CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



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Preparation: 



(1) Obtain the blood from the animal 

 whose tissue are to be cultivated or 

 another animal of the same species, 

 under aseptic conditions. Collect 

 the blood in sterile tubes cooled to 

 0°C., coated with sterile paraffin. 



(2) Cork the tubes immediately and 

 place in large tubes filled with ice and 

 centrifuge for 5 minutes, and place 

 in a small ice box at 0°C. 



(3) Remove the supernatant plasma with 

 sterile paraffin coated pipettes. It is 

 generally used immediately. 



(4) Take the tissue to be cultivated 

 directly from the living animal or 

 immediately after death. Cut the 

 tissue in small pieces and transfer 

 either to a cover glass or larger con- 

 tainer. The cutting must be done 

 quickly for desiccation kills the 

 tissue. 



(5) Cover the tissue with the plasma. 



Sterilization: See preparation. 



Use: Cultivation of tissue. Author re- 

 ported that citrated blood might be used. 

 Sufficient blood was added to a 1.0% 

 sodium oxalate solution making the 

 solution 0.1%. At the time of use pre- 

 cipitate the sodium oxalate quantitively 

 from the plasma by the addition of CaCU. 

 This does not give as good results as does 

 pure plasma. 



Variants: Carrel prepared the medium as 

 follows: 



(1) Dilute plasma with about 2/5 of its 

 volume of dist. water. 



(2) In order to accelerate the coagulation 

 of the plasma, embryonal extracts or 

 serum may be added. 



(3) Pour (1) over small pieces of tissue 

 to be cultivated. 



Reference: Carrel and Burrows (1911 p. 

 391), Carrel (1912 p. 393). 



2403. Ball's Coagulated Blood Medium 



Constituents: 1. Blood. 



Preparation: (1) Coagulate blood (not the 



serum only) in test tubes. 

 Sterilization: Not specified. 

 Use: General culture medium. 

 Reference: Ball (1919 p. 80). 



2404. Cobbett's Glucose Serum Medium 



Constituents : 



1. Serum 1000.0 cc. 



2. Glucose 20.0 g. 



Preparation: 



(1) Collect beef blood from a slaughter 

 house and store the blood in a cool 

 place to allow the serum to separate. 



(2) To 100.0 cc. of serum add 2.0 g. of 

 glucose and 1.75 cc. of 10% NaOH 

 solution. 



(3) Distribute into test tubes (or Petri 

 dishes). 



(4) Slant the tubes and place in the auto- 

 clave. In order to avoid air bubbles 

 in the medium close the stop cock of 

 the autoclave before all the air is 

 removed. This gives a high pressure. 



Sterilization: Sterilize for 20 minutes at 

 120°C. 



Use: Diagnosis of diphtheria. Author re- 

 ported that diptheria colonies were 

 flat, gray or colorless. After a few days 

 surface became irregular. Different sera 

 may require different amounts of alkali. 

 Old laboratory strains grew less luxuri- 

 antly than did freshly isolated strains. 



Variants: The author prepared a similar 

 medium as follows: 



(1) To 100.0 cc. of horse serum add 2.0 g. 

 dextrose, and 1.25 to 1.3 cc. of 10.0% 

 NaOH solution. 



(2) Distribute into tubes or Petri dishes. 



(3) Heat to 90°C. on two successive days 

 to sterilize. 



Reference: Cobbett (1898 p. 395), (1898 

 p. 362). 



2405. Costa's et al. Litmus Glucose Serum 



Constituents : 



1. Serum (horse) 100-0 cc. 



2. Glucose 



3. Litmus 



4. H2SO4 

 Preparation : 



(1) Prepare a solution of 30 parts glucose 

 to 100 of water. 



(2) Mix 10.0 cc. of sterile (1), 30 drops of a 

 sterile concentrated solution of tinc- 

 ture of litmus, 3.0 cc. of a sterile one 

 to one hundred H2SO4 and 100.0 cc. 

 of sterile horse serum. 



