202 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Use: Cultivation of Spirochaeta pallida. 

 Reference: Weiss and Wilkes-Weiss (1924 

 p. 222). 



724. Kelser's Blood Infusion Peptone 

 Solution 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Beef blood 500.0 g. 



3. Peptone 15.0 g. 



4. NaCl 7.5 g! 



Preparation : 



(1) Allow beef blood to clot in covered 

 vessel. Let stand in refrigerator to 

 allow serum to separate from clot. 



(2) Remove clot and chop finely in 

 chopping machine. 



(3) Mix serum and ground clot. Weigh. 



(4) Add two volumes of distilled water. 



(5) Boil gently for 5 minutes. 



(6) Filter thru cheese cloth. Take resi- 

 due and put thru fruit press to ex- 

 tract as much fluid as possible, 

 lining press with towel or heavy 

 cloth so that pulp does not pass thru. 



(7) Discard the residue. 



(8) Boil fluid, skimming off coagulated 

 proteins. 



(9) Add concentrated CH3COOH (0.5 cc. 

 per L) to cause flocculation and con- 

 tinue to boil 5 minutes. 



(10) Filter— first thru absorbent cotton 

 and two filter papers. 



(11) Ascertain volume and add 0.5% 

 NaCl, and 1.0% peptone. Heat to 

 get into solution. 



(12) Neutralize with NaOH. 

 Sterilization: Sterilize in autoclave i hour 



under 12 pounds pressure. 



Use: Nutrient medium for pathogenic 

 forms. The author reported that glucose 

 aided the growth of some organisms. 



Variants : 



(a) Kelser added 0.25% glucose. 



(b) Harvey prepared a medium, boiling 

 the water, clot and serum mixture 

 (see (5) above) for 10 minutes instead 

 of 5, specified glacial acetic acid, 

 steamed the peptone NaCl mixture 

 (see (11) above) for 45 minutes, and 

 adjusted the reaction to a definite 

 pH value or faintly alkaline to litmus, 

 or 1.0% acid to phenolphthalein. 



(c) Harvey prepared a medium by heat- 



ing the water, clot serum mixture 

 (see (4) above) at 50°C. for 20 minutes 

 and then boiling 10 minutes instead of 

 5, boiled 5 minutes after the addition 

 of the glacial acetic acid (see (9) 

 above), steamed the peptone NaCl 

 mixture (see (11) above) for 45 

 minutes instead of heating only until 

 solution was complete and adjusted 

 the reaction to a definite pH value, 

 faintly alkaline to litmus or 1.0% 

 acid to phenolphthalein. After ad- 

 justing the reaction the medium was 

 steamed for 30 minutes, filtered while 

 hot thru wet thick filter paper and 

 tubed. 

 References: Kelser (1916 pp. 615, 616)^ 

 Harvey (1921-22 pp. 69, 76). 



725. Wiens' Blood Peptone Solution 

 (Klimmer) 



Constituents: 



1. Water 1000.0 cc. 



2. Peptone (10.0%) 100.0 g. 



3. Glucose (1.0%) 10.0 g. 



4. Blood.. 100.0 cc. 



Preparation : 



(1) Prepare a 10.0% peptone solution in 

 water. 



(2) Make slightly alkaline. 



(3) Dissolve 1.0% glucose in (2). 



(4) Tube in 10.0 cc. quantities. 



(5) Add 1.0 cc. of blood to each tube. 

 Sterilization: Not specified. 



Use: Cultivation of pneumococci. 

 Reference: Klimmer (1923 p. 221). 



726. Kligler's Peptone Serum Blood 

 Medium 



Constituents : 



1. Saline 200.0 cc. 



2. Serum (horse or rabbit) 100.0 cc. 



3. Blood 



4. Peptone (1.0%) 3.0 g. 



Preparation: 



(1) Dilute horse or rabbit serum 1:2 with 

 saline. 



(2) Adjust to pH 7.0. 



(3) Add 1.0% peptone (1.0 cc. of a 10.0% 

 solution per 10.0 cc. medium). 



(4) Distribute in 3 to 4.0 cc. lots in Was- 

 sermann tubes. 



(5) Add 0.1 cc. of blood. (For initial 

 cultures, add infected blood. For 



