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(5) Add chloroform to each bottle until 

 0.5% chloroform has been added. 



(6) Add a drop of sterile oil to the stop- 

 per and fasten a dust cover tightly 

 over the stopper. 



(7) Place in the air incubator at 37°C. 

 for 24 hours shaking constantly. 



(8) After this time, remove a sample 

 under aseptic conditions and test its 

 sterility by mixing it with agar and 

 incubating. (Time not specified.) 

 If sterile, do not heat longer for 

 heating tends to darken the medium. 



(9) Collect horse or ox blood in tall 

 sterile jars from the slaughter house. 



(10) Allow to stand until all the cor- 

 puscles have deposited. It may re- 

 quire 14 days for this to take place. 

 After five days examine the jars and 

 pipette or siphon off the clear serum. 

 Do not disturb the sediment. 



(11) Distribute the clear serum in 

 200.0 cc. lots in perfectly fitting glass 

 stoppered flasks. 



(12) Add chloroform to each bottle until 

 0.5% chloroform has been added. 



(13) Add a drop of sterile oil to the stop- 

 per and fasten a dust cover tightly 

 over the stopper. 



(14) Place the bottles in a water bath for 

 one hour at 45"C. Shake occa- 

 sionally. 



(15) When the bottles have cooled, re- 

 move a sample under aseptic condi- 

 tions and mix with agar. Incubate 

 the mixture at 37° for five days to 

 test sterility. 



(16) Prepare a 2.5 to 3.0% agar solution in 

 water containing salt and Morson's 

 peptone only (meat infusion or 

 derivatives not employed to decrease 

 color). 



(17) Add 1.0 cc. of (8) to 50.0 cc. of (15) 

 under aseptic conditions, and add 

 this mixture to 400.0 cc. of (16) and 

 mix well. 



(18) Tube thru a "hooded pipette." 

 Sterilization: Method of sterilization of 



agar not specified. 

 Use : Cultivation of meningococci and other 

 pathogenic forms, pneumococci and strep- 

 tococci. The author reported that this 

 medium was very good for routine work 

 with the meningococci. Pneumococci 



and streptococci developed abnormally 

 large colonies on this medium. 

 Reference: Fildes (1917 p. 492). 



1642. GolowkofE's Blood Peptone Agar 

 (Uche) 



Constituents: 



1. Water 500.0 cc. 



2. Agar (2.0%) 10.0 g. 



3. Peptone 5.0 g. 



4. NaCl 2.5 g. 



5. Blood 500.0 cc. 



6. Sugar (0.5%) 2.5 g. 



Preparation : 



(1) Prepare a 2.0% agar solution in water. 



(2) Dissolve 3 and 4 in (1). 



(3) Add an equal volume of blood (with- 

 out any preparation) heated to 

 80°C., to (2). 



(4) Boil for 15-20 minutes. 



(5) While hot, separate the coagulum by 

 pressing thru marly (a type of em- 

 broidery cloth). 



(6) Filter. 



(7) Add 0.5% sugar. 

 Sterilization: Not specified. 

 Use: Diagnosis of diphtheria. 

 Reference: Uche (1899 p. 393). 



1643. Harvey's Blood Serum Peptone Agar 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Peptone 10.0 g. 



3. NaCl ,,..... 5.0 g. 



4. Lactose 10.0 g. 



5. Neutral red (sat. soln.).... 10.0 cc. 



6. Agar to solidify 



7. Blood (defibrinated ox) 



8. Serum (ox) 

 Preparation : 



(1) Prepare a medium from 1, 2, 3 and 4 

 (see medium 607) and solidify with 

 agar. 



(2) Mix one part chloroform with 200.0 

 to 250.0 cc. of ox serum obtained 

 under as nearly aseptic conditions 

 as possible. 



(3) Place in incubator 48 hours with 

 occasional shaking. 



(4) Test sterility. 



(5) Mix equal parts distilled water and 

 defibrinated ox blood obtained under 

 as nearly aseptic conditions as 

 possible. 



